Pharmaceutical composition for treating fibrosis comprising SGLT-2 inhibitor and angiotensin receptor blocker

Abstract

The present invention relates to a pharmaceutical composition for treating fibrosis comprising an SGLT-2 inhibitor and an angiotensin receptor blocker. Specifically, when renal fibrosis was induced in C57BL / 6 mice by administering folic acid, and the test substances dapagliflozin and olmesartan medoxomil were administered alone and in combination for 4 weeks, the albumin to creatinine ratio (ACR) results and histological analysis of kidney tissue in the dapagliflozin alone and dapagliflozin + olmesartan combination administration groups showed superior renal fibrosis and renal protective efficacy compared to the dapagliflozin or olmesartan medoxomil alone administration groups.

Description

Technology Field

[0001] The present invention relates to a pharmaceutical composition for treating fibrosis comprising an SGLT-2 inhibitor and an angiotensin receptor blocker. Background Technology

[0002] Although there is a negative view that developing combination drugs for the simultaneous treatment of hypertension and diabetes is practically difficult because there are so many drugs for each disease in terms of class, ingredients, and dosage, it is a market with sufficient potential for combination drug development given that there are over 2 million patients suffering from both diseases simultaneously.

[0003] It is understood that appropriate combinations are difficult to achieve for diabetes and hypertension treatments due to the wide variety of classes and dosages available. For diabetes, there are various drugs such as metformin, DPP-4 inhibitors, SGLT-2 inhibitors, sulfonylureas, and TZDs, while for hypertension, there are numerous classes including ARBs, CCBs, beta-blockers, and diuretics. Consequently, since dosages vary widely within each class, there are many variables when creating combination drugs, which presents a limitation in selecting drug combinations.

[0004] Meanwhile, fibrosis is a disease characterized by functional impairment resulting from the abnormal fibrosis of tissues, manifesting in various forms such as renal fibrosis, hepatic fibrosis, and pulmonary fibrosis. The progression of such fibrosis is closely associated with oxidative stress, inflammatory responses, and the excessive accumulation of the extracellular matrix. In particular, oxidative stress is known to be one of the major causes of cell damage and fibrosis, making the development of therapeutic agents targeting it a critical challenge.

[0005] SGLT-2 inhibitors, used as treatments for type 2 diabetes, primarily regulate blood sugar by inhibiting the renal reabsorption of glucose. Angiotensin receptor blockers (ARBs), used to treat hypertension, primarily lower blood pressure and provide vascular and renal protective effects by inhibiting the action of angiotensin II. However, existing studies have not sufficiently analyzed the synergistic effects of co-administering SGLT-2 inhibitors and ARBs. Despite the benefits of each drug, the understanding of the efficacy and mechanisms of combination therapy remains in its early stages. Based on this background, the present invention aimed to verify the potential of a pharmaceutical composition containing an SGLT-2 inhibitor and an ARB in the treatment of fibrosis.

[0006] Several studies using rodents have shown that the administration of high doses or repeated administration of folic acid can cause fibrosis in certain tissues, such as the liver and kidneys. The mechanism by which folic acid induces tissue fibrosis is the induction of oxidative stress, and it is known that fibrosis progresses following tissue damage caused by lipid peroxidation induced by folic acid. Tissue fibrosis caused by folic acid's oxidative stress has been reported in the kidneys and liver in rodent studies and is also being evaluated as a disease model. Prior art literature

[0007] Animal Model Exp Med. 2021 Nov 24;4(4):329-342.Exp Biol Med (Maywood). 2008 Jan;233(1):38-47. The problem to be solved

[0008] Accordingly, the present invention aims to provide a pharmaceutical composition for the prevention or treatment of fibrosis comprising an SGLT-2 inhibitor and an angiotensin receptor blocker. means of solving the problem

[0009] The present invention provides a pharmaceutical composition for the prevention or treatment of fibrosis comprising an SGLT-2 inhibitor and an angiotensin receptor blocker. Effects of the invention

