Pharmaceutical for preventing or treating nephropathy, noninfective inflammation and vascular disease

Abstract

To provide means that can prevent or treat a disease, symptom or disorder involved in the formation of calciprotein particles.SOLUTION: One embodiment of the present invention relates to a pharmaceutical for preventing or treating nephropathy, noninfective inflammation and vascular disease, comprising a bisphosphonate preparation or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a medicine for preventing or treating renal disorders, non-infectious inflammation, and vascular diseases. In particular, one aspect of the present invention relates to a medicine for preventing or treating renal disorders, non-infectious inflammation, or vascular diseases, comprising, as an active ingredient, one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations, or pharma- ceutical acceptable salts thereof, or pharma-ceutical acceptable solvates thereof. [Background technology]

[0002] Chronic kidney disease, a type of kidney disorder, often occurs not only as a renal complication of kidney diseases such as chronic glomerulonephritis, but also as a renal complication of lifestyle-related diseases such as diabetes and hypertension. In order to suppress the progression of chronic kidney disease, there is no other way than to identify the disease causing the kidney disorder in each patient and thoroughly treat that disease. Therefore, there is a need to identify a universal therapeutic target common to all patients with chronic kidney disease and develop a treatment method.

[0003] The main cause of death in patients with chronic kidney disease is cardiovascular events, and vascular calcification has been identified as a risk factor for these events. For example, Non-Patent Document 1 describes that alendronate, a bisphosphonate drug, has a therapeutic effect on vascular calcification. However, an effective treatment for vascular calcification has not been established at present. In addition, a pathology in which inflammatory findings are observed despite the absence of obvious infection is called non-infectious inflammation. It has been pointed out that non-infectious inflammation is associated with lifestyle-related diseases and / or aging, and accelerates these. However, the "pathogen" that causes non-infectious inflammation has not been identified.

[0004] A pathological condition commonly observed during the progression of chronic kidney disease is a decrease in the number of functional nephrons. A nephron is a functional unit of the kidney consisting of a renal tubule and a glomerulus. When the number of nephrons decreases, a compensatory mechanism is activated that increases the amount of substances excreted in urine per nephron. The problem at this time is phosphorus. When the amount of phosphorus excreted per nephron increases, the phosphorus concentration in the primary urine increases, and fine particles containing calcium phosphate crystals (hereinafter also referred to as "calciprotein particles") are formed in the lumen of the renal tubule. Calciprotein particles are complex nanoparticles of calcium phosphate crystals and fetuin A, a serum protein. Calciprotein particles are known to have the activity of damaging renal tubule cells. For this reason, the formation of calciprotein particles causes kidney damage.

[0005] It is also known that calciprotein particles appear in the blood, and thus the formation of calciprotein particles is a cause of vascular diseases such as vascular endothelial disorder and vascular calcification, as well as non-infectious inflammation.

[0006] For example, Patent Document 1 describes a method for measuring calciprotein particles and a method for assisting in the examination of chronic kidney disease. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Patent No. 6566697 [Non-patent literature]

[0008] [Non-Patent Document 1] Okamoto M, Yamanaka S, Yoshimoto W, Shigematsu T, Alendronate as an effective treatment for bone loss and vascular calcification in kidney transplant recipients. Journal of transplantation. 2014, Volume 2014, p. 269613 Summary of the Invention [Problem to be solved by the invention]

[0009] As described above, although knowledge has been gained about diseases, symptoms, or disorders associated with the formation of calciprotein particles, no means have been developed to date that can prevent or treat these diseases, symptoms, or disorders.

[0010] Therefore, an object of the present invention is to provide a means capable of preventing or treating diseases, symptoms or disorders associated with the formation of calciprotein particles. [Means for solving the problem]

[0011] The present inventors have studied various means for solving the above problems. The present inventors have found that administration of a bisphosphonate preparation or a blood coagulation factor Xa inhibitor can reduce inflammation and fibrosis in the kidney. The present inventors have also found that administration of a blood coagulation factor Xa inhibitor can suppress inflammation and vascular calcification in the aorta. The present inventors have completed the present invention based on the above findings.

[0012] That is, the present invention includes the following aspects and embodiments. (1) A pharmaceutical for preventing or treating renal disorders, non-infectious inflammation, or vascular diseases, comprising, as an active ingredient, one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations, or pharma- ceutical acceptable salts thereof, or pharma-ceutical acceptable solvates thereof. (2) The pharmaceutical composition according to the above embodiment (1), wherein the renal disorder is one or more disorders selected from the group consisting of chronic kidney disease, renal tubular disorder, renal fibrosis, inflammatory cell infiltration into the interstitium, and nephrocarcinosis. (3) The pharmaceutical according to embodiment (1), wherein the non-infectious inflammation is non-infectious chronic inflammation associated with one or more diseases, symptoms, or disorders selected from the group consisting of lifestyle-related diseases, cancer, and aging. (4) The pharmaceutical according to embodiment (1), wherein the vascular disease is one or more disorders selected from the group consisting of arteriosclerosis, vascular endothelial disorder, vascular calcification, atherosclerosis, inflammatory response in the aorta or coronary artery, and osteoblast-like transformation of smooth muscle. (5) The medicine according to any one of the above embodiments (1) to (4), wherein the one or more compounds or pharma- ceutical acceptable salts thereof, or pharma-ceutical acceptable solvates thereof inhibit binding between a Toll-like receptor and calciprotein particles, thereby preventing or treating nephropathy, non-infectious inflammation, and vascular disease. Effect of the Invention

