Application of medicine composition in preparation of medicine for treating diabetic nephropathy

Abstract

The invention relates to the technical field of biological medicine, in particular to application of a pharmaceutical composition in preparation of a medicine for treating diabetic nephropathy, the pharmaceutical composition is composed of active ingredients and pharmaceutically acceptable auxiliary materials, and the pharmaceutical composition is characterized in that the active ingredients are selected from the group consisting of active ingredients and pharmaceutically acceptable auxiliary materials; the active component is selected from one or more of melibiose or pharmaceutically acceptable salt, isomer and precursor thereof. The melibiose disclosed by the invention has the advantages that the melibiose can obviously improve diabetic mouse glomerular endothelial cell injury, reduce proteinuria, improve inflammation and play a role in protecting kidneys.

Description

Technical Field

[0001] The present invention relates to the field of biopharmaceutical technology, and particularly to the use of a pharmaceutical composition in the preparation of a medicament for treating diabetic kidney disease. Background Art

[0002] Diabetic kidney disease (DKD) is one of the most common microvascular complications of diabetes. Approximately 40% of diabetic patients develop DKD. Its pathological mechanism is complex and involves chronic hyperglycemia, oxidative stress, and inflammatory responses. In recent years, studies have shown that the continuous activation of the inflammatory microenvironment plays a key role in the progression of DKD. However, existing anti-inflammatory therapies may interfere with the host innate immunity, and there is an urgent need to develop new therapeutic strategies for targeted regulation of metabolic inflammation.

[0003] Melibiose is a naturally occurring oligosaccharide that is widely distributed in plants. There are currently no reports on its use in the treatment of DKD. Summary of the Invention

[0004] The object of the present invention is to overcome the deficiencies in the prior art and provide the use of a pharmaceutical composition in the preparation of a medicament for treating diabetic kidney disease. The active ingredient of the present invention is selected from one or more of melibiose or its pharmaceutically acceptable salts, isomers, and precursors. Through its natural source and unique anti-inflammatory mechanism, compared with the prior art, melibiose can target and improve the chronic inflammatory microenvironment of diabetic kidney disease (DKD) without affecting the host innate immunity, significantly reduce the urine protein / creatinine ratio (ACR) and plasma inflammatory factor (TNF-α, IL-6) levels. At the same time, its precursor stachyose is efficiently converted into melibiose through intestinal flora metabolism, improving the bioavailability and providing a new therapeutic strategy with both safety and effectiveness for diabetic kidney disease, breaking through the technical bottleneck that traditional anti-inflammatory drugs are prone to interfere with the immune system.

[0005] The present invention is achieved through the following technical solutions: On the one hand, there is provided the use of a pharmaceutical composition in the preparation of a medicament for treating diabetic kidney disease. The pharmaceutical composition is composed of an active ingredient and a pharmaceutically acceptable excipient, and the active ingredient is selected from one or more of melibiose or its pharmaceutically acceptable salts, isomers, and precursors.

[0006] Further, the pharmaceutical composition treats diabetic kidney disease by inhibiting the levels of TNF-α and IL-6 in the renal inflammatory microenvironment, repairing the damage of glomerular endothelial cells in diabetic kidney disease, and reducing the urine protein / creatinine ratio (ACR).

[0007] Further, the melibiose precursor is stachyose, which is metabolized by intestinal flora to generate melibiose, and the plasma melibiose concentration is positively correlated with the administration dose.

[0008] Furthermore, the dosage of the pharmaceutical composition administered to diabetic mice is 0.5 - 2 g·kg -1 ·d -1 , and after continuous administration for at least 8 weeks, the ACR is reduced by ≥30%.

[0009] Preferably, the pharmaceutical composition is selected from oral preparations, and the dosage form is solid powder or oral liquid.