[0010] The present invention relates to a pharmaceutical composition for treating fibrosis comprising an SGLT-2 inhibitor and an angiotensin receptor blocker. Specifically, when renal fibrosis was induced in C57BL / 6 mice by administering folic acid, and the test substances dapagliflozin and olmesartan medoxomil were administered alone and in combination for 4 weeks, the albumin to creatinine ratio (ACR) results and histological analysis of kidney tissue in the dapagliflozin alone and dapagliflozin + olmesartan combination administration groups showed superior renal fibrosis and renal protective efficacy compared to the dapagliflozin or olmesartan medoxomil alone administration groups. Brief explanation of the drawing

[0011] Figure 1 shows the effect on the urinary albumin to creatinine ratio (ACR) of folic acid-induced chronic renal fibrosis C57BL / 6 mice treated with the test substance during the experimental period compared to the control group. Data are expressed as mean ± SD. Significant difference from G2 Folic acid Vehicle control group by One-way ANOVA followed by the Dunnett's test: * p<0.05, ** p<0.01. Figure 2 shows the results for renal fibrosis area in folic acid-induced chronic renal fibrosis C57BL / 6 mice treated with the test substance during the experimental period and the control group. Data are expressed as mean ± SD. Significant difference from G2 Folic acid Vehicle control group by One-way ANOVA followed by the Dunnett's test: * p<0.05, ** p<0.01. Significant difference between G4 Folic acid Olmesartan medoxomil group and G5 Folic acid Dapagliflozin + Olmesartan medoxomil group by the Tukey's test: # p<0.05, ## p<0.01. Figure 3 shows the results for the relative mean glomerular area within the kidneys of folic acid-induced chronic renal fibrosis C57BL / 6 mice treated with the test substance during the experimental period and the control group. Data are expressed as mean ± SD. Significant difference between G2 Folic acid Vehicle control group and G3 Folic acid Dapagliflozin group, G4 Folic acid Olmesartan medoxomil group by student t-test: * p<0.05. Significant difference between G3 Folic acid Dapagliflozin group and G5 Folic acid Dapagliflozin + Olmesartan medoxomil group by student t-test: # p<0.05. Figure 4 shows the glomerular area of ​​the kidneys based on H&E staining of the control group and folic acid-induced chronic renal fibrosis C57BL / 6 mice treated with the test substance during the experimental period (dotted lines representing the glomerular area of ​​each kidney). Specific details for implementing the invention

[0012] In the present invention, the efficacy of the test substance against renal fibrosis was confirmed by oral administration of the test substance for 4 weeks to C57BL / 6 mice in a folic acid-induced chronic kidney disease model, urine analysis, and histological analysis of the kidney.

[0013] After separating the groups to ensure similar average body weight measurements, renal fibrosis was induced by administering folic acid, and urine analysis, autopsy, and tissue analysis were performed after oral administration of the test substance for 4 weeks. Clinical symptoms were checked during the administration period, and no notable clinical symptoms were observed. On the last day of administration, urine samples were collected and analyzed, revealing that urinary albumin levels in the olmesartan administration group were significantly reduced compared to the G2 Folic acid Vehicle control group, and the albumin-to-creaatinine ratio (ACR) decreased significantly in all test substance administration groups.

[0014] In addition, the fibrosis area of ​​the kidney tissue excised during autopsy was significantly reduced in the dapagliflozin administration group and the dapagliflozin + olmesartan administration group compared to the G2 Folic acid Vehicle control group, and the fibrosis area was significantly reduced in the dapagliflozin + olmesartan administration group compared to the olmesartan administration group. As a result of measuring the glomerular area within the kidney, it was confirmed that the glomerular area in the dapagliflozin + olmesartan administration group was significantly reduced compared to the G2 Folic acid Vehicle control group and the dapagliflozin administration group, respectively, and the present invention was completed.

