WO2019165622A1 - Use of niclosamide ethanolamine salt and pharmaceutical composition thereof - Google Patents

Abstract

Provided is the use of a niclosamide ethanolamine salt and a pharmaceutical composition thereof. The salt and the composition play an uncoupling role for renal mitochondria, reduce proteinuria excretion in mice with kidney diseases, improve glomerulosclerosis and renal tubule lesions, and decrease serum creatinine levels. Therefore, the niclosamide ethanolamine salt and the pharmaceutical composition thereof have a certain protective effect on the kidneys and can be used for preventing and treating kidney diseases.

Description

Application of niclosamide ethanolamine salt and its pharmaceutical composition Technical field

The present invention relates to the technical field of kidney drugs, and more particularly to the use of a niclosamide ethanolamine salt and a pharmaceutical composition thereof.

Background technique

Kidney disease is a kind of major chronic non-infectious disease that threatens human health. The cause of this disease is complicated, the treatment of diseases is difficult, the condition is difficult to heal, and the medical expenses are huge. Proteinuria, progressive elevation of serum creatinine, glomerular sclerosis, and tubular damage are major clinical features of the early stage. At present, there are no effective drugs for delaying the progression of kidney disease in the clinic.

Niclosamide is a snail-killing and anthelmintic drug recommended by WHO. Niclosamide ethanolamine salt (NEN) is a derivative with better water solubility. Recent studies have shown that these drugs are obvious. Anti-tumor and hypoglycemic effects.

Summary of the invention

It is an object of the present invention to provide an application of a niclosamide ethanolamine salt and a pharmaceutical composition thereof, which solves the problem of the prior art which does not have an effective drug for delaying the progression of kidney disease.

The technical solution adopted by the present invention to solve the technical problem is: the application of niclosamide ethanolamine salt and a pharmaceutical composition thereof for preparing a medicament for treating and preventing kidney diseases, respectively.

The use of niclosamide ethanolamine salt and its pharmaceutical composition for the preparation of a medicament for uncoupling kidney mitochondria, respectively.

The use of niclosamide ethanolamine salt and a pharmaceutical composition thereof for the preparation of a medicament for reducing the proteinuria excretion rate of the kidney, respectively.

The use of niclosamide ethanolamine salt and a pharmaceutical composition thereof for preparing a medicament for reducing renal pathological damage, respectively.

The use of niclosamide ethanolamine salt and a pharmaceutical composition thereof for preparing a medicament for reducing the level of serum creatinine in renal diseases and delaying renal failure, respectively.

In the application of the present invention, the pharmaceutical composition is an orally administered preparation comprising an effective active ingredient niclosamide ethanolamine salt and a pharmaceutically acceptable adjuvant, an injection administration preparation, a mucosal administration preparation or a transdermal administration preparation.

In the application of the present invention, the pharmaceutical composition is an orally administered preparation, the orally administered preparation is a tablet, a capsule, a granule or an oral solution; or the pharmaceutical composition is a mucosal administration preparation, The preparation for mucosal administration is a patch or a gel; or the pharmaceutical composition is a preparation for transdermal administration, and the preparation for transdermal administration is a patch or a gel.

The use of the niclosamide ethanolamine salt of the present invention and the pharmaceutical composition thereof have the following beneficial effects: the niclosamide ethanolamine salt of the present invention and the pharmaceutical composition thereof can uncouple the renal tissue mitochondria and reduce the kidney disease Excretion of proteinuria in mice, improvement of glomerular sclerosis and tubular damage, and reduction of serum creatinine levels, so niclosamide ethanolamine salt and its pharmaceutical composition have a protective effect on kidney disease and can be used for the prevention and treatment of kidneys. disease.

