Reagent for treating acute kidney injury caused by contrast

Abstract

The present invention provides a pharmaceutical composition for protecting the kidney or for treating an acute kidney injury caused by a contrast agent by reducing renal toxicity caused by a contrast agent, comprising a compound of Chemical Formula 1, which is a pyrazole derivative or a pharmaceutically acceptable salt thereof.

Description

[0001] This application is a divisional application. The application number of the original application is 202180027048X, the application date is March 23, 2021, and the name of the invention is "Agents for treating angiography-induced acute kidney injury." Technical Field

[0002] This application claims the benefit of Korean Patent Application No. 10-2020-0042899 filed on April 8, 2020, and Korean Patent Application No. 10-2021-0036838 filed on March 22, 2021, in the Korean Intellectual Property Office, the disclosures of which are incorporated herein by reference in their entirety.

[0003] The present invention relates to a pyrazole derivative for protecting kidneys by reducing nephrotoxicity caused by contrast agents, or for preventing and treating acute kidney injury caused by contrast agents; a method for preparing the pyrazole derivative; and a pharmaceutical composition of the pyrazole derivative. Background Art

[0004] A contrast medium (CM) is a drug introduced into the stomach, intestines, blood vessels, cerebrospinal cavity, joint cavity, etc. to increase the contrast of the image by artificially increasing the difference in X-ray absorption in various tissues so that the tissue or blood vessels can be clearly seen in radiological examinations such as magnetic resonance imaging (MRI) or computed tomography (CT) imaging. By using contrast agents, the diagnostic value is improved by being able to well distinguish biological structures or lesions from their surroundings.

[0005] Contrast agents are generally divided into negative contrast agents and positive contrast agents, wherein negative contrast agents transmit more X-rays than surrounding tissues to display images. Positive contrast agents include iodine-containing contrast agents, barium sulfate, etc., and negative contrast agents include air, gas, carbon dioxide, etc.

[0006] Some people are sensitive to contrast agents and may experience allergic reactions, leading to rash, itching, fever, nausea, vomiting, joint pain, bleeding tendency, etc.

[0007] As side effects of contrast agents, shock or allergic reactions may rarely occur, hypersensitivity reactions such as urticaria, flushing, rash, and itching may occur, and in severe cases, acute kidney injury may occur.

[0008] In particular, although the exact mechanism of contrast-induced acute kidney injury is unclear, it occurs after the administration of iodine-based contrast media and is known to be the leading cause of hospital-acquired acute kidney injury.

[0009] Acute kidney injury due to contrast media is defined as an increase of 25% or more or 0.5 mg / dL in the blood creatinine level over the existing level within 48 hours of use, and in this case, the patient should not have other causes of kidney injury, such as a drop in blood pressure, other nephrotoxicity, etc.

[0010] Typically, the creatinine level in the blood reaches a peak on the 3rd to 5th day after contrast medium administration, and then returns to the previous level within 7 to 10 days. Acute kidney injury caused by contrast agents accounts for about 12% of hospital-acquired acute kidney injury, and is one of the three major causes of acute kidney injury in hospitalized patients, along with ischemic acute kidney injury (42%) and acute kidney injury caused by urinary tract obstruction (18%). It is reported that the incidence of acute kidney injury caused by contrast agents is low (0% to 5%) in patients with normal renal function, but the incidence increases to 12% to 27% in patients with impaired renal function. In particular, it is reported that the incidence increases to 50% in high-risk patients such as patients with dehydration, diabetic nephropathy, renal damage, insufficient blood volume or congestive heart failure, and elderly patients, and 15% of patients are reported to require dialysis.

[0011] Despite these risks, especially in elderly patients at high risk and with severe comorbidities, the use of radiocontrast agents for computed tomography and vascular interventions has rapidly increased to more than 20% in patients over the age of 50. Therefore, in the context of the increasing number of elderly patients who are susceptible to contrast-induced renal injury and patients with diabetes, hypertension, and heart failure who are undergoing examinations with contrast agents, it is of great urgency to develop preventive and therapeutic approaches to reduce contrast-induced nephrotoxicity.

[0012] Although the pathophysiology of contrast-induced renal disease has not been clearly elucidated, the major mechanisms are hypothesized to be ischemic injury caused by decreased renal blood flow and direct injury to tubular cells caused by the contrast agent.

[0013] In the case of ischemic injury, it is known that due to changes in hormones regulating renal blood flow after contrast medium administration, vasoconstrictor hormones adenosine and endothelin increase, while vasodilator hormones nitric oxide (NO) and prostaglandins decrease, causing damage particularly in the region of the renal medulla due to decreased blood flow and hypoxia. Characteristically, ischemic injury and tubular cell damage are known to ultimately increase the synthesis of oxygen free radicals in renal tissue, causing widespread oxidative stress, which in turn causes renal cell damage due to increased inflammation and apoptosis caused by increased cytokines.

