TW201219370A - Agent for regulating the formation of nitrogen monoxide - Google Patents

Abstract

In order to provide a novel nitric oxide generation regulator for effectively regulating nitric oxide generation from vascular endothelial cells, macrophage cells including microglial cells, and the like that can be safely used in vivo, disclosed is a nitric oxide generation regulator that has the trinuclear pentamethine cyanine dye represented by the belowmentioned general formula (1) as the active ingredient. (In the general formula (1), R represents a potentially branched alkyl group having 2-4 carbon atoms, and X- represents an appropriate counteranion.)

Description

201219370 VI. Description of the Invention: [Technical Field of the Invention] The present invention relates to the use of a trinuclear pentamethine-based cyanine dye, and more particularly to a nitric oxide production regulator comprising a trinuclear five-type _ Alkyne cyanine pigment, and one or more other ingredients that can be tolerated by the formulation. [Prior Art] Nitric oxide (NO) is known to play an important role in various physiological reactions in vivo (see, for example, International Publication WO 95/03 1 98 7 pamphlet). Specifically, for example, a function related to blood pressure adjustment by a relaxation action of a smooth muscle system of a blood vessel, an action of inhibiting blood coagulation by a platelet aggregation inhibiting action, and the like. Other important roles include factors that act as an inflammatory process and are activated by macrophage toxic activity. An abnormal balance of nitric oxide in an organism can cause serious diseases or disorders. That is, excess nitric oxide is produced during septic or hemorrhagic shock, which causes a pathologically large blood pressure drop. Further, examples of diseases caused directly or indirectly by the decrease in the concentration of nitric oxide include arterial hypertension, septicemia, and heart disease. In addition to the above, diseases or disorders associated with abnormal balance of nitric oxide in the living body are known: rheumatoid arthritis, deformed arthritis, ulcerative colitis, tissue disorder after transplanting of a sputum, transplant rejection, viral infection Myocarditis and cardiomyopathy caused by myocarditis, inflammatory diseases such as nephritis, pancreatitis, aging, atherosclerosis, and vascular endothelium (including tiny blood vessels) Endothelial damage, etc. - The role of nitric oxide in organisms is clear. It is understood that the relationship between local or systemic imbalances and various diseases is caused by the prevention and treatment of nitric oxide balance in order to inhibit the production of nitric oxide in living organisms. Diseases as described above have been proposed in the literature as prophylactic or therapeutic agents for various diseases or disorders associated with abnormal nitric oxide balance, which are acridine derivatives (International Publication WO 95/03 1 987 pamphlet, Japanese special table 1 0 - 504 023, condensed piperidine compound (Japanese Patent Publication No. Hei 11-171866), a cyclic olefin derivative (JP-A-2005-232168), and Loxoprofen (Japanese Patent Laid-Open No. 2007-) A plant-derived component or the like as the active ingredient, such as a compound or a palm kernel-treated product (Japanese Laid-Open Patent Publication No. 2005-53864). However, in the current situation, many of these compounds have not yet been put into practical use. SUMMARY OF THE INVENTION The object of the present invention is to provide a novel nitric oxide production regulating agent which can be safely applied to an organism for effectively regulating macrophage cells or vascular endothelial cells derived from microglia. The formation of nitric oxide is equal. In order to solve the problem, the present inventors have focused on the polymethine-based cyanine dye, and found that the trinuclear pentamethine-based cyanine having the salino skeleton represented by the following general formula 1 has been found. The pigment has an effect of regulating the production of nitric oxide from microglia, and it is confirmed that this cyanine pigment does not exhibit toxicity or serious side effects even if it is directly applied to an organism, and the present invention is completed. invention. That is, the present invention is mainly constituted by a nitric oxide production regulator which is a trinuclear pentamethine-based cyanine dye represented by the following general formula as an active ingredient. [Chemical 1] - General 1

R (in the general formula 1, R represents an alkyl group which may have a branch and a carbon number of 2 to 4, and X' represents a suitable relative anion). The nitric oxide production regulating agent of the present invention containing the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 as an active ingredient can be regulated from microglia containing cells by parenteral or oral administration. Nitric oxide production in macrophage cells or vascular endothelial cells, etc., while regulating the abnormal balance of nitric oxide. Further, the dye represented by the general formula 1 as an active ingredient is extremely safe. Further, it is presumed that one of the nitrogen oxide production regulators of the present invention causes a decrease in blood pressure caused by a balance abnormality generated by nitric oxide, various serious inflammatory diseases such as myocarditis, or vascular tissues such as heart (including The prevention of tissues or cells such as microvascular endothelium, prevention of arterial hypertension, and septicemia is effective in terms of treatment. [Embodiment] As described above, the present invention relates to a nitric oxide production regulator which comprises the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 as an active ingredient. The trinuclear pentamethine-based cyanine dye itself is known as a known substance (see, for example, Japanese Laid-Open Patent Publication No. Hei. No. Hei. No. Hei. No. Hei. No. Hei. No. Hei. No. Hei. Role (refer to, for example, Masaru Sugawara, "Photosensitive Pigment", published on October 17, 1997, Industrial Book Co., Ltd., to 30 pages and 138 to 154 pages). However, in these documents, there is no suggestion or suggestion that the trinuclear pentamethine-based cyanine dye has a function of regulating the production of nitric oxide from macrophage cells or vascular endothelial cells including microglia. Therefore, the nitric oxide production regulator containing the trinuclear pentamethine-based cyanine dye represented by the general formula 1 as an active ingredient was first discovered by the present inventors. One of the nitric oxide production regulators of the present invention is a trinuclear pentamethine-based cyanine dye represented by the above general formula 1 as an active ingredient. The alkyl group represented by R in the general formula 1 has 2 to 4 carbon atoms, specifically, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a second butyl group, and a third butyl group. From the viewpoint of adjusting the intensity of the action of nitric oxide production from macrophage cells or vascular endothelial cells including microglia, the carbon number of the alkyl group represented by R in the general formula 1 is 2 Further, a trinuclear pentamethine-based cyanine dye having a linear alkyl group and a carbon number of 3 is preferable, and a dye having a carbon number of 2 in the alkyl group (R) is particularly preferable. Incidentally, as revealed by the experiments described later, the alkyl (R) has a trinuclear pentamethine-based cyanine pigment having a carbon number of 1 or 6, and -6 - 201219370 is not derived from microglia-containing cells. Nitric oxide production by macrophage cells or vascular endothelial cells exhibits a marked regulatory effect. The χ_ in the general formula 1 represents a suitable relative anion, and is usually selected, for example, from a fluorine anion, a chloride anion, a bromine anion, an iodine anion, a perchlorate anion, a periodic acid anion, a hexafluorophosphate anion, a hexafluoroantimonic acid. Anionic, sulphur hexafluoride anion, phosphate anion, borofluoride anion, tetrafluoroborate anion, etc., or thiocyanate anion, benzenesulfonate anion, naphthalenesulfonate anion, naphthalene disulfonate anion, P-toluene Sulfonic acid anion 'alkyl sulfonate anion, benzene carboxylate anion, alkyl carboxylate anion, trihaloalkyl carboxylate anion, alkyl sulfate anion, trihaloalkyl sulfate anion, nicotinic acid anion, aspartic acid An organic acid anion such as an anion. The regulation obtained by the nitric oxide modulator according to the present invention is derived from the action of nitric oxide production of macrophage cells or vascular endothelial cells including microglia, and basically depends on the general composition as an active ingredient. The alkyl group (R) represented by Formula 1 has a cation portion of a trinuclear pentamethine-based cyanine dye having 2 to 4 carbon atoms, and thus the relative anion is not particularly limited as long as it can be directly applied to a living body. From the viewpoint of the strength of action, it is preferable to use iodine anion and chloride anion, and it is particularly preferable to use an iodine anion. In the case of the trinuclear pentamethine-based cyanine dye having a carbon number of 2 to 4 in the alkyl group (R) represented by the general formula 1, the pigment having a relative anion of a chlorine anion is compared with the pigment of the iodine anion. The solubility in an aqueous solvent is excellent. Therefore, when it is used in the form of oral administration, it is excellent in absorption and absorption rate in the living body. 有效 An active ingredient of a nitrogen oxide production regulator of the present invention. Specifically, 201219370, a trinuclear pentamethine-based cyanine dye represented by the following Chemical Formulas 1 to 8 can be exemplified. Further, it may be a compound having the same cationic portion as the dye represented by Chemical Formulas 4 and 5 and a relative anion from an iodine anion to a chlorine anion. From the viewpoint of adjusting the intensity of action of nitric oxide production from macrophage cells or vascular endothelial cells including microglia, it is preferable to use a dye represented by Chemical Formula 1 or 2, which is represented by Chemical Formula 1. The pigment is especially good. [Chemical 2] Chemical Formula 1

