Pharmaceutical composition for preventing and treating diabetic nephropathy and application thereof

Abstract

The invention belongs to the technical field of medicines, and relates to a pharmaceutical composition for preventing and treating diabetic nephropathy and application thereof. The pharmaceutical composition for preventing and treating diabetic nephropathy is composed of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N '-hydroxyformamido)-4-(2-guanidino ethyl) amino)-1, 2, 5-oxadiazole or a pharmaceutically acceptable salt thereof, and anhydroicaritin. The pharmaceutical composition for preventing and treating the diabetic nephropathy can effectively control blood sugar and improve kidney functions, and the two medicines have a synergistic effect and have a remarkable prevention and treatment effect on the diabetic nephropathy.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to a pharmaceutical composition for the prevention and treatment of diabetic nephropathy and its application. Background Technology

[0002] Diabetic nephropathy (DN) is a common chronic kidney disease and one of the most dangerous chronic complications of diabetes. Its clinical features include proteinuria, progressive kidney function impairment, hypertension, edema, and severe kidney failure in the late stage. It is one of the leading causes of death in diabetic patients.

[0003] The etiology and pathogenesis of diabetic nephropathy are unclear. It is currently believed to be caused by multiple factors, resulting from a certain genetic background and the combined effects of several risk factors. Early signs of diabetic nephropathy include abnormal renal hemodynamics, manifested as glomerular hyperperfusion and hyperfiltration, with increased renal blood flow and glomerular filtration rate, which are more pronounced with increased protein intake. It is often accompanied by metabolic abnormalities caused by hyperglycemia, hypertension, and abnormal metabolism of vasoactive substances.

[0004] Based on the course and pathophysiological evolution of diabetic nephropathy, it can be divided into five stages: (I) glomerular hyperfiltration and renal hypertrophy stage; (II) normoalbuminuria stage; (III) early diabetic nephropathy stage; (IV) clinical diabetic nephropathy stage; and (V) end-stage renal failure. Diabetic nephropathy is characterized by vascular dysfunction and chronic inflammation. Current research has found that immune cell infiltration, especially macrophages, plays an important role in the occurrence and development of diabetic nephropathy. The degree of macrophage accumulation in kidney tissue is closely related to the severity of kidney tissue damage. Hyperglycemia activates circulating monocytes to differentiate into macrophages, causing them to adhere to endothelial cells and migrate to cardiovascular and renal tissues. Once localized, activated macrophages develop into foam cells and produce peroxides, as well as pro-inflammatory and pro-fibrotic cytokines, thereby affecting almost all cells in the kidney tissue, leading to extracellular matrix accumulation, fibrosis, cell dysfunction, and proteinuria.

[0005] Effective prevention and control of diabetic nephropathy is of particular urgency, and the search for drugs that can improve kidney damage in diabetic nephropathy has important clinical application value. Summary of the Invention

[0006] This invention provides a pharmaceutical composition for the prevention and treatment of diabetic nephropathy and its application. The pharmaceutical composition for the prevention and treatment of diabetic nephropathy of this invention comprises (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinylethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof and icariin. The pharmaceutical composition for the prevention and treatment of diabetic nephropathy of this invention can effectively control blood glucose and improve kidney function. The two drugs have a synergistic effect, exhibiting a significant preventive and therapeutic effect on diabetic nephropathy.

[0007] Specifically, the technical objective of this invention is achieved through the following technical solutions:

[0008] A pharmaceutical composition for the prevention and treatment of diabetic nephropathy, wherein the weight ratio of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof to icariin is 1.3-2.2:1.

[0009] Preferably, the weight ratio of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof to icariin in the pharmaceutical composition is 1.3:1, 1.7:1 or 2.2:1.

[0010] The present invention provides the use of a pharmaceutical composition containing (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof and icariin in the preparation of a medicament for treating diabetic nephropathy.

[0011] The present invention provides the use of a pharmaceutical composition containing (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof and icariin in the preparation of a medicament for improving the urinary microalbumin excretion rate in patients with diabetic nephropathy.

[0012] This invention provides the use of a pharmaceutical composition comprising (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof and icariin in the preparation of a medicament for improving serum creatinine and blood urea nitrogen levels in patients with diabetic nephropathy.

