JP4355578B2 - Medicaments containing riboflavin compounds - Google Patents

Description

表１に示すように、５’−ＦＭＮ−ＮａとＡＰＣを併用して投与することにより、極めて高い生存率を示すことが分かった。
〔実施例２：エキソトキシン誘発ショックマウスモデルに対する効果〕
まず、一群各１０例の雄ＢＡＬＢ／ｃマウス（６週齢）にＳＥＢ０．７５ｍｇ／ｋｇを腹腔内投与するとともに、Ｄ−ガラクトサミン１．８ｇ／ｋｇを腹腔内投与して、エキソトキシン（ＳＥＢ）誘発ショックマウスを作成した。
ＳＥＢの投与６時間後から、生理食塩液に溶解し濾過滅菌した５’−ＦＭＮ−Ｎａ（投与量：２０ｍｇ／ｋｇ）と、生理食塩液に溶解し濾過滅菌したＡＰＣ（投与量：３００ｕｎｉｔｓ／ｋｇ）を各群についてそれぞれ静脈内に６時間持続投与した。また、生理食塩液に溶解し濾過滅菌した５’−ＦＭＮ−Ｎａ（投与量：２０ｍｇ／ｋｇ）とＡＰＣ（投与量：３００ｕｎｉｔｓ／ｋｇ）とを混合して静脈内に６時間持続投与した。コントロールグループには、薬剤の代わりに３．５ｍｌの生理食塩液を静脈内に６時間持続投与した。
ＳＥＢを投与してから７日間経過後におけるマウスの生存率を調べた。薬効はＳｔｅｅｌ検定により判定した。その結果を表２に示す。

表２に示すように、本実施例においても、５’−ＦＭＮ−ＮａとＡＰＣを併用して投与することにより、極めて高い生存率を示すことが分かった。
〔実施例３：エンドトキシン誘発ショックマウスモデルに対する効果〕
実施例１と同様にして一群各１０例のＬＰＳ誘発ショックマウスを作成した。
ＬＰＳの投与６時間後から、生理食塩液に溶解し濾過滅菌した５’−ＦＭＮ−Ｎａ（投与量：１０ｍｇ／ｋｇ）と、生理食塩液に溶解し濾過滅菌したバリン（投与量：２００ｍｇ／ｋｇ）を各群についてそれぞれ静脈内に６時間持続投与した。また、生理食塩液に溶解し濾過滅菌した５’−ＦＭＮ−Ｎａ（投与量：１０ｍｇ／ｋｇ）とバリン（投与量：２００ｍｇ／ｋｇ）とを混合して静脈内に６時間持続投与した。コントロールグループには、薬剤の代わりに３．５ｍｌの生理食塩液を静脈内に６時間持続投与した。
ＬＰＳを投与してから７日間経過後におけるマウスの生存率を調べた。コントロールと５’−ＦＭＮ−Ｎａ投与グループについては、各１０例につき２回実験を行った。薬効はＳｔｅｅｌ検定により判定した。その結果を表３に示す。

表３に示すように、５’−ＦＭＮ−Ｎａとバリンを併用して投与することにより、極めて高い生存率を示すことが分かった。
〔実施例４：エンドトキシン誘発ショックマウスモデルに対する効果〕
実施例１と同様にして一群各１０例のＬＰＳ誘発ショックマウスを作成した。
ＬＰＳの投与６時間後から、生理食塩液に溶解し濾過滅菌した５’−ＦＭＮ−Ｎａ（投与量：１０ｍｇ／ｋｇ）と、アミノ酸注射液（投与量：３．５ｍｌ／マウス）を各群についてそれぞれ静脈内に６時間持続投与した。また、５’−ＦＭＮ−Ｎａ（投与量：１０ｍｇ／ｋｇ）とアミノ酸注射液（投与量：３．５ｍｌ／マウス）とを混合して静脈内に６時間持続投与した。コントロールグループには、薬剤の代わりに３．５ｍｌの生理食塩液を静脈内に６時間持続投与した。
ＬＰＳを投与してから７日間経過後におけるマウスの生存率を調べた。コントロールと５’−ＦＭＮ−Ｎａ投与グループについては、各１０例につき２回実験を行った。薬効はＳｔｅｅｌ検定により判定した。その結果を表４に示す。

表４に示すように、５’−ＦＭＮ−Ｎａとアミノ酸注射液を併用して投与することにより、極めて高い生存率を示すことが分かった。
産業上の利用可能性
本発明の医薬組成物は、免疫賦活剤又は感染防御治療剤、高サイトカイン血症、全身性炎症反応症候群、敗血症又は敗血症性ショックの予防又は治療剤等として有用である。Technical field
The present invention relates to riboflavin, riboflavin derivatives, or pharmacologically acceptable salts thereof (hereinafter sometimes referred to as riboflavin compounds), protein C, activated protein C, or derivatives thereof (hereinafter referred to as protein C systems). And / or a pharmaceutical composition containing valine as an active ingredient.
