Composition for alleviating or treating cystic kidney disease, and use thereof

Abstract

The present invention relates to a renal disease treatment effect obtained by the co-administration of desloratadine, which is an antihistamine, and duloxetine, which is a serotonin reuptake inhibitor. It has been identified that, when desloratadine and duloxetine are co-administered, the formation and growth of cysts were inhibited in an ADPKD animal model, the renal function was also maintained at a normal value, liver cysts and liver fibrosis caused by ADPKD were treated, renal fibrosis was significantly treated in in vitro and animal renal fibrosis models, and, in particular, the effects were synergistically increased.

Description

Composition for improving or treating cystic kidney disease and use thereof

[0001] The present invention relates to the therapeutic effect of renal disease through combined administration of an antihistamine and a selective serotonin reuptake inhibitor.

[0002] The kidneys are vital organs that maintain homeostasis in the body. They regulate the amount of body fluids, ion concentrations and pH in the blood, excrete metabolic waste products, toxins, and drugs, and perform blood pressure control and other metabolic and endocrine functions. They also activate vitamin D, which helps calcium be absorbed in the small intestine and is involved in the synthesis of various hormones. Kidney disease is a condition in which the kidneys fail to perform their excretory, regulatory, metabolic, and endocrine functions normally, resulting in overall decline or abnormality in their function. Decline in kidney function due to damage can lead to enlargement of the kidney and related structures, renal atrophy, changes in body fluid volume, electrolyte imbalances, metabolic acidosis, impaired gas exchange, impaired anti-infective function, and accumulation of uremic toxins.

[0003] Chronic kidney disease (CKD) (chronic kidney failure) is recognized as a serious condition worldwide. The main causes are diabetes and high blood pressure, but other causes include urinary tract blockage (obstruction), certain kidney abnormalities (such as polycystic kidney disease and glomerulonephritis), and autoimmune diseases (such as systemic lupus erythematosus [lupus]) in which antibodies damage the small blood vessels (glomeruli) and small tubes (tubules) of the kidneys. When irreversible kidney damage caused by these various conditions or when kidney function does not improve after treatment and persists for more than three months, acute kidney injury becomes chronic kidney disease. Symptoms of chronic kidney disease include nocturia, fatigue, nausea, itching, muscle twitching and cramps, loss of appetite, confusion, shortness of breath, and swelling of the body (most commonly in the legs). The condition of patients with chronic kidney disease requiring renal replacement therapy such as dialysis or transplant is called end-stage renal disease (ESRD), and currently, there is no effective treatment other than renin-angiotensin-aldosterone system (RASS) inhibitors, such as angiotensin receptor blockers (ARBs) and angiotensin-converting enzyme (ACE) inhibitors alone or in combination. However, even these treatments only delay the onset of end-stage renal disease (ESRD) or suppress the decline in glomerular filtration rate (GFR) in some CKD patients, but the effect is minimal in the majority of CKD patients. If various chronic kidney diseases persist, they ultimately lead to end-stage renal failure.Chronic renal failure is caused by chronic glomerulonephritis, diabetes, high blood pressure, urinary tract obstruction, renal tuberculosis, and hereditary kidney diseases. Damaged kidney function does not recover even if the cause is treated, and the rate of progression to renal failure can be delayed but not prevented.

[0004] Polycystic kidney disease (PKD), one of the causes of chronic kidney disease, is caused by genetic defects. There are several genetic defects that cause PKD, some types of which are caused by dominant genes, and one rare type is caused by a recessive gene. When the gene is dominant, it is called autosomal dominant polycystic kidney disease (ADPKD). It is the most common human genetic disease of the kidney, and is mainly caused by germline mutations in the PKD1 or PKD2 genes, which encode the ciliary cyst proteins polycystin 1 and 2 (PC1 and PC2). It is known that 85% of patients diagnosed with ADPKD are caused by defects in the PKD1 gene, and 15% are caused by defects in the PKD2 gene. PC1 and PC2 proteins coexist throughout the cell membranes of renal epithelial cells, the extracellular matrix, and primary cilia, and deficiency of the PC1-PC2 complex in cilia is known to play a role in cyst formation. Patients with ADPKD develop multiple cysts in both kidneys, increasing in size and number until renal failure and death occur in their 50s or 60s. Cyst progression occurs as cyst epithelial cells proliferate, followed by cyst fluid filling the cyst, leading to increased cyst size. ADPKD accounts for approximately 2% of dialysis cases in Korea and is the third most common cause after diabetes, hypertension, and glomerulonephritis. ADPKD not only causes structural and functional defects in both kidneys but can also be accompanied by various renal complications, necessitating management from a relatively early stage of the disease. There is no cure for ADPKD, and tolvaptan is the only FDA-approved medication to alleviate the symptoms of ADPKD. Therefore, there is an urgent need for more effective drugs that slow the progression of ADPKD and show significant therapeutic effects.

[0005] The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating kidney disease.

[0006] In addition, another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of at least one selected from the group consisting of liver cysts and liver fibrosis.

[0007] In addition, another object of the present invention is to provide a pharmaceutical composition for preventing or improving kidney disease.

[0008] In addition, another object of the present invention is to provide a pharmaceutical composition for the prevention or improvement of at least one selected from the group consisting of liver cysts and liver fibrosis.

[0009] In addition, another object of the present invention is to provide a food composition for preventing or improving kidney disease.

[0010] In addition, another object of the present invention is to provide a food composition for preventing or improving at least one selected from the group consisting of liver cysts and liver fibrosis.

[0011] In addition, another object of the present invention is to provide a health functional food composition for preventing or improving kidney disease.

[0012] In addition, another object of the present invention is to provide a health functional food composition for preventing or improving at least one selected from the group consisting of liver cysts and liver fibrosis.

[0013] In addition, it is an object of the present invention to provide a method for preventing or treating kidney disease.

[0014] In addition, it is an object of the present invention to provide a method for preventing or treating at least one selected from the group consisting of liver cysts and liver fibrosis.

[0015] To achieve the above purpose, the present invention provides a pharmaceutical composition for preventing or treating kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

[0016] In addition, to achieve the above other purposes, the present invention provides a pharmaceutical composition for the prevention or treatment of at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer, a pharmaceutically acceptable salt and a solvate thereof.

[0017] In addition, to achieve the above-mentioned other object, the present invention provides an over-the-counter drug composition for preventing or improving kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

[0018] In addition, to achieve the above-mentioned other object, the present invention provides an over-the-counter pharmaceutical composition for preventing or improving at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer, a pharmaceutically acceptable salt and a solvate thereof.

[0019] In addition, to achieve the above-mentioned other object, the present invention provides a food composition for preventing or improving kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate; and duloxetine, a stereoisomer thereof, a food-based acceptable salt and solvate.

[0020] In addition, to achieve the above-mentioned other object, the present invention provides a food composition for preventing or improving at least one selected from the group consisting of liver cysts and liver fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate; and duloxetine, a stereoisomer thereof, a food-based acceptable salt and solvate.

[0021] In addition, to achieve the above-mentioned other object, the present invention provides a health functional food composition for preventing or improving kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate; and duloxetine, a stereoisomer thereof, a food-based acceptable salt and solvate.

[0022] In addition, to achieve the above-mentioned other object, the present invention provides a health functional food composition for preventing or improving at least one selected from the group consisting of liver cysts and liver fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate; and duloxetine, a stereoisomer thereof, a food-based acceptable salt and solvate.

[0023] In addition, to achieve the above-described further object, the present invention provides a method for preventing or treating kidney disease, comprising the step of administering to a subject at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

[0024] In addition, to achieve the above another object, the present invention provides a method for preventing or treating at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising the step of administering to a subject at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

[0025] In the present invention, a drug with a novel use was selected by screening FDA-approved drugs for a new use using a kidney organoid that mimics polycystic kidney disease, and the antihistamine desloratadine and the serotonin reuptake inhibitor duloxetine selected in this way inhibited the formation and growth of cysts when administered together, inhibited the formation and growth of cysts in an ADPKD animal model, and maintained renal function at normal levels, and treated liver cysts and liver fibrosis caused by ADPKD. In particular, the above effects were synergistically increased, and in particular, it was confirmed that the synergistic effect was superior to that of other drug combinations, and thus, it can be usefully used for the prevention or treatment of kidney disease.

[0026] Figure 1 is PKD1 - / - and PKD2 - / - This is a schematic diagram showing the process of inducing a defective mutation.

[0027] Figure 2 is PKD1 - / -This is the result of confirming PKD1 expression in mutant cells. Figure 2A shows the result of confirming PKD1 protein expression, and Figure 2B is a graph quantifying PKD1 protein expression.

[0028] Figure 3 is PKD1 - / - and PKD2 - / - PKD1 using defective mutant cells - / - This is the result of manufacturing renal organoids and confirming cyst formation. Figure 3A is a diagram showing the manufacturing process of renal organoids, and Figure 3B is PKD1 - / - and PKD2 - / - This is a diagram confirming cyst formation in renal organoids. In Figure 3B, the black bar is 500 μm.

[0029] Figure 4 is PKD1 - / - This is a diagram showing cyst formation after administering desloratadine and duloxetine alone or in combination at a concentration of 10 μM to renal organoids. In Figure 4, the black bar represents 500 μm.

[0030] Figure 5 is PKD1 - / - This graph quantifies cyst formation after administration of desloratadine and duloxetine alone or in combination at a concentration of 10 μM to renal organoids.

[0031] Figure 6 is PKD1 - / - This figure shows cyst formation after administering desloratadine and duloxetine alone or in combination at a concentration of 30 μM to renal organoids. In Figure 6, the black bar represents 500 μm.

[0032] Figure 7 is PKD1 - / - This graph quantifies cyst formation after administration of desloratadine and duloxetine alone or in combination at a concentration of 30 μM to renal organoids.

[0033] Figure 8 is PKD2 - / -This is a diagram showing cyst formation after desloratadine and duloxetine were administered alone or in combination at a concentration of 10 μM to renal organoids. In Figure 8, the black bar represents 500 μm.

[0034] Figure 9 is PKD2 - / - This graph quantifies cyst formation after administration of desloratadine and duloxetine alone or in combination at a concentration of 10 μM to renal organoids.

[0035] Figure 10 is PKD2 - / - This is a diagram showing cyst formation after desloratadine and duloxetine were administered alone or in combination at a concentration of 30 μM to renal organoids. In Figure 10, the black bar represents 500 μm.

[0036] Figure 11 is PKD2 - / - This graph quantifies cyst formation after administration of desloratadine and duloxetine alone or in combination at a concentration of 30 μM to renal organoids.