[0013] The present invention also provides a pharmaceutical formulation comprising the pharmaceutical composition as described above and a pharmaceutically acceptable carrier.

[0014] Preferably, the dosage form of the pharmaceutical preparation is an oral preparation or an injectable preparation.

[0015] More preferably, the oral preparation is a tablet, capsule, powder, pill, granule, oral liquid, syrup, or decoction.

[0016] Preferably, the pharmaceutical preparation is used to prepare a drug for the prevention and / or treatment of diabetic nephropathy.

[0017] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological potency of the free acid and free base of the specified compound and has no adverse effects in biological or other respects. The compounds of this application also include pharmaceutically acceptable salts. A pharmaceutically acceptable salt is defined as a salt formed by converting a base group in a parent compound into its salt form. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts containing base groups such as amine (amino) groups. Pharmaceutically acceptable salts of this application can be synthesized from a parent compound by reacting a basic group in the parent compound with 1-4 equivalents of an acid in a solvent system. Suitable salts are listed in one or more of Remingtong's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977).

[0018] As used herein, the term "treatment" and other similar synonyms include relieving, reducing, or improving symptoms of a disease or condition; inhibiting a disease or condition, such as preventing its progression; alleviating a disease or condition; improving a disease or condition; relieving symptoms caused by a disease or condition; or stopping symptoms of a disease or condition; preventing other symptoms; improving or preventing the underlying metabolic causes of symptoms; and, moreover, the term includes preventative purposes. The term also includes achieving therapeutic and / or preventative effects. A therapeutic effect refers to the cure or improvement of the underlying disease being treated. Furthermore, the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect; for example, an improvement is observed in a patient even though they may still be affected by the underlying disease. In terms of preventative effects, the composition may be administered to patients at risk of developing a specific disease, or to patients exhibiting one or more physiological symptoms of a disease, even if no disease diagnosis has been made.

[0019] The present invention utilizes icariin and (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof in combination to significantly reduce body weight and fasting blood glucose in diabetic nephropathy mice, improve urinary microalbumin excretion rate, and reduce increases in serum creatinine and blood urea nitrogen, thereby achieving hypoglycemic and renal protective effects. The two drugs have a synergistic effect, significantly enhancing the prevention and treatment of diabetic nephropathy.

[0020] Compared with the prior art, the present invention has the following advantages:

[0021] The present invention comprises a pharmaceutical composition consisting of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof and icariin, which can lower blood sugar and improve kidney condition, effectively control blood sugar and improve kidney function.

[0022] The icariin of this invention is a natural product monomer extracted from the traditional Chinese medicine Epimedium. It has low toxicity and side effects on humans, significantly improving patient safety and medication adherence. Furthermore, the pharmaceutical composition of this invention, consisting of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof and icariin, exhibits a synergistic effect in the prevention and treatment of diabetic nephropathy, reducing dosage, lowering toxicity, and improving medication safety. Detailed Implementation

[0023] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0024] Pharmacodynamic Example 1: Effect of the pharmaceutical composition of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole and icariin on a mouse model of diabetic nephropathy.

[0025] 1. Laboratory animals

[0026] Eighty 7- to 8-week-old SPF male C57BLKS / J-Leprdb / Leprdb (db / db) mice and eight C57BLKS / J-Leprdb / Leprm (db / m) mice were purchased from Jiangsu Jicui Yakang Biotechnology Co., Ltd., with the production license number: SCXK (Su) 2023-0009. All mice were housed in a standard room with a 12h / 12h light / dark cycle, at a constant temperature of (24±1°C) and a relative humidity of (50±10%). The mice had free access to food and water.

[0027] 2. Main reagents

[0028] (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole (hereinafter referred to as "Compound A") was provided by Shandong Xinsdai Pharmaceutical Co., Ltd.

[0029] Icaritin was provided by Shandong Xinsdai Pharmaceutical Co., Ltd.

[0030] 3. Experimental methods and experimental groups

[0031] After 1 week of adaptive feeding of the animals, the db / db mice were randomly divided into 10 groups, namely the model group, the low-dose icariin group, the medium-dose icariin group, the high-dose icariin group, the low-dose Compound A group, the medium-dose Compound A group, the high-dose Compound A group, the Composition A group, the Composition B group, and the Composition C group, with 8 mice in each group. The db / m mice served as the normal group.