[0014] In the case of contrast-induced nephropathy, as with other acute kidney injuries, because the method of interrupting the progression of established kidney damage is unclear and the timing of induction is well-defined, efforts to interrupt its occurrence are primarily through active prevention, and multiple studies are ongoing to reduce contrast-induced kidney injury.

[0015] It is known that the conventional treatment for preventing contrast-induced nephrotoxicity is to try to prevent electrolyte imbalance while giving adequate fluid supply, but there is no clear treatment or prevention method to prevent the progression of kidney damage, and in some cases, dialysis is necessary, so more research is needed to develop a prevention method to prevent it from occurring in the early stages.

[0016] Recently, at the laboratory level, it has been reported that endopeptidase inhibitors or some aromatic-cationic peptides can prevent acute kidney injury induced by contrast agents.

[0017] However, most studies are observational studies in laboratories or small groups of patients, and to date, it is unclear whether contrast-induced kidney injury can be effectively prevented by using these drugs, nor is there a specific treatment for contrast-induced kidney injury.

[0018] Therefore, the present inventors have completed the present invention by determining that despite various studies, there is still a need to develop a preventive and therapeutic agent for acute kidney injury caused by contrast media.

[0019] On the other hand, none of the prior art documents discloses the effectiveness of the pyrazole compounds of the present invention in preventing and treating acute kidney injury caused by contrast agents.

[0020] [Prior art literature]

[0021] (Patent Document 1) Korean Patent No. 10-1280160

[0022] (Patent Document 2) Korean Patent No. 10-1886894

[0023] (Patent Document 3) U.S. Patent Publication No. US2019-0388492

[0024] (Non-patent document 1) Persson PB, Hansell P, Liss P. Pathophysiology of contrast medium-induced nephropathy. Kidney Int 2005; 68: 14-22.

[0025] (Non-patent document 2) Heyman SN, Reichman J, Brezis M. Pathophysiology of radiocontrast nephropathy: a role for medullary hypoxia. Invest Radiol 1999; 34: 685-691. Summary of the invention

[0026] Technical issues

[0027] An object of the present invention is to provide a pharmaceutical composition comprising the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0028] Another object of the present invention is to provide a pharmaceutical composition for protecting kidneys by reducing nephrotoxicity caused by contrast agents, comprising the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0029] Another object of the present invention is to provide a pharmaceutical composition for treating and preventing contrast agent-induced acute kidney injury, comprising the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0030] Another object of the present invention is to provide contrast agent-induced kidney damage reducing effect, blood creatinine level improving effect and renal tubular damage improving effect using the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0031] Another object of the present invention is to provide a method for protecting the kidney by reducing contrast-induced nephrotoxicity or a method for preventing or treating contrast-induced acute kidney injury by administering the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof to an individual.

[0032] Another object of the present invention is to provide a use of the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof for preventing or treating acute kidney injury induced by contrast agents.

[0033] Technical Solution

[0034] In order to achieve the above-mentioned object, the present invention provides a composition for protecting the kidney by reducing the nephrotoxicity caused by contrast agents or a pharmaceutical composition for preventing, improving or treating acute kidney injury caused by contrast agents, which comprises a pyrazole compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0035] <Chemical Formula 1>

[0036]

[0037] Here, R is a straight or branched chain alkyl group having 1 to 10 carbon atoms.

[0038] Beneficial Effects

[0039] The pyrazole compounds and pharmaceutically acceptable salts thereof according to the present invention can effectively alleviate the symptoms of acute kidney injury caused by contrast agent administration, and thus can be effectively used to protect the kidney by reducing contrast agent-induced nephrotoxicity, or to prevent or treat contrast agent-induced acute kidney injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 Shown are the results of analyzing the effects of the compounds of the present invention in an animal model with contrast-induced acute kidney injury.

[0041] Figure 2 Shown are the results of analyzing the effects of the compounds of the present invention on improving blood urea nitrogen (BUN) levels in an animal model with contrast-induced acute kidney injury.

[0042] Figure 3 Shown are the results of analyzing the effect of the compound of the present invention on improving blood creatinine levels in an animal model with contrast-induced acute kidney injury.

[0043] Figure 4 Shown are the results of analyzing the effects of the compounds of the present invention on reducing kidney injury markers NGAL, KIM-1 and urine microprotein (albumin) in an animal model with contrast-induced acute kidney injury.

[0044] Figure 5 Shown are the results of analyzing the effect of the compound of the present invention on improving renal tubular damage in an animal model with contrast-induced acute kidney injury.

[0045] Figure 6 Shown are the results of analyzing the effect of the compound of the present invention on improving inflammation of renal tissue in an animal model with contrast agent-induced acute kidney injury.