[Chemical 3] Chemical Formula 2

201219370 【化4】 Chemical Formula 3

[Chemical Formula 5] Chemical Formula 4

Ch3 ,CH \ ch3 h3c CH

[Chemical Formula 6] Chemical Formula 5

201219370 【化7】 Chemical formula 6

/CH2CH3 [Chemical 8] Chemical Formula 7

[Chemical Formula 9] Chemical Formula 8

CH2CH2CH2CH3 The so-called nitric oxide production regulation of the present invention means that the production of nitric oxide derived from macrophage cells or vascular endothelial cells including microglia cells is abnormal, and the local or systemic nitric oxide balance is abnormal. -10- 201219370 , inhibits the production of nitric oxide, reduces the amount of nitric oxide to a normal level, or reduces the production of nitric oxide from macrophage cells or vascular endothelial cells containing microglia, local or An abnormality in the balance of nitric oxide throughout the body causes the formation of nitric oxide to increase, and the amount of nitric oxide rises to a normal level. Next, the use of the nitric oxide production regulator obtained according to the present invention will be described. As described above, the trinuclear pentamethine-based cyanine dye as an active ingredient of the present invention has an adjustment from a small inclusion. The nature of nitric oxide production by macrophages or vascular endothelial cells of glial cells, even if directly applied to an organism, does not exhibit toxicity or serious side effects, and thus can be directly applied to an organism and can be used for The production of nitric oxide from macrophage cells or vascular endothelial cells containing microglia is regulated. When a nitrogen oxide production regulator of the present invention is applied to an organism, the formation of nitric oxide can be regulated, and the balance abnormality can be adjusted, so that various diseases or disorders caused by abnormal balance of nitric oxide can be presumed. Prevention is effective in terms of treatment. The various diseases or disorders caused by the abnormal balance of nitric oxide in the living body according to the present invention include, for example, sepsis or hemorrhagic shock, treatment of malignant tumor by interleukin, or cirrhosis. Pathological hypotension, rheumatoid arthritis, osteoarthritis, ulcerative colitis, tissue disorders after organ transplantation, transplant rejection, arteriosclerosis, viral infection, etc., myocarditis and cardiomyopathy, Tissues such as nephritis, pancreatitis, burns, inflammatory diseases, viral infections, cytopathic factors, or inflammatory reactions, such as spheroid nephritis, are caused by blood -11 - 201219370 endothelium (including microvascular endothelium) Injury or cell disorder (cell death), arterial hypertension, septicemia, heart disease, etc. The nitric oxide production-regulating agent of the present invention may be administered in a predetermined amount per day, once a day or in divided doses, in accordance with the degree of abnormality of the nitric oxide balance in the living body. The amount of administration per day is not particularly limited as long as it can obtain the desired effects of the present invention, and it is usually administered intravenously (including drip), subcutaneous, intradermal or intraperitoneal administration. The trinuclear pentamethine-based cyanine pigment represented by the above general formula 1 is 〇1 mg/kg body weight/day or more, more preferably 〇_1 to 20 mg/kg body weight/day, and particularly desirably 0.5 to 5 mg/kg body weight/day. When the dosage is less than 〇.〇1 mg/kg body weight/day, there is a case where the desired effect is not observed. Further, even if it is administered at a dose of 20 mg/kg body weight/day or more, an effect enhanced by the dose is not observed. In the case of oral administration, it is desirable to be 〇.1 mg/kg body weight/day or more, more desirably 0.5 to 100 mg/kg body weight/day, and particularly desirably 0.5 to 50 mg/kg body weight/day. Further, in consideration of the case where the nitric oxide production regulating agent of the present invention is used in the form of an oral preparation, the trinuclear pentamethine-based cyanine dye absorption used in the present invention is compared with subcutaneous or intraperitoneal administration. The absorption into the living body is low, and in order to obtain the desired effect of the present invention, it is necessary to increase the amount of administration more than the above-mentioned subcutaneous or intraperitoneal administration amount. In the case of the oral administration, in the case where the relative anion of the trinuclear pentamethine-based cyanine dye represented by the general formula 1 is a pigment of a chloride anion and a pigment of an iodine anion, it is considered to be absorbed into the living body. The absorption, or the intensity of the regulation of the formation of nitric oxide from macrophages or vascular endothelial cells containing microglia, such as macrophage cells or vascular endothelial cells, may be adjusted to vary the amount of administration. Further, the administration period of one of the nitrogen oxide production regulators of the present invention may be adjusted according to the degree of abnormality of the balance of nitric oxide. In an acute case, the degree of nitric oxide administered to the living body may be lowered, or the cause may be inferred. It is sufficient to improve or disappear the symptoms of abnormal nitric oxide balance. In the chronic case, it is hoped that the symptoms will continue to be observed until the symptoms are observed to disappear. In addition, the abnormality of the nitric oxide balance in the living body can be inferred from the disease in which the cause of the imbalance of nitric oxide is abnormal and the symptoms thereof, but it is desirable to directly confirm the degree of nitric oxide in the living body as much as possible. The method for quantifying nitric oxide for confirming the degree of nitric oxide in a living body, for example, after treating the blood taken according to the conventional method or the biological fluid derived from other tissues with nitric acid reducing enzyme, A method in which 3-diaminonaphthalene nucleates (fluoresces) a nitrite anion. Further, a commercially available nitrogen oxide analysis system (e.g., manufactured by Eicom Co., Ltd., trade name "NO system (ENO-20)") can be used for quantification. The nitric oxide production regulating agent of the present invention is usually provided in the form of a liquid preparation for parenteral administration or a dissolved powder at the time of use. Further, one of the nitrogen oxide production regulators of the present invention can also be provided in the form of an oral preparation. The dosage form of the oral preparation may, for example, be a powder, a granule, a tablet, a capsule, a syrup, a liquid or the like. In addition to an injection or an oral preparation, the nitric oxide production regulator of the present invention may be in the form of a paste or a sucking spray for lungs or the like, or may be in the form of a sputum preparation which is embedded in a subcutaneous body or the like. Further, one of the nitric oxide production regulating agents of the present invention further comprises a pharmaceutical agent in the form of a pharmaceutical unit of the formula -13 - 201219370. The medicinal agent of the present invention means that the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 as an active ingredient of the present invention contains, for example, a daily amount or an integral multiple (to 4 times) thereof or The approximate number (to 1/4) is 'a dosage form suitable for administration and capable of being physically separated. The nitric oxide production regulator of the present invention can also be used for animals other than humans such as livestock, poultry, and pets, for oxidizing macrophage cells or vascular endothelial cells derived from microglia. Nitrogen production is regulated. Next, a method for producing a nitrogen oxide generation regulator according to the present invention, a trinuclear pentamethine-based cyanine dye represented by the above general formula as an active ingredient of one of the nitrogen oxide generation regulators of the present invention, There is no limitation on the origin or the production method, and the desired amount can be obtained by a known method or a well-known method. For example, it is possible to obtain a desired amount by the method described in pages 240 to 30 of the publication of "Sensitivity Pigment", "Sensory Pigment" by Susukino Masahiro, October 17, 1997, and the publication of Industrial Book Co., Ltd. . In the case where the pigment is commercially available, it may be used as long as it is appropriately purified, and the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 may be used. Commercial products of anionic pigments include "NK-4" (a pigment having a carbon number of 2 in the alkyl group (R) in the general formula 1: a dye represented by the above chemical formula 1), and "NK-234" (general formula 1) The alkyl group (R) is a linear form, and has a carbon number of 3; the dye represented by the above Chemical Formula 2) and "NK-26" (the alkyl group (R) in the general formula 1 is linear) And the pigment having a carbon number of 4; the pigment shown in Tables 14 to 201219370 of the above Chemical Formula 3) (any of which is manufactured by Linyuan Biochemical Research Institute Co., Ltd.). Further, a commercially available product of a trinuclear pentamethine-based cyanine dye represented by the general formula 1 and a relative anion of a chloride anion may be exemplified by "NK-9" (alkyl group in general formula 1). a pigment having a carbon number of 2; a dye represented by the above Chemical Formula 6), and a dye having a carbon