[0010] Beneficial effects

[0011] The present invention for the first time reveals the mechanism by which melibiose and its precursor stachyose improve diabetic nephropathy through an anti-inflammatory pathway, breaking through the limitations of traditional direct anti-inflammatory therapies. As a natural oligosaccharide, melibiose has high safety and a wide source, avoiding the potential toxic and side effects of synthetic drugs and filling the gap in the application of natural products in this field.

[0012] The present invention targets the regulation of chronic metabolic inflammation rather than comprehensively inhibiting immunity, which can not only effectively reduce the levels of key inflammatory factors such as TNF-α and IL-6, but also avoid damaging the host's innate immune function. This selective regulation mechanism shows a dose-dependent reduction effect of urinary protein in animal experiments, solving the technical problem of the balance between immunosuppression and efficacy in traditional anti-inflammatory treatments.

[0013] The present invention uses stachyose as a prodrug and generates melibiose in situ through the metabolism of intestinal flora, overcoming the defect that melibiose is easily destroyed by gastric acid when directly administered. This biotransformation strategy significantly improves the bioavailability of the drug and prolongs the duration of drug efficacy through a sustained-release effect, providing a new technical path for the delivery of low-stability active ingredients. Description of the drawings

[0014] Figure 1 It is A. Changes in the ACR levels of mice in each group treated with melibiose (n = 8); B - C. Changes in the plasma TNF-α (B) and IL-6 (C) levels of mice in each group treated with melibiose (n = 8); D - F. Changes in the kidney TNF-α (D), IL-1β (E) and IL-6 (F) levels of mice in each group treated with melibiose (n = 3); G. Changes in the ultrastructure of glomerular endothelial cells of mice in each group treated with melibiose (n = 8). The abscissa is the grouping of mice, and the ordinate is the name of the relevant index; it shows that melibiose improves diabetic nephropathy and reduces ACR; melibiose can significantly improve the fenestrated structure of glomerular endothelial cells in DKD mice, showing that: in the NC group, the fenestrated structure of glomerular endothelium is regular, clear and evenly arranged; in the DKD group, the fenestrated structure of glomerular endothelium is almost completely lost, the endothelial surface is rough and the structure is severely damaged; after melibiose intervention, the fenestrated structure is partially restored, especially in the high-dose group, where the fenestrated structure of endothelium is close to the NC group state.

[0015] Figure 2A. Detect the cytotoxicity of melibiose on normally cultured glomerular endothelial cells using CCK8; B-C. The levels of TNF-α (B) and IL-6 (C) secreted by glomerular endothelial cells intervened with melibiose in each group; D-H. The protein expression levels and quantification of ET-1, OCLN, ICAM-1 and ZO-1 in cells of each group. The abscissa is the treatment of cells in each group, and the ordinate is the name of the relevant index; it shows that melibiose improves glomerular endothelial cell injury and tight junctions.

[0016] Figure 3 A. The metabolic pathway diagram of stachyose and melibiose; B. The plasma melibiose concentration of mice in each group treated with stachyose (n = 6); C. The ACR level of mice in each group treated with stachyose (n = 6); D-E. The plasma TNF-α (D) and IL-6 (E) levels of mice in each group treated with stachyose (n = 6). The abscissa is the grouping of mice in each group, and the ordinate is the name of the relevant index; it shows that stachyose can increase the plasma melibiose level and decrease ACR. Detailed implementation manners

[0017] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0018] Those skilled in the art can understand that, unless otherwise defined, all terms (including technical terms and scientific terms) used here have the same meaning as the general understanding of those of ordinary skill in the field to which the present invention belongs.

[0019] It should also be understood that terms such as those defined in a general dictionary should be understood to have a meaning consistent with the meaning in the context of the prior art, and will not be interpreted with an idealized or overly formal meaning unless defined as here.

[0020] The present invention relates to the use of a pharmaceutical composition in the preparation of a drug for treating diabetic nephropathy, the pharmaceutical composition is composed of an active ingredient and a pharmaceutically acceptable excipient, and the active ingredient is selected from one or more of melibiose or its pharmaceutically acceptable salts, isomers, precursors.