Related technology
An inflammatory reaction is involved in many diseases, and it has been found that the inflammatory reaction has an important influence not only in so-called inflammatory diseases but also in Alzheimer's dementia, heart disease and the like.
Among diseases involving an inflammatory reaction, a systemic inflammatory response syndrome, which is a pathological condition accompanied by systemic inflammatory signs, is particularly important as a sign for grasping a reaction of an invaded organism. In addition, in the state of this systemic inflammatory response syndrome, when symptoms progress or complications occur, adult respiratory distress syndrome (ARDS), disseminated intravascular blood coagulation (DIC), disseminated intravascular blood coagulation (DIC). coagulation), multiple organ failure (MOF) and other organ dysfunction syndrome (MODS), symptom of consciousness disorder, dyspnea, hypotension, etc. . Causes of this systemic inflammatory response syndrome include various invasions such as trauma, burns, pancreatitis, and surgery in addition to infections.
Conventionally, it has been reported that the administration of activated protein C to severe septic patients reduces the mortality rate (Efficacy and Safety of Recombinant Human Activated Protein Cever Severe in Japan 4). 699-709 March 8, 2001 Number 10, Gordon R. Bernard, MD et al).
On the other hand, the present inventors have obtained the knowledge that riboflavin compounds have an immune function stimulating action (Japanese Patent Laid-Open No. 5-201864), and that riboflavin compounds are used as a preventive agent for toxin shock. Knowledge that it is useful has been obtained (Japanese Patent Laid-Open No. 10-29941).
Disclosure of the invention
However, it is desired to provide an immunostimulant or an infection-protecting therapeutic agent that has a higher drug efficacy and that is further superior in improving the prognosis.
Accordingly, an object of the present invention is to provide a pharmaceutical composition and the like that are more excellent as an immunostimulant or an anti-infection therapeutic agent.
As a result of intensive studies, the present inventors have found that administration of a riboflavin compound and a protein C compound in combination can provide an extremely excellent immune function activation effect and infection protection treatment effect.
That is, the present invention provides a pharmaceutical comprising as an active ingredient at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof, and at least one of protein C, activated protein C, or a derivative thereof. A composition is provided.
This is considered to be caused by some synergistic action in vivo by administering a riboflavin compound and a protein C compound in combination. Since it is administered in combination, it is possible to obtain an excellent immune function activation effect and infection protection treatment effect while reducing the dose of each of the riboflavin compound and protein C compound compared to the case of single administration.
“At least one” means that any one or more of riboflavin, riboflavin derivatives, or pharmacologically acceptable salts thereof may be used as an active ingredient, and protein C, activity It means that any one or a plurality of modified protein C or derivatives thereof may be used as active ingredients.
The “pharmaceutical composition” is not limited to the case where each active ingredient is physically mixed, but includes the case where each active ingredient is dissolved in a solution.
The term “active ingredient” means a component that exhibits pharmacological activity in vivo, or a component that does not exhibit pharmacological activity alone, but enhances the pharmacological activity of other components when used in combination or coexists with other components.
In addition, as a result of intensive studies, the present inventors have found that an extremely excellent immune function activation effect can be obtained by administering a riboflavin compound and valine in combination or in combination.
That is, the present invention provides a pharmaceutical composition comprising as an active ingredient at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof, and valine.
This is considered to be caused by some synergistic action in vivo by administering a riboflavin compound and valine in combination or mixed.
“At least one” means that any one or more of riboflavin, riboflavin derivatives, or pharmacologically acceptable salts thereof may be used as an active ingredient.
The meanings of “pharmaceutical composition” and “active ingredient” are the same as described above. For example, valine may be added as an additive.
In the present invention, (1) at least one of riboflavin, riboflavin derivative, or a pharmacologically acceptable salt thereof and (2) valine are uniformly mixed in powder or solution, or contains (1) The powder, granule and powder containing (2) and the powder, granule and powder containing (2) may be mixed and tableted and filled as necessary to form tablets and capsules.
In the above, the riboflavin compound and valine are preferably contained in a weight ratio (riboflavin compound: valine) of 1: 1 to 1: 1000, more preferably 1: 5 to 1: 100. Preferably, it is contained at 1: 5 to 1:50.
In addition, as a result of intensive studies, the present inventors have found that an extremely excellent immune function activation effect can be obtained by administering a riboflavin compound, a protein C compound and valine in combination.
That is, the present invention comprises at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof, at least one of protein C, activated protein C, or a derivative thereof, and valine. A pharmaceutical composition is provided.