DRAWINGS

Figure 1 is a graph showing the effect of NEN on renal mitochondrial oxygen consumption in the absence of oligomycin;

Figure 2 is a graph showing the effect of NEN on renal mitochondrial oxygen consumption in the presence of oligomycin;

Figure 3 is a comparison of the effects of NEN on proteinuria excretion in mice with adriamycin-induced nephropathy;

Figure 4 is a comparison of the effects of NEN on glomerular sclerosis in mice with adriamycin-induced nephropathy;

Figure 5 is a comparative photograph of a stained section of NEN on glomerular sclerosis in mice with adriamycin-induced nephropathy;

Figure 6 is a comparison of the effects of NEN on renal tubular injury in mice with adriamycin-induced nephropathy;

Figure 7 is a photograph of a stained section of NEN on renal tubular injury in mice with adriamycin-induced nephropathy;

Figure 8 is a comparison of the effects of NEN on serum creatinine in mice with adriamycin-induced nephropathy.

Detailed ways

The pharmaceutical composition of the calcium sensitive receptor-related peptide of the present invention and its use will be further described below with reference to the accompanying drawings and examples:

Kidney disease is mainly caused by increased urinary protein excretion caused by kidney damage. Proteinuria can also aggravate kidney damage, leading to glomerular sclerosis and tubular damage, which in turn leads to a decrease in glomerular filtration function and a progressive increase in serum creatinine. Eventually toward renal failure, the cause of this disease is complex, the pathogenesis is not completely clear, and the treatment is difficult.

The present invention provides the use of a pharmaceutical composition comprising niclosamide ethanolamine salt (NEN) and niclosamide ethanolamine salt (NEN) for the preparation of a medicament for the treatment and prevention of kidney disease. Among them, niclosamide ethanolamine salt (NEN) is a structural compound numbered by CAS NO 1420-04-8, and its structural formula is as follows:

Among them, the pharmaceutical composition is an orally administered preparation containing an effective active ingredient niclosamide ethanolamine salt and a pharmaceutically acceptable adjuvant, an injection administration preparation, a mucosal administration preparation or a transdermal administration preparation. Such as tablets, capsules, granules, oral liquids, patches or gels, and the like. The above dosage forms of niclosamide ethanolamine salt are all prior art and will not be described in detail herein.

The application of niclosamide ethanolamine salt (NEN) and a pharmaceutical composition containing niclosamide ethanolamine salt (NEN) will be described in detail below.

First, the experimental method

1. Animal model: Male BALB/c mice (body weight 20-25g) were purchased from the Guangdong Medical Laboratory Animal Center. Animal experiments were carried out in strict accordance with animal ethics guidelines and regulations, and were raised at the Peking University Shenzhen Graduate School Animal Center. The experimental animals were controlled to freely ingest and drink at a constant room temperature of 23 ± 1 ° C, 12 hours light and 12 hours dark cycle. Eighteen mice were randomly divided into normal control group (ie, control group in the figure, n=6), adriamycin nephropathy (AN, n=6) and adriamycin nephropathy+NEN treatment group (AN+). NEN, n=6). The mouse model of adriamycin nephropathy was induced by a single tail vein injection of 10.4 mg/kg doxorubicin (sigma, USA), and the normal control group was given a single intravenous injection of the same amount of normal saline. Interventions began 2 weeks after modeling, in which normal control and AN mice were fed conventional food, and AN+NEN group was fed with NEN supplemented for 4 weeks. NEN was purchased from Hubei Shengtian Hengchuang Biotechnology Co., Ltd. (China) and was added to the food of mice in a standard ratio of 2g/kg.

2. Determination of mitochondrial function in the kidney: After extracting the kidney mitochondria from normal male BALB/c mice, the oxygen consumption of mitochondria was measured using an oxygen electrode (Hansatech Instrument, UK) under the conditions of no oligomycin and oligomycin. Mitochondria concentration was 1 mg/ml, substrate concentration: glutamate/malate, 2.5 mM/2.5 mM; adenosine diphosphate (ADP), 200 μM; oligomycin, 5 μg/ml; NEN, 10 μM.