[0021] Melibiose (MEL for short) involved in the present invention is a kind of oligosaccharide and naturally exists in many plants. There is currently no relevant research on whether melibiose can be used to improve diabetic nephropathy. In the present invention, it is verified that melibiose can improve diabetic nephropathy, providing a solid basic research for the application of melibiose in the treatment of diabetic nephropathy, so that melibiose can be used to prepare a drug for treating diabetic nephropathy.

[0022] In the present invention, melibiose can be used to improve diabetic nephropathy. Improving diabetic nephropathy can also be referred to as treating diabetic nephropathy.

[0023] In some examples, melibiose can be used to prepare a medicament for treating diabetic nephropathy. Among them, melibiose can include melibiose, and / or its isomers, and / or its pharmaceutically acceptable salts, and / or its precursors. That is to say, the melibiose described in the present invention can refer to any one or more of melibiose, melibiose isomers, pharmaceutically acceptable salts of melibiose, and melibiose precursors. In some examples, melibiose precursors can include raffinose, stachyose, mannotriose, etc. Among them, preferably, the melibiose precursor can be stachyose (ST). Stachyose is a precursor of melibiose and is also a naturally occurring oligosaccharide with good stability. Higher-purity melibiose can be obtained from stachyose. One molecule of stachyose can decompose into one molecule of melibiose, and this decomposition occurs in the human body, which helps melibiose to be directly absorbed into the blood and play its role. In the subsequent experiments of the present invention, it was verified that melibiose and its precursor stachyose can be used to improve diabetic nephropathy.

[0024] In some examples, melibiose can improve diabetic nephropathy by improving inflammation in the body. The present invention has studied and verified the regulatory mechanism of melibiose in treating diabetic nephropathy, and proved that melibiose can significantly reduce proteinuria in diabetic mice, improve inflammation, relieve glomerular endothelial cell injury, and play a role in protecting the kidneys.

[0025] As mentioned above, in the present invention, a preparation for treating diabetic nephropathy is provided, which includes melibiose. In some examples, the preparation can be a medicament, food, or health product. Thus, the application scenario of the preparation can be selected according to needs. Subsequently, taking the preparation as a medicament as an example, a detailed description will be given.

[0026] In some examples, melibiose can be used as the sole active ingredient in the preparation of a medicament for treating diabetic nephropathy. In other words, melibiose can be the sole active ingredient in the medicament. The sole active ingredient refers to the only ingredient that has a significant impact on the technical effect of the medicament. Therefore, the present invention can also provide an application of melibiose as the sole active substance in the preparation of a medicament for treating diabetic nephropathy.

[0027] In some examples, the medicament and / or the preparation can also include excipients permitted in pharmacy, ordinary food, health food, or special medical food. Among them, the excipients do not have a significant impact on the overall technical effect of the medicament. Adding excipients can be beneficial to improving other properties of the medicament and making the medicament more suitable for clinical needs.

[0028] In some examples, the preparation can consist of melibiose and excipients. In some examples, the drug for treating diabetic nephropathy can be a pharmaceutical composition comprising melibiose. Among them, the pharmaceutical composition refers to a pharmaceutical composition composed of melibiose and pharmaceutically acceptable excipients.

[0029] In some examples, the excipient can be any one or more of diluents, excipients, binders, fillers, solubilizers, sustained / controlled release agents, flavoring agents, and sweetening agents. Thus, it can be beneficial to improve the performance of the pharmaceutical composition and make the pharmaceutical composition more meet the clinical needs.

[0030] In the present invention, "administering" and "drug administration" can refer to providing a drug to a subject through known routes. In some examples, the drug administration methods can include oral, intra-arterial / venous, parenteral, topical, transdermal, rectal, intramuscular, subcutaneous, intraosseous, transmucosal, or intraperitoneal administration routes. In some examples, preferably, the drug administration method can be oral.

