Medicine for preventing or treating kidney disorders, non-infectious inflammation and vascular diseases

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, containing at least one compound selected from the group consisting of a blood coagulation Xa factor inhibitor and a bisphosphonate 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 pharmaceutical for preventing or treating renal disorders, non-infectious inflammation, and vascular diseases. In particular, one aspect of the present invention relates to 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 bisphosphonates, or pharmaceutically acceptable salts thereof, or pharmaceutically acceptable solvates thereof. [Background technology]

[0002] Chronic kidney disease, a type of kidney disorder, often occurs as a renal complication of lifestyle-related diseases such as diabetes and hypertension, as well as kidney diseases such as chronic glomerulonephritis. The only way to prevent the progression of chronic kidney disease is to identify the underlying disease in each patient and thoroughly treat that disease. Therefore, there is a need to identify universal therapeutic targets common to all patients with chronic kidney disease and develop treatments.

[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 example, Non-Patent Document 1 describes that the bisphosphonate drug alendronate has a therapeutic effect on vascular calcification. However, no effective treatment for vascular calcification has been established at present. In addition, a pathological condition in which inflammatory findings are observed despite the absence of obvious infection is called non-infectious inflammation. Non-infectious inflammation is associated with lifestyle-related diseases and / or aging and has been shown to accelerate these. However, the "pathogen" that causes non-infectious inflammation has not been identified.

[0004] A common pathological condition observed during the progression of chronic kidney disease is a decrease in the number of functional nephrons. Nephrons are functional units of the kidney, consisting of tubules and glomeruli. When the number of nephrons decreases, a compensatory mechanism is activated that increases the amount of substances excreted in urine per nephron. Phosphorus becomes a problem in this situation. When the amount of phosphorus excreted per nephron increases, the phosphorus concentration in the primary urine increases, leading to the formation of microparticles containing calcium phosphate crystals (hereinafter also referred to as "calciprotein particles") within the lumen of the renal tubules. Calciprotein particles are nanoparticles composed of complexes of calcium phosphate crystals and the serum protein fetuin A. Calciprotein particles are known to have the ability to damage renal tubule cells. Therefore, the formation of calciprotein particles can cause kidney damage.

[0005] Calciprotein particles are also known to appear in the blood, and their formation is thought to be a cause of vascular diseases such as vascular endothelial damage 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 mentioned 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 for preventing or treating diseases, symptoms or disorders associated with the formation of calciprotein particles. [Means for solving the problem]

[0011] The present inventors have investigated various means for solving the above problems. They have found that administration of a bisphosphonate preparation or a blood coagulation factor Xa inhibitor can reduce inflammation and fibrosis in the kidney. They 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 these 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, which contains, as an active ingredient, one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations, or pharmaceutically acceptable salts thereof, or pharmaceutically 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 composition according to embodiment (1), wherein the vascular disease is one or more disorders selected from the group consisting of arteriosclerosis, vascular endothelial damage, vascular calcification, atherosclerosis, inflammatory reactions 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 pharmaceutically acceptable salts thereof, or pharmaceutically acceptable solvates thereof inhibit the binding of a Toll-like receptor to calciprotein particles, thereby preventing or treating nephropathy, non-infectious inflammation, and vascular disease. [Effects of the Invention]

[0013] The present invention makes it possible to provide a means for preventing or treating diseases, symptoms or disorders associated with the formation of calciprotein particles. [Brief explanation 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. [Figure 2] Figure 2 is a graph showing the relative mRNA levels of markers of renal tubular injury or genes known to be decreased in expression due to renal 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 represent the results for the normal diet, and filled bars represent the results for the high-phosphate diet. Data in the figure represent 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, open bars represent the results for the normal diet, and filled bars represent the results for the high-phosphate diet. Data in the figure represent 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, open bars represent the results for the normal diet, and filled bars represent the results for the high-phosphate diet. Data in the figure represent the mean ± standard deviation. [Figure 4] Figure 4 shows photographs showing the results of SDS-PAGE analysis of calciprotein particles isolated from serum samples of healthy subjects and 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. [Figure 5] Figure 5 shows photographs showing the results of SDS-PAGE analysis of calciprotein particles separated from plasma or serum samples from dialysis patients after phosphoric acid treatment in Test III. In the figure, A is a photograph of a silver-stained gel, B is a photograph of a gel subjected to Western blotting using an anti-fetuin A antibody (F-180, rabbit), C is a photograph of a gel subjected to Western blotting using an anti-blood coagulation factor X antibody (AMX-5050, mouse), D is a photograph of a gel subjected to Western blotting using an anti-blood coagulation factor X antibody (H-120, rabbit), E is a photograph of a gel subjected to Western blotting using an anti-prothrombin antibody (ab184015, rabbit), and F is a photograph of a gel subjected to Western blotting using an anti-blood coagulation factor VII antibody (MAB2338, mouse). [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 INVENTION