[0013] According to the present invention, it is possible to provide a means for preventing or treating diseases, symptoms or disorders associated with the formation of calciprotein particles. [Brief description of the drawings]

[0014] [Figure 1] Figure 1 is a graph showing the relative mRNA levels of various markers in mice fed a high-phosphate diet and administered a bisphosphonate in Study I. Data in the figure show the mean ± standard deviation. * indicates P<0.05 vs. vehicle by Mann-Whitney U test. [Diagram 2] Figure 2 is a graph showing the relative mRNA levels of tubular injury markers or genes whose expression is known to be decreased in tubular injury in mice fed a normal diet or a high-phosphate diet and administered a blood coagulation factor Xa inhibitor in Study II. In the figure, open bars show the results for the normal diet, and filled bars show the results for the high-phosphate diet. Data in the figure show the mean ± standard deviation. [Figure 3-1] Figure 3-1 is a graph showing the relative mRNA levels of inflammatory markers in mice fed a normal diet or a high-phosphate diet and administered a blood coagulation factor Xa inhibitor in Study II. In the figure, the open bars show the results for the normal diet, and the closed bars show the results for the high-phosphate diet. The data in the figure show the mean ± standard deviation. [Figure 3-2] Figure 3-2 is a graph showing the relative mRNA levels of inflammatory markers in mice fed a normal diet or a high-phosphate diet and administered a blood coagulation factor Xa inhibitor in Study II. In the figure, the open bars show the results for the normal diet, and the closed bars show the results for the high-phosphate diet. The data in the figure show the mean ± standard deviation. [Figure 4] Figure 4 shows photographs of the results of SDS-PAGE analysis of calciprotein particles isolated from serum samples of healthy subjects or dialysis patients in Test III. In the figure, A is a photograph of a silver-stained gel, and B is a photograph of a Western-blotted gel. [Diagram 5] Figure 5 is a photograph showing the results of SDS-PAGE analysis of calciprotein particles separated from plasma or serum samples of dialysis patients after phosphoric acid treatment in Test III. In the figure, A is a photograph of a gel stained with silver, B is a photograph of a gel subjected to Western blotting using anti-fetuin A antibody (F-180, rabbit origin), C is a photograph of a gel subjected to Western blotting using anti-blood coagulation factor X antibody (AMX-5050, mouse origin), D is a photograph of a gel subjected to Western blotting using anti-blood coagulation factor X antibody (H-120, rabbit origin), E is a photograph of a gel subjected to Western blotting using anti-prothrombin antibody (ab184015, rabbit origin), and F is a photograph of a gel subjected to Western blotting using anti-blood coagulation factor VII antibody (MAB2338, mouse origin). [Figure 6]Figure 6 is a photograph showing the results of SDS-PAGE analysis of calciprotein particles isolated from serum samples from healthy individuals or dialysis patients treated with recombinant protein in Test III, followed by Western blotting using an anti-GST antibody. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The present inventors have found that administration of a bisphosphonate preparation or a blood coagulation factor Xa inhibitor can reduce inflammation and fibrosis in the kidney. The present inventors have also found that administration of a blood coagulation factor Xa inhibitor can suppress inflammation and vascular calcification in the aorta. Therefore, one aspect of the present invention relates to a medicament for preventing or treating renal disorders, non-infectious inflammation, and vascular diseases, which comprises one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations, or pharma- ceutically acceptable salts thereof, or pharma- ceutically acceptable solvates thereof, as an active ingredient. The medicament of this aspect can prevent or treat diseases, symptoms, or disorders associated with the formation of calciprotein particles, such as renal disorders, non-infectious inflammation, or vascular diseases.

[0016] In each embodiment of the present invention, calciprotein particles refer to nanoparticles of a complex of calcium phosphate crystals and a serum protein, fetuin A. As the number of functional nephrons decreases, which is a pathological condition commonly observed during the progression of chronic kidney disease, the amount of phosphorus excreted per nephron increases, increasing the phosphorus concentration in the primary urine, and calciprotein particles are formed in the lumen of the renal tubules.