[0046] Figure 7 Shown are the results of analyzing the effect of the compound of the present invention on reducing the infiltration of inflammatory cells in renal tissue in an animal model with contrast-induced acute kidney injury.

[0047] Figure 8 Shown are the results of analyzing the effects of the compounds of the present invention on reducing reactive oxygen species, which are indicators of oxidative stress in renal tissue, in an animal model with contrast medium-induced acute kidney injury.

[0048] Fig. 9 Shown are the results of analyzing the effects of the compounds of the present invention on reducing nitrotyrosine, a marker of oxidative stress in renal tissue, in an animal model with contrast-induced acute kidney injury. DETAILED DESCRIPTION

[0049] Hereinafter, the present invention will be described in more detail with reference to embodiments.

[0050] However, the present invention is not limited to the embodiments shown by way of example, but is defined only by the scope of the appended claims. In addition, even if it is a necessary component to implement the present invention, a detailed description of a component that can be easily implemented by a person skilled in the art will be omitted.

[0051] The terms and words used in this specification and claims should not be understood as limited to the conventional or dictionary meanings, but should be understood as meanings and concepts consistent with the technical concept of the present invention based on the principle that the inventor can appropriately define the concept of the terms to describe his invention in the best manner.

[0052] The terms used in the present invention are only used for the purpose of describing specific embodiments and are not intended to limit the present invention. Unless the context clearly indicates otherwise, singular expressions include plural expressions. In the present invention, terms such as "comprising" and "having" are intended to indicate the presence of features, numbers, steps, operations, components, parts or combinations thereof described in this specification, and it should be understood that these terms do not preclude the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts or combinations thereof.

[0053] As the number of diagnostic methods using contrast media increases, the use of contrast media has increased, and at the same time, the side effects caused by contrast media have also become a problem. In addition to allergic reactions such as allergies caused by contrast media, in severe cases, acute kidney injury may occur. In addition, it is known that an increasing number of elderly patients suffer from major underlying diseases such as diabetes and hypertension, and the increasing use of radiocontrast media in these patients is also one of the main causes of hospital-acquired acute kidney injury.

[0054] However, as treatments for contrast-induced acute kidney injury, only adequate fluid replacement and treatments to prevent electrolyte disturbances have been reported, and no specific methods or definitive treatments for preventing contrast-induced acute kidney injury have been reported.

[0055] Therefore, the present invention provides a pharmaceutical composition capable of preventing or treating acute kidney injury caused by contrast agents, comprising one or more compounds selected from the pyrazole compounds represented by Chemical Formula 1 or pharmaceutically acceptable salts thereof.

[0056] The pyrazole compound used in the present invention is represented by the following chemical formula 1:

[0057] <Chemical Formula 1>

[0058]

[0059] Here, R is a straight or branched chain alkyl group having 1 to 10 carbon atoms.

[0060] The pharmaceutically acceptable salt of the pyrazole compound contained in the pharmaceutical composition of the present invention refers to a salt that retains the biological effectiveness and performance of the parent compound and is not harmful to biology or other aspects when administered in a single dose. In addition, it refers to a salt commonly used in the pharmaceutical industry.

[0061] Specifically, pharmaceutically acceptable base addition salts can be prepared from inorganic bases and organic bases. Salts from inorganic bases can include, but are not limited to, salts of sodium, potassium, lithium, ammonium, calcium and magnesium. Salts from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines; substituted amines, including naturally occurring substituted amines; and isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hepamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucosamine, theobromine, purine, piperazine, piperidine and / or cyclic amines including N-ethylpiperidine.

[0062] It should also be understood that other carboxylic acid derivatives, specifically, carboxylic acid amides, including formamide, lower alkyl carboxamides, di(lower alkyl) carboxamides, and the like, may also be used in the practice of the present invention.

[0063] In addition, pharmaceutically acceptable acid addition salts can be prepared from inorganic acids and organic acids. Salts from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, iodic acid, tartaric acid, etc. Salts from organic acids can include, but are not limited to, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, lactic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid and / or salicylic acid, etc.

[0064] The pharmaceutically acceptable salt may be a hydrochloride.

[0065] The pyrazole compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof contained in the pharmaceutical composition of the present invention is specifically exemplified as follows:

[0066] 3-phenyl-4-methyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride;

[0067] 3-phenyl-4-ethyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride;

[0068] 3-phenyl-4-n-propyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride;

[0069] 3-phenyl-4-isopropyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride;

[0070] 3-phenyl-4-n-butyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride;

[0071] 3-phenyl-4-tert-butyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride;

[0072] 3-phenyl-4-n-pentyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride;

[0073] 3-phenyl-4-n-hexyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride.

[0074] Specifically, the pyrazole compound contained in the pharmaceutical composition of the present invention can be 3-phenyl-4-n-propyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride.

[0075] The compound of Chemical Formula 1 of the present invention can inhibit the generation of active oxygen in kidney tissue.