number of 3 in the general formula 1 wherein the sulfhydryl group (R) is linear and has a carbon number of 3; (a pigment represented by 7) and "NK-46" (a dye in which the alkyl group (R) in the general formula 1 is linear" and having a carbon number of 4; the dye represented by the above chemical formula 8) It is manufactured by Linyuan Biochemical Research Institute Co., Ltd.). The nitric oxide production regulating agent of the present invention may be used alone as a trinuclear pentamethine-based cyanine dye as an active ingredient, and can be generally tolerated in a blending formulation without departing from the scope of the present invention. One or two or more types of preparations which can be used in the pharmaceutical field, the quasi-drug field, the food field, or the cosmetics field can be provided. The pharmaceutically acceptable components may, for example, be additives such as pharmaceuticals, quasi drugs, and the like, excipients, disintegrating agents, lubricants, stabilizers, surfactants, preservatives (antibacterial agents), perfumes, and tackifiers. , antioxidants, chelation 6. Agents, vitamins, amino acids, aqueous solvents, saccharides, water-soluble polymers, pH adjusters, foaming agents, active ingredients for pharmaceutical or quasi-drugs, cosmetic raw materials, etc. One or two or more of these components may be appropriately combined and blended, and the target dosage form may be produced according to a usual method. Further, a nitrogen oxide production regulator of the present invention and a compound other than the trinuclear pentamethine-based cyanine dye used in the present invention are used as a nitric oxide production regulator effective for -15 to 201219370, or presumed. It is also advantageous to use a prophylactic or therapeutic agent for various diseases or disorders to which the nitric oxide production regulating agent of the present invention is applicable. These agents can be administered in the form of a mixture with the active ingredient of the present invention, or each of the preparations can be administered separately. The pigment to be used in the present invention may be blended into one of the nitric oxide production-modulating agents of the present invention as long as the composition of the subject preparation or the step of using it in the raw material stage to the completion of the product is considered. The method can be appropriately selected, for example, mixing, kneading, dissolving, melting, dispersing, suspending, emulsifying, demulsifying, infiltrating, crystallizing, spreading, coating, adhering, spraying, coating, injecting, dipping, curing. One or more methods, such as one or more. In the case of preparing a parenteral preparation such as an injection preparation, the nitric oxide production regulator of the present invention is usually dissolved in an aqueous solvent containing no pyrogen and administered to the skin, subcutaneous, intramuscular, or intravitreal ( Intrathoracic, intra-abdominal, etc., intravascular or other tissues or organs, so the form of the preparation may be a dry preparation or may be a liquid preparation. In the case of a dry preparation, it may be used by dissolving in an aqueous solvent such as purified water for injection, physiological saline, phosphate buffered physiological saline or glucose solution at the time of use. In addition, the powder component and the aqueous solvent may be separately stored in a plastic container which can be sealed separately, and the sealed portion may be opened, and the two components may be mixed and dissolved at the time of use. use. The liquid can be administered directly or can be added to an infusion solution, a perfusate, a peritoneal dialysate or the like. Further, in the case of preparing a releasable preparation or blending a lipophilic component, an amphiphilic solvent such as propylene glycol, polyethylene glycol or olive oil may be optionally used as an oily substrate or an emulsifier such as Tween 80. . Alternatively, the ribosome or the like can be optionally encapsulated and administered at -16 to 201219370. The aqueous solvent of the present invention generally means that water is an essential element, and if necessary, an alcohol such as ethanol, propanol or isopropanol, a ketone such as acetone or an ether such as diethyl ether or a dimethyl ester is blended therein. An aqueous solvent obtained by using one or two or more kinds of hydrophilic organic solvents containing a sulfur compound, such as a sulphur compound (hereinafter referred to as "DMS 0"). According to the liquid preparation obtained by the present invention, the aqueous solvent can be used alone as purified water for injection, physiological saline, Ringer's solution, etc., and can also optionally use purified water for injection and, for example, ethanol, propanol or isopropanol. A mixture of hydrophilic organic solvents that are physiologically acceptable, such as diethyl ether or DMSO. Alternatively, a pH adjuster such as lactic acid, hydrochloric acid, sulfonic acid, methanesulfonic acid, sodium hydroxide, potassium hydroxide, sodium hydrogencarbonate or a phosphate buffer solution may be added to adjust the pH to the solubility or stability of the formulated pigment. The highest pH is 6.5 to 8.0, preferably 6.8 to 7.4. In addition, the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 which is an active ingredient of the nitric oxide production-controlling agent of the present invention has a low stability to an aqueous solvent depending on the type of the relative anion or the like. In this case, it is desirable to use a dissolution-form preparation form at the time of use, and it is further desirable to use a pigment having an anion of an iodine anion. In the case of such a liquid preparation, the trinuclear pentamethine-based cyanine dye used in the present invention may be unstable due to dissolved oxygen or the like. Therefore, in this case, for example, such a dye solution may be used. The dissolved oxygen concentration can be lowered. The liquid composition can usually be prepared by the following steps: a step of dissolving the pigment in an aqueous solvent; and preparing the aqueous solvent to reduce the oxygen concentration in an atmospheric environment at normal temperature and pressure. The steps. In order to dissolve the pigment in -17-201219370 in an aqueous solvent, for example, if a predetermined amount of the pigment is added to an appropriate amount of the aqueous solvent, it is necessary to add and dissolve the aqueous solvent to the concentration of the dye. The degree is enough. Further, the dye may be dissolved in a hydrophilic organic solvent in advance, and then added to an aqueous solvent to be diluted until the concentration of the dye becomes a predetermined level. In order to make the dissolved oxygen concentration of the aqueous solvent lower than the concentration in an atmospheric environment at normal temperature and normal pressure, for example, a solution of the trinuclear pentamethine-based cyanine dye used in the present invention is prepared and stored under reduced pressure, or dissolved. A method in which oxygen of the dye solution is replaced by another gas or a method in which the dye solution is brought into contact with a deoxidizer is suitable. In order to replace the oxygen dissolved in the liquid composition with other gases, a relatively inert gas such as nitrogen or a rare gas such as helium, argon, neon or xenon may be foamed in the liquid composition. In order to reduce the oxygen concentration by using a deoxidizing agent, it is necessary to add, for example, L-ascorbic acid, L-ascorbyl stearate, sodium sulfite, sodium hydrogen sulfite, alpha thioglycerol, sodium edetate, and hydrochloric acid in an appropriate amount to the liquid composition. The reducing saccharide such as cystine, citric acid, lecithin, sodium thioglycolate, sodium sulphate, sodium metabisulfite, butyl hydroxyanisole, glucose or maltose may be used. These methods can be applied to a dye solution or an aqueous solvent which can be applied before the pigment is dissolved. In this case, the dissolved oxygen concentration in the aqueous solvent is usually set to 〇 4 ppm or less, and desirably 1 MPa or less. Further, in order to stabilize the trinuclear pentamethine-based cyanine dye used in the present invention, an appropriate amount such as tocopherol, carotene, histidine, tryptophan, tyrosine, methionine, Cysteamine, dopamine, thiocalcic acid, hypotaurine, bilirubin, cholesterol, quinoline, quercetin, musk-18-201219370 glucoside or its saccharide derivative, citavidin or a saccharide derivative, a catechin, an anthocyanin, a thiamine, or the like, which has an active component for eliminating singlet oxygen, or an adhesion promoter such as an alkyl cellulose, a carboxyvinyl polymer or a polytrimium. It is also advantageous to prepare a formulation by using a surfactant such as Triton X, Tween 80, deoxycholic acid or a salt thereof, or a bile acid or a salt thereof. The solution of the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 obtained in this manner may be stored in a state in which it is sealed in an appropriate container as long as the oxygen can be isolated. The material of the container is not particularly limited as long as it can maintain the liquid composition in principle, and the oxygen can be effectively isolated. It is desirable to use a brown bottle or a brown ampule or a plastic container as a light-shielding container. Usually, the liquid composition is sterilized by filtration sterilization before being dispensed into a glass ampoule, a sample bottle, a plastic container, or the like, or in the case of a glass ampoule or a sample bottle, after being dispensed and sealed, by autoclaving Sterilize, etc., however, depending on the