Application of lupinus luteus White ketone in preparation of medicine for preventing or treating kidney diseases

Abstract

The invention provides an application of lupinus luteus (Lupilightteone, Lup) in preparation of a medicine for preventing or treating kidney diseases, and relates to the field of medicines for preventing or treating kidney diseases. The Lup provided by the invention can protect renal tubular epithelial cells, significantly increase the activity of acute kidney injury cells, significantly improve the glomerular filtration function of mice, improve the pathological injury degree of kidneys and reduce renal fibrosis. The invention provides a new thought for developing medicines for preventing or treating kidney diseases.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical technology, and particularly relates to application of lupiwighteone in preparing drugs for preventing or treating kidney diseases. BACKGROUND

[0002] Acute kidney injury (AKI) is a clinical critical syndrome characterized by a rapid decline in renal function within hours to days, the core pathophysiological change of which is a rapid decrease in glomerular filtration rate (GFR), resulting in an increase in serum creatinine (SCr) concentration and retention of nitrogenous metabolic products, and often accompanied by water, electrolyte and acid-base balance disorders. According to the cause, AKI can be divided into three categories: pre-renal (insufficient perfusion), renal (renal parenchymal injury) and post-renal (urinary obstruction). At present, the clinical treatment of AKI lacks specific radical treatment, mainly relying on supportive treatment, such as correcting water and electrolyte disorders, eliminating the cause (such as stopping the use of nephrotoxic drugs) and renal replacement therapy (dialysis). Although these methods can maintain the vital signs of patients, they cannot fundamentally prevent the damage and death of renal tubular epithelial cells, nor can they effectively block the vicious pathological process initiated after injury.

[0003] Renal fibrosis is a common pathological change in almost all end-stage chronic kidney diseases (CKD), characterized by excessive deposition of extracellular matrix (ECM) in the renal interstitium, leading to destruction of normal nephron structure and loss of function. At present, there is no specific drug for anti-renal fibrosis. Moreover, most drugs target a single pathological link, and single-target treatment is difficult to comprehensively curb the complex network of diseases.

[0004] Lupiwighteone (Lup) is a natural isoprenylated isoflavone product, which is widely found in the pods and seeds of Lupinus luteus, the roots of Lupinus luteus and the leaves of Glycyrrhiza glabra. In recent years, research has found that Lup has various biological activities, such as antioxidant, antibacterial, anticancer, etc. In the study of anti-tumor, Lup can induce caspase-dependent and independent apoptosis of human breast cancer cells by inhibiting the PI3K / Akt / mTOR pathway, showing good anticancer potential. Its antioxidant properties can effectively scavenge excess reactive oxygen species in the body, reducing the damage of oxidative stress to cells. In addition, some studies have shown that some isoflavone compounds have potential therapeutic effects on kidney diseases, however, there is no report on the research of Lup in the treatment of acute kidney injury and renal fibrosis and other kidney diseases, and its mechanism of action and therapeutic effect still need to be further explored. SUMMARY

[0005] The purpose of this invention is to provide the use of lupiwighteone (Lup) in the preparation of medicaments for the prevention or treatment of kidney diseases.

[0006] The Lup described in this invention can be prepared by methods such as plant extraction or chemical synthesis (CN103936706A), and its structural formula is as follows: .

[0007] This invention provides the use of Lup in the preparation of drugs for the prevention or treatment of kidney diseases.

[0008] The kidney diseases described in this invention include acute kidney injury and chronic kidney disease.

[0009] Furthermore, the chronic kidney disease mentioned refers to chronic kidney disease with pathological features including renal fibrosis.

[0010] Furthermore, the chronic kidney disease mentioned includes diabetic nephropathy.

[0011] Pharmacological experiments have confirmed that Lup of this invention has the following pharmacological effects: (a) Lup can significantly increase the activity of cells in acute kidney injury.

[0012] (b) Lup can reduce blood urea nitrogen (BUN) and serum creatinine (CRE) in db / db mice and improve glomerular filtration function.

[0013] (c) Lup can significantly improve the degree of kidney pathological damage in db / db mice, significantly improve kidney damage, and slow down the fibrosis process of the kidneys in mice.

[0014] The present invention further provides a pharmaceutical composition for the prevention or treatment of kidney disease, comprising Lup.

[0015] The present invention further provides a pharmaceutical formulation for the prevention or treatment of kidney disease, comprising Lup and a pharmaceutically acceptable carrier.