[0076] In the present invention, oxidative stress refers to tissue damage caused by relatively excessive reactive oxygen species (ROS) due to the imbalance between the generation of reactive oxygen species (ROS) and the antioxidant defense mechanisms of biomolecules, cells and tissues. In this case, "reactive oxygen" may refer to reactive oxygen, activated oxygen and activated oxygen, which refer to the same substance.

[0077] "Contrast agents" of the present invention are known to those skilled in the art. Contrast agents enable a specific part of the body to be distinguished from surrounding parts of similar density. Preferably, the contrast agent in the context of the present invention is an opaque or positive contrast agent, i.e. a contrast agent with a greater attenuation density than the surrounding tissue, thereby enhancing the absorption of X-rays.

[0078] Positive contrast agents are well known in the art and are non-iodine based contrast agents, iodine based contrast agents, ie iodinated contrast agents. Examples of non-iodine based contrast agents and iodine based contrast agents are known to the person skilled in the art.

[0079] Monomeric contrast agents are divided into ionic and non-ionic types. Specific ionic monomeric contrast agents include ioglycate (Rayvist), iodamide (Uromiro), acetophenone (Diaginol, Urokon), diatrizoate (Angiografin; Hypaque; Renografin; Urografin; Urovison) and metrizoate (Isopaque; Triosil).

[0080] Specific non-ionic monomeric contrast agents include mepanematide (Amipaque), iohexol (Omnipaque), iopamiro (Iopamiro), iopentol (Imagopecque), iopromide (Ultravist), and ioversol (Optiray).

[0081] The diiodinated contrast agents preferably contain two triiodobenzene rings. They can be divided into ionic intravenous cholangiographic contrast agents, monoacid ionic contrast agents and non-ionic contrast agents. The diiodinated contrast agents are preferably iodixanol (Visipaque; Optiprep), iotrolan (Isovist) and iodixanol (Hexabrix).

[0082] Non-iodinated contrast media usually contain barium, primarily in the form of insoluble barium sulfate. They are preferably used for examinations of the gastrointestinal tract.

[0083] The contrast medium can be administered by any method deemed appropriate. It will be appreciated by those skilled in the art that the method selected for administration may depend on the purpose of the examination to which the contrast medium is to be administered and / or the contrast medium. Specifically, barium-based contrast media are administered by swallowing or enema. Iodine-based contrast media are preferably administered by injection into a vein, spinal canal or artery. A catheter for administration of iodine-based contrast media may be used.

[0084] Administration of iodine-based contrast agents is used for computed tomography and most preferably for angiography. Administration of contrast agents suitable for ultrasonography or magnetic resonance imaging (MRI) is further contemplated in the present invention.

[0085] "Acute kidney injury" or "AKI" or "acute renal failure (ARF)" of the present invention is well known in the art. As used in the present invention, the term refers to the rapid loss of renal function. The rapid loss of renal function is caused by damage to the kidneys. The diagnosis and classification criteria of AKI are based on changes in serum creatinine levels and urine excretion. The term is defined in the KDIGO guidelines (KDIGO, Kidney International Supplements (2012) 2, 69-88), which are incorporated herein by reference in their entirety.

[0086] Contrast-induced acute kidney injury is defined as an increase in blood creatinine level of more than 25% or more than 0.5 mg / dL above the existing level within 48 hours after the administration of contrast agent, not due to decreased renal function caused by other reasons such as hypotension, use of other nephrotoxic drugs, urinary tract obstruction, and embolism.

[0087] Generally, contrast media may show side effects such as decreased renal function in some cases even when administered to a person with normal renal function, but may more easily show worsening of renal function and aggravate decreased renal function particularly when administered to a person with decreased renal function.

[0088] Although the exact mechanism of acute kidney injury caused by contrast agents has not been fully elucidated, it is known that contrast agents increase osmotic pressure, reduce renal blood flow, and cause renal artery constriction. In addition, tubular necrosis or decreased renal perfusion is known to be the cause of acute kidney injury caused by contrast agents. It is known that in this state, the production of reactive oxygen species is promoted and ischemic tubular damage is caused, which may be the direct cause of tubular toxicity. In addition, inflammation is known to be one of the causes.

[0089] Therefore, to prevent or treat contrast-induced acute kidney injury, although the renoprotective effect of reducing reactive oxygen species by adequate fluid replacement and treatment with antioxidants such as bicarbonate, N-acetylcysteine, or ascorbic acid has been reported, there is currently no established treatment, and prevention is the only therapeutic approach.

[0090] The pharmaceutical compositions of the present invention, in particular all of 3-phenyl-4-ethyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol, 3-phenyl-4-n-propyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol and 3-phenyl-4-n-butyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol, exhibit an inhibitory effect on the formation of reactive oxygen species.