The pharmaceutical composition of the present invention is useful as an immunostimulant or an infection protection therapeutic agent.
“Immunostimulation” means enhancing the immune function of a human or animal.
The pharmaceutical composition of the present invention is particularly useful for the prevention or treatment of hypercytokinemia.
Hypercytokinemia is a disease accompanied by a condition in which the concentration of cytokines in the blood is high, such as Alzheimer's dementia, Castleman's disease, rheumatoid arthritis, osteoarthritis, multiple sclerosis, Behcet's disease, lupus erythematosus, alloteme Sclerosis, heart disease, atrial myxoma, Crohn's disease, ulcerative colitis, inflammatory colitis, gout, contact dermatitis, psoriasis, pulmonary fibrosis, acute glomerulonephritis, mesangial proliferative nephritis, atherosclerosis Shock, nodular arteriosclerosis, vasculitis, bronchial asthma, chronic gingivitis, periodontal disease, hemorrhagic shock, traumatic shock, brain tumor, malignant tumor, atopic dermatitis, allergic rhinitis, allergic Allergies such as dermatitis, sepsis, septic shock, multiple organ failure, multiple organ failure after sepsis, systemic inflammatory response syndrome, insulin dependence Diabetes, uveitis, chronic inflammation, blood disease, collagen disease, preoperative and postoperative infections, malaria disease, amyotrophic lateral sclerosis, idiopathic thrombocytopenic purpura, myasthenia gravis, hay fever Autoimmune diseases such as cerebral infarction and myocardial infarction, ischemia / reperfusion injury, photosensitivity, empyema, uterine empyema, otitis media, peritonitis, infective endocarditis, endocarditis, acute renal failure, acute Examples include liver damage and diarrhea.
The pharmaceutical composition of the present invention is particularly useful for the prevention or treatment of systemic inflammatory response syndrome.
Systemic inflammatory response syndrome, which is one of hypercytokinemia, is characterized by SIRS (systemic intensity response syndrome) in which inflammatory cytokines predominate in blood and inflammatory reactions occur, and anti-inflammatory cytokines predominate in blood It is thought that there are two pathological conditions, CARS (compensatory anti-inflammatory response syndrome) in which an anti-inflammatory reaction (for example, severe infection) occurs. By administering the pharmaceutical composition of the present invention, SIRS or CARS symptoms can be prevented or treated.
“Prevention or treatment of systemic inflammatory response syndrome” of the present invention refers to adult respiratory distress syndrome (ARDS) caused by progression of the above-mentioned SIRS or CARS symptoms, disseminated intravascular blood coagulation (DIC), multiple organs This also includes prevention or treatment of dysfunction (MODS) such as failure (MOF), and the pharmaceutical composition of the present invention is extremely useful for prevention or treatment of pathological conditions in which SIRS or CARS has progressed.
The pharmaceutical composition of the present invention is particularly useful for the prevention or treatment of sepsis or septic shock.
Sepsis refers to a state in which microorganisms themselves, exotoxins excreted by microorganisms, or endotoxins excreted when microorganisms are destroyed are spread throughout the body by infecting microorganisms.
Septic shock is the result of the progression of sepsis, the dysfunction of one or more of the internal organs such as heart, lung, liver, kidney, spleen, brain, spinal cord, or the result of organ dysfunction It means a state of symptom such as weakness, dizziness, difficulty in standing, low blood pressure, hypothermia, arrhythmia, ventricular fibrillation, dyspnea, hypothermia, convulsions, turbidity, unconsciousness.
The riboflavin derivative or a pharmacologically acceptable salt thereof includes flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, riboflavin sodium phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leuco A flavin phosphate, leucoflavin mononucleotide, or leucoflavin adenine dinucleotide, or a pharmaceutically acceptable salt thereof is preferable.
Protein C is an inactive glycoprotein dependent on vitamin K produced in the liver, and is one of important blood coagulation factors.
The activated protein C is a serine protease in which protein C is activated by a thrombin-thrombomodulin complex or the like on vascular endothelial cells. Activated protein C decomposes both blood coagulation factor Va and factor VIIIa to specifically inactivate them and has profibrinolytic activity and anticoagulant activity.
Although it does not specifically limit as protein C, activated protein C, or these derivatives, Natural type human protein C, recombinant human protein C, natural type human activated protein C (For example, dry concentrated human activated protein C) ), Recombinant human activated protein C (for example, drotrecogin α) and the like. Examples of the recombinant human protein C and its activated protein C include European Patent Application No. 88311421.7 (EP0319312A2), European Patent Application No. 91301450.2 (LIN derivatives and FLIN derivatives), and European Patent Application No. 91301446.0. (Q313 and Q329), European Patent Application No. 88312201.2 (F167), Characterization and novel purification of recombinant human protein cell lines, S.M. C. Betty Yan et al, BIOTECHNOLOGY Lol. 8 July 1990 and the like.