[0032] The dosing doses for each group were as follows:

[0033] Low-dose icariin group: Intragastric administration of icariin at 6 mg / (kg·d);

[0034] Medium-dose icariin group: Intragastric administration of icariin at 12 mg / (kg·d);

[0035] High-dose icariin group: Intragastric administration of icariin at 24 mg / (kg·d);

[0036] Low-dose Compound A group: Intragastric administration of Compound A at 13 mg / (kg·d);

[0037] Medium-dose Compound A group: Intragastric administration of Compound A at 20 mg / (kg·d);

[0038] High-dose Compound A group: Intragastric administration of Compound A at 30 mg / (kg·d);

[0039] Composition A group: Intragastric administration of icariin at 6 mg / (kg·d) and Compound A at 13 mg / (kg·d);

[0040] Composition B: Icariin 12 mg / (kg·d) and compound A 20 mg / (kg·d) were administered by gavage;

[0041] Composition C: Icariin 24 mg / (kg·d) and compound A 30 mg / (kg·d) were administered by gavage.

[0042] Each treatment group was administered the drug via gavage at a preset dose, while the normal group and the model group were given an equal volume of purified water, once daily for 24 weeks.

[0043] Experimental data are expressed as mean ± standard deviation. Statistical analysis was performed using GraphPad Prism 8.0.2 software. One-way ANOVA was used for comparisons between groups, and multi-way ANOVA was used for repeated measures data. A p-value < 0.05 was considered statistically significant.

[0044] 4. Measurement of mouse body weight and blood glucose levels

[0045] After administration, the weight of mice in each group was recorded. After fasting for 12 hours but not water, the fasting blood glucose level of mice was measured by tail tip sampling.

[0046] 5. Mouse renal function test

[0047] One week before the end of administration, 24-hour urine samples were collected from mice. After administration, the mice were dissected and serum was separated. The urinary microalbumin excretion rate and serum creatinine and urea nitrogen levels were detected using a fully automated biochemical analyzer.

[0048] 6. Experimental Results

[0049] 6.1 Mouse body weight and blood glucose levels

[0050] Table 1 shows that the body weight of mice in the model group was significantly higher than that of the normal group. Compared with the model group, the body weight of mice was significantly reduced by each dose of icariin, each dose of compound A, and each combination group. The combination groups were more effective in reducing body weight than the icariin and compound A dose groups. This indicates that icariin and compound A have a significant synergistic effect in reducing body weight in diabetic nephropathy mice.

[0051] Compared with the normal group, the fasting blood glucose level of mice in the model group was significantly increased. Compared with the model group, the various doses of icariin, compound A, and the combined composition significantly reduced fasting blood glucose levels in mice. The combined composition group showed a better effect in reducing blood glucose levels, significantly superior to the various doses of icariin and compound A. This indicates that icariin and compound A have a significant synergistic effect in reducing fasting blood glucose in diabetic nephropathy mice.

[0052] Table 1 Comparison of body weight and fasting blood glucose levels in mice of different groups

[0053] Group Dosage (mg / kg) Body weight (g) Fasting blood glucose (mmol / L) normal group - 28.76±1.16 4.13±0.59 Model group - <![CDATA[66.66±3.05 ### ]]> <![CDATA[31.46±1.54 ### ]]> Low-dose group of icariin 6 <![CDATA[57.56±1.47 ** ]]> <![CDATA[28.43±0.81 ** ]]> Medium dose group of icariin 12 <![CDATA[56.35±1.48 ** ]]> <![CDATA[27.94±0.94 ** ]]> High-dose group of icariin 24 <![CDATA[55.21±1.70 ** ]]> <![CDATA[26.64±0.36 ** ]]> Low-dose group of compound A 13 <![CDATA[55.53±2.74 ** ]]> <![CDATA[28.63±0.70 ** ]]> Medium dose group of compound A 20 <![CDATA[53.80±1.72 ** ]]> <![CDATA[26.71±0.63 ** ]]> High-dose group of compound A 30 <![CDATA[55.45±1.57 ** ]]> <![CDATA[26.76±0.48 ** ]]> Group A of compositions 6+13 <![CDATA[44.54±1.39 *** ]]> <![CDATA[23.84±0.72 *** ]]> Group B of Compositions 12+20 <![CDATA[42.40±0.90 *** ]]> <![CDATA[22.45±0.31 *** ]]> Composition C group 24+30 <![CDATA[42.38±0.51 *** ]]> <![CDATA[20.25±0.53 *** ]]>

[0054] Note: Compared with the normal group ### P<0.001; compared with the model group, ** P<0.01, *** P<0.001.