[0091] As confirmed by the animal experiments in the embodiments of the present invention, the serum creatinine concentration as an indicator of acute kidney injury caused by contrast agents is significantly reduced, and the degree of renal tubular injury is also reduced. In addition, neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1) and urine microprotein, which are markers of kidney injury, are reduced, and the infiltration of inflammatory cells in kidney tissue and reactive oxygen and nitrotyrosine, which are indicators of oxidative stress, are reduced.

[0092] Therefore, in the contrast agent-induced acute kidney injury model, it was confirmed that the inflammatory response was inhibited by inhibiting the production of reactive oxygen species in renal tissue, thereby preventing or alleviating acute kidney injury.

[0093] In addition to the compound of Chemical Formula 1 or a salt thereof, the pharmaceutical composition of the present invention may further include bicarbonate and / or an antioxidant for removing active oxygen.

[0094] The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier within the range not impairing the effects of the present invention.

[0095] The "pharmaceutically acceptable carrier" includes any and all kinds of solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (antibacterial or antifungal agents), isotonic agents, diluents, absorption delaying agents, salts, preservatives, stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavorings, dyes, etc., and combinations thereof, as known to those skilled in the art.

[0096] The diluent may be selected from, but is not limited to, microcrystalline cellulose, lactose monohydrate, anhydrolactose, lactose, starch, mannitol, carboxymethylcellulose, sorbitol, and combinations thereof.

[0097] The disintegrant may be selected from, but not limited to, low-substituted hydroxypropyl cellulose, crospovidone, croscarmellose sodium, sodium starch glycolate, F-melt, and combinations thereof.

[0098] The binder may be selected from, but is not limited to, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose, polyvinyl acetate, povidone, polyvinyl pyrrolidone, copovidone, polyethylene glycol, sodium lauryl sulfate, light anhydrous silicic acid, synthetic aluminum silicate, silicate derivatives such as calcium silicate or magnesium aluminate metasilicate, phosphates such as calcium hydrogen phosphate, carbonates such as calcium carbonate, pregelatinized starch, gums such as gum arabic, gelatin, cellulose derivatives such as ethyl cellulose, and mixtures thereof.

[0099] The lubricant may be selected from, but not limited to, magnesium stearate, silicon dioxide, talc, light anhydrous silicic acid, sodium stearyl fumarate, and combinations thereof.

[0100] As the pH adjuster, acidulants such as acetic acid, adipic acid, ascorbic acid, sodium ascorbate, sodium etherate, malic acid, succinic acid, tartaric acid, fumaric acid and citric acid, and alkalizing agents such as ammonia water, sodium carbonate, magnesium oxide, magnesium carbonate, sodium citrate and tricalcium phosphate can be used.

[0101] As the antioxidant, butylated hydroxytoluene, butylated hydroxyanisole, tocopherol acetate, tocopherol, propyl gallate, sodium hydrogen sulfite, sodium pyrosulfite and the like can be used.

[0102] Furthermore, the agent of the present invention can be formulated by selectively using various additives selected from coloring agents and flavoring agents as pharmaceutically acceptable additives.

[0103] In the present invention, the range of the additives is not limited to the use of the additives, and can be formulated to include a dose within a normal range by selectively using the additives.

[0104] The pharmaceutical composition according to the present invention can be formulated and used by conventional methods in the form of oral preparations, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories or sterile injections.

[0105] In one aspect of the present invention, it is a pharmaceutical composition for preventing, improving or treating acute kidney injury induced by contrast agents, comprising an active ingredient in a range of 0.00001 wt % to 100 wt %, 0.0001 wt % to 95 wt % or 0.001 wt % to 90 wt %, based on the total weight of the pharmaceutical composition.

[0106] In the preventive or therapeutic agent for contrast agent-induced acute kidney injury according to the present invention, the dosage of the pyrazole compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof can be appropriately changed according to the patient's age and weight, symptoms, administration route, etc.

[0107] The dosage of the pyrazole compound represented by Chemical Formula 1 of the present invention or a pharmaceutically acceptable salt thereof may be 0.00001 mg / kg / day to 2000 mg / kg / day, 0.0001 mg / kg / day to 1000 mg / kg / day, 0.001 mg / kg / day to 800 mg / kg / day, 0.001 mg / kg / day to 500 mg / kg / day, 0.001 mg / kg / day to 100 mg / kg / day, 0.001 mg / kg / day to 80 mg / kg / day or 0.01 mg / kg / day to 70 mg / kg / day.

[0108] The content of the pyrazole compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof of the present invention may be 0.00001 to 100 weight %, 0.0001 to 95 weight %, 0.0001 to 90 weight %, 0.001 to 70 weight % or 0.001 to 50 weight % per unit dosage form.