application. The nitric oxide production regulating agent of the present invention produced in this manner has no serious side effects even if it is continuously used for a long period of time, and is a safe preparation. The present invention will be further described in detail below by experiments. <Experiment 1: Effect of trinuclear pentamethine-based cyanine dye on nitric oxide production> Primary culture of rat fetal brain cells widely used in an in vitro model produced as human nitric oxide The prepared microglia' was evaluated for the effect of the trinuclear pentamethine cyanine pigment on the production of nitric oxide in the organism -19-201219370 by the method shown below. The nitric oxide-inducing agent uses lipopolysaccharide (LPS) which is commonly used as an inducing model substance in an inflammatory state or sepsis. In addition, in the following experiment, any of the trinuclear pentamethine-based cyanine pigments was synthesized by the Linyuan Biochemical Research Institute Co., Ltd., and purified to a purity of 99% by mass or more. <Test sample> The relative anion (X-) of the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 is an iodine anion, and the alkyl group (R) has a carbon number of 1 (hereinafter referred to as "compound 1"), a dye having a carbon number of 2 (a dye represented by the above Chemical Formula 1; hereinafter referred to as "Compound 2"), and an alkyl group (R) are linear, and a pigment having a carbon number of 3 (a dye represented by the above Chemical Formula 2; hereinafter referred to as "Compound 3") and a dye having a carbon number of 4 (a dye represented by the above Chemical Formula 3; hereinafter referred to as "Compound 4") And a pigment having a carbon number of 6 (hereinafter referred to as "Compound 5"), which is dissolved in DMSO at a concentration of 5 mg/ml (sold by SIGMA, product number "D8418"), and then subjected to membrane filtration (sold by Millipore) , DMSO-resistant film of the product name "Millex-LG SLLG025SS". Before using this solution, use volume% bovine fetal serum (FBS) plus Dulbecco's MEM medium (sold by Rishui Pharmaceutical Co., Ltd., DMEM medium containing 20 mM glucose, 10 mM HEPES, 50μ8/ηι1 streptomycin and 50 The unit (U)/ml penicillin, hereinafter abbreviated as "10% FBS plus DMEM medium", was diluted to a final concentration of the compound to have the concentration shown in Table 1, and a test sample was prepared for testing. In addition, -20-201219370 was pre-confirmed. When the test standard dissolved in DMS 0 was diluted to the concentration used in the test with 10% FBS plus D Μ E Μ medium, the concentration of DMSO contained therein did not affect the following. Test system. <Modulation of microglial cells> Weiss rats (sold by Charles River, Inc., Japan, 10 weeks old, female) on the 16th to 18th day of pregnancy were dissected under ether anesthesia, and the uterus was After being immersed in a 70% by volume ethanol solution, it was immediately immersed in ice-cold sterilized Hanks buffered saline solution (HBSS), and the fetus was removed. The extracted fetus is immersed in fresh ice-cold sterilized HBSS, the brain is removed from the fetus under a stereoscopic microscope, and the cortex is partially excised, the medulla is removed, and shredded. Fine sections of the cerebral cortex were recovered, and 0.01% by mass of DNaseI (containing 0.01% by mass of DNaseI containing L-cysteine hydrochloride at 0.3 mg/m丨 was added to the cerebral cortex extracted from each mouse fetus. 20 units (U)/ml papaya enzyme solution (manufactured by Worthington Co., Ltd.) manufactured by Worthington Co., Ltd., 1 ml, was treated at 37 t for 30 minutes to disperse the cells. 0.5 ml of horse serum (manufactured by Cell Culture Tech Co., Ltd.) was added to the cerebral cortex extracted from the mouse fetus, and after the enzyme reaction was stopped, the cell suspension was recovered to a centrifuge tube used for centrifugation, and centrifuged at 800 rpm at room temperature. 3 minutes. After centrifugation, the supernatant was removed, and the pellets of the cells were added to Neurobasal medium (manufactured by Invitrogen; containing 25 μL of glutamic acid, 〇5 mM glutamic acid, 1% by mass of B27 supplement (Invitrogen) , 5 (^g/ml streptomycin and 50 units (U) / ml penicillin), by using the i〇mi liquid pipette, slowly sucking in a way that does not produce a - 21 - 19, 19,370 foam After suspending the cells, 'centrifuged at 80 °C for 3 minutes at room temperature. After removing the supernatant, 're-add the nerve cell basal medium again, using a 1 ml adjustable pipette' to slowly reduce the foam. The granules were inoculated to suspend the cells' and then passed through a sieve of a pore size of 4 μm (cell filtration cap tube 'Becton Dickinson Co., Ltd.) to prepare a cerebral cortical cell fraction. Further cerebral cortical cell isolates were prepared. Suspension in 1% FBS plus DMEM medium, and inoculation into a 150 cm 2 culture flask (manufactured by Becton Dickinson Co., Ltd.) in an amount of 3 χ 107 / 40 ml / flask 'Change medium with the same medium, in After about one month of culture, microglia were prepared by slightly vibrating the flask to recover the cells floating on the glial layer and the culture supernatant. <Measurement of LPS-inducible nitric oxide production amount from microglial cells> The above microglia cells were suspended in 10% FBS plus DMEM medium at 3x10 5 /ml, and inoculated into tissues at 100 μl/well. The 96-well microplate was cultured. After the culture for 1 day, the supernatant was removed, and any of the test samples diluted with 10% FBS plus DMEM medium was added as 50 μl/well, and further dissolved in 10% FBS after 1 hour. The lipopolysaccharide (LPS, manufactured by Difco) of DMEM medium was added to the well to which the test sample was previously added in an amount of 50 μl/well (the final concentration of LPS was 30 ng/ml). 24 hours after the addition of LPS, the Griess reagent was added to the culture supernatant at a dose of 50 μl/well, reacted, and chromogenic, and then measured by absorbance at 540 nm using a -22-201219370 multi-function microplate analyzer. The amount of nitric oxide released into the culture supernatant. The amount of nitric oxide produced when only LPS was added to the culture medium was determined to be 100, and the amount of nitric oxide produced in the case where any of the test samples were added before the addition of LPS was determined to be one. The rate of formation of nitrogen oxides (%) is disclosed in Table 1. In Table 1, the lower the nitric oxide production rate, the stronger the inhibition of nitric oxide production. In addition, in this experimental system, it is presumed that when the microglial cells are modulated, the stellate cells which are detached from the glial layer attached to the culture flask and slightly mixed into the microglial cells coexist even at 30 ng/ml of LPS. After 24 hours of culture, no nitric oxide was induced, so it was judged that the nitric oxide observed in this experiment was produced by microglia. •23- 201219370 [Table i] The concentration of the compound in the test sample (//g/ml) LPS addition (30 ng/ml) Nitric oxide production rate (%) — — 0 — — + 100 Compound 1 0.016 + 98 0.08 + 99 0.4 + 100 2 + 100 10 + 97 25 + 96 Compound 2 0.016 + 88 0.08 + 70 0.4 + 59 2 + 48 10 + 41 25 + 35 Compound 3 0.016 + 99 0.08 + 96 0.4 + 87 2 + 72 10 + 62 25 + 47 0.016 + 99 0.08 + 95 0.4 + 89 Redundant cut # 2 + 80 10 + 70 25 + 61 0.016 + 99 0.08 + 99 0.4 + 99 TL port cut 2 + 98 10 + 96 25 + 95 -24-201219370 It is apparent from the results of Table 1 that the carbon number of the alkyl group (R) of the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 is 1 (compound 1) and 6 (Compound 5), and the pigment having a relative anion of iodine anion, the concentration of nitric oxide derived from microglia induced by LPS was 95 to 100% at the concentration used in this test, and did not appear. Significant regulation of nitric oxide production. On the other hand, the alkyl group (R) of the trinuclear pentamethine-based cyanine dye represented by the general formula 1 has a carbon number of 2 to 4, and the pigment having an anion of an iodine anion (compounds 2 to 4). In the case of the pigment of Compound 2, at a concentration of p.〇8 pg/ml or more, the concentration-dependently, apparently exhibits a regulation of nitric oxide production from microglia induced by LPS, at 25 pg/ At a concentration of ml, the rate of production is reduced to 35 %. In the case of the dyes of the compound 3 and the compound 4, at a concentration of 2 pg/ml or more, the concentration-dependent expression of nitric oxide derived from LPS-induced small glial cells is exhibited at 25 pg/ml. The concentration rate was reduced to 47% and 61%, respectively. The degree of nitric oxide production regulation is compared with the concentration of each compound when the rate of production is reduced to 50%, then compound 2 (a pigment having an alkyl group of 2 (R) is 2) and a compound 3 (alkyl ( R) which is linear and has a carbon number of 3) exhibits a remarkable effect even at a low concentration, and is particularly remarkable with the compound 2. This result shows that the alkyl group (R) of the trinuclear pentamethine-based cyanine dye represented by the general formula 1 has a carbon number of 2 to 4, and its relative anion is an iodide anion pigment, especially an alkyl group (R). A dye having a linear shape and having a carbon number of 2 or 3, particularly a dye having a carbon number of 2 in the alkyl group (R), is an effective nitric oxide production regulator in the living body. -25- 201219370 <Experiment 2: Effect of the degree of nitric oxide in the organism to which the trinuclear pentamethine-based cyanine dye is administered> With the inflammatory reaction in