[0015] The present inventors have found that administration of a bisphosphonate 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 pharmaceutical composition for preventing or treating renal disorders, non-infectious inflammation, and vascular diseases, which comprises, as an active ingredient, one or more compounds selected from the group consisting of a blood coagulation factor Xa inhibitor and a bisphosphonate, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof. This pharmaceutical composition 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 the serum protein fetuin A. As the number of functional nephrons decreases, a pathological condition commonly observed during the progression of chronic kidney disease, the amount of phosphorus excreted per nephron increases, leading to an increase in the phosphorus concentration in the primary urine and the formation of calciprotein particles in the lumen of the renal tubules.

[0017] In each embodiment of the present invention, the term "Toll-like receptor" refers to a membrane-bound receptor (hereinafter also referred to as "TLR") involved in innate immunity. Examples of Toll-like receptors expressed in the proximal tubule include TLR4. TLR4 is known as a pattern recognition receptor that mediates the inflammatory response 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, T.M., et al. Am J Physiol Renal Physiol 295, F534-544 (2008)).

[0018] The compound serving as 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, alendronic acid, bisphosphonate, ibandronic acid, risedronic acid, minodronic acid, or zoledronic acid, and more preferably etidronic acid or alendronic acid. The blood coagulation factor Xa inhibitor is preferably rivaroxaban, abixaban, fondavarinux sodium, or edoxaban tocil. As shown in the following examples, administration of a bisphosphonate preparation or a blood coagulation factor Xa inhibitor can reduce inflammation and fibrosis in the kidney. Furthermore, 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-exemplified 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 kidney disorders, non-infectious inflammation, or vascular diseases.

[0019] The reasons why each aspect of the present invention exhibits the above-described effects can be explained as follows. However, each aspect of the present invention is not limited to the following effects and principles. Calciprotein particles bind to Toll-like receptors and are endocytosed, thereby exhibiting pathogenic activity such as cell injury, vascular calcification, or inflammation. As described above, calciprotein particles are composite nanoparticles of calcium phosphate crystals and fetuin A. Calcium phosphate exists in two distinct phases: calcium phosphate crystals with a crystalline structure and amorphous calcium phosphate without a crystalline structure. Calciprotein particles with pathogenic activity contain calcium phosphate crystals. Bisphosphonates are known to inhibit the phase transition from amorphous calcium phosphate to calcium phosphate crystals. Therefore, it is speculated that administration of bisphosphonates inhibits the phase transition to calcium phosphate crystals, thereby preventing calciprotein particles from acquiring pathogenic activity. Furthermore, calciprotein particles may contain blood coagulation factor Xa, an activated form of blood coagulation factor X, in addition to fetuin A as a protein component. Therefore, it is presumed that administration of a blood coagulation factor Xa inhibitor inhibits the enzymatic activity of blood coagulation factor Xa contained in pathogenic calciprotein particles, thereby changing the activity of the calciprotein particles and / or their binding affinity to Toll-like receptors. Due to this mechanism of action, one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonate preparations used as active ingredients of the medicament of this embodiment are presumed to suppress diseases, symptoms, or disorders associated with the formation of calciprotein particles. Therefore, 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 associated with the formation of calciprotein particles, such as kidney damage, non-infectious inflammation, or vascular disease, by suppressing the pathogenic activity of calciprotein particles or inhibiting the binding of Toll-like receptors to 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, tubular damage, 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 damage, vascular calcification, atherosclerosis, inflammatory responses 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-exemplified disease, symptom, or disorder, for example, a human patient having the disease, symptom, or disorder, the disease, symptom, or disorder associated with calciprotein particle formation 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 kidney damage, 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, can be determined by using, for example, the amounts of blood fibroblast growth factor 23 (FGF23), blood creatinine, blood urea nitrogen, urinary L-fatty acid binding protein (L-FABP), urinary N-acetylglucosaminidase (NAG), urinary beta2-microglobulin, urinary creatinine, or the like as indicators.