[0109] The pyrazole compound represented by Chemical Formula 1 of the present invention or a pharmaceutically acceptable salt thereof may be administered at a concentration of 0.0001 μM to 500 μM, 0.001 μM to 300 μM, 0.001 μM to 150 μM, 0.001 μM to 130 μM, 0.001 μM to 100 μM, 0.001 μM to 80 μM or 0.01 μM to 70 μM.

[0110] The pharmaceutical composition of the present invention can be administered together with a contrast agent or separately with a contrast agent by conventional routes, and can be specifically formulated for intramuscular, intrathecal, intradigestive, intracardiovascular, intrarenal or intravenous administration. The formulation method adopts conventional methods known to those skilled in the art.

[0111] Conventional compositions for intramuscular or intrathecal administration may consist of, but are not limited to, for example, an active ingredient and a sterile isotonic aqueous solvent containing glucose, sodium chloride, or both glucose and sodium chloride. Other examples include, but are not limited to, Lactated Ringer's Injection, Lactated Ringer's Injection + Dextrose Injection, Normosol-M and Dextrose, Isolyte E, Acylated Ringer's Injection, and the like. Optionally, the formulations of the present invention may include, but are not limited to, cosolvents such as polyethylene glycol; chelating agents such as ethylenediaminetetraacetic acid; and antioxidants such as sodium metabisulfite. Optionally, but not limited to, the solution may be lyophilized and then reconstituted with a suitable solvent just prior to administration.

[0112] Preferred embodiments are provided to help understand the present invention. The following embodiments are provided not to limit the present invention but to facilitate understanding of the present invention.

[0113] <Synthesis example>

[0114] <Synthesis Example 1> Synthesis of 3-phenyl-4-ethyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol

[0115]

[0116] In a round bottom flask, 2-ethyl-3-oxo-3-phenylpropionic acid ethyl ester (10.7 g, 49 mmol) and 2-hydrazinopyridine (5.6 g, 51.4 mmol) were heated to reflux under nitrogen for 1 day without solvent. The obtained solid was purified with hexane and ethyl acetate and then dried in vacuo to obtain the title compound in 70% yield.

[0117] 1H NMR (300MHz, DMSO-d6) δ8.25-8.24 (1H, d), 8.00-7.97 (1H, d), 7.84-7.82 (1H, t), 7.73-7.71 (2H, m ), 7.46-7.37 (3H, m) 7.12-7.11 (1H, t), 2.62-2.57 (2H, m), 1.23-1.17 (3H, m); ESI (m / z) 266.1 [M+H] +

[0118] <Synthesis Example 2> Synthesis of 3-phenyl-4-butyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol

[0119]

[0120] In a round-bottom flask, 2-butyl-3-oxo-3-phenylpropionic acid ethyl ester (12.1 g, 49 mmol) and 2-hydrazinopyridine (5.6 g, 51.4 mmol) were heated to reflux under nitrogen for 1 day without solvent. The obtained solid was purified with hexane and ethyl acetate and then dried in vacuo to obtain the title compound in 75% yield.

[0121] 1H NMR (300MHz, DMSO-d6) δ8.25-8.24(1H,d), 8.03-8.02(1H,d), 7.85-7.83(1H,t), 7.70-7.69(2H,m), 7.44-7.35(3H,m )7.12-7.11(1H,t),2.56-2.53(2H,t),1.58-1.52(2H,m),1.38-1.24(2H,m),0.89-0.86(3H,t);ESI(m / z)294.0[M+H] +

[0122] <Synthesis Example 3> Synthesis of 3-phenyl-4-propyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol

[0123]

[0124] 2-propyl-3-oxo-3-phenylpropionic acid ethyl ester (2.52 g, 10.7 mmol) and 10 mL of ethanol were placed in a round-bottom flask, and then a solution of 2-hydrazinopyridine (1.29 g, 1.18 mmol) diluted in 3 mL of ethanol was slowly added dropwise at 0° C. The mixture was heated to reflux for 3 days at 100° C. The solvent was removed by distillation under reduced pressure, and the obtained solid was washed with hexane and ethyl acetate, and then dried in vacuo to obtain the title compound in a yield of 82%.

[0125] 1H NMR (300MHz, CDCl3) δ12.50 (1H, s), 8.27-8.25 (1H, m), 8.01 (1H, d, J = 8.5Hz), 7.81 (1H, m), 7. 69 (2H, m), 7.48-7.34 (3H, m), 7.12-7.10 (1H, m), 2.54 (2H, d, J = 7.5Hz), 1.64 (2H, m), 0.93 (3H,

[0126] t, J=7.3Hz); EIMS (70eV) m / z (rel intensity) 279 (M+, 37), 250 (100)

[0127] <Synthesis Example 4> Synthesis of 3-phenyl-4-propyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol hydrochloride (Compound 1)

[0128]

[0129] 3-phenyl-4-propyl-1-(pyridin-2-yl)-1H-pyrazole-5-ol (280 mg, 1.0 mmol) prepared in the above-mentioned synthesis example 3 was dissolved in 4 mL of ether in a round-bottom flask, and then 0.55 mL of ether dissolved in 2M HCl was slowly added dropwise thereto at 0° C. The solid produced by the reaction solution was filtered under reduced pressure, the solvent was removed, washed with hexane and ethyl acetate, and then dried in vacuo to obtain the title compound (270 mg, 0.85 mmol).