[0017] In each embodiment of the present invention, the Toll-like receptor refers to a Toll-like receptor (hereinafter also referred to as "TLR"), which is a membrane-bound receptor involved in innate immunity. An example of a Toll-like receptor expressed in the proximal tubule is TLR4. TLR4 is known as a pattern recognition receptor that mediates inflammatory responses induced by calciprotein particles in macrophages (Koppert, S., et al. Front Immunol 9, 1991 (2018)). In addition, TLR4 is expressed on the apical side of the proximal tubule distributed at the corticomedullary junction (El-Achkar, TM, et al. Am J Physiol Renal Physiol 295, F534-544 (2008)).

[0018] The compound that is the active ingredient of the pharmaceutical of this embodiment is one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations. The bisphosphonate preparation is preferably etidronic acid, alendronate, bisphosphonate, ibandronic acid, risedronic acid, minodronic acid or zoledronic acid, more preferably etidronic acid or alendronate. The blood coagulation factor Xa inhibitor is preferably rivaroxaban, avixaban, fondavarinux sodium or edoxaban tocil. As shown in the following examples, administration of a bisphosphonate preparation or blood coagulation factor Xa inhibitor can reduce inflammation and fibrosis in the kidney. In addition, administration of a blood coagulation factor Xa inhibitor can suppress inflammation and vascular calcification in the aorta. Therefore, by administering the pharmaceutical of this embodiment containing the above-mentioned compound as an active ingredient to a subject, for example a human patient, it is possible to prevent or treat diseases, symptoms or disorders associated with the formation of calciprotein particles, such as renal disorders, non-infectious inflammation or vascular diseases.

[0019] The reason why each aspect of the present invention exhibits the above-mentioned action and effect can be explained as follows. Note that each aspect of the present invention is not limited to the following action and principle. Calciprotein particles express pathogenic activity such as cell injury, vascular calcification, or inflammation by binding to Toll-like receptors and being endocytosed. As described above, calciprotein particles are complex nanoparticles of calcium phosphate crystals and fetuin A. Calcium phosphate has two different phases, calcium phosphate crystals having a crystalline structure and amorphous calcium phosphate not having a crystalline structure, but calciprotein particles having pathogenic activity contain calcium phosphate crystals. Bisphosphonate preparations are known to have the activity of inhibiting the phase transition from amorphous calcium phosphate to calcium phosphate crystals. For this reason, it is speculated that administration of a bisphosphonate preparation inhibits the phase transition to calcium phosphate crystals and suppresses the calciprotein particles from acquiring pathogenic activity. In addition, calciprotein particles may contain blood coagulation factor Xa, which is an activated blood coagulation factor X, in addition to fetuin A as a protein component. Therefore, it is presumed that the administration of blood coagulation factor Xa inhibitor inhibits the enzyme activity of blood coagulation factor Xa contained in calciprotein particles having pathogenic activity, and changes the activity of calciprotein particles and / or the binding affinity to Toll-like receptors.By this mechanism of action, it is presumed that one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations used as the active ingredient of the medicament of this embodiment suppresses diseases, symptoms, or disorders related to the formation of calciprotein particles.For this reason, by administering the medicament of this embodiment to a subject, for example, a human patient, it is possible to prevent or treat diseases, symptoms, or disorders related to the formation of calciprotein particles, such as kidney disorders, non-infectious inflammation, or vascular diseases, by suppressing the pathogenic activity of calciprotein particles or inhibiting the binding between Toll-like receptors and calciprotein particles.

[0020] In each aspect of the present invention, the renal disorder to be prevented or treated is preferably one or more disorders selected from the group consisting of chronic kidney disease, renal tubular disorder, renal fibrosis, inflammatory cell infiltration into the interstitium, and nephrocarcinosis. The non-infectious inflammation to be prevented or treated is preferably non-infectious chronic inflammation associated with one or more diseases, symptoms, or disorders selected from the group consisting of lifestyle-related diseases, cancer, and aging. The vascular disease to be prevented or treated is preferably one or more disorders selected from the group consisting of arteriosclerosis, vascular endothelial disorder, vascular calcification, atherosclerosis, inflammatory reaction in the aorta or coronary artery, and osteoblast-like transformation of smooth muscle. By administering the pharmaceutical agent of this aspect to a subject having the above-mentioned disease, symptom, or disorder, for example, a human patient having the disease, symptom, or disorder, the disease, symptom, or disorder associated with the formation of calciprotein particles can be prevented or treated.

[0021] In each aspect of the present invention, the preventive or therapeutic effect of a disease, symptom or disorder associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation or vascular disease, in a subject having such a disease, symptom or disorder, for example, a human patient having such a disease, symptom or disorder, can be determined, for example, by using the amount of blood fibroblast growth factor 23 (FGF23), blood creatinine, blood urea nitrogen, urinary L-type fatty acid binding protein (L-FABP), urinary N-acetylglucosaminidase (NAG), urinary beta 2-microglobulin or urinary creatinine as an indicator.