[0015] The present invention provides a pharmaceutical composition for the prevention or treatment of fibrosis comprising an SGLT-2 inhibitor and an angiotensin receptor blocker.

[0016] Preferably, the SGLT-2 inhibitor may be selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin, sotagliflozin, ipragliflozin, ertugliflozin, and pharmaceutically acceptable salts thereof, but is not limited thereto.

[0017] Preferably, the angiotensin receptor blocker may be selected from the group consisting of olmesartan, fimasartan, telmisartan, candesartan, azilsartan, and pharmaceutically acceptable salts or prodrugs thereof, but is not limited thereto.

[0018] Preferably, the weight ratio of the SGLT-2 inhibitor and the angiotensin receptor blocker may be 1:(1 to 4), but is not limited thereto.

[0019] Preferably, the fibrosis may be selected from renal fibrosis, renal failure, hepatic fibrosis, or pulmonary fibrosis, and more preferably, the fibrosis may be oxidative stress-induced fibrosis, but is not limited thereto.

[0020] Preferably, the pharmaceutical composition may be administered orally, but is not limited thereto.

[0021] The pharmaceutical composition of the present invention may be prepared using pharmaceutically suitable and physiologically acceptable adjuvants in addition to the active ingredient, and the adjuvants may include solubilizing agents such as excipients, disintegrants, sweeteners, binders, coating agents, leavening agents, lubricants, lubricants, or flavoring agents. The pharmaceutical composition of the present invention may preferably be formulated as a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the active ingredient for administration. As acceptable pharmaceutical carriers for compositions formulated as liquid solutions, which are sterile and biocompatible, saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture of one or more of these components may be used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed. In addition, by additionally adding diluents, dispersants, surfactants, binders, and lubricants, it can be formulated into injectable formulations such as aqueous solutions, suspensions, and emulsions, as well as pills, capsules, granules, or tablets.

[0022] The pharmaceutical formulation forms of the pharmaceutical composition of the present invention may be granules, powders, coated tablets, tablets, capsules, suppositories, syrups, juices, suspensions, emulsions, drops, or injectable liquids, as well as sustained-release formulations of the active compound. The pharmaceutical composition of the present invention may be administered in a conventional manner via intravenous, intra-arterial, intraperitoneal, intramuscular, intra-arterial, intraperitoneal, intrasternal, transdermal, nasal, inhalation, topical, rectal, oral, intraocular, or intradermal routes. The effective amount of the active ingredient in the pharmaceutical composition of the present invention refers to the amount required for the prevention or treatment of a disease. Accordingly, it may be adjusted according to various factors including the type of disease, the severity of the disease, the type and content of the active ingredient and other ingredients contained in the composition, the type of formulation, the patient's age, weight, general health status, gender and diet, the time of administration, the route of administration and the secretion rate of the composition, the duration of treatment, and concurrently used drugs.

[0023] Hereinafter, the present invention will be described in detail with reference to examples to aid in understanding. However, the following examples are merely illustrative of the content of the present invention and the scope of the present invention is not limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to those with average knowledge in the art.

[0024] <Experimental Example>

[0025] The following experimental examples are intended to provide experimental examples that are commonly applied to each embodiment according to the present invention.

[0026] 1. Purpose of the test

[0027] This study aimed to confirm the efficacy of the test substance against renal fibrosis by orally administering the test substance to C57BL / 6 mice for 4 weeks in a folic acid-induced chronic kidney disease model, confirming urine analysis, and histological analysis of the kidney.

[0028] 2. Preparation of test substance

[0029] 1) Preparation of Dapagliflozin propanediol hydrate

[0030] Dapagliflozin was weighed in its free form (since 1 mg of Dapagliflozin corresponds to 1.23 mg of Dapagliflozin propanediol hydrate, the dose was corrected), excipients were added to the weighed substance, vortexing was performed for 1 minute, and sonication was performed in an ultrasonic cleaner for 10 minutes to clear the substance before administration.