[0130] 1H NMR (300MHz, CDCl3) δ8.44 (1H, d, J = 4.2Hz), 8.0-8.03 (2H, m), 7.66-7.64 (2H, m), 7.48-7.42 (3H , m), 7.34-7.30 (1H, m), 2.49 (2H, brs), 2.43 (2H, t, J = 7.5Hz), 1.48 (2H, m), 0.48 (3H, t, J = 7.3Hz)

[0131] <Example> Evaluation of the therapeutic effect of contrast agent-induced acute kidney injury in mice animal model

[0132] As experimental animals, 6-week-old male C57BL / 6 mice were purchased from LionBio. All mice were maintained under standard conditions. After about 2 weeks of breeding, at 8 weeks of age, compound 1 (3-phenyl-4-n-propyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol hydrochloride) was orally administered at a dose of 60 mg / kg / day for 5 days before contrast medium administration.

[0133] In order to induce acute renal injury, after fasting for 16 hours, the anti-inflammatory analgesic indomethacin (10 mg / kg) and NO blocker N-nitro-L-arginine (L-NAME, 10 mg / kg) were intraperitoneally administered 15 minutes before contrast agent administration, and contrast agent (Iohexol, 4000 mg iodine / kg) was intraperitoneally administered 15 minutes later. After 24 hours of food and water intake, the animal test substances were divided into three groups: a vehicle administration group (control, Control) and a contrast agent administration group (iohexol), and a contrast agent and compound 1 simultaneous administration group (iohexol + compound 1).

[0134] After the experiment, the animals were anesthetized, kidney tissue was removed from each individual, and blood was drawn for examination. The tissue was fixed in 10% neutral formalin buffer solution. The fixed tissue was sliced ​​to a certain thickness and embedded in paraffin by a conventional tissue processing process to prepare 4 μm to 5 μm tissue sections, and then hematoxylin & eosin staining (H&E staining) as a conventional staining method was performed to observe the degree of renal damage in the renal cortex and renal medulla regions in histopathology, and F4 / 80 staining, Tunel staining, DHE staining and nitrotyrosine staining were performed to observe the effects of inflammation and oxidative stress in renal injury tissue.

[0135] Blood was collected and centrifuged at 3000 rpm and 4°C for 10 minutes, and the upper serum was taken. The blood urea nitrogen (BUN) and creatinine levels were measured using an automatic blood biochemical analyzer (AU480, Beckman Coulter, USA) to confirm the index of renal injury. The histological evaluation of the degree of renal injury was calculated as follows: no tubular injury was 0 points, mild injury was 1 point, tubular vacuolar lesions less than 25% to 60% were 2 points, and tubular vacuolar lesions of more than 60% were 3 points, thereby obtaining the average value of tubular injury under the microscope.

[0136] Figure 1 The displayed photographs show the effect of reducing tubular damage in kidney tissue in the group administered with 60 mg / kg of compound 1 once a day for 5 days in the evaluation of the animal model of acute kidney injury in mice induced by contrast agent. Figure 1 As shown, as a result of collecting renal tissue and confirming the degree of renal damage in the renal cortex and renal medulla areas, it was found that, compared with the normal control group, in the group treated with contrast agent, vacuolar changes occurred due to tubular cell damage, and severe tubular tissue damage occurred due to the disappearance of the brush border, while the damage to the renal medulla and renal cortex in the group treated with compound 1 was significantly reduced.

[0137] Figure 2 and Figure 3 The figure shows the effect of reducing kidney damage in a blood test in a group treated with compound 1 at 60 mg / kg once a day for 5 days in an animal model of acute kidney injury in mice induced by contrast agent. Figure 2 and Figure 3 As shown, it can be found that the concentrations of BUN and creatinine (serum Cr) in the group treated with contrast agent were significantly increased compared with the normal control group, while the concentrations of BUN and creatinine in the group treated with compound 1 were decreased, especially the concentration of creatinine was decreased, which reduced kidney damage.

[0138] Figure 4The figure shows the effect of reducing the kidney injury markers NGAL, KIM-1 and urine microprotein (albumin) in urine in the group administered with compound 1 once a day, 60 mg / kg each time, for 5 days in the evaluation of the mouse acute kidney injury animal model caused by contrast agent. Figure 4 As shown, it can be found that the concentrations of NGAL, KIM-1 and urine microprotein (albumin) in the group treated with contrast agent were significantly increased compared with the normal control group, while the concentrations of NGAL, KIM-1 and urine microprotein (albumin) in the group treated with compound 1 were decreased, which reduced kidney damage.