[0022] As used herein, "prevention" means to substantially prevent the occurrence (onset or manifestation) of a disease, symptom or disorder in a subject having a disease, symptom or disorder associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation or vascular disease, for example, a human patient having said disease, symptom or disorder. Also, as used herein, "treatment" means to suppress (e.g., suppress progression), alleviate, repair and / or cure a disease, symptom or disorder that has occurred (onset or manifestation) in a subject having a disease, symptom or disorder associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation or vascular disease, for example, a human patient having said disease, symptom or disorder.

[0023] In each aspect of the present invention, the compound used as an active ingredient includes not only the compound itself but also its salt. When the compound used as an active ingredient is in the form of a salt, it is preferably a pharma- ceutically acceptable salt. Counterions of the salt of the compound used as an active ingredient include, but are not limited to, cations such as sodium ion, potassium ion, calcium ion, magnesium ion, or substituted or unsubstituted ammonium ion, or chloride ion, bromide ion, iodide ion, phosphate ion, nitrate ion, sulfate ion, carbonate ion, bicarbonate ion, perchlorate ion, formate ion, acetate ion, trifluoroacetate ion, propionate ion, lactate ion, maleate ion, hydroxymaleate ion, methylmaleate ion, fumarate ion, adipate ion, benzoate ion, 2-acetoxybenzoate ion, p-aminobenzoate ion, benzoic acid ... Anions such as benzoate, nicotinate, cinnamate, ascorbate, pamoate, succinate, salicylate, bismethylenesalicylate, oxalate, tartrate, malate, citrate, gluconate, aspartate, stearate, palmitate, itaconate, glycolate, glutamate, benzenesulfonate, cyclohexylsulfamate, methanesulfonate, ethanesulfonate, isethionate, benzenesulfonate, p-toluenesulfonate, or naphthalenesulfonate are preferred. When the compound used as an active ingredient is in the form of a salt with the above counter ion, its pharmacological activity can be substantially equivalent to that of the compound itself.

[0024] In each aspect of the present invention, the compound used as an active ingredient includes not only the compound itself but also a solvate of the compound or a salt thereof. When the compound or a salt thereof used as an active ingredient is in the form of a solvate, it is preferably a pharma- ceutically acceptable solvate. The solvent capable of forming a solvate with the compound or a salt thereof is not limited, but is preferably, for example, water or an organic solvent such as methanol, ethanol, 2-propanol (isopropyl alcohol), dimethyl sulfoxide (DMSO), acetic acid, ethanolamine, acetonitrile or ethyl acetate. When the compound or a salt thereof used as an active ingredient is in the form of a solvate with the above-mentioned solvent, its pharmacological activity can be substantially equivalent to that of the compound itself.

[0025] In each embodiment of the present invention, the compounds used as active ingredients also include mixtures of stereoisomers of the compounds, such as individual enantiomers and diastereomers of the compounds, as well as racemates.

[0026] In the pharmaceutical of this embodiment, the compound used as an active ingredient may be used alone or in combination with one or more pharma- ceutically acceptable ingredients. The pharmaceutical of this embodiment may be formulated into various dosage forms commonly used in the art according to the desired administration method. Thus, the pharmaceutical of this embodiment may also be provided in the form of a pharmaceutical composition containing one or more compounds selected from the group consisting of bisphosphonates and blood coagulation factor Xa inhibitors and one or more pharma- ceutically acceptable carriers. In this embodiment, the pharmaceutical composition may contain, in addition to the ingredients, one or more pharma- ceutically acceptable vehicles (e.g., solvents such as sterilized water or solutions such as saline), excipients, binders, vehicles, solubilizers, preservatives, stabilizers, disintegrants, disintegration inhibitors, swelling agents, lubricants, surfactants, emulsifiers, oily liquids (e.g., vegetable oils), suspending agents, buffers, soothing agents, antioxidants, sweeteners, and flavoring agents.

[0027] The dosage form of the pharmaceutical of this embodiment is not particularly limited, and may be a preparation for use in parenteral administration, a preparation for use in transmucosal (e.g., nasal, sublingual, or oral mucosa), transdermal, transanal (enema), or transvaginal administration, or a preparation for use in oral administration. The dosage form of the pharmaceutical of this embodiment may be a preparation in a unit dose form or a preparation in a multiple dose form. Preparations for use in parenteral administration include, for example, injections such as sterile solutions or suspensions with water or other pharma- ceutical acceptable liquids. Examples of additives that can be mixed with the injection include, but are not limited to, vehicles such as isotonic solutions containing physiological saline, glucose, or other auxiliary agents (e.g., D-sorbitol, D-mannitol, or sodium chloride), solubilizing agents such as alcohols (e.g., ethanol or benzyl alcohol), esters (e.g., benzyl benzoate), and polyalcohols (e.g., propylene glycol or polyethylene glycol), nonionic surfactants such as polysorbate 80 or polyoxyethylene hydrogenated castor oil, oily liquids such as sesame oil or soybean oil, buffers such as phosphate buffer or sodium acetate buffer, pain-relieving agents such as benzalkonium chloride or procaine hydrochloride, stabilizers such as human serum albumin or polyethylene glycol, preservatives, and antioxidants. The prepared injection is usually filled into a suitable container (e.g., a vial or an ampoule) and stored under an appropriate environment until use.