[0037] Figure 12 is PKD1 - / - IC of each drug in renal organoids 50 This is the result of confirming. Figure 12A shows the IC of desloratadine. 50 The results are as follows, and Figure 12B shows the IC of duloxetine. 50 This is the diagram that was confirmed.

[0038] Figure 13 is PKD1 - / - This is a diagram showing cyst formation after desloratadine and fluoxetine were administered alone or in combination at a concentration of 30 μM to renal organoids. In Figure 13, the black bar is 500 μm.

[0039] Figure 14 is PKD1 - / - This graph quantifies cyst formation after administration of desloratadine and fluoxetine alone or in combination at a concentration of 30 μM to renal organoids.

[0040] Figure 15 is PKD1 - / -This figure shows cyst formation after azelastine and fluoxetine were administered alone or in combination at a concentration of 30 μM to renal organoids. In Figure 15, the black bar represents 500 μm.

[0041] Figure 16 is PKD1 - / - This graph quantifies cyst formation after administration of azelastine and fluoxetine alone or in combination at a concentration of 30 μM to renal organoids.

[0042] Figure 17 is PKD1 - / - This figure shows cyst formation after azelastine and duloxetine were administered alone or in combination at a concentration of 10 μM to renal organoids. In Figure 17, the black bar represents 500 μm.

[0043] Figure 18 is PKD1 - / - This graph quantifies cyst formation after administration of azelastine and duloxetine alone or in combination at a concentration of 10 μM to renal organoids.

[0044] Figure 19 is PKD1 - / - This figure shows cyst formation after azelastine and duloxetine were administered alone or in combination at a concentration of 30 μM to renal organoids. In Figure 19, the black bar represents 500 μm.

[0045] Figure 20 is PKD1 - / - This graph quantifies cyst formation after azelastine and duloxetine were administered alone or in combination at a concentration of 30 μM to renal organoids.

[0046] Figure 21 is PKD1 - / - This figure shows cyst formation after ketotifen and fluoxetine were administered alone or in combination at a concentration of 30 μM to renal organoids. In Figure 21, the black bar represents 500 μm.

[0047] Figure 22 is PKD1 - / - This graph quantifies cyst formation after administering ketotifen and fluoxetine alone or in combination at a concentration of 30 μM to renal organoids.

[0048] Figure 23 is PKD1 - / - This is a diagram analyzing the mechanism of the cyst formation inhibitory effect of desloratadine in renal organoids.

[0049] Figure 24 is PKD1 - / - This is a diagram analyzing the mechanism of duloxetine's inhibitory effect on cyst formation in renal organoids.

[0050] Figure 25 is PKD1 - / - This is the result of confirming the effect on cyst growth when desloratadine metabolites and duloxetine metabolites were administered to renal organoids, respectively. Figure 25A is an image confirmed by bright-field microscopy after administration of 3-hydroxy desloratadine, a desloratadine metabolite, and (S)-5-hydroxy-6-methoxy duloxetine, a duloxetine metabolite. Figure 25B is the result of quantifying the cyst area of ​​the renal organoid, and Figure 25C is the structural formula of 3-hydroxy desloratadine, a desloratadine metabolite, and (S)-5-hydroxy-6-methoxy duloxetine, a duloxetine metabolite. Values ​​are expressed as mean ± standard error (SEM), * indicates p<0.05 vs PKD1 - / - It is a comparison with . In the above Fig. 25A, the black bar is 500 μm.

[0051] Figure 26 shows the results of examining the synergistic effect of co-administration of desloratadine and duloxetine in reducing the growth of renal cysts. Figure 26A shows the CI (combination index) calculation method using the Chou-Talalay method. Figure 26B shows the PKD1 - / - IC of desloratadine and duloxetine based on cyst formation rate in renal organoids 50 This is the result of measurement. The values ​​are expressed as the mean ± standard error. Figure 26C shows the CI value calculated using the above CI calculation method.

[0052] Figure 27 shows the results of confirming the effect of co-administration of desloratadine and duloxetine on receptors involved in the pathological mechanism in an ADPKD mouse model. Figure 27A shows an ADPKD mouse model, Pkd1 flox / flox ;R26CreER + / - A schematic diagram showing the manufacturing process of the mouse, and Figure 27B shows Pkd1 flox / flox ;R26CreER + / - A schematic diagram of the in vivo experimental design for administering drugs to mice. Figure 27C shows Pkd1 flox / flox ;R26CreER + / - Confocal images of histamine H1 receptors and serotonin 1A (5-HT1A) receptors in mouse kidney. Figure 27D is an IHC (Immunohistochemistry) image for the histamine H1 receptor. Figure 27E is an IHC image for 5-HT1A. Figure 27F is an image confirming the expression of histamine H1 receptor, histamine H3 receptor, 5-HT1A, and 5-HT2A receptor proteins. Figure 27G is a graph quantifying histamine H1 receptor protein expression. Figure 27H is a graph quantifying histamine H3 receptor protein expression. Figure 27I is a graph quantifying 5-HT1A protein expression. Figure 27J is a graph quantifying 5-HT2A protein expression. In the above Figures 27G to 27J, post hoc Mann-Whitney U test was performed, and the values ​​are expressed as mean ± standard error (SEM). *, p<0.05 vs. wild type; #, p<0.05 vs. vehicle-treated Pkd1 flox / flox ;R26CreER + / -This is a comparison with the military. In the above Fig. 27C, the white bar is 200 μm. In Fig. 27D, the upper black bar is 1000 μm, and the lower black bar is 100 μm. In Fig. 27E, the upper black bar is 1000 μm, and the lower black bar is 100 μm.

[0053] Figure 28 shows the results of examining the effects of co-administration of desloratadine and duloxetine on renal cyst growth and renal function in an ADPKD mouse model. Figure 28A shows the kidney of a wild-type (WT) mouse and a vehicle-treated Pkd1 mouse. flox / flox ;R26CreER + / - Mouse kidney, Pkd1 co-administered with desloratadine and duloxetine flox / flox ;R26CreER + / - The results of staining the mouse kidney with H&E are shown in Fig. 28B. The results of confirming the kidney weight per body weight of each experimental group are shown in Fig. 28C. The results of confirming the renal cyst index of each experimental group are shown in Fig. 28D. The results of confirming the number of renal cysts of each experimental group are shown in Fig. 28E. The graph quantifying the serum BUN of each experimental group is shown in Fig. 28F. The IHC images of the kidneys of each experimental group stained with PCNA are shown in Fig. 28G. The results of quantifying the PCNA-positive area are shown in Fig. 28H. The results of confirming the PCNA protein expression of each experimental group are shown in Fig. 28I. In Fig. 28, a post hoc Mann-Whitney U test was performed, and the values ​​are expressed as the mean ± standard error (SEM). *, p<0.05 vs wild type; #, p<0.05 vs vehicle-treated Pkd1 flox / flox ;R26CreER + / - This is a comparison with the military. In the above Fig. 28A, the black bar is 1 mm. In Fig. 28F, the upper black bar is 1000 μm, and the lower black bar is 100 μm.

[0054] Figure 29 shows the results of examining the effects of co-administration of desloratadine and duloxetine on renal fibrosis in an ADPKD mouse model. Figure 29A shows the kidney of a wild-type (WT) mouse and a Pkd1 mouse treated with vehicle. flox / flox ;R26CreER + / - Mouse kidney, Pkd1 co-administered with desloratadine and duloxetine flox / flox ;R26CreER + / - This is the result of Masson's trichrome staining of the mouse kidney, and Fig. 29B is a graph quantifying the renal fibrosis area measured by Masson's trichrome staining in each experimental group, Fig. 29C is a confocal image of fibronectin in each experimental group, Fig. 29D is the result of confirming the fibronectin protein expression in each experimental group, Fig. 29E is a graph quantifying the fibronectin protein expression in each experimental group, Fig. 29F is the result of IHC staining with α-SMA in each experimental group, Fig. 29G is the result of confirming the α-SMA protein expression in each experimental group, Fig. 29H is a graph quantifying the α-SMA protein expression in each experimental group, Fig. 29I is the result of IHC staining with TGF-β in each experimental group, Fig. 29J is the result of confirming the TGF-β and Smad4 protein expression in each experimental group, and Fig. 29K is the result of confirming the TGF-β protein expression in each experimental group. Figure 29L is a graph quantifying Smad4 protein expression in each experimental group, Figure 29M is a graph confirming the expression of IL-1β, YAP, and C-myc proteins in each experimental group, Figure 29N is a graph quantifying IL-1β protein expression, Figure 29O is a graph quantifying YAP protein expression, and Figure 29P is a graph quantifying C-myc protein expression. In Figure 29, post hoc Mann-Whitney U test was performed, and the values ​​are expressed as the mean ± standard error (SEM). *, p<0.05 vs. wild type; #, p<0.05 vs. Pkd1 treated with vehicle. flox / flox ;R26CreER+ / - This is a comparison with the military. In the above Fig. 29A, the upper black bar is 1000 μm, and the lower black bar is 100 μm. In Fig. 29C, the white bar is 200 μm. In Fig. 29F, the upper black bar is 1000 μm, and the lower black bar is 100 μm. In Fig. 29I, the upper black bar is 1000 μm, and the lower black bar is 100 μm.

[0055] Figure 30 shows the results of confirming the effect of co-treatment with desloratadine and duloxetine on a renal fibrosis-inducing cell model (in vitro). Figure 30A is a schematic diagram of the experimental design for the preparation of a renal fibrosis-inducing cell model using human kidney proximal tubular epithelial cells (HK-2) and drug administration. Figure 30B shows the results of confirming the appropriate drug administration concentration for the renal fibrosis-inducing cell model by confirming the expression of renal fibrosis markers (Collagen I, CTGF) after administering desloratadine at 5, 10, or 15 μM. Figure 30C shows the results of confirming the appropriate drug administration concentration for the renal fibrosis-inducing cell model by confirming the expression of renal fibrosis markers (Collagen I, CTGF) after administering doloxetine at 5, 10, or 15 μM. Figure 30D shows the results of confirming the expression of renal fibrosis markers (Collagen I, CTGF) after single and co-administration of desloratadine (10 μM) and duloxetine (15 μM) in a renal fibrosis-induced cell model. In Figure 30D, *; p<0.05 vs. CTR (control group), and #; p<0.05 vs. TGF treatment group. Figure 30E is a schematic diagram of the process for preparing an adenine diet animal model, which is an animal model with reduced renal function as an in vivo chronic kidney disease (CKD) animal model, and the experimental design for drug administration. Figure 30F shows the results of confirming the serum BUN (blood urea levels) levels after co-administration of desloratadine and duloxetine to a wild-type animal model and an adenine diet animal model. In the above Figure 30F, * is p<0.05 vs. comparison with the adenine diet animal model (23 weeks) group.