the living body caused by infection or immune abnormality, etc. When nitric oxide is produced from macrophage cells or vascular endothelial cells containing microglia, it is rapidly metabolized to nitrous acid and nitric acid to increase the concentration of nitrogen oxides (NOx) in the blood. Then, with respect to the compounds 1 to 5 used in the experiment 1, the effects on the degree of nitric oxide in the living body when the compounds were directly administered to the living body were investigated. Namely, the same compounds 1 to 5 as used in Experiment 1 were dissolved in DMSO at a concentration of 5 mg/ml (sold by SIGMA, Inc., product number "D8418j", and membrane filtration was carried out (Millipore is sold under the trade name "". Millex-LG SLLG025 SS"). Before the solution was administered to mice, it was diluted to 50 pg/ml with phosphate buffered physiological saline (PBS) to prepare a test sample. A DMSO solution was used as a control group, and diluted with PBS to the same concentration as the test sample. <Test method> 36 BALB/c mice (sold by Charles River, Japan, 6-week old 'female) were randomly divided into 6 groups of 6 per group after body weight measurement (average weight 2 〇g) . For the mice of 6 groups of 6 mice, the administration amount of the compound was 500 ng/kg body weight and the method of tail vein administration (about 200 μl/only) was administered to the test containing any of the compounds 1 to 5. Sample. For the remaining 1 group, only DMSO solution (control group) was administered at 200 μl/to the tail -26-201219370 intravenously. After 15 minutes from the test sample or the control, the LPS used in the experiment 1 was dissolved in PBS for 6 mice of all the mice, and administered to the peritoneal cavity at a dose of 10 mg/kg (about 400 μl). /only). Six hours after the administration of LPS, blood was collected from the heart of each mouse, and a nitrate anion and a blood in the blood were measured by a commercially available nitrogen oxide analysis system (manufactured by Eicom Co., Ltd., trade name "NO system (ENO-20)"). The total concentration of nitrate anions was set to 1 血 when the blood concentration was administered to the control group, and the relative enthalpy of the blood concentration of the test sample was determined, and the nitric oxide production rate (%) was determined. Revealed in Table 2. In Table 2, the lower the nitric oxide production rate (%), the stronger the regulation of nitric oxide production. Incidentally, if nitric oxide is produced in the living body, it will be rapidly metabolized to nitrous acid and nitric acid. Therefore, in this experiment, the total concentration of nitrate anion and nitrite anion in the blood is determined as the amount of nitric oxide. . [Table 2] Test sample - Nitric oxide production rate (%) DMSO solution (control group) 100 Compound 1 100 Compound 2 58 Compound 3 74 Compound 4 86 Compound 5 100 It is apparent from the results of Table 2 that the above general formula 1 The alkyl group (R) of the trinuclear pentamethine-based cyanine dye has a carbon number of 1 (compound and 6 (compound 5), and its relative anion is an iodine anion pigment') in -27-201219370 Among the doses tested, the nitric oxide production rate of mice induced by LP S was 100%, which did not show significant regulation of nitric oxide production compared with the control group. When the alkyl group (R) of the trinuclear pentamethine-based cyanine dye represented by the general formula 1 is a carbon number of 2 to 4 and the relative anion is an iodine anion pigment (compounds 2 to 4), Nitric oxide production rates were 58%, 74%, and 86%, respectively, and it was confirmed that any of the compounds had a regulation of nitric oxide production. This regulation showed the strongest strength with Compound 2, and Compound 3 was second. As a result, with the compounds in Experiment 1, for from the LP S The presence or absence of the regulation of nitric oxide production in induced microglia is very consistent with the strength and weakness. Further, this result shows that compounds 2 to 4 are effective in the production of nitric oxide when administered to an organism. Conditioner. <Experiment 3: Effect of relative anion of trinuclear pentamethine-based cyanine dye on absorption absorbed into living body upon oral administration> Compounds 2 to 4 of these trinuclear pentadecyne Since the cyanine dye is highly hydrophobic, when it is administered orally, the absorption into the living body is low, and it is assumed that a sufficient blood concentration which can be expected by the desired effect of the present invention is transferred to the treatment. In the case of the target tissue, it was tested in the following manner to confirm whether the compounds are among the trinuclear pentamethine-based cyanine dyes, and the relative anion which is considered to have high solubility in the aqueous solvent is a chloride anion. The pigment is different in absorption absorbed into the living body when administered orally. That is, in Experiments 1 and 2, respectively, the compound 2 which is judged to be particularly strong in the regulation of nitric oxide production was orally administered (the carbon number of the alkyl group (R) in the general formula 1 of the above-mentioned -28-201219370 a trinuclear pentamethine-based cyanine dye having a relative anion of an iodine anion; a dye represented by the above Chemical Formula 1; or a carbon number of the alkyl group (R) in the above general formula 1 is 2, and is relatively The trinuclear pentamethine-based cyanine dye having an anion of a chlorine anion (the dye represented by the above Chemical Formula 6; hereinafter referred to as "Compound 6") is used to evaluate the absorption and transfer in a living body by the following evaluation method. Transferability to the organization. In addition, the organ to be evaluated is highly likely to be selected to be most relevant to the absorption, decomposition, and discharge of such a pigment, the liver 'kidney, and the presence of a blood-brain barrier (BBB). brain. <Test sample> The compound 2 and the compound 6 were suspended or dissolved in the mass/vol% of the polytriglucose so as to be 9.2 mg/ml (Linyuan Biochemical Research Institute Co., Ltd. sold, commodity The name "Japanese Pharmacopoeia polytriglyceride" was prepared as a test sample. <Test method> 24 ddY mice (sold by Japanese SLC company, 8 weeks old, male, average body weight 37 g) were randomly divided into 2 groups of 12 per group. From the administration of the test sample, the decidation was started 1 hour before the stagnation of sputum, and the test sample containing the compound 2 was orally administered to a group of 12 mice. The test sample containing Compound 6 was orally administered to the remaining 1 group of 12 using a gastric cannula at 0.4 ml/head. After the administration of the test sample 1, 2, 8 and -29-201219370 24 hours, 3 mice each randomly administered the test sample containing Compound 2 or Compound 6 were under ether anesthesia, Blood is taken from the large abdomen and EDTA is added as a blood coagulation inhibitor. Acetonitrile was added to the blood to be taken and added to 70% by volume, and the respective supernatants after centrifugation were concentrated under reduced pressure, dried, and re-dissolved in 70% by volume of an acetonitrile solution. , centrifuge again, and recover the supernatant. The supernatant was supplied to a high-performance liquid chromatography (HP LC) analysis under the following conditions, and was obtained in a chromatogram obtained by measuring an absorbance at wavelengths of 7 76 nm and 2 4 4 nm. The area of the peak was determined by the amount of Compound 2 or Compound 6. The mice after the blood collection were perfused with heart by using 15 ml of phosphate buffered saline (PBS(-)), and after the blood was released, the whole brain (brain parenchyma) containing the olfactory bulb, the liver, and the kidney were removed. After measuring the wet mass, the whole brain was measured, and twice the mass of distilled water was added and homogenized, and then acetonitrile was added to make it 70% by volume and mixed, and centrifuged at 12,00 rpm at 4 ° C. Minutes and recover the supernatant. The whole amount of the recovered supernatant was concentrated under reduced pressure, dried, and re-dissolved in 70% by volume of acetonitrile solution, and then centrifuged again, and the supernatant was recovered, in the same manner as blood, by HP. The amount of Compound 2 or Compound 6 was determined by LC method. After measuring the wet mass, the liver and kidney were stored frozen at -80 °C. The frozen liver and kidney were thawed in an ice-cold environment, and a mass of twice the amount of PBS (-) was added and homogenized, and then acetonitrile was added to make it 70% by volume and mixed, and the supernatant after centrifugation was recovered. The amount of Compound 2 or Compound 6 was determined by HPLC in the same manner as blood. In the case of any organ, the operations after homogenization are carried out under ice-cooling and shading. -30- 201219370 The results are shown in Table 3. <Quantification method of Compounds 2 and 6> Compounds 2 and 6 as standards were each diluted with a 70 volume% acetonitrile solution to 25 to 1,000 ng/tn1, and supplied to HPLC under the following conditions. System: HITACHI Co., Ltd. (trade name "La Chrom Elite") Pipe column: Cosmosil C8 (manufactured by Nacalai Tesque Co., Ltd., φ 4.6 mm x 250 mm, unprotected column) Mobile phase: 70 vol% acetonitrile, 0.2 vol% triethylamine, 0.2 Volume % Acetic acid flow rate: 0.8 ml / min Column temperature: 40 ° C Sample cooler: 5 ° C Sample injection amount: 0.1 ml Analysis time: 25 minutes Measurement wavelength: 776 nm, 254 nm -31 - 201219370 [Table 3 ]