3. Before the modeling, 4 weeks and 6 weeks after the modeling, the mouse metabolic cage (Tecniplast, Italy) was used to collect the 24h urine of each group of mice, using the Bradford protein assay kit (Bio-Rad, USA). Quantification of 24 h total urine protein in each group of mice was performed.

4. Organizational preparation: After 6 weeks of modeling, 4 weeks after NEN intervention, after the mice in each group were sacrificed, serum samples were taken, the kidneys were removed, and a certain amount of kidney tissue was cut along the longitudinal section with 10% formalin. After fixation, dehydrated paraffin was embedded, and the remaining kidney tissue was immediately frozen in liquid nitrogen and stored at -80 ° C for subsequent experimental studies. Serum creatinine was measured using an automated biochemical analyzer (Roche, Switzerland).

5. Evaluation method of glomerular sclerosis index: 4 samples were randomly selected from each group. Under the PAS staining section, each slice was randomly photographed under the 40× objective lens for 20-30 glomeruli in the renal cortex. The main manifestations of glomerular sclerosis For mesangial matrix expansion, capillary occlusion, hyaline degeneration and balloon adhesion. If the glomerulus is basically normal (or lesion <5%), it will accumulate 0 points. If the lesion accumulates glomerulus 5 to 25%, it will accumulate 1 point, 25 to 50% product 2 points, 50 to 75% product 3 points. 75 to 100% is accumulated for 4 points, and then the average value of each slice is calculated.

6. Evaluation method of renal tubular injury index: 4 samples were randomly selected from each group. Under the PAS staining section, each section was randomly photographed under the 20× objective lens for 10-20 renal tubules in the renal cortex. The renal tubular injury was mainly characterized by small tubules. Tubular, atrophic, dilated, and inflammatory cell infiltration. If the renal tubule is basically normal (or lesion <5%), it will accumulate 0 points. If the lesion accumulates in the 5-25% area of the renal tubule, it will accumulate 1 point, 25-50% of the product will be 2 points, 50-100% will accumulate 3 points, then Calculate the average of each slice.

7, statistical analysis

Measurement data are expressed as mean ± standard deviation. Statistical differences between the two groups of samples were analyzed using independent sample t-test. Comparisons between groups of samples were performed using one-way analysis of variance, and statistical analysis was performed using SPSS 16.0 statistical software. A statistical difference was considered to be significant at P < 0.05.

Second, the results

1. Niclosamide ethanolamine salt (NEN) has obvious uncoupling effect on mitochondria of BALB/c mice.

As shown in Figure 1, NEN increases renal mitochondrial oxygen consumption in the absence of oligomycin; as shown in Figure 2, NEN increases renal mitochondrial oxygen consumption in the presence of oligomycin. The mitochondrial concentration was 1 mg/ml. Substrate concentrations: glutamate/malate, 2.5 mM / 2.5 mM; adenosine diphosphate (ADP), 200 μM; oligomycin, 5 μg/ml; NEN, 10 μM.

2, niclosamide ethanolamine salt (NEN) can significantly reduce the excretion of proteinuria in mice with adriamycin-induced nephropathy.

As shown in Figure 3, it is the effect of NEN on proteinuria excretion in mice with adriamycin-induced nephropathy (n=6). ^* P < 0.05, ^*** P < 0.001 compared with the normal group; ^# P < 0.05, ^## P < 0.01 compared with the AN group.

At the beginning of doxorubicin modeling (at 0 weeks), there was no significant difference in urinary protein excretion rates between the control group, the AN group, and the AN+NEN group. Four weeks after model establishment (that is, NEN treatment for 2 weeks), compared with the control group, the urine protein of the AN group increased significantly. After 2 weeks of NEN treatment, the urine protein of the AN+NEN group was significantly lower than that of the AN group. Six weeks after model establishment (ie, NEN treatment for 4 weeks), the urine protein of the AN group decreased compared with the previous 4 weeks, but it was still significantly higher than that of the control group. After 4 weeks of NEN treatment, the urine protein of the AN+NEN group was still significantly lower. In the AN group.