[0056] Figure 31 shows the results of confirming the therapeutic effect of co-administration of desloratadine and duloxetine on hepatic cysts and fibrosis caused by ADPKD in an ADPKD mouse model. Figure 31A shows the liver of a wild-type (WT) mouse and Pkd1 treated with vehicle. flox / flox ;R26CreER + / - Pkd1 in mice co-administered with desloratadine and duloxetine flox / flox ;R26CreER + / - H&E staining results of mouse liver, Fig. 31B shows the results of confirming the liver cyst index of each experimental group, Fig. 31C shows the results of confirming the number of liver cysts of each experimental group, Fig. 31D is a graph quantifying serum SGOT of each experimental group, Fig. 31E is a graph quantifying serum SGPT of each experimental group, Fig. 31F is an IHC image stained with PCNA of each experimental group, Fig. 31G is a graph quantifying the PCNA positive area of ​​each experimental group, and Fig. 31H shows wild-type (WT) mouse liver and Pkd1 treated with vehicle. flox / flox ;R26CreER + / - Pkd1 in mice co-administered with desloratadine and duloxetine flox / flox ;R26CreER + / - Masson's trichrome staining results of mouse livers, Figure 31I is a graph quantifying the fibrotic area in the livers of each experimental group, Figure 31J is the IHC results stained with α-SMA in the livers of each experimental group, and Figure 31K is a graph quantifying the α-SMA-positive area in the livers of each experimental group. In Figure 31, post hoc Mann-Whitney U test was performed, and the values ​​are expressed as the mean ± standard error (SEM). *, p<0.05 vs. wild type; #, p<0.05 vs. vehicle-treated Pkd1 flox / flox ;R26CreER + / -This is a comparison with the military. In the above Fig. 31A, the black bar is 1 mm. In Fig. 31F, the upper black bar is 1000 μm, and the lower black bar is 500 μm. In Fig. 31H, the upper black bar is 1000 μm, and the lower black bar is 100 μm. In Fig. 31J, the upper black bar is 1000 μm, and the lower black bar is 100 μm.

[0057] Hereinafter, the present invention will be described in more detail.

[0058] The present invention provides a pharmaceutical composition for preventing or treating kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

[0059] The above desloratadine may be 8-chloro-6,11-dihydro-11-(4-piperidylidene)-5H-benzo[5,6]cyclohepta-[1,2-b]pyridine, and may be a compound represented by the following chemical formula 1:

[0060]

[0061] In addition, the above duloxetine may be (+)-(S)-N-methyl-3-(naphthalen-1-yloxy)-3-(thiophen-2-yl)propan-1-amine, and may be a compound represented by the following chemical formula 2:

[0062]

[0063] According to one embodiment of the present invention, PKD1- / - When desloratadine and duloxetine were co-administered to renal organoids, a more significant cyst growth inhibition effect was observed than when azelastine, an antihistamine, and fluoxetine, an SSRI, were co-administered, when azelastine, an antihistamine, and duloxetine, an SSRI, were co-administered, or when ketotifen fumarate, a histamine receptor antagonist, and fluoxetine, an SSRI, were co-administered.

[0064] In addition, in the present invention, the desloratadine, its stereoisomer, pharmaceutically acceptable salt or solvate and the duloxetine, its stereoisomer, pharmaceutically acceptable salt or solvate may be mixed in a weight ratio of 1:1 to 20, but is not limited thereto. Preferably, the desloratadine, its stereoisomer, pharmaceutically acceptable salt or solvate may be mixed in a weight ratio of 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18 or 1:19, but is not limited thereto. More preferably, it may be mixed in a weight ratio of 1:1 to 5, even more preferably, it may be mixed in a weight ratio of 1:1 or 1:2, and even more preferably, it may be mixed in a weight ratio of 1:2, but is not limited thereto.

[0065] In addition, according to one embodiment of the present invention, desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate showed a significant therapeutic effect on hepatic cysts or liver fibrosis caused by renal disease.

[0066] Accordingly, the present invention provides a pharmaceutical composition for the prevention or treatment of at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer, a pharmaceutically acceptable salt and a solvate thereof.

[0067] The pharmaceutically acceptable salts may be selected from the group consisting of alkali metal salts, alkaline earth metal salts, salts with inorganic acids, salts with organic acids and salts with acidic amino acids, and as salts, acid addition salts formed by pharmaceutically acceptable free acids are useful. Acid addition salts are obtained from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and non-toxic organic acids such as aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates and alkanedioates, aromatic acids, aliphatic and aromatic sulfonic acids. These pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, Contains phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate or mandelate.

[0068] The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compound in an excess of an aqueous acid solution and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Alternatively, the salt can be prepared by evaporating the solvent or excess acid from the mixture and drying it, or by suction filtration of the precipitated salt. In addition, a pharmaceutically acceptable metal salt can be prepared using a base. An alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. In this case, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt. In addition, the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0069] The present invention encompasses not only the above compounds, but also all possible solvates, hydrates, racemates or stereoisomers that can be prepared therefrom.

[0070] In one embodiment, the renal disease may be one or more selected from the group consisting of cyst formation or renal disease associated with cyst formation, polycystic kidney disease (PKD), renal fibrosis, acute kidney injury, end-stage renal disease (ESKD), renal failure, systemic lupus erythematosus, diabetic nephropathy (DN), IgA nephritis (IgAN), HIV-associated nephropathy, chronic kidney disease (CKD), nephrosclerosis, focal segmental glomerulosclerosis (FSGS), minimal change nephrotic syndrome (MCD), xanthine oxidase deficiency, abetalipoproteinemia, familial hypobetalipoproteinemia (FHBL), chylomicrons retention disease (CRD), sitosterolemia, glomerular hyperfiltration, and pruritus of renal failure, and may be accompanied by, but is not limited to, hepatic cysts and hepatic fibrosis.

[0071] In the present invention, the renal disease is more preferably polycystic kidney disease (PKD) or renal fibrosis. Furthermore, liver cysts and liver fibrosis may be caused by polycystic kidney disease (PKD), but are not limited thereto.

[0072] In one embodiment, the polycystic kidney disease (PKD) can be autosomal dominant polycystic kidney disease (ADPKD) or autosomal recessive polycystic kidney disease (ARPKD), more preferably a dominant polycystic kidney disease, most preferably a dominant polycystic kidney disease having a deletion mutation at position 36 of the PKD1 gene or a deletion mutation at exon 4 of the PKD2 gene.

[0073] The term "cyst" used in the present invention is a general term for a pouch-shaped structure with a membrane that separates it from the surrounding tissues in the body. Cysts contain liquid, air, or semi-solid substances. Cysts can be congenital due to genetic causes, or can be caused by tumors, infections, chronic inflammation, or blood vessel damage. Congenital cysts include liver cysts, kidney cysts, and dermoid cysts that appear on the skin and ovaries, while acquired cysts include pancreatic cysts caused by blockage of the glandular drainage duct, and echinococcal cysts caused by parasitic infections.

[0074] In one embodiment, the composition of the present invention can inhibit cystogenesis or growth, for example, by reducing the size of cysts formed in the kidney by 10 to 100%, 10 to 90%, 10 to 80%, 10 to 70%, 10 to 60%, 10 to 50%, 10 to 40%, 10 to 30%, 10 to 20%, 20 to 100%, 20 to 80%, 20 to 60%, 20 to 40%, 30 to 100%, 30 to 80%, 30 to 60%, 30 to 40%, 40 to 100%, 40 to 80%, 40 to 60%, 50 to 100%, 50 to 80%, 50 to 60 %, 60 to 100%, 60 to 80%, 70 to 100%, 70 to 80%, 80 to 100%, or 90 to 100%.

[0075] In addition, the composition of the present invention can maintain renal function, reduce the increased BUN level at the time of onset of polycystic kidney disease, or maintain it at the level before the onset of the disease.

[0076] Additionally, the composition of the present invention can increase phosphorylation of Akt.

[0077] Additionally, the composition of the present invention can down-regulate the B-Raf-MEK-ERK signaling pathway or the Wnt signaling pathway.

[0078] Additionally, the composition of the present invention can reduce phosphorylation of p38 MAPK, ERK(1 / 2), GSK3β, β-catenin, or mTOR.

[0079] Additionally, the composition of the present invention can reduce the expression of B-Raf or c-Myc.

[0080] The composition of the present invention may further comprise at least one therapeutic agent selected from the group consisting of known renal diseases or liver cysts and liver fibrosis, in addition to at least one selected from the group consisting of desloratadine, stereoisomers, pharmaceutically acceptable salts or solvates thereof; and duloxetine, stereoisomers, pharmaceutically acceptable salts and solvates thereof, and may be used in combination with other known treatments for the treatment of these diseases.

[0081] As used herein, the term "prevention" means any act of inhibiting or delaying the occurrence, spread, and recurrence of any one or more diseases selected from the group consisting of kidney disease, liver cysts, and liver fibrosis by administering the pharmaceutical composition according to the present invention, and "treatment" means any act of improving or beneficially changing any one or more symptoms selected from the group consisting of kidney disease, liver cysts, and liver fibrosis by administering the composition of the present invention. Anyone of ordinary skill in the art to which the present invention pertains will be able to know the exact criteria for diseases to which the composition of the present invention is effective and determine the degree of improvement, enhancement, and treatment by referring to materials presented by the Korean Medical Association, etc.

[0082] The term "therapeutically effective amount" used in combination with the active ingredient in the present invention means an amount effective in preventing or treating at least one selected from the group consisting of kidney disease, liver cysts, and liver fibrosis, and the therapeutically effective amount of the composition of the present invention may vary depending on various factors, such as the administration method, target site, and patient condition. Therefore, the dosage for use in humans should be determined as an appropriate amount by taking both safety and efficacy into consideration. It is also possible to estimate the amount to be used in humans from the effective amount determined through animal testing.

[0083] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects. The effective dosage level may be determined based on factors including the patient's health condition, type of kidney disease, cause of kidney disease, severity, activity of the drug, sensitivity to the drug, method of administration, time of administration, route of administration and excretion rate, duration of treatment, combination or concurrent use of drugs, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by those skilled in the art.