[0016] Furthermore, the pharmaceutical preparation may be granules, tablets, powders, capsules, oral liquids, injections, sprays, aerosols, powder inhalers, emulsions, nasal drops, suppositories, pills, microspheres, or tinctures, etc.

[0017] This invention is the first to discover that Lup has a protective effect on renal tubular epithelial cells, improves renal function, reduces renal fibrosis, and slows the progression of the disease. This invention provides new insights into the development of drugs for the prevention or treatment of kidney diseases such as acute kidney injury and renal fibrosis. Attached Figure Description

[0018] Figure 1The effect of Lps on HK-2 (human proximal tubular cell line) cytotoxicity; each bar represents the mean ± standard deviation (SD) of the sample. p < 0.0001.

[0019] Figure 2 The effect of Lup on HK-2 (human proximal tubular cell line) cytotoxicity; each bar represents the mean ± standard deviation (SD) of the sample. p < 0.0001.

[0020] Figure 3 The effect of Lup on the viability of HK-2 (human proximal renal tubule cell line) cells; each bar represents the mean ± standard deviation (SD) of the sample. p < 0.0001.

[0021] Figure 4 The effect of Lup on blood urea nitrogen (BUN) levels in mouse serum; each bar represents the mean ± standard deviation (SD) of the sample. p < 0.05 p < 0.01.

[0022] Figure 5 The effect of Lup on serum creatinine (CRE) levels in mice; each bar represents the mean ± standard deviation (SD) of the sample. p < 0.05 p < 0.001.

[0023] Figure 6 Effects of Lup on mouse kidney histopathology: Mouse kidney sections were stained with H&E, Sirius Red, Masson, and PAS, respectively. Detailed Implementation

[0024] The following examples are intended to enable those skilled in the art to more fully understand the present invention, but do not limit the invention in any way.

[0025] Example 1: Effect of Lup on the viability of HK-2 (human proximal renal tubule cell line) cells

[0026] Lipopolysaccharide (LPS)-induced human proximal renal tubular cell line (HK-2 cells) serves as an in vitro model of septic acute kidney injury. The aim of this study was to investigate whether Lup (Lipopolysaccharide) has a cytoprotective effect in LPS-induced HK-2 cells. Figure 1 As shown, HK-2 cells were treated with 30 µg / ml LPS for 24 hours to establish an in vitro model of acute kidney injury. Figure 2 , 3Experimental results showed that, compared with the model group, Lup treatment at different concentration gradients could enhance the activity of damaged HK-2 cells in a concentration-dependent manner, reverse the cell morphological changes and decreased viability caused by LPS, and confirm that Lup has a direct protective effect on renal tubular epithelial cells.

[0027] Example 2: Effects of Lup on physiological and biochemical indicators related to nephropathy in db / db mice

[0028] 1. Laboratory animals

[0029] This study used db / db mice as the model group and db / m mice as littermates of db / db mice to eliminate the interference of genetic background and feeding conditions on the experimental results. The model group mice were treated with Lup, and various biochemical assays and staining methods were used to evaluate the preventive and therapeutic effects of Lup on acute kidney injury.

[0030] db / m and db / db mice were housed in an SPF-grade barrier facility with a temperature of 23±2℃, humidity of 55±5%, and 12 hours of sunlight.

[0031] 2. Establishment, grouping, and administration of experimental animal models

[0032] Male db / m and db / db mice (approximately 12 weeks old) were used and divided into a blank control group (db / m), a model group (db / db), and a model-treated group receiving compound Lup (20 mg / kg). All mice were administered the compound via gavage in a saline suspension for one week, followed by daily gavage for two weeks. The control group and the db / db model group received the same volume of saline.

[0033] 3. Experimental Data Processing and Analysis

[0034] Statistical analysis and output of experimental results were performed using GraphPad Prism 9 software. Results are expressed as mean ± standard deviation (SD). Differences between groups were analyzed using one-way ANOVA, and p < 0.05 was considered statistically significant.

[0035] Biochemical reagent kits were used to measure physiological and biochemical indicators related to kidney injury in mice: blood urea nitrogen (BUN) and serum creatinine (CRE). The effects of Lup on renal function were as follows: Figure 4 and Figure 5 As shown.

[0036] A prolonged high blood sugar environment can damage the kidneys, leading to a decrease in glomerular filtration rate and a reduced ability of the kidneys to excrete urea nitrogen, thereby causing elevated BUN levels in the blood.Figure 4 As can be seen, the blood BUN level in the db / db model group mice was significantly higher than that in the control group; while the BUN level in the db / db mice treated with Lup was significantly lower.