3, niclosamide ethanolamine salt (NEN) can significantly reduce glomerular sclerosis in mice with adriamycin nephropathy.

As shown in Figures 4 and 5, the effect of NEN on glomerular sclerosis in mice with adriamycin-induced nephropathy (n = 4). Compared with the normal group, ^*** P <0.001; compared with the AN group, ^# P < 0.05. After 6 weeks of modeling (NEN treatment for 4 weeks), the glomerular sclerosis index of the AN group was significantly increased, and the glomerular sclerosis index of the AN+NEN group was significantly lower than that of the AN group.

4, niclosamide ethanolamine salt (NEN) can significantly reduce renal tubular damage in mice with adriamycin nephropathy.

As shown in Figures 6 and 7, the effect of NEN on renal tubular injury in mice with adriamycin-induced nephropathy (n = 4). Compared with the normal group, ^*** P <0.001; compared with the AN group, ^## P < 0.01. After 6 weeks of modeling (4 weeks after NEN treatment), the renal tubular injury index of the AN group was significantly increased. Compared with the AN group, the renal tubular injury index of the AN+NEN group was significantly reduced.

5, niclosamide ethanolamine salt (NEN) can significantly reduce serum creatinine levels in mice with adriamycin nephropathy.

As shown in Figure 8, it is the effect of NEN on serum creatinine in mice with adriamycin-induced nephropathy (n=6). Compared with the normal group, ^** P <0.01; compared with the AN group, ^# P <0.05. After 6 weeks of modeling (NEN treatment for 4 weeks), serum creatinine was significantly increased in the AN group, and serum creatinine was significantly decreased in the AN+NEN group compared with the AN group.

The above experimental results show that the prior art niclosamide ethanolamine salt (NEN), as an anthelmintic and anti-tumor drug, can uncouple the kidney mitochondria, reduce proteinuria excretion of kidney disease, and improve Glomerular sclerosis and tubular damage reduce serum creatinine levels, thereby delaying the progression of kidney disease to renal failure. Therefore, the pharmaceutical composition containing niclosamide ethanolamine salt (NEN) can also uncoupling the kidney mitochondria, reducing proteinuria excretion of kidney disease, improving glomerular sclerosis and tubular damage, and lowering serum creatinine level. , thereby delaying the progression of kidney disease to the progression of kidney failure.

It is to be understood that those skilled in the art can make modifications and changes in the above-described description, all of which are within the scope of the appended claims.

Claims (7)

The use of niclosamide ethanolamine salt and a pharmaceutical composition thereof for the preparation of a medicament for the treatment and prevention of kidney diseases, respectively. The use of niclosamide ethanolamine salt and its pharmaceutical composition for the preparation of a medicament for uncoupling kidney mitochondria, respectively. The use of niclosamide ethanolamine salt and a pharmaceutical composition thereof for the preparation of a medicament for reducing the proteinuria excretion rate of the kidney, respectively. The use of niclosamide ethanolamine salt and a pharmaceutical composition thereof for preparing a medicament for reducing renal pathological damage, respectively. The use of niclosamide ethanolamine salt and a pharmaceutical composition thereof for preparing a medicament for reducing the level of serum creatinine in renal diseases and delaying renal failure, respectively. The use according to any one of claims 1 to 5, wherein the pharmaceutical composition is an orally administered preparation comprising an effective active ingredient niclosamide ethanolamine salt and a pharmaceutically acceptable adjuvant, and an injection preparation A mucosal administration preparation or a preparation for transdermal administration. The use according to claim 6, wherein the pharmaceutical composition is an orally administered preparation, and the orally administered preparation is a tablet, a capsule, a granule or an oral solution; or the pharmaceutical composition is A preparation for mucosal administration, which is a patch or a gel; or the pharmaceutical composition is a preparation for transdermal administration, and the preparation for transdermal administration is a patch or a gel.

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