[0084] The pharmaceutical composition of the present invention may include a carrier, diluent, excipient, or a combination of two or more thereof commonly used in biological preparations. The term "pharmaceutically acceptable" as used herein means that the composition exhibits non-toxic properties to cells or humans exposed to the composition. The carrier is not particularly limited as long as it is suitable for in vivo delivery of the composition. For example, compounds, saline solution, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components may be mixed and used, and other common additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into a main-use dosage form such as an aqueous solution, suspension, or emulsion, pills, capsules, granules, or tablets. Furthermore, the composition may be preferably formulated according to each disease or component using an appropriate method in the art.

[0085] In one embodiment, the pharmaceutical composition may be in one or more dosage forms selected from the group consisting of oral dosage forms, topical preparations, suppositories, sterile injectable solutions and sprays, with oral or injectable dosage forms being more preferred.

[0086] The term "administration" used in the present invention means providing a predetermined substance to an individual or patient by any appropriate method, and may be administered parenterally (for example, intravenously, subcutaneously, intraperitoneally, or locally in the form of an injection) or orally depending on the intended method, and the dosage range varies depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease. Liquid preparations for oral administration of the composition of the present invention include suspensions, oral solutions, emulsions, syrups, etc., and may include various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. in addition to commonly used simple diluents such as water and liquid paraffin. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, etc. The pharmaceutical composition of the present invention may be administered by any device that allows the active substance to move to target cells. Preferred administration methods and formulations include intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, and drip injection. Injections can be manufactured using aqueous solvents such as saline solution and Ringer's solution, non-aqueous solvents such as vegetable oil, higher fatty acid ester (e.g., ethyl oleate, etc.), alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.), and pharmaceutical carriers such as stabilizers to prevent deterioration (e.g., ascorbic acid, sodium bisulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers to adjust pH, and preservatives to inhibit microbial growth (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

[0087] The term "subject" as used in the present invention means any animal, including a human, a monkey, a cow, a horse, a sheep, a pig, a chicken, a turkey, a quail, a cat, a dog, a mouse, a rat, a rabbit or a guinea pig, which has developed or may develop at least one disease selected from the group consisting of kidney disease, liver cysts and liver fibrosis, and the above diseases can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to the subject. The pharmaceutical composition of the present invention can be administered in combination with an existing therapeutic agent.

[0088] The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable additive. At this time, the pharmaceutically acceptable additive may include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, maltose, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol, and talc. The pharmaceutically acceptable additive according to the present invention is preferably included in the composition in an amount of 0.1 to 90 parts by weight, but is not limited thereto.

[0089] The present invention also provides an over-the-counter drug composition for preventing or improving kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

[0090] In addition, the present invention provides an over-the-counter pharmaceutical composition for preventing or improving liver cysts and liver fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof.

[0091] In the present invention, the above "quasi-drug" means a fiber, rubber product or similar product used for the purpose of treating, alleviating, managing or preventing a disease of humans or animals; a product that has a weak effect on the human body or does not directly affect the human body, is not an apparatus or machine or similar product; and is one of the preparations used for sterilization, insecticide and similar purposes to prevent infection. It means a product used for the purpose of diagnosing, treating, alleviating, managing or preventing a disease of humans or animals, which is not an apparatus, machine or device, and is used for the purpose of exerting a pharmacological effect on the structure and function of humans or animals, and includes external skin preparations and personal hygiene products.

[0092] The present invention may be used as is, including at least one selected from the group consisting of desloratadine, stereoisomers thereof, pharmaceutically acceptable salts or solvates thereof, and duloxetine, stereoisomers thereof, pharmaceutically acceptable salts and solvates thereof, or may be used together with other quasi-drug ingredients, and may be used appropriately according to a conventional method. The mixing amount of the active ingredients may be appropriately determined depending on the intended use.

[0093] The quasi-drug of the present invention is not particularly limited thereto, but can be manufactured and used in the form of, for example, a cream, a lotion, an aerosol, a gel, or a pack. In the case of a cream, an ointment, a gel, or a pack, a base such as white petrolatum, yellow petrolatum, linolenic acid, bleached beeswax, cetanol, stearyl alcohol, stearic acid, hydrogenated oil, gelling hydrocarbon, polyethylene glycol, liquid paraffin, or squalane; a solvent and a solubilizing agent such as oleic acid, isopropyl myristate, glycerin triisooctanoate, crotamiton, diethyl sebacate, diisopropyl adipate, hexyl laurate, fatty acids, fatty acid esters, aliphatic alcohols, or vegetable oils; an antioxidant such as a tocopherol derivative, L-ascorbic acid, dibutylhydroxytoluene, or butylhydroxyanisole; a preservative such as parahydroxybenzoic acid ester; There are moisturizers such as glycerin, propylene glycol, and sodium hyaluronate; surfactants such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, and lecithin; and thickeners such as carboxyvinyl polymers, xanthan gum, carboxymethylcellulose, carboxymethylcellulose sodium salts, hydroxypropylcellulose, and hydroxypropylmethylcellulose.

[0094] In the case of aerosols, bases such as white petrolatum, yellow petrolatum, lanolin, bleached beeswax, cetanol, stearyl alcohol, stearic acid, hydrogenated oils, gelling hydrocarbons, polyethylene glycol, liquid paraffin, and squalane used in the preparation of ointments, creams, gels, suspensions, emulsions, solutions, and lotions; solvents and solubilizing agents such as oleic acid, isopropyl myristate, diisopropyl adipate, isopropyl sebacate, glycerin triisooctanoate, crotamiton, diethyl sebacate, hexyl laurate, fatty acids, fatty acid esters, aliphatic alcohols, and vegetable oils; antioxidants such as tocopherol derivatives, L-ascorbic acid, dibutylhydroxytoluene, and butylhydroxyanisole; preservatives such as parahydroxybenzoic acid esters; Humectants such as glycerin, propylene glycol, and sodium hyaluronate; surfactants such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, and lecithin; thickeners such as carboxyvinyl polymers, xanthan gum, carboxymethylcellulose, carboxymethylcellulose sodium salts, hydroxypropylcellulose, and hydroxypropylmethylcellulose; In addition, various stabilizers, buffers, coagulants, suspending agents, emulsifiers, fragrances, preservatives, solubilizers, and other suitable additives can be blended. In addition, stabilizers, preservatives, absorption promoters, pH adjusters, and other suitable additives can be blended as needed.

[0095] For the pharmaceutical product of the present invention, a more specific description is the same as the description for the above pharmaceutical composition.

[0096] The present invention also provides a food composition for preventing or improving kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a food-acceptable salt and solvate thereof.

[0097] In addition, the present invention provides a food composition for preventing or improving at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate; and duloxetine, a stereoisomer thereof, a food-based acceptable salt and solvate.

[0098] In addition, the present invention provides a health functional food composition for preventing or improving kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a food-acceptable salt and solvate thereof.

[0099] In addition, the present invention provides a health functional food composition for preventing or improving at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate; and duloxetine, a stereoisomer thereof, a food-based acceptable salt and solvate.

[0100] In the present invention, the food-wise acceptable salt may include a salt derived from a food-wise acceptable organic acid, inorganic acid, or base.

[0101] When the composition of the present invention is used as a food composition, the compound may be added as is or used in combination with other foods or food ingredients, and may be used appropriately according to conventional methods. In addition to the active ingredient, the composition may include a food-related acceptable food additive, and the amount of the active ingredient mixed may be appropriately determined depending on the intended use (prevention, health, or therapeutic treatment).

[0102] The term "food" used in the present invention means a natural product or processed product containing one or more nutrients, preferably a product that has gone through a certain degree of processing to become directly edible, and in its general meaning includes all health functional foods, beverages, food additives, and beverage additives.

[0103] The term "food supplement additive" used in the present invention means a component that can be added to food as an auxiliary, and can be appropriately selected and used by a person skilled in the art as added in the manufacture of health functional foods of each formulation. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc., but the types of food supplement additives of the present invention are not limited by the above examples.

[0104] The food composition of the present invention may include a health functional food. The term "health functional food" as used in the present invention refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients having functionality useful to the human body. Here, "functionality" means obtaining a useful effect for health purposes, such as regulating nutrients for the structure and function of the human body or physiological effects. The health functional food of the present invention can be manufactured by a method commonly used in the art, and during the manufacturing process, raw materials and ingredients commonly added in the art can be added. In addition, the formulation of the health functional food can also be manufactured without limitation as long as it is a formulation recognized as a health functional food. The health functional food of the present invention can be taken as a supplement to enhance the effect of a kidney disease treatment agent.

[0105] In addition, there is no limitation on the type of health food in which the composition of the present invention can be used. In addition, the composition of the present invention can be manufactured by mixing other appropriate auxiliary ingredients that can be included in health functional foods and known additives according to the selection of a person skilled in the art. Examples of foods to which it can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and it can be manufactured by adding it to juice, tea, jelly, and juice manufactured using the extract according to the present invention as a main ingredient.

[0106] The term "improvement" as used in the present invention means any action that reduces a parameter related to a desired disease, for example, the severity of a symptom, by administration of a composition according to the present invention.

[0107] In addition, the present invention provides a method for preventing or treating kidney disease, comprising the step of administering to a subject at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

[0108] In addition, the present invention provides a method for preventing or treating at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising the step of administering to a subject at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof, and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

[0109] In one embodiment, the renal disease may be one or more selected from the group consisting of cyst formation or renal disease associated with cyst formation, polycystic kidney disease (PKD), renal fibrosis, acute kidney injury, end-stage renal disease (ESKD), renal failure, systemic lupus erythematosus, diabetic nephropathy (DN), IgA nephritis (IgAN), HIV-associated nephropathy, chronic kidney disease (CKD), nephrosclerosis, focal segmental glomerulosclerosis (FSGS), minimal change nephrotic syndrome (MCD), xanthine oxidase deficiency, abetalipoproteinemia, familial hypobetalipoproteinemia (FHBL), chylomicrons retention disease (CRD), sitosterolemia, glomerular hyperfiltration, and pruritus of renal failure, and may be accompanied by, but is not limited to, hepatic cysts and hepatic fibrosis.

[0110] In the present invention, the renal disease is more preferably polycystic kidney disease (PKD) or renal fibrosis. Furthermore, liver cysts and liver fibrosis may be caused by polycystic kidney disease (PKD), but are not limited thereto.

[0111] In one embodiment, the polycystic kidney disease (PKD) can be autosomal dominant polycystic kidney disease (ADPKD) or autosomal recessive polycystic kidney disease (ARPKD), more preferably a dominant polycystic kidney disease, most preferably a dominant polycystic kidney disease having a deletion mutation at position 36 of the PKD1 gene or a deletion mutation at exon 4 of the PKD2 gene.