Valine is not necessarily used in the form of a free amino acid, and may be in the form of an inorganic acid salt, an organic acid salt, an ester, or an N-substituted product.
The subject to which the pharmaceutical composition according to the present invention is administered is a human or an animal. The medicament of the present invention is particularly useful for prevention or treatment in humans.
In the present invention, animals refer to industrial animals, companion animals, and experimental animals. Industrial animals include domestic animals such as cattle, horses, pigs, goats, and sheep, poultry such as chickens, ducks, quails, turkeys, and ostriches, and fish such as yellowtail, yellowtail, red sea bream, red sea bream, carp, rainbow trout, and eels. It is an animal that is needed to do. Companion animals refer to companion animals such as dogs, cats, marmosets, birds, hamsters, and goldfish, and experimental animals refer to rats, guinea pigs, beagle dogs, miniature pigs, rhesus monkeys, and cynomolgus monkeys. This refers to animals used for research in fields such as agriculture and pharmacy.
The dosage form of the pharmaceutical composition according to the present invention is not particularly limited, and varies depending on the disease state and its progress, and other conditions, but is an injection, tablet, granule, powder, fine granule, capsule, pill, Or it is preferable to contain in the form of an oral solution (including syrup).
When administered in the form of an injection, it can be administered intravenously, intraperitoneally, intramuscularly, subcutaneously, transdermally, intraarticularly, within the bursa, intravesicular, intraperiosteally, sublingually, orally. In particular, intravenous administration or intraperitoneal administration is preferred. Intravenous administration may be either drip administration or bolus administration (bolus).
The dose (active ingredient amount) is not particularly limited, and the pathological condition and its progress, other conditions (type of subject to be administered, symptoms, age, weight, sex, complications, administration time, administration method, dosage form, sensitivity) Depending on the difference, etc., when the injection is administered intravenously, the riboflavin compound is preferably administered at 0.01 to 8 mg / kg / h for several minutes to one week. More preferably, it is administered at -1 mg / kg / h for 6 hours to 4 days. The protein C compound is preferably administered at 1-1000 μg / kg / h for several minutes to one week, more preferably at 10-400 μg / kg / h for 6 hours to 4 days. Valine is preferably administered at 0.01 to 100 mg / kg / h for several minutes to one week, more preferably at 1 to 50 mg / kg / h for 6 hours to 4 days.
In the case of intravenous bolus administration, the riboflavin compound is 0.1 to 50 mg / kg, preferably 0.3 to 20 mg / kg, and the protein C compound is 6.01 to 10 mg / kg, preferably Is 0.1 to 1 mg / kg, and valine is 1 to 2000 mg / kg, preferably 10 to 500 mg / kg.
When administered intraperitoneally, the riboflavin compound is 0.1 to 50 mg / kg, preferably 0.3 to 20 mg / kg, and the protein C compound is 0.01 to 10 mg / kg, preferably 0. 0.1-1 mg / kg, and for valine, 10-500 mg / kg.
When administered intramuscularly, the riboflavin compound is 0.1 to 50 mg / kg, preferably 3 to 20 mg / kg, and the protein C compound is 0.01 to 10 mg / kg, preferably 0.1. It is ˜1 mg / kg, and for valine it is 10 to 500 mg / kg.
When administered orally, the riboflavin compound is 1-1000 mg / kg, preferably 10-500 mg / kg, and the protein C compound is 0.1-100 mg / kg, preferably 1-50 mg / kg. Yes, for valine it is 10-2000 mg / kg.
In the present invention, a riboflavin compound and a protein C compound and / or valine are administered in combination, so that each dose can be reduced as compared with the case of single administration.
The pharmaceutical composition according to the present invention may be administered as it is, or an injection (for intravenous administration (for infusion or bolus administration) or intraperitoneal administration) by a known method with the addition of commonly used formulation additives. , Intramuscular administration, subcutaneous administration, etc.), oral preparations (tablets, granules, powders, fine granules, capsules, pills, oral solutions, syrups, etc.), transdermal preparations, eye drops, eye drops A nasal agent, a suppository, an inhalant (aerosol, powder inhaler, liquid inhaler, etc.) and an external preparation (ointment, cream, liquid, etc.) can be used. It can also be mixed with food, feed, drinking water, etc.
When producing injections, add solubilizers, pH adjusters, buffers, suspending agents, antioxidants, preservatives, tonicity agents, etc., as necessary, and manufacture by conventional methods. can do. Intravenous solutions may be composed of a solubilizing agent or the like as an infusion solution, or instillations may be composed of an infusion solution obtained by adding a pharmacologically acceptable modification thereto. These injections can be administered intravenously, intraperitoneally, intramuscularly, subcutaneously, and the like. Alternatively, it may be lyophilized to obtain a freeze-dried preparation that is dissolved at the time of use.