[0022] As used herein, "prevention" means substantially preventing 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 damage, non-infectious inflammation, or vascular disease, for example, a human patient having the disease, symptom, or disorder. Also, as used herein, "treatment" means suppressing (e.g., suppressing progression), alleviating, repairing, and / or curing 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 damage, non-infectious inflammation, or vascular disease, for example, a human patient having the 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 pharmaceutically acceptable salt. Counter ions 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, and the like. Preferred anions include 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, and naphthalenesulfonate. When the compound used as an active ingredient is in the form of a salt with the above counter ions, 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 solvates of the compound or a salt thereof. When the compound used as an active ingredient or a salt thereof is in the form of a solvate, it is preferably a pharmaceutically acceptable solvate. Solvents that can form solvates with the compound or a salt thereof include, but are not limited to, water or organic solvents such as methanol, ethanol, 2-propanol (isopropyl alcohol), dimethyl sulfoxide (DMSO), acetic acid, ethanolamine, acetonitrile, or ethyl acetate. When the compound used as an active ingredient or a salt thereof is in the form of a solvate with the above 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 composition of this embodiment, the compound used as the active ingredient may be used alone or in combination with one or more pharmaceutically acceptable ingredients. The pharmaceutical composition of this embodiment can be formulated into various dosage forms commonly used in the art depending on the desired administration method. Therefore, the pharmaceutical composition of this embodiment can 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 pharmaceutically acceptable carriers. In this embodiment, the pharmaceutical composition may contain, in addition to the ingredients, one or more pharmaceutically acceptable additives such as one or more pharmaceutically acceptable vehicles (e.g., solvents such as sterile water or solutions such as physiological saline), excipients, binders, vehicles, solubilizers, preservatives, stabilizers, disintegrants, disintegration inhibitors, bulking 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 formulation for use in parenteral administration, a formulation for use in transmucosal (e.g., nasal, sublingual, or oral mucosal), transdermal, transanal (enema), or transvaginal administration, or a formulation for use in oral administration. Furthermore, the dosage form of the pharmaceutical of this embodiment may be a formulation in a unit dose form or a formulation in a multiple dose form. Examples of formulations for use in parenteral administration include injections such as sterile solutions or suspensions with water or other pharmaceutically acceptable liquids. Additives that can be mixed into injections 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), solubilizers 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 solutions such as sesame oil or soybean oil, buffers such as phosphate buffer or sodium acetate buffer, soothing agents such as benzalkonium chloride or procaine hydrochloride, stabilizers such as human serum albumin or polyethylene glycol, preservatives, and antioxidants. Prepared injections are usually filled into appropriate containers (e.g., vials or ampoules) and stored under appropriate conditions until use.

[0028] The medicament of this embodiment can also be used in combination with one or more other medicaments useful as a medicine. In this case, the medicament of this embodiment may be provided in the form of a single medicine 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, the respective preparations can be administered simultaneously or separately (for example, sequentially).

[0029] In the pharmaceutical of this embodiment, the compound used as the active ingredient includes not only the compound itself but also pharmaceutically acceptable salts of the compound and pharmaceutically acceptable solvates thereof. Pharmaceutically acceptable salts of the compound used as the active ingredient and pharmaceutically acceptable solvates thereof are not limited, but are preferably, for example, the salts or solvates exemplified above. When the compound used as the active ingredient is in the form of the salt or solvate, the compound can be used for the desired pharmaceutical application.

[0030] When one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonates are used for pharmaceutical purposes, the compounds encompass not only the compounds themselves but also prodrug forms of the compounds. As used herein, "prodrug" refers to a compound that is converted into a parent drug in vivo. In this embodiment of the pharmaceutical, examples of prodrug forms of the compound used as an active ingredient include, but are not limited to, esters of the hydroxyl group of the compound with any carboxylic acid, amides of the hydroxyl group with any amine, and amides of the amino group of the compound with any carboxylic acid. When the compound used as an active ingredient is in the prodrug form, the pharmacokinetics of the prodrug form upon administration to a subject can be improved without substantially reducing the activity of the parent drug.