[0112] The above-mentioned subject is preferably a mammal, including a human, and includes all patients who are being treated, have been treated, or need to be treated for a disease such as kidney disease or liver cysts or liver fibrosis, and may also include patients who have undergone surgical operation for the treatment of kidney disease or liver cysts or liver fibrosis.

[0113]

[0114] Throughout this specification, '%' used to indicate the concentration of a particular substance means (w / w)% for solid / solid, (w / v)% for solid / liquid, and (v / v)% for liquid / liquid, unless otherwise stated.

[0115] The terminology used in this specification is intended to appropriately express preferred embodiments of the present invention, and may vary depending on the intent of the user or operator, or the customs of the field to which the present invention pertains. Therefore, the definitions of these terms should be determined based on the contents of this specification as a whole. Throughout this specification, when a part is said to "include" a certain component, unless specifically stated otherwise, this does not mean that other components are excluded, but rather that other components may be included.

[0116] Unless otherwise defined, all technical terms used in this invention have the same meaning as commonly understood by those skilled in the art. While preferred methods and samples are described herein, similar or equivalent methods are also included within the scope of the present invention. The contents of all publications cited as references herein are incorporated herein by reference.

[0117] Hereinafter, the present invention will be described in detail with reference to the attached drawings and exemplary embodiments thereof. However, the following exemplary embodiments are provided as illustrative examples of the present invention. If a detailed description of a technology or configuration well known to those skilled in the art is judged to unnecessarily obscure the gist of the present invention, such detailed description may be omitted, and the present invention is not limited thereby. The present invention is capable of various modifications and applications within the scope of the following claims and equivalents interpreted therefrom.

[0118]

[0119] Example 1. In vitro modeling of polycystic kidney disease

[0120] 1-1. Generation of PKD1 and PKD2 deletion iPSCs

[0121] To model the disease and screen for substances that alter the disease phenotype, human induced pluripotent stem cells (iPSCs) containing deletions in the PKD1 and PKD2 genes, which have been reported to be associated with autosomal dominant polycystic kidney disease (ADPKD), were generated using CRISPR-Cas9 technology to introduce deletion mutations in exon 36 of the PKD1 gene and exon 4 of the PKD2 gene. Specifically, PKD1 and PKD2 gene oligos (PKD1 gRNA: GTG GGT GCG AGC TTC CCC CC (SEQ ID NO: 1); and PKD2 gRNA: CGA ATA CGG CAA CTC CGA GT (SEQ ID NO: 2)) were constructed, cloned into an all-in-one vector (All-in-one CRISPR / Cas9) (Cat. A21174, GeneArt CRISPR Nuclease Vector Kit, ThermoFisher scientific), and sequenced to verify. The cloned all-in-one Cas9 nuclease reporter vector DNA was transfected into iPSCs (WTC11) by electroporation, and one day later, cells expressing OPF were selected with FACs and cultured for 7 to 10 days. Only cells in which colonies were formed were selected, DNA was extracted, and sequenced. To verify PKD1 and PKD2 mutant alleles, single-cell-derived clones of PKD1 and PKD2 targeted hPSC cell lines were expanded and genomic DNA was extracted using the AccuPrep Genomoic DNA Extraction Kit (Bioneer, K-3032).The CRISPR-Cas9 targeting region was amplified with PCR Master Mix (PKD1 primer set: 5'-AGGTTAACATGGGCTTGGCT-3' (SEQ ID NO: 3) and 5'- GAGACAAGAGACGGAGGTGG-3' (SEQ ID NO: 4); and PKD2 primer set: 5'-GTAAAACGACGGCCAGT-3' (SEQ ID NO: 5) and 5'- CAGGAAACAGCTATGAC-3' (SEQ ID NO: 6)) and Sanger sequencing was used to confirm the CRISPR mutation in each allele, thereby generating clones of the human iPSC WTC11 cell line containing deletion mutations in exon 36 of the PKD1 gene and exon 4 of the PKD2 gene (Fig. 1).

[0122]

[0123] 1-2. Check for loss of function

[0124] PKD1 manufactured in the above Example 1-1 - / - To verify the loss-of-function mutation in iPSCs, the expression level of PKD1 protein was analyzed by immunoblotting. Specifically, PKD1 - / -iPSCs were disrupted to isolate total proteins, and the concentration of the isolated proteins was quantified using Bradford Assay reagent (Bio-Rad. Hercules, CA, USA). The same amount of protein was electrophoresed in NuPAGETM 3–8% Tris-Acetate protein Gel (Invitrogen, EA03785) with a high-molecular-weight protein marker (Invitrogen, Cat. no #LC5699), and then transferred to a PVDF membrane (Bio-Rad, Hercules, CA, USA). The membrane was blocked with 5% skim milk in PBS-T (80 mM Na2HPO4, 20 mM NaH2PO4, 100 mM NaCl, 0.1% Tween-20, pH 7.5) for 1 h and reacted with rabbit polyclonal anti-PKD1 antibody (MyBioSource, Catalog no. a173900) overnight at 4°C. Subsequently, the samples were reacted with horseradish peroxidase (HP) (Jackson ImmunoResearch, West Grove, PA)-conjugated goat anti-rabbit antibody (Catalog no. 111-035-003) as a secondary antibody, visualized by enhanced chemiluminescence (WEST-ZOL®Plus, Intron Biotechnology, Sungnam, Korea), and the density of protein bands was quantified using a laser scanner and Quantity One software (Version 5.2, Bio-Rad Laboratory, Berkeley, CA).

[0125] As a result, wild-type PDK1 with a molecular weight of 462 kDa was expressed in the control cell, WTC11, whereas it was not expressed in clones #1, #2, and #3 of the human iPSC WTC11 cell line containing a deletion mutation in exon 36 of the PKD1 gene selected in Example 1-1 (Fig. 2A and Fig. 2B). This confirmed that the PKD1 deletion mutation was successfully induced in the human iPSC cell line by the CRISPR-Cas9 gene editing system.

[0126]

[0127] 1-3. Disease Modeling Kidney Organoid Manufacturing

[0128] PKD1 - / - and PKD2 - / - To determine whether the PKD1 produced in Example 1-1 can form a cyst, which is a phenotype of ADPKD, one of the polycystic kidney diseases (PKD). - / - Human iPSC cell lines WTC11 and PKD2 - / - Human iPSCs cell line WTC11 were induced to differentiate into kidney organoids (Fig. 3A). Specifically, PKD1 at passages 30 to 60 - / - Human iPSC cell lines WTC11 and PKD2 - / -Human iPSCs cell line WTC11 were seeded at a density of 15,000 cells / well on 1% GelTrex (Thermo Fisher Scientific)-coated glass plates (LabTek) placed in 6-well plates containing mTeSR1 medium (Stem Cell Technologies) containing 10 μM Y27632 (LC Laboratories) (~day 0). The cells were cultured for 3 days (~day 3) by replacing the medium with mTeSR1 medium containing 1.5% GelTrex, and then cultured for 1.5 days (~day 4.5) by replacing the medium with RPMI medium (Thermo Fisher Scientific) containing 12 μM CHIR99021 (Tocris). Afterwards, the medium was replaced with RPMI medium containing B27 supplement (Thermo Fisher Scientific) and cultured with the medium changed every 2-3 days (day 4.5~). PKD1 cells were differentiated by culturing for 16 days in this manner. - / - and PKD2 - / - Each kidney organoid was transferred using a needle to a 6-well plate containing RPMI medium containing B27 supplement and exposed to laminar fluid shear stress in a waving plate device. Cystogenesis was induced by shaking culture in the waving plate for 1 week, with medium replacement every 3 days, and confirmed by high-resolution imaging.

[0129] As a result, PKD1 - / - Human iPSCs and PKD2 - / -Both human iPSCs differentiated into translucent, tubular kidney organoids. From day 16 after differentiation into cardiac organoids, cystogenesis, a swelling of kidney tubules to form balloon-like fluid sacs, was observed in both kidney organoids (Fig. 3B). Furthermore, the formed cysts continued to expand during subsequent culture, reaching a diameter of more than 1 mm in the culture plate after 2 weeks of culture. This confirmed that deletion mutants of PKD1 or PKD2 recapitulate the cyst formation phenotype of polycystic kidney disease, particularly ADPKD, in kidney organoids.

[0130]

[0131] Example 2. Screening for drugs that inhibit cyst formation

[0132] In renal organoids exhibiting the cyst formation phenotype prepared in Examples 1-3 above, drugs from an FDA-approved drug library were administered, and inhibitors that suppress cyst formation were screened. Specifically, 2,570 FDA-approved drugs (Selleckchem, FDA-approved Drug Library, Cat. L1300) selected from the antihistamine drug and SSRI (Selective serotonin reuptake inhibitor) clusters were diluted with DMSO or water to make a 10 mM stock, and then PKD1 - / - Kidney organoids and PKD2 - / - Renal organoids were administered with final concentrations of 10 or 30 μM, respectively, and cultured on shaking plates for 3 days. Approximately 30 cysts per well were selected and imaged using brightfield microscopy. Cyst size was quantified using ImageJ. Statistically significant changes in size were analyzed using a paired t-test, and drugs that reduced cyst size by approximately 20–40% were screened.

[0133] As a result, antihistamines and SSRI drugs were selected as drugs that suppressed cyst occurrence and size.

[0134]

[0135] Example 3. Confirmation of the inhibitory effect of antihistamines and SSRI drugs on cyst formation.

[0136] 3-1. Desloratadine and duloxetine

[0137] 3-1-1. PKD1 - / - Effects on renal organoids

[0138] In order to confirm the cyst formation inhibitory effect of desloratadine, an antihistamine derived from screening in the above example, and duloxetine, an SSRI drug, on day 21, PKD1 - / - Desloratadine and duloxetine, alone or in combination, were administered to renal organoids at concentrations of 10 μM or 30 μM, respectively, and the inhibitory effect on cyst growth in renal organoids was analyzed by live imaging for 3 days. At this time, tolvaptan, an AVPR2 inhibitor and the first FDA-approved drug for the treatment of ADPKD, was administered as a positive control at concentrations of 10 μM or 30 μM.

[0139] As a result, when tolvaptan, a positive control, was administered, cysts were found to expand in renal organoids, whereas in organoids administered with 10 μM desloratadine or duloxetine alone, cyst size was found to decrease (Fig. 4). In particular, when desloratadine (10 μM) and duloxetine (10 μM) were administered together, cyst growth was found to be significantly reduced compared to when each drug was administered alone (Fig. 5). In addition, when administered at a concentration of 30 μM, the cyst growth inhibitory effect of desloratadine or duloxetine was found to increase in a dose-dependent manner, and the cyst growth inhibitory effect was found to be significantly increased when administered together compared to when administered alone (Figs. 6 and 7).