Compound 6 j 24 hours csi 4586 1422 8 hours CO CNJ &lt;q 4878 2086 2 hours σ&gt; cd CO CO oi 5270 3922 1 hour if CM O) c\j 5110 4436 Compound 2 24 hours 〇〇CO iq 1910 S CJ&gt; 1 1 — i 8 hours CO S 1 2 hours C\4 CO CO CO CSJ CO O) CO 1 hour 5 〇C\j 00 CM in r· Time after the test sample is administered to the test sample (ng /ml blood) Brain tissue (ng/g tissue) Kidney (ng/g tissue) Liver (ng/g tissue) Kai 璨璁锴g flap M -32- 201219370 It is apparent from the results in Table 3 that it is contained in oral administration. In the case of the test sample of the compound 2, the concentration of the compound 2 in the blood reached a peak 2 hours after the administration, and the concentration at this time was 13.2 ng/ml, and then decreased, and the concentration was approximately 8 hours and 24 hours after the administration. On the other hand, in the case where the test sample containing the compound 6 is orally administered, the blood concentration of the compound 6 has reached a peak one hour after the administration, and the concentration is 206 ng/ml, and the compound 2 is administered. The peak concentration is more than 15 times. At approximately 8 hours and 24 hours, the sputum was approximately present, however this concentration was reduced to about one-third of the case of administration of Compound 2. Further, it has been observed that, in any of the whole brain, kidney, and liver, the case where the test sample containing the compound 6 is administered orally is compared with the case where the test sample containing the compound 2 is orally administered, Will happen in a shorter time. The case where the test sample containing the compound 2 was administered orally was compared with the case where the test sample containing the compound 6 was administered, the amount transferred to each organ was small, but in the case of administration for 24 hours, the transfer was performed. The amount shows a tendency to increase. This result shows that, in the case of administration of such a compound orally or via a digestive tract, the difference in relative anions has a significant influence on the amount of absorption and the rate of absorption absorbed into the living body. This result further shows that, when the compound is administered orally, the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 is compared when the concentration of the compound at the peak in the blood is compared and the peak time is reached. When the carbon number of the alkyl group (R.) is the same, the absorption and absorption rate of the absorption to the living body is superior to that of the pigment having the opposite anion of the chlorine anion and the pigment of the iodine anion, that is, the bioavailability rate. More excellent. In addition, when compared with the kidney or liver sputum, a difference in the amount is observed. However, since the compound to be administered is also detected in the brain, it can be presumed that the alkyl group is represented by the above general formula 1 ( R) a trinuclear pentamethine-based cyanine pigment having a carbon number of 2 to 4 and a relative anion of an iodine anion or a chloride anion, either of which can pass the blood-brain barrier (BBB) ° &lt;Experiment 4: General formula The difference in solubility in an aqueous solvent caused by the difference in relative anions in the trinuclear pentamethine-based cyanine dye represented by 1&gt; In Experiment 3, the trinuclear type five represented by the above general formula 1 The difference in relative anions of the acetylene-based cyanine pigment was observed to be significantly different in the absorption to the organism. The reason for this is because the difference in solubility between the two compounds in the digestive tract is caused by the difference in the anion. Therefore, an aqueous solvent is used in consideration of the environment inside the digestive tract, and a test for confirming the difference in solubility of the chromophors is performed. That is, according to the solubility test method of the Japanese Pharmacopoeia, the same compounds 2 to 4 as used in Experiment 1, the same compound 6 used in Experiment 3, and the alkyl group (R) are linear and have carbon. The pigment having a number of 3 and a relative anion of a chlorine anion (the dye represented by the above Chemical Formula 7; hereinafter referred to as "Compound 7"), the alkyl group (R) is linear, and its carbon number is 4, and the relative anion The pigment which is a chlorine anion (the dye represented by the above Chemical Formula 8; hereinafter referred to as "Compound 8") is micronized in advance using hydrazine. Based on the pre-reviewed solubility test of each compound for distilled water, each compound of the scale is about 2.5 to 20 mg. A transparent centrifuge tube (Spitz tube) with a volume of 15 or 50 ml is gradually added with a small amount of distilled water according to the solubility of each pigment. Stirring with a blender for 5 - 34 - 2012 19370 minutes ' After the addition of purified water, it was visually confirmed whether or not the insoluble matter (precipitate) remained within 30 minutes. In the case where insoluble matter was observed, a small amount of distilled water was further added' and stirred with a stirrer for 5 minutes. After adding distilled water, it was visually confirmed whether or not insoluble matter (precipitation) was observed within 30 minutes. The insoluble matter was observed to determine the minimum amount of distilled water in which insoluble matter could not be observed. The mass of each compound used in the test was divided by the minimum added mass of distilled water to determine the solubility. The results are disclosed in Table 4. In addition, the alkyl group (R) of the trinuclear pentamethine-based cyanine dye represented by the above general formula 1 is a linear form, and the pigment having a carbon number of 6 is also used as a pigment having a relative anion of an iodine anion ( The compound 5) and the chloride anion pigment (hereinafter referred to as "compound 9") were subjected to the same test. The results are disclosed in Table 4. The experiment was carried out 3 times for each pigment and the average enthalpy was obtained. [Table 4] Test sample alkyl (R) carbon number relative ion to water, solubility (mg / ml) Compound 2 2 Γ &lt;0. 01 Compound 6 2 C plant 8.3 Compound 3 3 " &lt; 0. 01 Compound 7 3 cr 20.3 Compound 4 4 Γ &lt;0. 01 Compound 8 4 cr 4.7 Compound 5 6 I- &lt; 0. 01 Compound 9 6 cr 0.01 It is apparent from the results of Table 4 that 'the general formula 1 above The solubility of the pigment of the alkyl group (R) of the trinuclear pentamethine-based cyanine dye (R) of 2 to 4 (chemical-35-201219370 compound 2 to 4) for distilled water, the relative anion of which is a chloride anion In any case, the pigment is higher than the pigment of the iodine anion, wherein the alkyl group (R) of the trinuclear pentaacetylene cyanine dye represented by the general formula 1 is linear and has a carbon number of 3 In the case of (Compounds 3 and 7), the difference in solubility in water is remarkable depending on the relative anion. In contrast, the alkyl group of the trinuclear pentamethine-based cyanine dye represented by the general formula 1 is used. (R) is a linear form of a pigment having a carbon number of 6 and a relative anion of a chloride anion (compound 9) The solubility of the pigment with respect to the anion of the iodine anion (Compound 5) was also low, and no difference in solubility caused by the difference in the relative anions was observed. This result can be compared with the result of Experiment 3: In the case where the alkyl group (R) of the trinuclear pentaacetylene cyanine dye has a carbon number of 2, when it is used as a pigment having a relative anion of a chlorine anion, it is absorbed into the living body when administered orally. The absorbability is superior to that of the anionic dye, and is well integrated. Further, the carbon number of the alkyl group (R) of the trinuclear pentaacetylene cyanine dye represented by the general formula 1 can be presumed. In the case of a dye of 3 or 4, the use of a dye having a relative anion of a chlorine anion is superior to the case of using a relative anion of an iodine anion, and is preferably absorbed into a living body when administered orally. When a dye having a carbon number of 6 or more of the alkyl group (R) of the dye represented by the general formula 1 is presumed, when the compound is orally administered, the carbon number of the alkyl group (R) is 2 to 4 Pigment In contrast, the difference in bioavailability caused by the difference in relative anions is not so large. -36- 201219370 &lt;Experiment 5: Safety of trinuclear pentamethine cyanine pigment&gt; The alkyl group of the trinuclear pentamethine-based cyanine dye (R) has a carbon number of 2 to 4', and the relative anion is an iodine anion or a chlorine anion. The compound is confirmed to be administered to a living body. For the safety, the test was carried out as follows: (1) Oral, single administration test &lt;test sample&gt; The same compound 2, compound 3, compound 4, and compound 6, as used in Experiment 4, respectively. Compound 7 and Compound 8 were dissolved in a carboxymethylcellulose solution suspended in 1 mass/vol% in a manner of 90 mg/ml to prepare test samples (test samples 1 to 6). The control group used a 1 mass/vol% carboxymethylcellulose solution. &lt;Evaluation Method&gt; The amount of 2,000 mg/kg body weight of the limit amount of the test using the caries was set to 1 dose based on the pharmaceutical toxicity test specification. 35 CD1 (ICR) mice (sold by Charles River, Japan, 6 weeks old, male) were randomly divided into 7 groups of 5 per group. All the mice were domesticated for 6 days in a solid feed (sold by Oriental Yeast, trade name "NMF"). The drinking water system uses a water supply bottle to allow it to freely consume tap water. The body weight of the mice that had been fasted for 1 day after the domesticated feeding was measured, and the test sample was smashed to 6 to be administered to the group of 5 for the use of the gastric cannula, respectively, in such a manner that the dose of the compound was 2,000 mg/kg of body weight. Oral administration (about 55mi/only). -37-201219370 For the remaining 1 group of 5, a gastric cannula was used for forced oral administration (0.5 ml/mouse) of 1 mass/vol% carboxymethylcellulose solution (control group). Further, CD5 (ICR) mice (sold by Charles River, 6 weeks old, female) were used, and the same test as in the case of using males was carried out. <Observation item> The test sample was administered. Thereafter, the state of the mouse was visually observed once a day for 14 days. The body weight was measured before the administration, and after the first, third, eighth, sixth, eighth, 10, 13 and 14 days after the administration, and the amount of the feed and the amount of water were measured. After the observation on the 14th day and the measurement of the body weight, the entire individual was anesthetized with ether, and the blood was excised, and the abnormality of the organ was confirmed by visual observation, and the minimum lethal dose was determined based on the observation results. The results are disclosed in Table 5. In addition, no difference was observed between the male and the female in the test results, so the results of the female and male are disclosed in Table 5. (2) subcutaneous, 28-day continuous administration test &lt;test sample&gt; Compound 2, Compound 3, Compound 4, Compound 6, Compound 7, and Compound 8 were dissolved in DMSO at a concentration of 5 mg/ml, respectively (SIGMA Corporation) After the sale, membrane filtration (using a DMSO-resistant film sold under the trade name "Millex-LGSLLG025SS" by Millipore Corporation) was carried out. Before the administration of the mouse, the compound solution was diluted with PBS by adding about 0.1 ml/kg to 5 〇mg/kg body weight to prepare a test sample (tested -38-201219370) Samples 1 to 6). &lt;Evaluation Method&gt; 35 CD 1 (ICR) mice (sold by Charles River, Japan, 6 weeks old, male) were randomly divided into 7 groups of 5 per group. All the mice were domesticated for 6 days in a solid feed (sold under the trade name "NMF" by the company of the company "Neriental Yeast"). The drinking water system uses a water bottle to make it free to ingest tap water. The body weight of the mice that had been fasted for one day after the domesticated feeding was measured, and one of the samples 1 to 6 was administered in such a manner that the compound was administered in an amount