[0031] 2) Preparation of Olmesartan medoxomil

[0032] Olmesartan medoxomil was weighed as Olmesartan medoxomil (salt factor: 1.00), and after adding an excipient to the weighed substance, vortexing was performed for 1 minute, and sonication was performed in an ultrasonic cleaner for 10 minutes to become a suspension before administration.

[0033] 3. Experimental animals

[0034] I purchased the animal (Mouse, C57BL / 6 (naive), male) from Coatec Co., Ltd. through Peer Bio Co., Ltd.

[0035] In numerous studies, the C57BL / 6 mouse is known as a representative animal model suitable for research on acute and chronic kidney disease using cisplatin, gentamicin, folic acid, etc., and was selected because a large amount of comparable basic data has been accumulated.

[0036] 1) Number of animals and age at the start of administration

[0037] Mouse, C57BL / 6 - Male, 31 individuals, 15 weeks old

[0038] 2) Quarantine and Acclimation

[0039] Upon receiving the animals, inspection and quarantine were conducted by referring to the microbiological monitoring report of the test system provided by the supplier and observing their appearance. Animals without external abnormalities were brought into the rearing area and acclimatized for 7 weeks in the animal testing facility. Healthy animals were selected after verifying their health and suitability for the experiment during the 7-week quarantine and acclimatization period.

[0040] 3) Identification of individuals and breeding boxes

[0041] During the acclimatization period, individual markings were made on the tails of the animals using a red permanent marker upon entry, and temporary individual identification cards (test name, individual number, entry time) were attached to the rearing boxes during the quarantine and acclimatization period. When separating the groups, individual markings were made on the tails of the animals using a black permanent marker, and individual identification cards (test name, group information, individual number, sex, entry time, administration period) were attached to each rearing box.

[0042] 4) Group separation

[0043] Group separation was performed when the average body weight of the animals reached 26 g or more during the acclimatization period. After the acclimatization period ended, the body weight of all animals was measured, and group separation was carried out to ensure that there was no difference in the average body weight measurements between the groups. In addition, group separation was performed after consultation with the client, and after the separation was completed, individual identification cards (indicating test number, test group, individual number, sex, test period, etc.) were attached to each rearing box. Administration of the test substance began on the day following group separation.

[0044] 5) Induction of renal fibrosis by folic acid administration

[0045] After the end of the acclimatization period, renal fibrosis was induced in the remaining animals, excluding the normal group, by administering folic acid (Sigma, USA, 250 mg / kg in 0.3M NaHCO3) once intraperitoneally at a dose of 250 mg / kg.

[0046] 6) Feed and Water

[0047] It was provided to all animals for free intake during the acclimatization and administration periods.

[0048] 7) Administration

[0049] Drug administration began the day after group separation, and the administration time was 10:00 AM every day. Oral administration was performed for all administration groups.

[0050] The dosage of the test substance was set at 10 mL / kg, and the dosage per individual was calculated based on the most recently measured body weight. It was forcibly administered into the stomach once a day for 4 weeks using a 1 mL disposable syringe equipped with an oral administration sonde every morning.

[0051] 8) Group composition and dosage

[0052] Groups were formed and dosages were determined as shown in Table 1.

[0053] Group Folic acid Test article Dose(mg / kg / day) Dose frequency Route Vol.(mL / kg) Head G1 X Vehicle - QD PO 10 3 G2 O Vehicle - QD PO 10 7 G3 O Dapagliflozin 5 QD PO 10 7 G4 O Olmesartan medoxomil 10 QD PO 10 7 G5 O Dapagliflozin + Olmesartan medoxomil Dapa. 5 + Olme. 10 QD PO 10 7

[0054] 4. Observation and Examination

[0055] For the following observation, measurement, and examination dates, the start date of administration was designated as Day 1, and the 7 days from the start date of administration were calculated as 1 week of administration.