[0028] The pharmaceutical of this embodiment can also be used in combination with one or more other pharmaceutical drugs. In this case, the pharmaceutical of this embodiment may be provided in the form of a single pharmaceutical drug containing one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations and one or more other drugs, or may be provided in the form of a pharmaceutical combination or kit containing multiple preparations in which one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations and one or more other drugs are separately formulated. In the case of the pharmaceutical combination or kit form, each preparation can be administered simultaneously or separately (for example, consecutively).

[0029] In the pharmaceutical of this embodiment, the compound used as an active ingredient includes not only the compound itself, but also pharma- ceutically acceptable salts of the compound and pharma-ceutically acceptable solvates thereof. The pharma-ceutically acceptable salts of the compound used as an active ingredient and pharma-ceutically acceptable solvates thereof are not limited, but are preferably, for example, the salts or solvates exemplified above. When the compound used as an active ingredient is in the form of the above-mentioned salt or solvate, the compound can be applied to the desired medicinal use.

[0030] When one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations are applied to medical use, the compounds include not only themselves but also the prodrug forms of the compounds. In this specification, "prodrug" means a compound that is converted to a parent drug in vivo. In the pharmaceutical of this embodiment, the prodrug form of the compound used as an active ingredient can be, but is not limited to, for example, an ester of the hydroxyl group of the compound and any carboxylic acid, an amide of the hydroxyl group of the compound and any amine, and an amide of the amino group of the compound and any carboxylic acid. When the compound used as an active ingredient is in the prodrug form, the pharmacokinetics of the prodrug form when administered to a subject can be improved without substantially reducing the activity of the parent drug.

[0031] The pharmaceutical of this embodiment can be applied to various subjects who need to prevent or treat diseases, symptoms, or disorders associated with the formation of calciprotein particles, such as kidney damage, non-infectious inflammation, or vascular disease. The subject is preferably a human or non-human mammalian subject or patient (e.g., a warm-blooded animal such as a pig, dog, cow, rat, mouse, guinea pig, rabbit, chicken, sheep, cat, monkey, Mandrill, or chimpanzee), more preferably a human patient. By administering the pharmaceutical of this embodiment to the subject, diseases, symptoms, or disorders associated with the formation of calciprotein particles, such as kidney damage, non-infectious inflammation, or vascular disease, in the subject can be prevented or treated.

[0032] When the pharmaceutical of this embodiment is administered to a subject, particularly a human patient, the exact dosage and administration should be ultimately determined by the attending physician in consideration of many factors, such as the subject's age, sex, symptoms to be prevented or treated, the exact state (e.g., severity) of the disease and / or disorder, and the route of administration. Therefore, in the pharmaceutical of this embodiment, one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations, which are active ingredients, are administered to the subject in a therapeutically effective dosage and administration (e.g., dosage and administration route). For example, when the pharmaceutical of this embodiment is administered to a subject, particularly a human patient, the dosage of the compound used as the active ingredient is usually in the range of 0.01 to 1000 μg / kg body weight / day, for example, in the range of 0.5 to 200 μg / kg body weight / day.

[0033] The medicament of this embodiment may be administered by any administration route. For example, the medicament of this embodiment may be administered by a route such as intravenous administration, enema administration, subcutaneous administration, intramuscular administration, oral administration, or intraperitoneal administration.

[0034] One or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations used as an active ingredient of the medicament of this embodiment can be used to prevent or treat a disease, symptom, or disorder associated with the formation of calciprotein particles, such as renal impairment, non-infectious inflammation, or vascular disease, in a subject having the disease, symptom, or disorder, for example, a human patient having the disease, symptom, or disorder. Therefore, another aspect of the present invention relates to a prophylactic or therapeutic agent for renal impairment, comprising one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations as an active ingredient. The prophylactic or therapeutic agent of this embodiment has the same characteristics as the medicament of this embodiment described above. In addition, the prophylactic or therapeutic agent of this embodiment can be used in the same manner and dosage as the medicament of this embodiment described above. The prophylactic or therapeutic agent of this embodiment can be used to prevent or treat a disease, symptom, or disorder associated with the formation of calciprotein particles, such as renal impairment, non-infectious inflammation, or vascular disease, in a subject having the disease, symptom, or disorder, for example, a human patient having the disease, symptom, or disorder. In the preventive or therapeutic agent of this embodiment, the renal disorder to be prevented or treated is preferably one or more disorders selected from the group consisting of chronic kidney disease, renal tubular disorder, renal fibrosis, inflammatory cell infiltration into the interstitium, and nephrocarcinosis. The non-infectious inflammation to be prevented or treated is preferably non-infectious chronic inflammation associated with one or more diseases, symptoms, or disorders selected from the group consisting of lifestyle-related diseases, cancer, and aging. The vascular disease to be prevented or treated is preferably one or more disorders selected from the group consisting of arteriosclerosis, vascular endothelial disorder, vascular calcification, atherosclerosis, inflammatory reaction in the aorta or coronary artery, and osteoblast-like transformation of smooth muscle. The above-mentioned diseases, symptoms, or disorders can be prevented or treated by administering an effective amount of one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations to a subject having a disease, symptom, or disorder associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation, or vascular disease, for example, a human patient having the disease, symptom, or disorder.