of 50 mg/kg of body weight. Give. For the remaining 1 group, 5 were subcutaneously administered to PBS (control group). &lt;Observation item&gt; The state of the mouse was visually observed once a day for a total of 42 days from the day when the test sample was administered. Before the start of the administration and after the start of the administration, the body weight was measured every two to three days, and the amount of the feed and the amount of water were measured. From the day of the start of the administration to the observation of the first day and the measurement of the body weight, the whole body was anesthetized with ether, and the blood was dissected, and the abnormality of the organ was confirmed by visual observation. The minimum lethal dose was determined based on the observation results. The results are shown in Table 5. (3) intraperitoneal, continuous administration for 2*70 days &lt; test sample&gt; Compound 2, Compound 3, Compound 4, Compound 6, Compound-39-201219370, and Compound 8 were respectively 5 mg/ml. After the concentration was dissolved in DMSO (sold by SIGMA Co., Ltd.), membrane filtration (a DMSO-resistant film sold under the trade name "Millex-LG SLLG025SS" by MilliP〇re Co., Ltd.) was carried out. At the time of use, each compound solution was diluted with PBS in such a manner that the dose of the test sample was about 0.5 ml/kg when the mice were administered about 2 ml/kg, and the test sample was prepared. Test samples 1 to 6). &lt;Evaluation method&gt; 35 CD1 (ICR) mice (sold by Charles River, Japan, 6 weeks old, male) were randomly divided into 7 groups of 5 per group. All the mice were domesticated for 6 days in a solid feed (sold by Oriental Yeast, trade name "NMF"). The drinking water system uses a water supply bottle to allow it to freely consume tap water. The body weight of the mice that had been fasted for one day after the domesticated feeding was measured, and one of the test samples 1 to 6 was administered to each of the five groups using a syringe at a dose of 0.5 mg/kg of body weight. In the abdominal cavity. For the remaining 1 group 5, PB S (control group) was administered to the abdominal cavity using a syringe. &lt;Observation item&gt; The state of the mouse was visually observed once a day for a total of 270 days from the day when the test sample was administered. The body weight is measured every 2 to 3 days before the start of the injection, and from the start of the injection, and the amount of the feed and the amount of water taken are measured. After the start of the administration, the observation of the 270th day and the measurement of the body weight were performed. The whole individual was anesthetized with ether, and the blood was dissected and dissected. The presence or absence of abnormality of the organ was confirmed by visual observation, and the minimum lethal dose was determined based on the observation results. . The results of -40-201219370 are disclosed in Table 5. [Table 5] Test sample administration route, etc., the minimum lethal dose (mg/kg body weight) Compound 2 Oral single 2000 &lt; Subcutaneous continuous administration for 28 days 50 &lt; Intra-abdominal continuous administration for 270 days 0.5 &lt; Compound 3 Oral single 2000 &lt; subcutaneous continuous administration for 28 days 50 &lt; intraperitoneal continuous administration for 270 days 0_5 &lt; compound 4 oral single 2000 &lt; subcutaneous continuous administration for 28 days 50 &lt; intraperitoneal continuous administration of 270 days 0.5 &lt; Compound 6 Oral single 2000 &lt; subcutaneous continuous administration for 28 days 50 &lt; intraperitoneal continuous administration for 270 days 0.5 &lt; Compound 7 oral single 2000 &lt; subcutaneous continuous administration for 28 days 50 &lt; intraperitoneal continuous administration for 270 days 0.5 &lt; Compound 8 Oral single 2000 &lt; subcutaneous continuous administration for 28 days 50 &lt; intraperitoneal continuous administration for 270 days 0.5 &lt; The carbon number of the alkyl (R) of the trinuclear pentamethine cyanine dye represented by the above general formula 1 is 2 to 4, the relative anions are 6 kinds of compounds of iodine anion or chloride anion. During the test, the administration dose, the administration route, and the administration period, no death or difference was observed in either case. In normal mice, there was no significant difference in weight, intake, and water intake from the control group. In addition, in the oral, single-dose test, no difference was observed between the male and female mice. Even in any of the tests, no abnormality of the organ was observed by visual inspection, so the case of any test -41 - 201219370 could not find the correct minimum lethal capacity. From this result, the trinuclear pentamethine-based cyanine dye which is an active ingredient of the present invention can be judged, and any of them is highly safe for long-term continuous administration to a living body even in a non-oral to oral manner. Compound. The invention is further illustrated by the following examples, but the invention is not limited by the examples. [Example 1] &lt;Liquid for injection&gt; 50 g of pyrogen-free aqueous crystal α,α-trehalose (manufactured by Hayashibara Co., Ltd.), 0.5 g of ascorbic acid, 1.25 were dissolved in 3 70 g of purified water for injection. g sodium hydrogencarbonate, a solution adjusted to pH 7.2, and 3 g of Tween 80 (trade name "Polysorbate 80" sold by Nippon Oil & Fat Co., Ltd.) and 120 mg of active compounds 2 to 4 in 177 g of purified water for injection and A solution of any one of the compounds 6 to 8 (all of which is manufactured by Iwahara Biochemical Research Institute Co., Ltd.) was separately mixed, filtered and sterilized, and then foamed with sterile nitrogen gas until the dissolved oxygen concentration became about 0.1 ppm. The ampoules were sealed under a stream of nitrogen under a nitrogen flow. Any of the articles is pyrogen-free and can be used as a nitric oxide generating regulator. In addition, this article can be used for pathological blood pressure reduction caused by abnormal balance of nitric oxide, tissue disorder after organ transplantation, transplant rejection, arteriosclerosis, myocarditis, cardiomyopathy, spheroidal nephritis Inflammatory diseases such as nephritis and pancreatitis; tissue damage or cell disorders (cell death) and arterial properties such as vascular endothelium (including microvascular endothelium-42-201219370) caused by viral infection, cytotoxic factors, and inflammatory reactions Prevention and treatment of diseases or disorders such as hypertension, septic disease, and heart disease. Further, one of the nitric oxide production regulators of the present invention can also be applied to animals other than humans. [Example 2] &lt;Dissolved powder at the time of use&gt; 30 g of purified maltose for injection (manufactured by Hayashibara Co., Ltd.), g5 g of ascorbic acid, lg sodium hydrogencarbonate, and pH were dissolved in 370 g of purified water for injection. A solution adjusted to 7.0, and a solution in which 200 mg of the compound 6 to 8 (any one of which is manufactured by Iwahara Biochemical Research Co., Ltd.) in which 100 mg of the active ingredient is dissolved is dissolved in 100 g of purified water for injection, and mixed separately After filtration and sterilization, the mixture was dispensed into a brown ampule at a rate of 1 〇ml, and lyophilized according to the usual method, and the ampoule was sealed under a nitrogen stream. Any one of the articles is pyrogen-free, and the purified water for injection is added to the ampoule to 2 to 10 ml of the physiological saline to dissolve it, and the method can be intravenously administered, subcutaneously administered, or intraperitoneally administered. This article can also be used as a nitric oxide production regulator for injection administration. In addition, this article can be used for pathological blood pressure reduction caused by abnormal balance of nitric oxide, tissue disorder after organ transplantation, transplant rejection, arteriosclerosis, myocarditis, cardiomyopathy, and spheroid nephritis Inflammatory diseases such as pyelitis and pancreatitis; tissue damage such as vascular endothelium (including microvascular endothelium) caused by viral infection, cytotoxic factors, and inflammatory reaction, or cell disorder (cell death), arterial hypertension Prevention, treatment of diseases or disorders such as stagnation, heart disease, etc. -43- 201219370. Further, one of the nitric oxide production regulators of the present invention can also be applied to animals other than humans. [Example 3] &lt;Dissolved powder at the time of use&gt; The solutions of Compounds 2 to 4 and Compounds 6 to 8 before encapsulation prepared in the same manner as in Example 1 were filtered and sterilized, respectively. It is freeze-dried, pulverized, and pulverized. Each of the powders is added to one of the accommodating portions of the light-shielding plastic container so that any of the freeze-dried powders of the compounds 2 to 4 and the compounds 6 to 8 is 1 mg/container in terms of the compound, and then sealed. The light-shielding plastic container has a structure in which the seal between the two housing portions is easily opened when the two housing portions are pressurized. The purified water for injection was dispensed into another accommodating portion of the same container in a manner of 25 ml/container, and then sealed. When the article is used, the powder is pressurized with a portion of the purified water, and the powder is mixed with purified water to be dissolved, and intravenously administered, intravenously administered, or intraperitoneally administered. This article can be used as a nitric oxide production regulator for injection administration. In addition, this article can be used for pathological blood pressure reduction caused by abnormal balance of nitric oxide, tissue disorder after organ transplantation, transplant rejection, arteriosclerosis, myocarditis, cardiomyopathy, spheroidal nephritis Inflammatory diseases such as pyelitis and pancreatitis, viral infections, cytopathic factors, inflammatory reactions, etc., tissue damage such as vascular endothelium (including microvascular endothelium) or cellular disorders (cell death), arterial hypertension, Prevention and treatment of diseases or disorders such as stagnation diseases and heart diseases. Further, -44-201219370 one of the nitric oxide production regulators of the present invention can also be applied to animals other than humans [Example 4] &lt; Oral use agent&gt; Compounds 2 to 4 for micronization using hydrazine in advance And 4 parts by mass of any of the compounds 6 to 8 (all of which are manufactured by Hayashi Biochemical Research Institute Co., Ltd.), uniformly mixing 4.25 parts by mass of sodium hydrogencarbonate, and aqueous crystalline α,α-trehalose (Linyuan 1.5 parts by mass of 0.25 parts by mass of magnesium stearate manufactured by the company, and ingots were prepared by ingoting into 〇5 g each according to the usual method. This article can be used as a nitric oxide production regulator for oral administration. In addition, this article can also be used for pathological blood pressure reduction caused by abnormal balance of nitric oxide, tissue disorder after organ transplantation, transplant rejection, arteriosclerosis, myocarditis, cardiomyopathy, spheroid pyelitis Inflammatory diseases such as nephritis and pancreatitis: tissue damage such as vascular endothelium (including microvascular endothelium) caused by viral infection, cytotoxic factors, and inflammatory reaction, or cell dysfunction (cell death), arterial hypertension Prevention and treatment of diseases or disorders such as diseases, stagnation diseases, and heart diseases. Further, a nitrogen oxide production regulator of the present invention can also be applied to animals other than humans. [Industrial Applicability] One of the nitric oxide production regulators of the present invention has a function of regulating the balance of nitric oxide in a living body, and even if administered to an organism, there is no toxicity or serious side effects of -45-201219370, and As a substance which is safe, it is possible to use an industry which produces pharmaceuticals, quasi-drugs and the like which are aimed at regulating the abnormality of the balance of nitric oxide in the living body. The present invention is an invention having significant significance in that the invention which exerts such a remarkable effect is greatly contributed to the field. -46-