[0037] Changes in CRE are typically associated with glomerular filtration function and the kidneys' ability to concentrate urine. Figure 5 As can be seen, compared with normal mice with db / m, db / db mice had higher levels of CRE in their serum. However, Lup treatment significantly downregulated the CRE levels. These results indicate that Lup treatment reduced the damage to glomerular filtration function in the model group and improved its ability to clear metabolic waste, suggesting that Lup has a certain effect on improving kidney function.

[0038] Pathological changes in the structure of kidney tissue are a direct cause of decreased kidney function. After dissecting the kidney tissues of mice in each group, H&E staining, Sirius Red staining, Masson staining, PAS staining, and scanning imaging were performed to observe the lesions in the kidney tissues. Results are as follows: Figure 6 As shown: (1) H&E staining: The kidney structure of the db / m control group mice was clearly visible and intact, with regular renal tubule morphology and no inflammatory cell infiltration. Compared with the control group, the db / db model group mice had thickened basement membrane, mesangial cell proliferation, obvious renal tubule dilation, and glomerular hypertrophy. After Lup treatment, the above lesions were significantly improved, the thickening of basement membrane and mesangial cell proliferation were reduced, the renal tubule morphology became more intact, and the glomerular hypertrophy was reduced.

[0039] (2) Masson staining: In the middle and late stages of kidney disease, glomerular sclerosis and renal interstitial fibrosis are key factors in the deterioration of renal function. Masson staining can specifically show collagen fibers stained blue. Figure 6 As can be seen, the kidneys of mice in the db / m control group were all normal tissues stained red. Compared with the control group, the kidney tissues of the db / db model group showed a large number of blue fibers, while the blue fibers in the Lup treatment group were significantly reduced.

[0040] (3) Sirius Red staining: Before Masson staining showed obvious blue collagen, Sirius Red staining detected scattered red fine fibers in the renal interstitium, which contrasted sharply with the background. The collagen fibers in the glomeruli and tubulointerstitium of the db / db model group mice were significantly increased compared with the db / m control group. Compared with the model group, the area of ​​renal interstitial fibrosis in the Lup treatment group was significantly reduced, and the collagen fibers were also reduced. The results showed that Lup can significantly improve kidney injury and slow down the fibrosis process of the kidneys in mice.

[0041] (4) PAS staining: PAS staining reveals the polysaccharide and glycoprotein components in kidney tissue, providing a direct view of pathological changes in kidney structure, especially lesions related to glucose metabolism disorders. In the kidney tissue of the db / m control group mice, positive areas were mainly distributed along the glomerular capillary walls and renal tubules, with no positive areas in the middle. Compared with the control group, the db / db model group showed significantly increased deposition of positive substances, thickened basement membrane, expanded mesangial region, and increased accumulation of cytoplasmic glycogen. After Lup treatment, the positive areas were significantly reduced, the renal tubular structure became relatively regular, the deposition of positive substances decreased, and the thickening of the basement membrane and mesangial proliferation were improved.

[0042] Therefore, the above staining results revealed the effects of Lup on the pathological changes of kidney tissue in db / db mice from different dimensions. The results showed that Lup can significantly improve the degree of kidney pathological damage, slow down the thickening of the basement membrane and mesangial proliferation, and reduce glycogen accumulation and fibrosis.

[0043] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Application of yellow lupin white ketone in the preparation of drugs for the prevention or treatment of kidney diseases.

2. The application according to claim 1, characterized in that, The kidney diseases mentioned include acute kidney injury and chronic kidney disease.

3. The application according to claim 2, characterized in that, The chronic kidney disease mentioned refers to chronic kidney disease with pathological features including renal fibrosis.

4. The application according to claim 2, characterized in that, The chronic kidney disease mentioned includes diabetic nephropathy.

5. A pharmaceutical composition for the prevention or treatment of kidney disease, characterized in that, Including yellow lupin white ketone.

6. A pharmaceutical preparation for the prevention or treatment of kidney disease, characterized in that, This includes yellow lupin white ketone, as well as pharmaceutically acceptable carriers.

7. The pharmaceutical preparation for the prevention or treatment of kidney disease according to claim 6, characterized in that, The dosage forms of the pharmaceutical preparations include granules, tablets, powders, capsules, oral liquids, injections, sprays, aerosols, powder inhalers, emulsions, nasal drops, suppositories, pills, microspheres, or tinctures.

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