[0056] 1) Inspection Schedule

[0057] Observation items Acclimation period (week) Administration period (week) Termination of administration 1~6 7 1 2 3 4 Military separation ● Folic acid administration ● administration ● ● ● ● Urine sampling ● Histological examination of the kidney ●

[0058] 2) Observation of clinical symptoms

[0059] General symptoms were observed once a day for all animals, and the presence of morbid or deceased animals was checked twice a day. Observation was conducted from the first day of administration until the end of administration.

[0060] 3) Spot urinary albumin / creatinine ratio (ACR) measurement

[0061] On the last day of administration of the test substance (D28), urine was collected from the test animals, and urinary albumin and creatinine were measured using a biochemical analyzer (AU480, Beckman Coulter, Germany) with the collected urine.

[0062] 4) Histological analysis of the kidney

[0063] For histological examination, kidney tissue was fixed in 10% neutral formalin, and the fixed tissue was prepared into paraffin blocks and sectioned through general tissue processing procedures such as trimming, dehydration, and paraffin embedding. The remaining tissue was stored in a 10% neutral formalin solution.

[0064] i) Analysis of fibrosis area in kidney tissue

[0065] Sirius-red stained slides were photographed at 200x magnification, and the area of ​​the stained region was measured using Image Analyzer (NIS-Elements BR 5.11.01, Nikon) software. Subsequently, the stained region was divided by the total area of ​​the region of interest (ROI), and the means between groups were compared based on this ratio. In cases where false positives could be observed due to background staining or artifacts, the threshold was adjusted for measurement.

[0066] ii) Analysis of glomerular area in kidney tissue

[0067] Hematoxylin & Eosin (H&E) staining was performed on sectioned kidney tissue sections, and images were taken at 40x magnification. The glomerular area of ​​the renal cortex was analyzed using the Image J program (1.54K, https: / / imagej.net / ij / ). All glomerular areas were measured on the slide images, and the average between groups was compared based on the average area value of the normal group (G1).

[0068] 5. Statistical Analysis

[0069] All experimental results obtained from the experiment were expressed as mean ± standard deviation and analyzed using SPSS (version 20, IBM SPSS Statistics, USA). One-way analysis of variance (ANOVA) was performed on all data; if significance was observed, Dunnett's test was conducted to identify the experimental group showing a significant difference from the control group. (Significance levels: 5% and 1% for two-sided studies)

[0070] The statistical significance of the glomerular area results was analyzed using GraphPad Prism 8 software, and a Student T-test was conducted to confirm the significance compared to the control group or the monotherapy group. (Significance level: two-sided 5%)

[0071] Based on the histological analysis results, statistics between the G4 Folic acid Olmesartan medoxomil administration group and the G5 Folic acid Dapagliflozin + Olmesartan medoxomil administration group were analyzed by the sponsor using GraphPrism 8. Tukey's test was performed to confirm significance. (Significance levels: two-sided 5% and 1%)

[0072] < Examples 1> Observation of clinical symptoms

[0073] No animals exhibited abnormal symptoms during the induction period following the admission of the test animals. After the induction period ended, renal fibrosis was induced in the remaining animals—excluding the G1 Vehicle control group—by intraperitoneal administration of folic acid at a dose of 250 mg / kg, followed by the initiation of administration of the test substance. No notable clinical symptoms were observed during the test period, nor were any adverse reactions that could be caused by the test substance observed.

[0074] < Examples 2> Spot urinary albumin / creatinine ratio ( ACR ) measurement

[0075] On the last day of test substance administration, urine was collected from test animals and urinary albumin, creatinine, and albumin to creatinine ratio (ACR) were measured. As a result, urinary albumin levels were significantly reduced in the G4 Folic acid olmesartan administration group (2.29 ± 0.93 μg / mL) compared to the G2 Folic acid Vehicle control group (8.17 ± 4.88 μg / mL). The ACR results were significantly reduced in all administration groups—the G3 Folic acid dapagliflozin administration group (18.14 ± 13.07 mg / g), the G4 Folic acid olmesartan administration group (14.95 ± 6.26 mg / g), and the G5 Folic acid dapagliflozin + olmesartan administration group (23.53 ± 10.34 mg / g)—compared to the G2 Folic acid Vehicle control group (58.97 ± 29.44 mg / g) (Figure 1 and Table 3).