[0035] Another aspect of the present invention is a method for preventing or treating a disease, symptom, or disorder associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation, or vascular disease, comprising administering an effective amount of one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations to a subject having the disease, symptom, or disorder, for example, a human patient having the disease, symptom, or disorder. The compound administered in the method of this aspect has the same characteristics as the compound used as the active ingredient of the medicament of this aspect described above. In addition, the method of this aspect can be carried out in the same manner and dosage as the medicament of this aspect described above. In the method of this aspect, the renal disorder to be prevented or treated is preferably one or more disorders selected from the group consisting of chronic kidney disease, renal tubular disorder, renal fibrosis, inflammatory cell infiltration into the interstitium, and nephrocarcinosis. The non-infectious inflammation to be prevented or treated is preferably non-infectious chronic inflammation associated with one or more diseases, symptoms, or disorders selected from the group consisting of lifestyle-related diseases, cancer, and aging. The vascular disease to be prevented or treated is preferably one or more disorders selected from the group consisting of arteriosclerosis, vascular endothelial disorder, vascular calcification, atherosclerosis, inflammatory reaction in the aorta or coronary artery, and osteoblast-like transformation of smooth muscle. By administering an effective amount of one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations to a subject having a disease, symptom, or disorder associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation, or vascular disease, for example, a human patient having the disease, symptom, or disorder, the above-mentioned diseases, symptoms, or disorders can be prevented or treated.

[0036] Another aspect of the present invention is one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonates, or their pharmaceutically acceptable salts, or their pharmaceutically acceptable solvates, for use in the prevention or treatment of diseases, symptoms, or disorders associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation, or vascular disease, in subjects, for example, human patients, having said diseases, symptoms, or disorders associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation, or vascular disease. Yet another aspect of the present invention is the use of one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonates, or their pharmaceutically acceptable salts, or their pharmaceutically acceptable solvates, in the manufacture of a medicament for the prevention or treatment of diseases, symptoms, or disorders associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation, or vascular disease, in subjects, for example, human patients, having said diseases, symptoms, or disorders associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation, or vascular disease. Yet another aspect of the present invention is the use of one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations, or pharma- ceutically acceptable salts thereof, or pharma- ceutically acceptable solvates thereof, for the prophylaxis or treatment of a disease, symptom or disorder associated with the formation of calciprotein particles, such as renal impairment, non-infectious inflammation or vascular disease, in a subject having such a disease, symptom or disorder, for example, a human patient having such a disease, symptom or disorder. The compound of this aspect has the same characteristics as the compound used as an active ingredient in the medicament of this aspect described above. In addition, the compound of this aspect can be used in the same manner and dosage as the medicament of this aspect described above. In the compound or use of this aspect, the nephropathy to be prevented or treated is preferably one or more disorders selected from the group consisting of chronic kidney disease, renal tubular disorder, renal fibrosis, inflammatory cell infiltration into the interstitium, and nephrocarcinosis. The non-infectious inflammation to be prevented or treated is preferably non-infectious chronic inflammation associated with one or more diseases, symptoms, or disorders selected from the group consisting of lifestyle-related diseases, cancer, and aging.The vascular disease to be prevented or treated is preferably one or more disorders selected from the group consisting of arteriosclerosis, vascular endothelial disorder, vascular calcification, atherosclerosis, inflammatory reaction in the aorta or coronary artery, and osteoblast-like transformation of smooth muscle. The above-mentioned diseases, symptoms, or disorders can be prevented or treated by administering an effective amount of one or more compounds selected from the group consisting of bisphosphonate preparations and blood coagulation factor Xa inhibitors to a subject having a disease, symptom, or disorder associated with the formation of calciprotein particles, such as renal disorder, non-infectious inflammation, or vascular disease, for example, a human patient having the disease, symptom, or disorder. EXAMPLES

[0037] The present invention will be described in more detail below with reference to examples, although the technical scope of the present invention is not limited to these examples.