[0031] The medicament of this embodiment can be applied to various subjects in need of prevention or treatment of 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, hamadryas baboon, or chimpanzee), more preferably a human patient. By administering the medicament 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, can be prevented or treated in the subject.

[0032] When administering the pharmaceutical of this embodiment to a subject, particularly a human patient, the exact dosage and administration method should ultimately be determined by the attending physician, taking into account many factors, such as the subject's age and sex, the 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, the active ingredient, one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonates, are administered to the subject at a therapeutically effective dosage and administration method (e.g., dosage amount and administration route). For example, when administering the pharmaceutical of this embodiment to a subject, particularly a human patient, the dosage of the compound used as the active ingredient is typically in the range of 0.01 to 1000 μg / kg body weight / day, for example, 0.5 to 200 μg / kg body weight / day.

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

[0034] The one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonates used as active ingredients in 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 such a disease, symptom, or disorder, for example, a human patient having such a disease, symptom, or disorder. Therefore, another embodiment of the present invention relates to a prophylactic or therapeutic agent for nephropathy, comprising as an active ingredient one or more compounds selected from the group consisting of blood coagulation factor Xa inhibitors and bisphosphonates. The prophylactic or therapeutic agent of this embodiment has the same characteristics as the medicament of this embodiment described above. Furthermore, the prophylactic or therapeutic agent of this embodiment can be used in the same dosage and administration 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 such a disease, symptom, or disorder, for example, a human patient having such a 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 damage, 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 damage, vascular calcification, atherosclerosis, inflammatory responses 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 bisphosphonates to a subject with a disease, symptom, or disorder associated with calciprotein particle formation, such as renal disorder, non-infectious inflammation, or vascular disease, for example, a human patient with 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 injury, 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 bisphosphonates to a subject suffering from the disease, symptom, or disorder, e.g., a human patient suffering from the disease, symptom, or disorder. The compounds administered in the method of this aspect have similar characteristics to the compounds used as the active ingredients of the medicament of this aspect described above. Furthermore, the method of this aspect can be carried out using the same dosage and administration method as the medicament of this aspect described above. In the method of this aspect, the renal injury to be prevented or treated is preferably one or more disorders selected from the group consisting of chronic kidney disease, renal tubular injury, 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 damage, vascular calcification, atherosclerosis, inflammatory reactions 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 bisphosphonates 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 such a disease, symptom, or disorder.

[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 pharmaceutically acceptable salts thereof, or pharmaceutically acceptable solvates thereof, for use in the prevention or treatment of diseases, symptoms, or disorders associated with the formation of calciprotein particles, such as renal damage, non-infectious inflammation, or vascular disease, in subjects having such diseases, symptoms, or disorders, for example, human patients having such diseases, symptoms, or disorders. 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 pharmaceutically acceptable salts thereof, or pharmaceutically acceptable solvates thereof, 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 damage, non-infectious inflammation, or vascular disease, in subjects having such diseases, symptoms, or disorders, for example, human patients having such diseases, symptoms, or disorders. 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 pharmaceutically acceptable salts thereof, or pharmaceutically acceptable solvates thereof, for the prevention or treatment of a disease, symptom, or disorder associated with the formation of calciprotein particles, such as renal injury, non-infectious inflammation, or vascular disease, in a subject, e.g., a human patient, having such a disease, symptom, or disorder. The compound of this aspect has similar characteristics to the compound used as the active ingredient in the medicament of this aspect described above. Furthermore, the compound of this aspect can be used in the same dosage and administration 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 injury, 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 damage, vascular calcification, atherosclerosis, inflammatory response 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 bisphosphonates 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. [Example]

[0037] The present invention will be described in more detail below using 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. Figure 1 shows the relative mRNA levels of various markers. Data in the figure represent mean ± standard deviation. * indicates a P<0.05 vs. vehicle by Mann-Whitney U test. αSMA is α-smooth muscle actin, NLRP3 is nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3, MCP1 is monocyte chemoattractant protein-1, and TGF-β1 is transforming growth factor-β1.