[0140]

[0141] 3-1-2. PKD2 - / - Effects on renal organoids

[0142] As in Example 3-1 above, the cyst formation inhibition effect by single or combined administration (10 μM or 30 μM) of desloratadine and duloxetine was measured on PKD2 - / - As confirmed in kidney organoids, PKD1 - / - Similar to the results in renal organoids, desloratadine or duloxetine significantly inhibited cyst growth in a dose-dependent manner compared to the positive control group, which did not inhibit cyst growth. Furthermore, co-administration of desloratadine and duloxetine significantly inhibited cyst growth compared to monotherapy (Figures 8 to 11).

[0143]

[0144] 3-1-3. IC50

[0145] PKD1 on day 21 - / -Desloratadine and duloxetine were administered alone to renal organoids at 2.5, 5, 10, 30, 50, and 100 μM, respectively, and IC was calculated by nonlinear regression and log(compound) versus normalized response-variable slope using GraphPad Prism software. 50 The value was calculated.

[0146] As a result, the IC of desloratadine 50 was found to be 23.95 μM (Figure 12A), and the IC of duloxetine 50 was found to be 15.6 μM (Figure 12B).

[0147]

[0148] 3-2. Desloratadine and fluoxetine

[0149] To determine the inhibitory effect of cyst formation by single or combined administration of desloratadine, an antihistamine, and fluoxetine, an SSRI drug, on PKD1 on day 21 - / - Desloratadine and fluoxetine, alone or in combination, were administered to renal organoids at a concentration of 30 μM, and the inhibitory effect on cyst growth in renal organoids was analyzed by live imaging for 3 days. Tolvaptan was administered as a positive control.

[0150] As a result, administration of tolvaptan, a positive control, showed enlargement of cysts in renal organoids, whereas 30 μM desloratadine reduced cyst size. In particular, co-administration of desloratadine and fluoxetine showed a significant reduction in cyst size compared to administration of each drug alone (Figs. 13 and 14).

[0151]

[0152] 3-3. Azelastine and fluoxetine

[0153] To determine the inhibitory effect of cyst formation by single or combined administration of azelastine, an antihistamine, and fluoxetine, an SSRI drug, on PKD1 on day 21 - / - Azelastine and fluoxetine were administered alone or in combination at a concentration of 30 μM to renal organoids, and the inhibitory effect on cyst growth in renal organoids was analyzed by live imaging for 3 days. Tolvaptan was administered as a positive control.

[0154] As a result, co-administration of azelastine and fluoxetine at a concentration of 30 μM was shown to suppress cysts more effectively than administration of each drug alone (Figs. 15 and 16).

[0155] However, it was confirmed that the effect was lower than that of the previously confirmed combined administration of desloratadine and duloxetine (Figures 5 and 7).

[0156]

[0157] 3-4. Azelastine and duloxetine

[0158] To determine the inhibitory effect of cyst formation by single or combined administration of azelastine, an antihistamine, and duloxetine, an SSRI drug, on PKD1 on day 21 - / - Azelastine and duloxetine were administered to renal organoids, either alone or in combination, at concentrations of 10 μM or 30 μM, and live images were taken for 3 days to analyze the inhibitory effect on cyst growth in renal organoids. Tolvaptan was administered as a positive control.

[0159] As a result, co-administration of azelastine and duloxetine at a concentration of 10 μM (Figs. 17 and 18) and co-administration of azelastine and duloxetine at a concentration of 30 μM were found to suppress cysts more than administration of each drug alone (Figs. 19 and 20).

[0160] However, it was confirmed that the effect was lower than that of the previously confirmed combined administration of desloratadine and duloxetine (Figures 5 and 7).

[0161]

[0162] 3-5. Ketotifen and fluoxetine

[0163] To determine the inhibitory effect of cyst formation by single or combined administration of ketotifen fumarate, a histamine receptor antagonist, and fluoxetine, an SSRI drug, on PKD1 on day 21. - / - Ketotifen and fluoxetine, alone or in combination, were administered to renal organoids at a concentration of 30 μM, and the inhibitory effect on cyst growth in renal organoids was analyzed by live imaging for 3 days. Tolvaptan was administered as a positive control.

[0164] As a result, co-administration of ketotifen and fluoxetine at a concentration of 30 μM was shown to suppress cysts more than administration of each drug alone (Figs. 21 and 22).

[0165] However, it was confirmed that the effect was lower than that of the previously confirmed combined administration of desloratadine and duloxetine (Figures 5 and 7).

[0166] Therefore, it was confirmed that the combination of desloratadine and duloxetine of the present invention showed a superior effect than other combinations.

[0167]

[0168] Example 4. Mechanistic analysis of desloratadine and duloxetine

[0169] 4-1. Mechanism of action of desloratadine

[0170] In normal renal epithelial cells, cAMP inhibits the Raf-1 / MEK / ERK pathway, but in the development of ADPKD, cAMP has been reported to stimulate B-Raf through cSrc-dependent phosphorylation and activate the MEK / ERK pathway, thereby increasing cell proliferation. Therefore, to determine the effect of desloratadine on intracellular signaling components including ERK / AKT / B-Raf / Raf-1 and MAPK, and the PI3K / Akt / mTOR / c-Myc pathway, which is abnormally activated in ADPKD and important for proliferation, PKD1 - / - Whether the expression of the above proteins changes due to desloratadine administration in renal organoids was confirmed by immunoblot analysis as described in Example 1-2. At this time, the primary antibodies used were rabbit monoclonal anti-Akt (Cell signaling, Catalog no. 9272S), mouse monoclonal anti-β-actin (Sigma Aldrich, Catalog no. A5441), abbit monoclonal anti-pSer473-Akt (Cell signaling, Catalog no. 9271S), and rabbit monoclonal. anti-pThr180 / Tyr182-p38 MAPK (Cell signaling, Catalog no. 4511S), rabbit monoclonal anti-p38 MAPK (Cell signaling, Catalog no. 9212S), rabbit monoclonal anti-pThr202 / Tyr204-Erk1 / 2 (Cell signaling, Catalog no. 9101S), rabbit monoclonal anti-Erk1 / 2 (Cell signaling, Catalog no. 4695S) and rabbit monoclonal anti-c-Myc (abcam, Catalog no. ab32072).

[0171] As a result, PKD1 - / -In renal organoids, the ratios of B-Raf, p-p38 MAPK / p38 MAPK, and pERK1 / 2 / ERK1 / 2 were significantly reduced by desloratadine administration, confirming that desloratadine reduces the phosphorylation of p38 and ERK1 / 2 proteins (Fig. 23). In addition, desloratadine was found to significantly reduce the expression levels of B-Raf and c-Myc proteins, which are increased during the onset of ADPKD (Fig. 23). Through this, it was confirmed that desloratadine inhibits cytogenesis by inducing the suppression of c-Myc protein, which leads to the downregulation of the B-Raf-MEK-ERK pathway.

[0172]

[0173] 4-2. Mechanism of Duloxetine

[0174] To confirm the β-catenin / mTOR / c-Myc pathway regulation effect by duloxetine in ADPKD, rabbit polyclonal anti-pSer9-GSK-3β (Cell signaling, Catalog no. 5558S), mouse monoclonal anti-GSK-3β (BD Biosciences, Catalog no. 610201), rabbit monoclonal anti-pThr41 / Ser45-beta catenin (Thermo Fisher, Catalog no. 702969), mouse monoclonal anti-beta catenin (BD Biosciences, Catalog no. 610154), rabbit polyclonal anti-pSer2448-mTOR (Cell signaling, Catalog no. 2971S), and rabbit monoclonal anti-mTOR (Cell signaling, Catalog no. 2983) and rabbit monoclonal anti-c-Myc (abcam, Catalog no. Immunoblot analysis was performed using the primary antibody (no. ab32072).

[0175] As a result, PKD1 was reduced by duloxetine administration. - / - In renal organoids, the expression levels of total GSK-3β and β-catenin were increased, and the ratio of phosphorylated protein / total protein was calculated. The phosphorylation of serine 9 of GSK3β, which is inversely correlated with the activity of GSK-3β, was reduced by duloxetine administration, and the phosphorylation of β-catenin and mTOR (mammalian target of rapamycin) was also reduced (Fig. 24). In addition, the protein expression level of c-Myc was reduced, indicating that mTOR phosphorylation was inhibited by duloxetine, resulting in a decrease in c-Myc protein expression (Fig. 24).

[0176]

[0177] The above results show that desloratadine downregulates the B-Raf-MEK-ERK signaling pathway during cyst formation, and duloxetine downregulates the Wnt signaling pathway, indicating that co-administration of these two drugs has a synergistic effect.

[0178]

[0179] Also PKD1 - / - The effects of administration of 3-hydroxy desloratadine, a desloratadine metabolite, and (S)-5-hydroxy-6-methoxy duloxetine, a duloxetine metabolite, on renal cyst growth in renal organoids were investigated. Desloratadine metabolites and duloxetine metabolites were found to have PKD1 - / - Cyst growth in renal organoids was reduced by 48.2% and 47%, respectively (Figures 25A and 25B). These results suggest that the use of desloratadine metabolites and duloxetine metabolites also exhibited therapeutic effects on renal cyst growth in the PKD mutant organoid model.

[0180]

[0181] Furthermore, we examined whether the combined administration of desloratadine and duloxetine resulted in a synergistic effect in reducing renal cyst growth. To this end, the combination index (CI) was calculated using the Chou-Talalay method (Fig. 26A). The calculated CI was 0.87, indicating that the combined administration of desloratadine and duloxetine exhibited a synergistic effect in reducing renal cyst growth (Figs. 26B and 26C).

[0182]

[0183] Example 5. Confirmation of the effects of desloratadine and duloxetine in an ADPKD mouse model.

[0184] 5-1. Drug selection and experimental design

[0185] Among the drugs screened in an in vitro platform using kidney organoids modeling ADPKD, desloratadine and duloxetine were selected for in vivo experiments. The synergistic effect of the combination of desloratadine and duloxetine was demonstrated in in vitro experiments in the previous examples. Based on this, an in vivo experiment was designed to demonstrate the therapeutic effect of the combination of desloratadine and duloxetine, rather than the specificity of the combination itself. The in vivo experimental design is illustrated in Figures 27A and 27B.