[0038] <Study I: Effect of bisphosphonates on phosphate-induced renal fibrosis> Mice (4-week-old C57BL / 6 males) were placed on a high-phosphate diet containing 2.0% inorganic phosphate for 8 weeks and subcutaneously injected with etidronate (100 mg / kg, N=9), alendronate (10 mg / kg, N=6), or vehicle (saline, N=7) every other day. Relative mRNA levels of the indicated markers were determined by quantitative RT-PCR. Relative mRNA levels of various markers are shown in Figure 1. Data in the figure show mean ± standard deviation. * indicates P<0.05 vs. vehicle by Mann-Whitney U test. αSMA, α-smooth muscle actin; Nlrp3, nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3; MCP1, monocyte chemotactic protein-1; TGFβ1, transforming growth factor-β1.

[0039] As shown in Figure 1, administration of bisphosphonates, etidronic acid or alendronic acid, significantly reduced the expression levels of markers associated with renal fibrosis. These results demonstrate that administration of bisphosphonates can reduce inflammation and fibrosis in the kidney.

[0040] <Study II: Effect of blood coagulation factor Xa inhibitor on phosphate-induced renal fibrosis> Mice (12-week-old C57BL / 6 males) were placed on a normal diet containing 0.35% inorganic phosphate or a high-phosphate diet containing 2.0% inorganic phosphate for 4 weeks and were administered rivaroxaban (12 ppm) or vehicle (physiological saline) daily in the diet. At 16 weeks of age, kidneys were harvested from each mouse. Relative mRNA levels of selected markers in the kidney were determined by quantitative RT-PCR. The relative mRNA levels of various markers are shown in Figures 2, 3-1, and 3-2. In the figures, open bars show the results of the normal diet, and filled bars show the results of the high-phosphate diet. Data in the figures show the mean ± standard deviation. Opn is osteopontin, Ngal is neutrophil gelatinase-associated lipocalin, Kim1 is kidney injury molecule-1, and Klotho is anti-aging hormone. Il-6 is interleukin-6, Il-1b is interleukin-1b, McP1 is monocyte chemotactic protein-1, Tnfa is tumor necrosis factor alpha, Tgfb is transforming growth factor beta, and C3 is the third component of complement. Opn, Ngal, and Kim1 are tubular injury markers, and Klotho is a gene whose expression is known to be decreased in tubular injury. Il-6, Il-1b, McP1, Tnfa, Tgfb, and C3 are inflammatory markers.

[0041] As shown in Figures 2, 3-1 and 3-2, the administration of rivaroxaban, a blood coagulation factor Xa inhibitor, significantly reduced the expression levels of markers related to renal tubular damage and inflammation. These results demonstrate that the administration of a blood coagulation factor Xa inhibitor can reduce inflammation and renal tubular damage in the kidney.

[0042] <Study III: Relationship between calciprotein particles and blood coagulation factor Xa in patients with renal impairment> To clarify the mechanism of action of the preventive or therapeutic effect of the administration of factor Xa inhibitors on diseases, symptoms, or disorders related to the formation of calciprotein particles, calciprotein particles in the blood of patients undergoing dialysis due to renal impairment were analyzed. Alendronate, a bisphosphonate preparation that binds to calcium phosphate crystals, was bound to magnetic beads to prepare solid-phase alendronate beads. When solid-phase alendronate beads are added to plasma or serum samples, the calcium phosphate crystals of the calciprotein particles bind to alendronate, allowing the calciprotein particles to be specifically separated. After collecting and washing the beads using a magnet, the beads can be added to an EDTA solution to elute the calciprotein particles from the beads.

[0043] Blood was collected from healthy subjects or patients undergoing dialysis due to renal impairment, and serum was prepared. Immobilized alendronate beads were added to serum samples from healthy subjects or dialysis patients, respectively, to separate calciprotein particles. Calciprotein particles were eluted from the beads using EDTA solution. The eluate was separated by SDS-PAGE, and the gel was silver stained. In addition, this gel was subjected to Western blotting using anti-fetuin A antibody (F-180, rabbit origin) or anti-blood coagulation factor X antibody (AMX-5050, mouse origin). The results of SDS-PAGE analysis of calciprotein particles separated from serum samples from healthy subjects or dialysis patients are shown in Figure 4. In the figure, A is a photograph of a silver-stained gel, and B is a photograph of a Western-blotted gel.

[0044] As shown in Figure 4A, a strong band was detected at approximately 50 kDa in serum samples from both healthy subjects and dialysis patients. Western blotting confirmed that this band was fetuin A. Furthermore, a blood coagulation factor X band was also detected in the serum sample from the dialysis patient (Figure 4B). The bands were excised from the gel and subjected to mass spectrometry and Edman analysis, and were identified as fetuin A and blood coagulation factor X.