[0039] As shown in Figure 1, administration of the bisphosphonates etidronic acid or alendronate 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 inhibitors 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 (saline) daily in the diet. At 16 weeks of age, kidneys were harvested from each mouse. Relative mRNA levels of selected markers in the kidneys 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 represent results for the normal diet, and filled bars represent results for the high-phosphate diet. Data in the figures represent the mean ± standard deviation. Opn: osteopontin, Ngal: neutrophil gelatinase-associated lipocalin, Kim1: kidney injury molecule-1, and Klotho: anti-aging hormone. Il-6 is interleukin-6, Il-1b is interleukin-1b, McP1 is monocyte chemotactic protein-1, Tnfa is tumor necrosis factor α, Tgfb is transforming growth factor-β, and C3 is the third component of complement. Opn, Ngal, and Kim1 are markers of renal tubular injury, and Klotho is a gene whose expression is known to decrease in renal 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, 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 administration of a blood coagulation factor Xa inhibitor can alleviate 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 factor Xa inhibitors in preventing or treating diseases, symptoms, or disorders associated with calciprotein particle formation, calciprotein particles were analyzed in the blood of patients undergoing dialysis due to renal failure. Alendronate, a bisphosphonate drug that binds to calcium phosphate crystals, was attached to magnetic beads to prepare immobilized alendronate beads. Adding the immobilized alendronate beads to plasma or serum samples allowed the calcium phosphate crystals on the calciprotein particles to bind to the alendronate, enabling specific isolation of the calciprotein particles. After collecting and washing the beads using a magnet, the calciprotein particles were eluted by adding them to an EDTA solution.

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

[0044] As shown in Figure 4A, a strong band at approximately 50 kDa was detected in serum samples from both healthy individuals and dialysis patients. Western blotting confirmed that this band was fetuin A. In addition, 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, which identified them as fetuin A and blood coagulation factor X.

[0045] A predetermined concentration of phosphoric acid was added to plasma or serum samples from dialysis patients to promote the formation of calciprotein particles. The concentration of added phosphoric acid was 0 (no phosphoric acid added), 3 (an amount that increases the phosphorus concentration in the sample by 3 mM), or 10 (an amount that increases the phosphorus concentration in the sample by 10 mM). Calciprotein particles were isolated from the treated plasma or serum samples using the same procedure as described above. The SDS-PAGE gel was silver stained. This gel was also subjected to Western blotting using various antibodies. The plasma or serum samples from dialysis patients were treated with phosphoric acid, and the calciprotein particles isolated from the samples were analyzed by SDS-PAGE. The results 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 factor VII, in addition to fetuin A and factor X.

[0047] To characterize the blood coagulation factor X contained in calciprotein particles in the blood of dialysis patients, we prepared a recombinant protein in which blood coagulation factor X was linked to a GST tag. Blood coagulation factor X was cleaved by serine protease to produce activated blood coagulation factor Xa, which has enzymatic activity. The recombinant protein was added to serum samples from healthy individuals or dialysis patients using the same procedure as described above. Calciprotein particles were isolated from the treated serum samples using the same procedure as described above. The SDS-PAGE gel was silver stained. Furthermore, this gel was subjected to Western blotting using an anti-GST antibody. After treating serum samples from healthy individuals or dialysis patients with the recombinant protein, the calciprotein particles isolated from the samples were analyzed by SDS-PAGE and Western blotting using an anti-GST antibody. The results are shown in Figure 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 demonstrate that 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 administered rivaroxaban (50 mg / kg, N=3) or vehicle (saline, N=3) daily in the diet. 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. The addition of rivaroxaban, a blood coagulation factor Xa inhibitor, reduced the expression levels of vascular calcification and inflammatory markers. These results demonstrate that administration of a blood coagulation factor Xa inhibitor can alleviate vascular disease.

[0050] The present invention is not limited to the above-described embodiments, but includes various modifications. 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 described configurations. Furthermore, 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 chronic kidney disease, renal tubular damage, or renal fibrosis, comprising as an active ingredient a blood coagulation factor Xa inhibitor selected from the group consisting of rivaroxaban, apixaban, fondaparinux sodium, and edoxaban tosilate, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

2. The pharmaceutical composition according to claim 1, for preventing or treating renal tubular damage or renal fibrosis.

3. 3. The pharmaceutical composition according to claim 1, wherein the blood coagulation factor Xa inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof prevents or treats the disease by inhibiting binding between a Toll-like receptor and calciprotein particles.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the blood coagulation factor Xa inhibitor is rivaroxaban.

Images