[0076] Group Albumin (μg / mL) Creatinine (mg / dL) ACR (mg / g) G1 Vehicle Control N Mean 2.06 * 25.80 9.62 ** 3 SD 0.52 15.80 5.16 G2Folic acid Vehicle control N Mean 8.17 14.75 58.97 7 SD 4.88 4.96 29.44 G3Folic acidDapagliflozin 5 mg / kg N Mean 4.23 25.56 18.14 ** 7 SD 3.13 11.60 13.07 G4Folic acidOlmesartan medoxomil 10 mg / kg N Mean 2.29 ** 16.19 14.95 ** 7 SD 0.93 4.35 6.26 G5Folic acidDapagliflozin 5 mg / kg +Olmesartan medoxomil 10 mg / kg N Mean 4.71 21.94 23.53 ** 7 SD 1.71 7.07 10.34

[0077] Significant difference from G2 Folic acid Vehicle control group by One-way ANOVA followed by the Dunnett' test: * p<0.05, ** p<0.01.

[0078] < Examples 3> Histological analysis of the kidney

[0079] Kidney tissues excised during autopsy were fixed and stained with Sirius-red to evaluate and analyze the fibrosis area. The fibrosis area was significantly reduced in the G3 Folic Acid Dapagliflozin administration group (1.83 ± 0.51%) and the G5 Folic Acid Dapagliflozin + Olmesartan administration group (1.44 ± 0.51%) compared to the G2 Folic Acid Vehicle control group (2.74 ± 0.46%), and significantly reduced in the G5 Folic Acid Dapagliflozin + Olmesartan administration group (1.44 ± 0.51%) compared to the G4 Folic Acid Olmesartan administration group (2.34 ± 0.70%) (Fig. 2 and Table 4).

[0080] Group H&E staining Sirius-red staining Tubulointerstitial damage index Fibrosis area mean(%) G1 Vehicle Control N Mean 0.00 ** 0.14 ** 3 SD 0.00 0.01 G2Folic acid Vehicle control N Mean 2.71 2.74 7 SD 0.49 0.46 G3Folic acidDapagliflozin 5 mg / kg N Mean 1.00** 1.83* 7 SD 0.00 0.51 G4Folic acidOlmesartan medoxomil 10 mg / kg N Mean 2.71 2.34 7 SD 0.76 0.70 G5Folic acidDapagliflozin 5 mg / kg +Olmesartan medoxomil 10 mg / kg N Mean 1.00 **## 1.44 **# 7 SD 0.58 0.51

[0081] Significant difference from G2 Folic acid Vehicle control group by One-way ANOVA followed by the Dunnett' test: * p<0.05, ** p<0.01. Significant difference between G4 Folic acid Olmesartan medoxomil group and G5 Folic acid Dapagliflozin + Olmesartan medoxomil group by the Tukey's test: # p<0.05, ##p<0.01.

[0082] As a result of measuring glomerular area, the G2 Folic acid Vehicle control group and the G3 Folic acid Dapagliflozin administration group showed a tendency for glomerular area to increase compared to the G1 Vehicle control group, but no significant difference was observed. On the other hand, the G4 Folic acid Olmesartan administration group or the G5 Folic acid Dapagliflozin + Olmesartan administration group each showed a statistically significant decrease in glomerular area compared to the G2 Folic acid Vehicle control group. In addition, the G5 Folic acid Dapagliflozin + Olmesartan administration group showed a significant decrease in glomerular area compared to the G3 Folic acid Dapagliflozin administration group (Figs. 3, 4, and Table 5).