When manufacturing solid preparations for oral use, excipients, binders, disintegrants, lubricants, colorants, flavoring agents, antioxidants, solubilizers, pills, etc. are added as necessary. However, it can be made into tablets, coated tablets, granules, powders, fine granules, capsules, pills and the like by conventional methods.
Although it does not specifically limit as a solubilizing agent, For example, physiological saline, phosphate buffered saline, lactated Ringer's solution, polyoxyethylene hydrogenated castor oil, polysorbate 80, nicotinamide, polyoxyethylene sorbitan monolaur Rate, macrogol, castor oil fatty acid ethyl ester and the like.
Although it does not specifically limit as a pH adjuster and a buffering agent, For example, an organic acid or an inorganic acid and / or its salt, sodium hydroxide, meglumine etc. are mentioned.
The suspending agent is not particularly limited, and examples thereof include methyl cellulose, polysorbate 80, hydroxyethyl cellulose, gum arabic, sodium carboxymethyl cellulose, polyoxyethylene sorbitan monolaurate, and tragacanth powder.
Examples of the antioxidant include, but are not limited to, ascorbic acid, α-tocopherol, ethoxyquin, dibutylhydroxytoluene, butylhydroxyanisole, and the like.
Examples of the preservative include, but are not limited to, methyl paraoxybenzoate, ethyl paraoxybenzoate, sorbic acid, phenol, cresol, chlorocresol, and the like.
Although it does not specifically limit as an isotonizing agent, For example, sodium chloride etc. are mentioned.
The excipient is not particularly limited. For example, starch, corn starch, dextrin, glucose, lactose, sucrose, sugar alcohol, hydrogenated oil, mannitol, erythritol, xylitol, crystalline cellulose, anhydrous silicic acid, calcium silicate, diphosphoric acid Examples thereof include calcium hydrogen.
The binder is not particularly limited. For example, polyvinylpyrrolidone, ethylcellulose, methylcellulose, pregelatinized starch, gum arabic, hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, propylene glycol, sodium polyacrylate, polyvinyl Alcohol etc. are mentioned.
Although it does not specifically limit as a disintegrating agent, For example, crospovidone, low substituted hydroxypropyl cellulose, bridge | crosslinking type sodium carboxymethylcellulose, carboxymethylcellulose, calcium carbonate, carboxymethylcellulose calcium etc. are mentioned.
The lubricant is not particularly limited, and examples thereof include magnesium stearate, talc, calcium stearate, sodium stearyl fumarate, polyethylene glycol 6000, and the like.
In the case of tablets, granules, and powders, a film coating such as hydroxypropylmethylcellulose may be applied as necessary.
In the case of producing an oral liquid, it can be produced by a conventional method by adding a colorant, a flavoring agent, an antioxidant, a solubilizing agent, etc. as necessary.
Further, as a result of earnest research, the present inventors have administered a riboflavin compound and a protein C compound simultaneously or at the same time, administered a riboflavin compound and valine at the same time or at another time, or a riboflavin system. It has been found that an extremely excellent immune function-stimulating action can be obtained also by administering a compound, a protein C compound and valine simultaneously or separately.
That is, the present invention is a pharmaceutical composition containing at least one of riboflavin, riboflavin derivatives, or pharmacologically acceptable salts thereof, wherein at least protein C, activated protein C, or derivatives thereof are contained. It is intended to provide a pharmaceutical composition for administration at the same time or at the same time as one or at the same time or at the same time as a valine.
“Simultaneous or separate administration” means when both pharmaceutical compositions are prepared separately and administered simultaneously, or when one pharmaceutical composition is administered and the other pharmaceutical composition is administered after a certain period of time It means to include. In this case, the administration forms of both pharmaceutical compositions may be the same or different, and the administration sites may be the same or different.
The present invention also includes a container containing a pharmaceutical composition containing at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof as an active ingredient, protein C, activated protein C, or these And a container containing a pharmaceutical composition containing at least one derivative as an active ingredient.
The present invention also includes a container containing a pharmaceutical composition containing at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof as an active ingredient, and a pharmaceutical composition containing valine as an active ingredient. And an apparatus including the container.
The “apparatus” may be, for example, a treatment kit including the container, an infusion apparatus including the container, or the like.
The present invention also relates to a pharmaceutical composition comprising at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof as an active ingredient, and at least protein C, activated protein C, or a derivative thereof. The present invention provides a pharmaceutical comprising a pharmaceutical composition comprising one as an active ingredient, wherein the pharmaceutical compositions are held in a state where they are not in contact with each other.