[0045] A certain concentration of phosphoric acid was added to plasma or serum samples from dialysis patients to promote the formation of calciprotein particles. The concentration of phosphoric acid added was 0 (no phosphoric acid added), 3 (amount that increases the phosphorus concentration in the sample by 3 mM), or 10 (amount that increases the phosphorus concentration in the sample by 10 mM). Calciprotein particles were separated from the treated plasma or serum samples in the same manner as above. The SDS-PAGE gel was silver stained. In addition, this gel was subjected to Western blotting using various antibodies. The results of SDS-PAGE analysis of calciprotein particles separated from plasma or serum samples from dialysis patients after phosphoric acid treatment are shown in Figure 5. In the figure, A is a photograph of a silver stained gel, B is a photograph of a gel subjected to Western blotting using anti-fetuin A antibody (F-180, rabbit origin), C is a photograph of a gel subjected to Western blotting using anti-blood coagulation factor X antibody (AMX-5050, mouse origin), D is a photograph of a gel subjected to Western blotting using anti-blood coagulation factor X antibody (H-120, rabbit origin), E is a photograph of a gel subjected to Western blotting using anti-prothrombin antibody (ab184015, rabbit origin), and F is a photograph of a gel subjected to Western blotting using anti-blood coagulation factor VII antibody (MAB2338, mouse origin).

[0046] As shown in Figure 5A, calciprotein particles from both plasma and serum samples contained several proteins, which Western blotting revealed to be the blood clotting factors prothrombin and blood clotting factor VII, in addition to fetuin A and blood clotting factor X.

[0047] In order to characterize blood coagulation factor X contained in calciprotein particles in the blood of dialysis patients, a recombinant protein was prepared in which blood coagulation factor X was linked to a GST tag. Blood coagulation factor X is cleaved by serine protease to become activated blood coagulation factor Xa having enzymatic activity. The recombinant protein was added to serum samples from healthy subjects or dialysis patients in the same manner as above. Calciprotein particles were separated from the treated serum samples in the same manner as above. The SDS-PAGE gel was silver stained. In addition, this gel was subjected to Western blotting using an anti-GST antibody. After treating serum samples from healthy subjects or dialysis patients with recombinant protein, the calciprotein particles separated from the samples were analyzed by SDS-PAGE, and the results of Western blotting using an anti-GST antibody are shown in FIG. 6.

[0048] As shown in Figure 6, only the full-length protein was detected in the serum samples from healthy subjects, whereas the truncated protein was also detected in the serum samples from dialysis patients. These results demonstrated that the calciprotein particles from dialysis patients contain activated blood coagulation factor Xa with enzymatic activity.

[0049] <Study IV: Effect of blood coagulation factor Xa inhibitors on phosphate-induced vascular disease> Klotho knockout mice (5 weeks old / female) were placed on a high phosphate diet containing 2.0% inorganic phosphate for 13 weeks and were fed rivaroxaban (50 mg / kg, N=3) or vehicle (saline, N=3) daily. At 18 weeks of age, aortas were harvested from each mouse. Relative mRNA levels of selected markers in the aorta were determined by quantitative RT-PCR. As a result, the addition of rivaroxaban, a blood coagulation factor Xa inhibitor, reduced the expression levels of vascular calcification markers and inflammatory markers. The above results revealed that administration of a blood coagulation factor Xa inhibitor can alleviate vascular diseases.

[0050] The present invention is not limited to the above-described embodiments, but includes various modified examples. For example, the above-described embodiments have been described in detail to clearly explain the present invention, and the present invention is not necessarily limited to those including all of the configurations described. In addition, it is possible to add, delete, and / or replace part of the configuration of each embodiment with other configurations.

Claims

1. A pharmaceutical for preventing or treating renal disorders, non-infectious inflammation, or vascular diseases, comprising a bisphosphonate preparation or a pharma- ceutically acceptable salt thereof, or a pharma- ceutically acceptable solvate thereof as an active ingredient.

2. The pharmaceutical composition according to claim 1, wherein the renal disorder is one or more disorders selected from the group consisting of chronic kidney disease, renal tubular disorder, renal fibrosis, inflammatory cell infiltration into the interstitium, and nephrocarcinosis.

3. The pharmaceutical according to claim 1, wherein the non-infectious inflammation is non-infectious chronic inflammation associated with one or more diseases, symptoms or disorders selected from the group consisting of lifestyle-related diseases, cancer and aging.

4. The pharmaceutical composition according to claim 1, wherein the vascular disease is one or more disorders selected from the group consisting of arteriosclerosis, vascular endothelial disorder, vascular calcification, atherosclerosis, inflammatory response in the aorta or coronary artery, and osteoblast-like transformation of smooth muscle.

5. The medicine according to any one of claims 1 to 4, wherein the bisphosphonate preparation or a pharma- ceutical acceptable salt thereof, or a pharma-ceutical acceptable solvate thereof prevents or treats renal disorders, non-infectious inflammation, and vascular diseases by inhibiting binding between a Toll-like receptor and calciprotein particles.

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