[0083] Group Relative mean glomerulus area (%) G1Vehicle control N Mean 100.01 3 SD 8.54 G2Folic acidVehicle control N Mean 110.17 7 SD 11.55 G3Folic acidDapagliflozin 5 mg / kg N Mean 120.71 7 SD 14.29 G4Folic acidOlmesartan medoxomil 10 mg / kg N Mean 96.31 * 7 SD 11.88 G5Folic acidDapagliflozin 5 mg / kg +Olmesartan medoxomil 10 mg / kg N Mean 95.77 *,# 7 SD 6.33

[0084] Significant difference from G2 Folic acid Vehicle control group by student t-test: * p<0.05. Significant difference between G3 Folic acid Dapagliflozin group and G5 Folic acid Dapagliflozin + Olmesartan medoxomil group by student t-test: # p<0.05.

[0085] In conclusion, the present invention aimed to confirm the efficacy of the test substance against renal fibrosis disease by orally administering the test substance to C57BL / 6 mice for 4 weeks in a folic acid-induced chronic kidney disease model, confirming urine analysis, and histological analysis of the kidney.

[0086] Clinical symptoms were checked during the period of administration of the test substance, and no notable clinical symptoms were observed in any of the administration groups until the end of administration.

[0087] As a result of analyzing urine collected on the last day of administration, the albumin to creatinine ratio (ACR) in the dapagliflozin, olmesartan, and dapagliflozin + olmesartan groups decreased significantly compared to the G2 Folic Acid Vehicle control group.

[0088] As a result of evaluating and analyzing the fibrosis area by staining the kidney tissue excised during autopsy with Sirius-red, the fibrosis area in the dapagliflozin administration group and the dapagliflozin + olmesartan administration group was significantly reduced compared to the G2 Folic acid Vehicle control group, and the fibrosis area in the dapagliflozin + olmesartan administration group was significantly reduced compared to the olmesartan administration group.

[0089] It is known that in animal models of rodent renal fibrosis, renal dysfunction is correlated with an increase in intrarenal glomerular area, and that intrarenal glomerular area decreases when renal function improves due to drug administration. To confirm this, intrarenal glomerular area was measured, and it was confirmed that the dapagliflozin + olmesartan administration group showed a decrease in glomerular area when compared to the G2 Folic Acid Vehicle control group and the dapagliflozin administration group, respectively.

[0090] Based on the above results, when renal fibrosis was induced in C57BL / 6 mice by administering folic acid and the test substances dapagliflozin and olmesartan medoxomil were administered alone and in combination for 4 weeks, the efficacy against renal fibrosis was confirmed by observing a significant reduction in the albumin to creatinine ratio (ACR) and the fibrosis area of ​​the kidney tissue in the dapagliflozin alone and dapagliflozin + olmesartan combination groups. Furthermore, histological analysis indicated that the dapagliflozin + olmesartan combination group showed superior renal fibrosis and renal protective efficacy compared to the groups administered dapagliflozin or olmesartan medoxomil alone.

[0091] Foregoing, specific parts of the present invention have been described in detail. It will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Accordingly, the actual scope of the invention is defined by the appended claims and their equivalents.

Claims

Claim 1 A pharmaceutical composition for the prevention or treatment of renal fibrosis comprising: dapagliflozin, an SGLT-2 inhibitor, or a pharmaceutically acceptable salt thereof; and olmesartan, olmesartan medoxomil, or a pharmaceutically acceptable salt thereof, which are angiotensin receptor blockers. Claim 2 delete Claim 3 delete Claim 4 A pharmaceutical composition according to claim 1, characterized in that the weight ratio of the SGLT-2 inhibitor dapagliflozin or a pharmaceutically acceptable salt thereof; and the angiotensin receptor blocker olmesartan, olmesartan medoxomil or a pharmaceutically acceptable salt thereof is 1:(1 to 4). Claim 5 delete Claim 6 A pharmaceutical composition according to claim 1, characterized in that the renal fibrosis is oxidative stress-induced renal fibrosis. Claim 7 A pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition is administered orally.

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