[0186] First, chronic-onset PKD1 mice, in which PKD1 is conditionally knocked out by tamoxifen administration, were generated as an animal model of ADPKD, and the cystic phenotype changes due to co-administration of desloratadine and duloxetine were observed. Specifically, for the generation of chronic-onset PKD1 mice, Pkd1 flox Mouse (C57BL / 6J.129S4-Pkd1 tm2Ggg / J)(Strain #: 010671, Jackson Laboratories) were crossed with C57 / B6 tamoxifen-Cre(B6;129-Gt(ROSA)26Sortm1(Cre / ERT)Nat / J) mice (Strain #: 004847, Jackson Laboratories) to obtain Pkd1 flox / flox R26CreER - / - A mouse was generated. The Pkd1 flox / flox R26CreER - / - Mouse and Pkd1 flox Pkd1 generated by crossbreeding mice flox / flox R26CreER + / - A chronic-onset PKD1 mouse model exhibiting a mild cystic phenotype with minimal renal function impairment was generated by intraperitoneally injecting tamoxifen (total dose of 200 mg / kg) into the mothers on postnatal day 6 (P6) to delete PKD1.

[0187] All animal experiments were conducted in compliance with the Animal Welfare Act, the Laboratory Animal Act, and the Guidelines for Use of Laboratory Animals, as well as the Guidelines and Policies for Rodent Experiments, provided by the Institutional Animal Care and Use Committee (IACUC) of the Catholic University of Korea College of Medicine (Approval Numbers: CUMS-2023-0311-02, CUMS-2023-0235-03). The Catholic University of Korea Institutional Animal Care and Use Committee (IACUC) and the Department of Laboratory Animals (DOLA) on the Seongui Campus were recognized as Korea Excellence Animal Laboratory Facilities by the Ministry of Food and Drug Safety in 2017 and received AAALAC International Full Accreditation in 2018.

[0188]

[0189] 5-2. Confirmation of histamine receptor and serotonin receptor expression

[0190] Pkd1 flox / flox R26CreER + / - When tamoxifen was administered orally to mother mice that gave birth to Pkd1 flox / flox R26CreER + / - The organism's Cre-flox regulatory system is induced, resulting in targeted inactivation of the Pkd1 gene. In vivo evaluations have shown that Pkd1 flox / flox R26CreER + / - Mice and control mice without Cre-flox system (Pkd1 flox / flox R26CreER + ) were administered tamoxifen on the 6th day after birth to induce sporadic cysts.

[0191] Afterwards, desloratadine 5 mg / kg and duloxetine 10 mg / kg were administered together, or the excipient normal mouse diet (Altromin diet) was administered for 20 weeks from the 21st day of birth (Figure 27B).

[0192] Pkd1 flox / flox ;R26CreER + / - We observed a decrease in histamine receptor expression and serotonin receptor expression in mouse kidneys. Immunofluorescence microscopy analysis revealed a decrease in Pkd1 expression compared to wild-type mouse kidneys. flox / flox ;R26CreER + / - In mouse kidneys, histamine H1 receptor expression was increased and serotonin 1A (5-HT1A) receptor expression was decreased (Fig. 27C).

[0193] In addition, co-administration of 5 mg / kg desloratadine and 10 mg / kg duloxetine resulted in Pkd1 flox / flox ;R26CreER + / -In the mouse kidney, the expression of histamine H1 receptors and histamine H3 receptors was decreased, and the expression of serotonin receptors, 5-HT1A, and 5-HT2A receptors was increased, showing a pattern similar to that of wild-type (WT) kidney in IHC (Figures 27D and 27E) and immunoblot analysis (Figures 27F to 27J). These results suggest that co-administration of desloratadine and duloxetine restores the expression levels of histamine receptors and serotonin receptors in the kidney of the ADPKD mouse model to those of wild-type mice.

[0194]

[0195] 5-3. Confirmation of the effect of suppressing renal cyst growth and improving renal function.

[0196] Pkd1 flox / flox ;R26CreER + / - Mice had kidneys with significantly larger, multiple renal cysts than wild-type mouse kidneys (Figure 28A). Pkd1 flox / flox ;R26CreER + / - In mice, the experimental group administered a combination drug consisting of 5 mg / kg desloratadine and 10 mg / kg duloxetine significantly reduced the height-to-body weight ratio, cyst index (cyst area percentage), and number of cysts compared to the control group (Figures 28A to 28D). Interestingly, Pkd1 flox / flox ;R26CreER + / - Increased blood urea nitrogen (BUN) levels in mice were reduced when administered a combination drug consisting of desloratadine and duloxetine (Fig. 28E).

[0197] Cystic epithelial cell proliferation confirmed by PCNA staining was Pkd1 flox / flox ;R26CreER + / - In the experimental group co-administered with desloratadine and duloxetine in the mouse kidney, the level was significantly reduced compared to the vehicle-treated experimental group (Figures 28F and 28G). The PCNA immunoblot analysis results were consistent with the immunohistochemical staining results (Figures 28H and 28I).

[0198] Through this, we confirmed that co-administration of desloratadine and duloxetine not only alleviated renal cyst growth by inhibiting cell proliferation, but also improved renal function in an ADPKD mouse model.

[0199]

[0200] 5-4. Pkd1 flox / flox ;R26CreER + / - Confirmation of the therapeutic effect of renal fibrosis in mice

[0201] Next, co-administration of desloratadine and duloxetine Pkd1 flox / flox ;R26CreER + / - The effect on renal fibrosis in mice was confirmed. Masson's trichrome staining results showed that Pkd1 flox / flox ;R26CreER + / - In mouse kidneys, the accumulation of extracellular matrix (ECM) in the tubulointerstitial space was significantly increased, and this accumulation was significantly reduced when desloratadine and duloxetine were co-administered (Figs. 29A and 29B). Similar results were observed in confocal images, immunohistochemical staining, and Western blot analysis for fibronectin (Figs. 29C to 29E). These results suggest that the combination drug consisting of desloratadine and duloxetine inhibits Pkd1 flox / flox ;R26CreER + / - It suggests that Pkd1 has a therapeutic effect on renal fibrosis in mouse kidneys. SMA (α-smooth muscle actin), an important marker of myofibroblasts, is expressed in the kidney. flox / flox ;R26CreER + / - It was markedly upregulated in mouse kidneys and significantly reduced by co-administration of desloratadine and duloxetine (Figures 29F to 29H). This suggests that co-administration of desloratadine and duloxetine reduced the accumulation of myofibroblasts in the renal interstitium in the ADPKD mouse model.

[0202] TGF-β is a key cytokine mediating renal fibrosis and induces the production of ECM proteins. TGF-β / Smad signaling is a major pathway that causes renal fibrosis. Based on the therapeutic effect of co-administration of desloratadine and duloxetine on renal fibrosis, Pkd1 flox / flox ;R26CreER + / - We investigated the effects of co-administration of desloratadine and duloxetine on the expression of the TGF-β / Smad pathway in mouse kidneys. Immunohistochemical staining revealed that Pkd1 flox / flox ;R26CreER + / - We confirmed that the expression of TGF-β was increased in the mouse renal interstitium (Fig. 29I). Pkd1 flox / flox ;R26CreER + / - TGF-β and Smad4 protein expression in mouse kidneys was significantly increased compared to WT mouse kidneys, as confirmed by Western blot analysis (Figs. 29J to 29L). Pkd1 flox / flox ;R26CreER + / - Increased TGF-β and Smad4 expression in mouse kidneys was significantly reduced by co-administration of desloratadine and duloxetine (Figures 29I to 29K).

[0203] Collectively, these data suggest that co-administration of desloratadine and duloxetine may enhance Pkd1 flox / flox ;R26CreER + / - Suggesting that modulating the Smad4-dependent TGF-β pathway in mouse kidney alleviates renal fibrosis.

[0204]

[0205] 5-5. Confirmation of the effect of regulating the expression of factors involved in renal cyst growth.

[0206] In particular, IL-1β is a key regulatory cytokine that influences multiple cell types to promote transcription of hundreds of genes, including those encoding both inflammatory cytokines and chemokines. It further amplifies inflammation by promoting the infiltration and activation of neutrophils, dendritic cells, monocytes, and lymphocytes. IL-1β enhances the stability of the Yes-associated protein (YAP) and activates its target transcriptional pathway. YAP is a key transcription factor in the Hippo signaling pathway, which is crucial for cell proliferation and regeneration. MYC is a transcriptional target of YAP, and the YAP-MYC signaling axis has been shown to mediate the development of cystic kidneys in a human-PKD mouse model.

[0207] Therefore, Pkd1 flox / flox ;R26CreER + / - The modulatory role of co-administration of desloratadine and duloxetine on the IL-1β-YAP-MYC signaling axis in renal cyst growth in mouse kidneys was investigated.

[0208] Pkd1 flox / flox ;R26CreER + / - IL1-β protein expression in mouse kidneys was significantly increased compared to WT mouse kidneys, whereas it was significantly decreased by co-administration of desloratadine and duloxetine (Fig. 29M and Fig. 29N).

[0209] In addition, Pkd1 compared to WT mouse kidney flox / flox ;R26CreER + / - YAP protein expression in mouse kidney was also significantly increased, whereas it was significantly decreased by co-administration of desloratadine and duloxetine (Fig. 29M and Fig. 29O).

[0210] In addition, Pkd1 compared to WT mouse kidney flox / flox ;R26CreER + / -c-MYC protein expression in mouse kidney was significantly increased, whereas it was significantly decreased by co-administration of desloratadine and duloxetine (Fig. 29M and Fig. 29P).

[0211] Through this, co-administration of desloratadine and duloxetine modulates Pkd1 through the regulation of the IL1-β / YAP / c-Myc signaling pathway. flox / flox ;R26CreER + / - It was confirmed to have a therapeutic effect on the growth of renal cysts in mice.

[0212]

[0213] 5-6. Therapeutic effect of co-administration of desloratadine and duloxetine on an in vitro renal fibrosis model.

[0214] We also investigated the effect of co-administration of desloratadine and duloxetine on renal fibrosis in an in vitro model. Human kidney proximal tubular epithelial cells (HK-2) were cultured in DMEM / F12 (Dulbecco's Modified Eagle Medium / Nutrient Mixture F-12) supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C in a humidified atmosphere containing 5% CO₂. To induce fibrosis, HK-2 cells were seeded in 12-well plates and cultured to approximately 70–80% confluence. The medium was then replaced with serum-free DMEM / F12 and cultured for 24 h to induce serum starvation of the cells. After starvation, TGF-β1 (5 ng / mL) was administered to the cells for 72 hours to induce fibrotic changes, thereby creating a renal fibrosis-induced cell model (Fig. 30A).