[0031] In some examples, the drug can be in liquid dosage forms (e.g., suspensions, gels, and pastes), solid dosage forms (e.g., tablets, pills, powders, and bulk powders), or gaseous dosage forms.

[0032] In some examples, the dosage of the drug can be 0.1 g·kg –1 ·d –1 to 2 g·kg –1 ·d –1 . In the present invention, the dosage of the drug can be adjusted based on different subject types. Specifically, regarding drug administration, when the subject is a mouse, a small dosage may be able to saturate the intracellular accumulation and metabolism of the mouse; while when the subject is a human, a higher dosage than that of the mouse may be required to reach saturation. Therefore, the dosage of the drug can be adjusted according to actual needs.

[0033] Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the scope of protection of the present invention. The experimental methods without specific conditions noted in the following embodiments are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, all percentages, ratios, proportions, or parts are by weight.

[0034] The reagents and raw materials used in the examples and comparative examples of the present invention can be obtained through commercial channels without special instructions.

[0035] Example 1 Treatment of Diabetic Nephropathy Mice with Melibiose

[0036] I. Experimental Steps

[0037] 1. Animal Experiment Treatment

[0038] Eight-week-old SPF male C57BLKS / J-db / db mice (24) and eight-week-old SPF male C57BLKS / J mice (8) were provided by Jiangsu Jicui Yakang (SCXK (Su) 2018-0008). They were housed in a SPF-class closed environment at the Henan Academy of Medical Sciences. The environmental temperature was 23 ± 1 °C, the environmental humidity was 40%, with a 12-hour day-night cycle, and free access to water. The mice were fed a normal diet and had free access to food and water. They were adaptively fed for 1 week before drug intervention. The db / db mice were randomly grouped and given intragastric administration of different concentrations of melibiose (MEL, 0.5 g·kg –1 ·d –1 、1 g·kg –1 ·d –1 ) / equal volume of normal saline.

[0039] The grouping was as follows: normal control (NC) group, diabetes (DM) group, diabetes + normal saline (DM + Saline) group, diabetes + low-dose melibiose (DM + MEL + L, melibiose concentration 0.5 g·kg –1 ·d –1 ) group, diabetes + high-dose melibiose (DM + MEL - H, melibiose concentration 1 g·kg –1 ·d –1 ) group. The mice in each group were weighed and their blood glucose was measured every 2 weeks, and urine specimens were collected every 4 weeks. This continued for 8 weeks, and plasma and kidney tissue specimens were collected according to the experimental requirements.

[0040] 2. Biochemical analysis of urine samples

[0041] Twenty-four-hour urine samples of mice in each group were collected. After centrifugation (3000 rpm, 10 min), the supernatant was taken for detection; uALB was determined by immunoturbidimetry (wavelength 600 nm), and uCr was determined by enzymatic method (wavelength 500 - 550 nm). The instrument parameters were set strictly according to the reagent instructions, and calibration and quality control were carried out. The calculation formula for the urinary microalbumin / creatinine ratio (ACR) is the ratio of uALB (mg / L) to uCr (g / L), and the normal reference range is <30 mg / g.

[0042] 3. Biochemical analysis of plasma samples

[0043] Blood from the orbital venous plexus of mice was collected into an EDTA anticoagulant tube. Plasma was separated by centrifugation at 3000 × g for 10 min at 4 °C and stored at -80 °C for later use. The detection of inflammatory factors was strictly carried out according to the instructions of commercial mouse TNF-α and IL-6 ELISA kits. The absorbance value at 450 nm was measured by an enzyme-labeled instrument, and the concentration of each index was calculated according to the standard curve.

[0044] 4. Transmission electron microscopy (TEM)

[0045] Fresh renal cortical tissues were fixed using 4°C glutaraldehyde and 1% osmium tetroxide. Subsequently, the samples were dehydrated successively in 50%-100% ethanol and finally dehydrated with acetone. The samples were presoaked in a resin / acetone mixture, immersed in pure resin overnight, baked at different temperatures after embedding. After making ultra-thin sections, double staining with uranyl acetate and lead citrate was performed for 30 minutes, and finally rinsed. The stained grid samples were placed under an electron microscope for observation and the ultrastructural images of glomerular endothelial cells were collected.