The present invention also provides a pharmaceutical composition comprising a pharmaceutical composition comprising as an active ingredient at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof, and a pharmaceutical composition comprising valine as an active ingredient. Then, a medicament is provided in which the pharmaceutical compositions are held in a state where they are not in contact with each other.
“Pharmaceuticals held in a state where the pharmaceutical compositions are not in contact with each other” means, for example, that each pharmaceutical composition in an injection is partitioned and filled in a syringe and / or in an injection bag via a partition wall, etc. In other words, the case where each pharmaceutical composition is partitioned and filled in a capsule tablet via a partition wall or the like.
In addition, the present invention provides at least one of riboflavin, riboflavin derivatives, or pharmacologically acceptable salts thereof, protein C, activated protein C, or It provides for the use of at least one of these derivatives. In other words, the present invention relates to an immunostimulator by using at least one of riboflavin, riboflavin derivatives, or pharmacologically acceptable salts thereof, and protein C, activated protein C, or derivatives thereof. Alternatively, the present invention provides a method for producing an infection-protecting therapeutic agent.
The present invention also provides the use of valine and at least one of riboflavin, riboflavin derivatives, or pharmacologically acceptable salts thereof for producing an immunostimulatory agent or an anti-infective therapeutic agent. . In other words, the present invention provides a method for producing an immunostimulatory or anti-infective therapeutic agent by using at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof, and valine. .
The present invention also relates to a pharmaceutical composition comprising at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof as an active ingredient, and at least protein C, activated protein C, or a derivative thereof. The present invention provides an immunostimulatory agent or an infection-protecting therapeutic agent comprising a combination of a pharmaceutical composition containing one as an active ingredient.
The present invention also includes a pharmaceutical composition comprising at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof as an active ingredient, and a pharmaceutical composition comprising valine as an active ingredient. An immunostimulant or an infection prevention / treatment agent is provided.
The present invention also relates to a pharmaceutical composition comprising at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof as an active ingredient, and at least protein C, activated protein C, or a derivative thereof. The present invention provides use of a pharmaceutical composition comprising one pharmaceutical composition as an active ingredient at the same time or at another time for immunostimulation or infection protection. In other words, the present invention relates to a pharmaceutical composition comprising at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof, and protein C, activated protein C, or a derivative thereof. A pharmaceutical composition containing at least one of the above as an active ingredient, and a disease (for example, inflammatory) that requires immunostimulation or infection protection, characterized by being administered to a subject (for example, an animal) simultaneously or separately The present invention provides a method for treating diseases, hypercytokinemia, and systemic inflammatory response syndrome.
The present invention also provides a pharmaceutical composition containing at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof as an active ingredient and a pharmaceutical composition containing valine as an active ingredient simultaneously or It is intended to provide use of the pharmaceutical composition to be administered at another time for immunostimulation or infection protection. In other words, the present invention provides a pharmaceutical composition comprising as an active ingredient at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof, and a pharmaceutical composition comprising valine as an active ingredient. Treatment of diseases (for example, inflammatory diseases, hypercytokinemia, systemic inflammatory response syndrome) that require immunostimulation or infection protection characterized by being administered to a subject (for example, an animal) simultaneously or separately A method is provided.
Example
Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.
[Materials used in the experiment]
5'-riboflavin sodium phosphate ester (riboflavin sodium phosphate) was prepared by synthesizing Japanese Pharmacopoeia riboflavin sodium phosphate ester (5'-FMN-Na).
APC (Activated Protein C: activated protein C) was used by purchasing Anact C (trade name of Teijin Limited, Lot No: SC002) from Teijin Limited.
LPS (Lipopolysaccharide) is available from Escherichia coli Escherichi. E. coli serotype O111: B4-derived lipopolysaccharide was purchased from Sigma (Co., St. Louis, MO, USA).
Valine was purchased from Wako Pure Chemical.
The 5′-FMN-Na, APC, LPS, and valine were used after being dissolved in physiological saline (Otsuka Pharmaceutical Co., Ltd., Tokyo, Japan).
The amino acid injection solution for improving hepatic encephalopathy (hereinafter referred to as amino acid injection solution) was purchased from Amino Levan (trade name) from Otsuka Pharmaceutical Co., Ltd. This aminolevane (trade name) contains 8.4 g of L-valine in 1000 mL, and contains multiple amino acids whose Fischer ratio (valine + leucine + isoleucine) / (tyrosin + phenylalanine) [molar ratio] is 37.05. It is an injection solution.
Male ICR mice (about 30 g / mouse) were 5 weeks old from Japan SLC Inc. (Shizuoka, Japan), light / dark cycle every 12 hours (lights at 7:00 am) under conditions of 23 ° C (acceptable range: 20-26 ° C) and relative humidity 55% (acceptable range: 40-70%) The one housed in (turned on and lighted off at 7 pm) was used. At this time, the mice were given sterilized tap water and pellet food (MF, Oriental Yeast Co., Tokyo, Japan). After acclimatization to this environment for one week, it was used for the experiment.