Claims (1)

201219370 VII. Patent Application Range: 1. A nitric oxide production regulator comprising: a trinuclear pentamethine-based cyanine dye represented by the general formula 1 as an active ingredient, and one or more preparations Learn from other ingredients that can be tolerated, [Chemical 1] General Formula 1 X. (In general formula 1, R represents an alkyl group which may have a branch and a carbon number of 2 to 4, and X_ represents a suitable relative anion). 2. The nitric oxide production regulator according to the first aspect of the patent application, wherein among the trinuclear pentamethine-based cyanine dye represented by the general formula 1, the anion is an iodine anion or a chloride anion. (3) The oxidation-nitrogen generation regulator according to the second aspect of the patent application, wherein the trinuclear pentamethine-based cyanine dye represented by the general formula 1 is any one of Chemical Formulas 1 to 5, -47-201219370 2] Chemical Formula 1 /CH2CH3 [Chemical 3] Chemical Formula 2 /CH2CH2CH3 Chemical Formula 3 CH2CH2CH2CH3 -48- 201219370 Chemical Formula 4 [Chemical Formula 6] Chemical Formula 5 4. The nitric oxide production regulator according to any one of claims 1 to 3, which is a preparation which is dissolved and used at the time of use. 5. A nitrous oxide production regulator according to any one of claims 1 to 4, which is administered orally or by injection. -49- 201219370 III. Abstracts of English: IV. Designated representative circle: (1) The representative representative of the case is: No (2) The representative symbol of the representative figure is a simple description: No. 5. If there is a chemical formula in this case, please reveal The chemical formula that best shows the characteristics of the invention: Chemical 1 N* X.