[0055] 6.2 Changes in urinary microalbumin excretion rate, serum creatinine, and blood urea nitrogen in mice

[0056] Table 2 shows that, compared with the normal group, the urinary microalbumin excretion rate of mice in the model group was significantly increased. Compared with the model group, the various doses of icariin, compound A, and the combined composition significantly reduced the urinary microalbumin excretion rate in mice. The combined composition group showed better effects, significantly superior to the individual doses of icariin and compound A. This indicates that icariin and compound A have a significant synergistic effect in reducing the urinary microalbumin excretion rate in diabetic nephropathy mice.

[0057] Compared with the normal group, the serum creatinine level in the model group mice was significantly increased. Compared with the model group, the various doses of icariin, compound A, and the combined composition significantly reduced serum creatinine levels in mice. The combined composition groups showed a better effect in reducing serum creatinine levels, significantly superior to the individual doses of icariin and compound A. This indicates that icariin and compound A have a significant synergistic effect in reducing serum creatinine levels in mice.

[0058] Compared with the normal group, the serum urea nitrogen level in the model group mice was significantly increased. Compared with the model group, the various dose groups of icariin, compound A, and the combination group significantly reduced serum urea nitrogen in mice. The combination group was more effective in reducing serum urea nitrogen in mice, significantly better than the various dose groups of icariin and compound A. This indicates that icariin and compound A have a significant synergistic effect in reducing serum urea nitrogen in diabetic nephropathy mice.

[0059] Table 2 Comparison of urinary microalbumin excretion rate, serum creatinine, and serum urea nitrogen levels in mice of different groups

[0060]

[0061] Note: Compared with the normal group ### P<0.001; compared with the model group, * P<0.01, ** P<0.01, *** P<0.001.

[0062] In summary, the combined use of icariin and (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole significantly reduced mouse body weight and fasting blood glucose, improved urinary microalbumin excretion rate, and reduced serum creatinine and blood urea nitrogen levels, demonstrating a significant synergistic effect in the treatment of diabetic nephropathy.

[0063] The above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description, and any obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A pharmaceutical composition for the prevention and treatment of diabetic nephropathy, characterized in that, The pharmaceutical composition for preventing and treating diabetic nephropathy comprises (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof with icariin.

2. The pharmaceutical composition for preventing and treating diabetic nephropathy according to claim 1, characterized in that, The weight ratio of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof to icariin in the pharmaceutical composition is 1.3-2.2:

1.

3. The pharmaceutical composition for preventing and treating diabetic nephropathy according to claim 2, characterized in that, The weight ratio of (Z)-3-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamido)-4-(2-guanidinoethyl)amino)-1,2,5-oxadiazole or a pharmaceutically acceptable salt thereof to icariin in the pharmaceutical composition is 1.3:1, 1.7:1 or 2.2:

1.

4. Use of the pharmaceutical composition for preventing or treating diabetic nephropathy according to any one of claims 1-3 in the preparation of a medicament for the prevention or treatment of diabetic nephropathy.

5. The use according to claim 4, characterized in that, The pharmaceutical composition improves the urinary microalbumin excretion rate in patients with diabetic nephropathy.

6. The use according to claim 4, characterized in that, The pharmaceutical composition improves serum creatinine and blood urea nitrogen levels in patients with diabetic nephropathy.

7. A pharmaceutical formulation comprising the pharmaceutical composition of any one of claims 1-3 and a pharmaceutically acceptable carrier.

8. The pharmaceutical preparation according to claim 7, characterized in that, The dosage form of the pharmaceutical preparation is an oral preparation or an injectable preparation.

9. The pharmaceutical preparation according to claim 8, characterized in that, The oral preparations are tablets, capsules, powders, pills, granules, oral liquids, syrups, or decoctions.

10. Use of the pharmaceutical preparation of claim 7 in the preparation of a medicament for treating diabetic nephropathy.

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