As SEB (Staphylococcus aureus toxin), Staphylococcus aureus enterotoxin B, BT-202 was used.
Male BALB / c mice (about 25 g each) were purchased from Nippon Charles River and were adapted for one week under the same conditions as the male ICR mice, and then used for the experiment.
[Example 1: Effect on endotoxin-induced shock mouse model]
First, LPS 12 mg / kg was intravenously administered to 10 male ICR mice (6 weeks old) in each group to prepare endotoxin (LPS) -induced shock mice.
Six hours after the administration of LPS, 5′-FMN-Na (dose: 10 mg / kg) dissolved in physiological saline and sterilized by filtration, and APC (dose: 75 units / kg) dissolved in physiological saline and sterilized by filtration. ) Was administered intravenously for 6 hours for each group. Further, 5′-FMN-Na (dose: 10 mg / kg) and APC (dose: 75 units / kg) dissolved in physiological saline and sterilized by filtration were mixed and continuously administered intravenously for 6 hours. In the control group, 3.5 ml of physiological saline was continuously administered intravenously for 6 hours instead of the drug. One unit of APC is considered to correspond to about 5 μg.
The survival rate of mice after 7 days from the administration of LPS was examined. The drug efficacy was determined by Steel test. The results are shown in Table 1.

As shown in Table 1, it was found that administration of 5′-FMN-Na and APC in combination showed an extremely high survival rate.
[Example 2: Effect on exotoxin-induced shock mouse model]
First, SEB 0.75 mg / kg was intraperitoneally administered to each group of 10 male BALB / c mice (6 weeks old), and D-galactosamine 1.8 g / kg was intraperitoneally administered. Induced shock mice were created.
6 hours after administration of SEB, 5′-FMN-Na (dose: 20 mg / kg) dissolved in physiological saline and sterilized by filtration, and APC (dose: 300 units / kg) dissolved in physiological saline and sterilized by filtration. ) Was administered intravenously for 6 hours for each group. Further, 5′-FMN-Na (dose: 20 mg / kg) dissolved in physiological saline and sterilized by filtration was mixed with APC (dose: 300 units / kg) and continuously administered intravenously for 6 hours. In the control group, 3.5 ml of physiological saline was continuously administered intravenously for 6 hours instead of the drug.
The survival rate of the mice 7 days after SEB administration was examined. The drug efficacy was determined by Steel test. The results are shown in Table 2.

As shown in Table 2, it was found that, even in this example, 5′-FMN-Na and APC were administered in combination to show an extremely high survival rate.
[Example 3: Effect on endotoxin-induced shock mouse model]
In the same manner as in Example 1, a group of 10 LPS-induced shock mice was prepared.
6 hours after the administration of LPS, 5′-FMN-Na (dose: 10 mg / kg) dissolved in physiological saline and sterilized by filtration, and valine (dose: 200 mg / kg) dissolved in physiological saline and sterilized by filtration. ) Was administered intravenously for 6 hours for each group. Further, 5′-FMN-Na (dose: 10 mg / kg) dissolved in physiological saline and sterilized by filtration was mixed with valine (dose: 200 mg / kg) and continuously administered intravenously for 6 hours. In the control group, 3.5 ml of physiological saline was continuously administered intravenously for 6 hours instead of the drug.
The survival rate of mice after 7 days from the administration of LPS was examined. For the control and 5′-FMN-Na administration groups, experiments were performed twice for each 10 cases. The drug efficacy was determined by Steel test. The results are shown in Table 3.

As shown in Table 3, it was found that administration of 5′-FMN-Na and valine in combination showed an extremely high survival rate.
[Example 4: Effect on endotoxin-induced shock mouse model]
In the same manner as in Example 1, a group of 10 LPS-induced shock mice was prepared.
6 hours after administration of LPS, 5′-FMN-Na (dose: 10 mg / kg) dissolved in physiological saline and sterilized by filtration, and amino acid injection solution (dose: 3.5 ml / mouse) for each group Each was administered intravenously for 6 hours. Further, 5′-FMN-Na (dose: 10 mg / kg) and amino acid injection solution (dose: 3.5 ml / mouse) were mixed and continuously administered intravenously for 6 hours. In the control group, 3.5 ml of physiological saline was continuously administered intravenously for 6 hours instead of the drug.
The survival rate of mice after 7 days from the administration of LPS was examined. For the control and 5′-FMN-Na administration groups, experiments were performed twice for each 10 cases. The drug efficacy was determined by Steel test. The results are shown in Table 4.