[0139] Figure 5 A graph showing the effect of reducing tubular damage in renal biopsy in the group dosed with Compound 1 at 60 mg / kg once a day for 5 days in the evaluation of the animal model of acute kidney injury in mice induced by contrast agent. Figure 5 The results of collecting kidney tissues and comparing the degree of reduction in tubular injury are shown, and it can be found that the tubular injury score in the group treated with contrast agent was significantly increased, while the tubular injury score in the group treated with compound 1 was significantly decreased compared with the normal control group.

[0140] Figure 6 A graph showing the effect of reducing inflammation in kidney tissue in the group administered with 60 mg / kg of Compound 1 once a day for 5 days in the evaluation of the animal model of acute kidney injury in mice induced by contrast agent. Figure 6 The analysis of the expression level of the monocyte chemoattractant protein-1 (MCP-1) gene in kidney tissue, which is an indicator of inflammation, shows that the expression of MCP-1 increased in the group treated with contrast agent compared with the normal control group, while the inflammation (MCP-1) was significantly reduced in the group treated with compound 1.

[0141] Figure 7 The analysis of the degree of infiltration of inflammatory cells (macrophages, F4 / 80 positive cells) in the kidney tissue of the group treated with compound 1 at 60 mg / kg once a day for 5 days in the evaluation of the animal model of acute kidney injury in mice induced by contrast agent showed that the infiltration of inflammatory cells in the group treated with contrast agent was significantly increased compared with the normal control group, and the infiltration of inflammatory cells in the group treated with compound 1 was significantly reduced compared with the group treated with contrast agent.

[0142] Figure 8The results show that in the evaluation of the contrast-induced acute kidney injury animal model in mice, the level of reactive oxygen species (DHE staining), which is an indicator of oxidative stress in kidney tissue, was analyzed in the group treated with compound 1 at 60 mg / kg once a day for 5 days. It was found that the reactive oxygen species in the group treated with contrast agents was significantly increased compared with the normal control group, and the reactive oxygen species in the group treated with compound 1 was significantly reduced compared with the group treated with contrast agents.

[0143] Fig. 9 The present invention shows an analysis of the level of nitrotyrosine, an indicator of oxidative stress, in the renal tissue of the group treated with compound 1 at 60 mg / kg once a day for 5 days in the evaluation of an animal model of acute kidney injury in mice induced by contrast agents. It was found that nitrotyrosine in the group treated with contrast agents was significantly increased compared with the normal control group, and nitrotyrosine in the group treated with compound 1 was significantly decreased compared with the group treated with contrast agents.

[0144] Therefore, it can be confirmed that the compound of the present application has a significant renal protective effect in the contrast agent-induced acute kidney injury model.

Claims

1. Use of a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof in the preparation of a pharmaceutical composition for protecting the kidney by reducing the nephrotoxicity caused by contrast agents or for preventing or treating acute kidney injury caused by contrast agents: Chemical formula 1 in, R is a linear or branched alkyl group having 1 to 10 carbon atoms.

2. The use according to claim 1, wherein In Chemical Formula 1, R is a linear or branched alkyl group having 1 to 6 carbon atoms.

3. The use according to claim 2, wherein The compound of the chemical formula 1 is: 3-phenyl-4-methyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride; 3-phenyl-4-ethyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride; 3-phenyl-4-n-propyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride; 3-phenyl-4-isopropyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride; 3-phenyl-4-n-butyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride; 3-phenyl-4-tert-butyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride; 3-phenyl-4-n-pentyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride; or 3-phenyl-4-n-hexyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride.

4. The use according to claim 3, wherein The compound of Chemical Formula 1 is 3-phenyl-4-n-propyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol or its hydrochloride.

5. The use according to any one of claims 1 to 4, wherein The contrast agent is selected from an ionic monomer contrast agent or a non-ionic monomer contrast agent.

6. The use according to claim 5, wherein The ionic monomer contrast agent is selected from the group consisting of iodoxyglycate, iodamide, acetophenate, diatrizoate and metrizoate.

7. The use according to claim 5, wherein The non-ionic monomeric contrast agent is selected from mepanema, iohexol, iopamidol, iopentol, iopromide and ioversol.

8. The use according to claim 7, wherein The non-ionic monomeric contrast agent is iohexol.

9. The use according to any one of claims 1 to 4, wherein The acute kidney injury is caused by tubular necrosis or decreased renal perfusion.

10. The use according to any one of claims 1 to 4, wherein The acute kidney injury is caused by oxidative stress or inflammation.

11. The use according to any one of claims 1 to 4, wherein The pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

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