[0215] After administering desloratadine or duloxetine at 5, 10, or 15 μM to the above renal fibrosis-induced cell model, the appropriate drug administration concentration for the renal fibrosis-induced cell model was determined. As a result, desloratadine at 10 μM and duloxetine at 15 μM showed the most effective inhibitory effect on the expression of renal fibrosis markers (Figures 30B and 30C).

[0216] Therefore, the fibrosis-induced cell model was treated with desloratadine (10 μM), duloxetine (15 μM), or a combination of desloratadine and duloxetine for an additional 72 h. Control groups included untreated cells and cells treated with TGF-β1 alone.

[0217] As a result, the increased gene expression of collagen I and connective tissue growth factor (CTGF) induced by TGF-β was reduced by desloratadine and duloxetine administration, respectively. Interestingly, co-administration of desloratadine and duloxetine reduced gene expression to a greater extent than administration of desloratadine or duloxetine alone (Fig. 30D). Therefore, this confirmed that co-administration of desloratadine and duloxetine exhibited a therapeutic effect on renal fibrosis.

[0218]

[0219] 5-7. Confirmation of the therapeutic effect on renal fibrosis in an animal model of chronic kidney disease (CKD) (in vivo)

[0220] Based on the previously confirmed renal fibrosis-inducing cell model (in vitro) in which desloratadine and duloxetine were combined to treat renal fibrosis, the effects of desloratadine and duloxetine were confirmed in an animal model of chronic kidney disease. To this end, an adenine-fed animal model with reduced renal function was prepared. To prepare the adenine-fed animal model, 6-week-old C57BL / 6J male mice were acclimated for 1 week and then divided into a normal diet control group and a diet group containing 0.3% adenine. A 0.3% adenine diet was provided to the adenine diet group for 10 days, followed by a maintenance diet supplemented with 0.2% adenine diet for 4 weeks. A 5-week period was used to create an animal model of renal fibrosis (chronic kidney disease, CKD) induced by adenine diet intake. After a 10-week rest period, desloratadine (0.6 mg / kg) and duloxetine (10 mg / kg) were co-administered for 8 weeks at the 15th week (Fig. 30E). After establishing the adenine diet renal fibrosis animal model, blood urea levels (BUN) were measured twice a week using an I-STAT1 Analyzer (Abbott, Ottignies, Belgium) to evaluate renal function. As a result, it was confirmed that the BUN level, which was significantly increased in the adenine diet renal fibrosis animal model compared to wild type mice, was significantly reduced by co-administration of desloratadine and duloxetine (Fig. 30F).

[0221]

[0222] 5-8. Confirming the effectiveness of liver cyst treatment

[0223] Liver cysts are the most common extrarenal manifestation of ADPKD, with a prevalence of liver cysts reaching 58% in adults aged 15 to 24 and 94% in those aged 35 and older. Liver cysts are typically mild or asymptomatic, but serious consequences, such as cyst infection, can occur. Additionally, some ADPKD patients develop polycystic liver disease (PLD), characterized by numerous liver cysts and liver enlargement, which can cause abdominal discomfort and hepatomegaly. Unfortunately, liver transplantation is the only curative treatment option for severe PLD.

[0224] Therefore, Pkd1 flox / flox ;R26CreER + / - The therapeutic effect of combined administration of desloratadine and duloxetine on hepatic cysts in mice was confirmed.

[0225] Pkd1 flox / flox ;R26CreER + / - Mice exhibited significantly larger liver multiple cysts than WT mice (Figures 31A-31C). Pkd1 flox / flox ;R26CreER + / - In mice, the group co-administered with desloratadine and duloxetine showed a significant decrease in the hepatic cyst index (cyst area percentage) and the number of hepatic cysts compared to the vehicle-treated group (Figures 31A to 31C). In addition, the group co-administered with desloratadine and duloxetine showed a significant decrease in the levels of serum glutamate pyruvate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT), which were associated with a decrease in Pkd1. flox / flox ;R26CreER + / - In mice, the levels were significantly increased beyond the normal limits (Fig. 31D and Fig. 31E).

[0226] Hepatic cystic epithelial cell proliferation confirmed by PCNA staining is Pkd1 flox / flox ;R26CreER + / -In the experimental group co-administered with desloratadine and duloxetine in mice, a significant decrease was observed compared to vehicle treatment (Figures 31F and 31G). These results suggest that co-administration of desloratadine and duloxetine inhibits Pkd1 flox / flox ;R26CreER + / - This means that it can treat liver cyst growth in mice.

[0227] Masson's trichrome staining results, Pkd1 flox / flox ;R26CreER + / - In the mouse liver, extracellular matrix deposition was significantly increased and significantly reduced by co-administration of desloratadine and duloxetine (Figures 31H and 31I). This suggests that co-administration of desloratadine and duloxetine inhibits Pkd1 flox / flox ;R26CreER + / - It was confirmed that there was a therapeutic effect on liver fibrosis in mice.

[0228] In addition, expression of α-SMA is regulated by Pkd1 flox / flox ;R26CreER + / - It was significantly upregulated in the mouse liver and significantly reduced when co-administered with desloratadine and duloxetine (Figures 31J and 31K). These results suggest that co-administration of desloratadine and duloxetine attenuated the accumulation of myofibroblasts in the liver.

[0229] Collectively, these data confirmed that co-administration of desloratadine and duloxetine could alleviate hepatic cyst growth and liver fibrosis caused by ADPKD and improve liver function.

Claims

1. A pharmaceutical composition for preventing or treating kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof; and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

2. A pharmaceutical composition for preventing or treating kidney disease, wherein the desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate and the duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate are mixed in a weight ratio of 1:1 to 20 in the first paragraph.

3. A pharmaceutical composition for preventing or treating kidney disease, wherein the kidney disease in claim 1 is at least one selected from the group consisting of cyst formation or kidney disease associated with cyst formation, polycystic kidney disease (PKD), renal fibrosis, acute kidney injury, end-stage renal disease (ESKD), renal failure, systemic lupus erythematosus, diabetic nephropathy (DN), IgA nephritis (IgAN), HIV-associated nephropathy, chronic kidney disease (CKD), nephrosclerosis, focal segmental glomerulosclerosis (FSGS), minimal change nephrotic syndrome (MCD), xanthine oxidase deficiency, abetalipoproteinemia, familial hypobetalipoproteinemia (FHBL), chylomicrons retention disease (CRD), sitosterolemia, glomerular hyperfiltration, and pruritus of renal failure.

4. A pharmaceutical composition for preventing or treating kidney disease, wherein the polycystic kidney disease (PKD) in the third paragraph is autosomal dominant polycystic kidney disease (ADPKD) or autosomal recessive polycystic kidney disease (ARPKD).

5. A pharmaceutical composition for preventing or treating kidney disease, wherein the composition according to claim 1 inhibits cystogenesis or growth.

6. A pharmaceutical composition for preventing or treating kidney disease, wherein the composition maintains kidney function in accordance with paragraph 1.

7. A pharmaceutical composition for preventing or treating kidney disease, wherein the composition increases phosphorylation of Akt in the first paragraph.

8. A pharmaceutical composition for preventing or treating kidney disease, wherein the composition reduces phosphorylation of p38 MAPK, ERK(1 / 2), GSK3β, β-catenin, or mTOR in the first paragraph.

9. A pharmaceutical composition for preventing or treating kidney disease, wherein the composition reduces the expression of B-Raf or c-Myc in claim 1.

10. A pharmaceutical composition for the prevention or treatment of at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof, and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof.

11. A pharmaceutical composition for the prevention or treatment of at least one selected from the group consisting of liver cysts and liver fibrosis, wherein the desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate and the duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate are mixed in a weight ratio of 1:1 to 20 in the 10th paragraph.

12. A pharmaceutical composition for prevention or treatment of at least one selected from the group consisting of liver cysts and liver fibrosis, wherein at least one selected from the group consisting of liver cysts and liver fibrosis is caused by polycystic kidney disease (PKD).

13. A pharmaceutical composition for the prevention or treatment of at least one disease selected from the group consisting of liver cysts and liver fibrosis, wherein the polycystic kidney disease (PKD) in claim 12 is autosomal dominant polycystic kidney disease (ADPKD) or autosomal recessive polycystic kidney disease (ARPKD).

14. A pharmaceutical composition for preventing or improving renal disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof, and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

15. A pharmaceutical composition for the prevention or improvement of at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof, and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof.

16. A food composition for preventing or improving kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate; and duloxetine, a stereoisomer thereof, a food-based acceptable salt and solvate.

17. A food composition for the prevention or improvement of at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate thereof, and duloxetine, a stereoisomer thereof, a food-based acceptable salt or solvate thereof.

18. A health functional food composition for preventing or improving kidney disease, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate, and duloxetine, a stereoisomer thereof, a food-based acceptable salt and solvate.

19. A health functional food composition for the prevention or improvement of at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a food-based acceptable salt or solvate, and duloxetine, a stereoisomer thereof, a food-based acceptable salt and solvate.

20. A method for preventing or treating kidney disease, comprising administering to a subject at least one selected from the group consisting of desloratadine, a stereoisomer thereof, a pharmaceutically acceptable salt or solvate thereof, and duloxetine, a stereoisomer thereof, a pharmaceutically acceptable salt and solvate thereof.

21. In paragraph 20, A method for preventing or treating a kidney disease, wherein the kidney disease is at least one selected from the group consisting of cyst formation or kidney disease associated with cyst formation, polycystic kidney disease (PKD), renal fibrosis, acute kidney injury, end-stage renal disease (ESKD), renal failure, systemic lupus erythematosus, diabetic nephropathy (DN), IgA nephritis (IgAN), HIV-associated nephropathy, chronic kidney disease (CKD), nephrosclerosis, focal segmental glomerulosclerosis (FSGS), minimal change nephrotic syndrome (MCD), xanthine oxidase deficiency, abetalipoproteinemia, familial hypobetalipoproteinemia (FHBL), chylomicrons retention disease (CRD), sitosterolemia, glomerular hyperfiltration, and pruritus of renal failure.

22. A method for preventing or treating at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, comprising the step of administering to a subject at least one selected from the group consisting of desloratadine, stereoisomers thereof, pharmaceutically acceptable salts or solvates thereof; and duloxetine, stereoisomers thereof, pharmaceutically acceptable salts and solvates thereof.

23. In paragraph 22, A method for preventing or treating at least one selected from the group consisting of hepatic cysts and hepatic fibrosis, wherein at least one selected from the group consisting of hepatic cysts and hepatic fibrosis is caused by polycystic kidney disease (PKD).

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