As shown in Table 4, it was found that administration of 5′-FMN-Na in combination with an amino acid injection showed extremely high survival rate.
Industrial applicability
The pharmaceutical composition of the present invention is useful as an immunostimulant or anti-infection therapeutic agent, a prophylactic or therapeutic agent for hypercytokinemia, systemic inflammatory response syndrome, sepsis or septic shock, and the like.

Claims (13)

Riboflavin, riboflavin derivative, or at least one of pharmacologically acceptable salts thereof, and at least one of protein C or activated protein C as active ingredients ,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or a pharmaceutical composition used for the prevention or treatment of systemic inflammatory response syndrome, sepsis or septic shock, which is at least one of leucoflavin adenine dinucleotide . Riboflavin, riboflavin derivative, or at least one of pharmacologically acceptable salts thereof, and valine as an active ingredient ,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or a pharmaceutical composition used for the prevention or treatment of systemic inflammatory response syndrome, sepsis or septic shock, which is at least one of leucoflavin adenine dinucleotide .   The pharmaceutical composition according to claim 2, wherein the valine is contained in an amount of 1 to 1000 parts by weight with respect to 1 part by weight of the riboflavin, riboflavin derivative, or pharmacologically acceptable salt thereof. Riboflavin, riboflavin derivative, or at least one of pharmacologically acceptable salts thereof, at least one of protein C or activated protein C, and valine as active ingredients ,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or a pharmaceutical composition used for the prevention or treatment of systemic inflammatory response syndrome, sepsis or septic shock, which is at least one of leucoflavin adenine dinucleotide . The pharmaceutical composition according to any one of claims 1 to 4, comprising the active ingredient in the form of an injection, a tablet, a granule, a powder, a fine granule, a capsule, a pill, or an oral liquid. A container containing a pharmaceutical composition containing at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof as an active ingredient, and a pharmaceutical containing at least one of protein C or activated protein C as an active ingredient A container containing the composition ,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or a device used for prevention or treatment of systemic inflammatory response syndrome, sepsis or septic shock, which is at least one of leucoflavin adenine dinucleotide . A container containing a pharmaceutical composition containing riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof as an active ingredient, and a container containing a pharmaceutical composition containing valine as an active ingredient ,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or a device used for prevention or treatment of systemic inflammatory response syndrome, sepsis or septic shock, which is at least one of leucoflavin adenine dinucleotide . A medicament used for the prevention or treatment of systemic inflammatory response syndrome, sepsis or septic shock ,
The pharmaceutical comprises a pharmaceutical composition containing at least one of riboflavin, riboflavin derivatives, or pharmacologically acceptable salts thereof as an active ingredient, and at least one of protein C or activated protein C as an active ingredient. A pharmaceutical composition,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or at least one of leucoflavin adenine dinucleotide,
A medicament in which the pharmaceutical compositions are held in a state where they are not in contact with each other. A medicament used for the prevention or treatment of systemic inflammatory response syndrome, sepsis or septic shock ,
The medicament includes a pharmaceutical composition containing as an active ingredient at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof, and a pharmaceutical composition containing valine as an active ingredient,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or at least one of leucoflavin adenine dinucleotide,
A medicament in which the pharmaceutical compositions are held in a state where they are not in contact with each other. A pharmaceutical composition comprising as an active ingredient at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof; and a pharmaceutical composition comprising at least one of protein C or activated protein C as an active ingredient; A combination of
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or a preventive or therapeutic agent for systemic inflammatory response syndrome, sepsis or septic shock, which is at least one of leucoflavin adenine dinucleotide . A pharmaceutical composition comprising as an active ingredient at least one of riboflavin, a riboflavin derivative, or a pharmacologically acceptable salt thereof, and a pharmaceutical composition comprising valine as an active ingredient ,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or a preventive or therapeutic agent for systemic inflammatory response syndrome, sepsis or septic shock, which is at least one of leucoflavin adenine dinucleotide . Riboflavin, riboflavin derivative, or at least one of pharmacologically acceptable salts thereof is dissolved in a solution containing valine as an active ingredient ,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or a preventive or therapeutic agent for systemic inflammatory response syndrome, sepsis or septic shock, which is at least one of leucoflavin adenine dinucleotide . Riboflavin, riboflavin derivative, or at least one of pharmacologically acceptable salts thereof and at least one of protein C or activated protein C is dissolved in a solution containing valine as an active ingredient ,
The riboflavin derivatives include flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetrabutyrate, sodium riboflavin phosphate, monodiethanolamine salt of riboflavin phosphate, leucoflavin, monohydroflavin, leucoflavin phosphate, leucoflavin mononucleotide, Or a preventive or therapeutic agent for systemic inflammatory response syndrome, sepsis or septic shock, which is at least one of leucoflavin adenine dinucleotide .