[0046] 5. Scanning electron microscopy (SEM)

[0047] Fresh renal cortical tissues were fixed with 4°C glutaraldehyde for 48 hours, rinsed and fixed with PBS and 1% osmium tetroxide respectively. The samples were dehydrated successively in 30%-100% ethanol and finally immersed in isoamyl acetate. After drying the samples with a CO 2 critical point dryer, the samples were fixed on the SEM sample stage and coated with a metal film in a vacuum sputtering instrument. Finally, the samples were placed in the SEM sample chamber for scanning and the ultrastructural images of the fenestrations of glomerular endothelial cells were collected.

[0048] 6. Cell culture and treatment

[0049] Human glomerular endothelial cells (HGECs) were cultured in RPMI-1640 medium containing 10% fetal bovine serum and 1% antibiotics (penicillin and streptomycin) in an incubator at a constant temperature of 37°C and a CO 2 concentration of 5%. When the cells reached 70-80% confluence, the cells were treated with 0.25% trypsin and passaged at a ratio of 1:2-4 every 3-5 days.

[0050] In vitro experiments were set up with different concentrations of melibiose (MEL) intervention groups, and the specific grouping was as follows: normal glucose group (5.6 mmol / L, NG group), high glucose group (30 mmol / L, HG group), high glucose + low-dose melibiose group (HG + L-MEL group), high glucose + high-dose melibiose group (HG + H-MEL group).

[0051] When the cells adhered and reached 70-80% confluence, drug treatment was given according to the experimental design. The NG group and the HG group were given an equal concentration and equal volume of DMSO as a control. The drug treatment time was 48 hours. After the treatment, the cells and culture supernatants were collected for subsequent experimental analysis.

[0052] 7. Real-time fluorescence quantitative PCR experiment

[0053] Total RNA was extracted from kidney tissue samples using Trizol. cDNA samples were synthesized according to the instructions, and real-time quantitative PCR (qRT-PCR) was performed using the SYBR Green I Kit. The relative quantification of gene expression was determined by the 2-^ΔΔCt method and normalized to β-actin. The results were expressed as fold change compared with the control group.

[0054] 8. Western Blot Experiment

[0055] Total protein in cells was extracted using RIPA lysis buffer, followed by SDS-PAGE electrophoresis. Subsequently, transfer to a PVDF membrane and blocking were carried out. After incubation with primary and secondary antibodies respectively, signal images were captured using a chemiluminescent imaging system, and the intensity and position of the target protein bands were recorded.

[0056] 9. Data Analysis

[0057] Statistical analysis and image construction were performed using GraphPad Prism 9.5 software. Data were expressed as mean ± standard deviation (Mean ± SDs), and one-way ANOVA two-way variance analysis was used for comparison between three or more groups.

[0058] Example 2 Treatment of Diabetic Nephropathy Mice by Stachyose Generating Melibiose through Metabolism

[0059] I. Experimental Procedures

[0060] 1. Animal Experiment Treatment

[0061] Thirty-two 8-week-old SPF-grade male C57BLKS / J-db / db mice and eight 8-week-old SPF-grade male C57BLKS / J mice were provided by Jiangsu Jicui Yakang (SCXK (Su) 2018-0008). They were housed in a SPF-grade closed environment at the Henan Academy of Medical Sciences, with an environmental temperature of 23 ± 1°C, an environmental humidity of 40%, a 12-hour day-night cycle, and free access to water. The mice were fed a normal diet and had free access to food and water. They were adaptively fed for 1 week before drug intervention. The db / db mice were randomly grouped and gavaged with different concentrations of stachyose (ST, 0.5 g·kg–1·d–1, 1 g·kg–1·d–1, 2 g·kg–1·d–1) / equal volume of normal saline.

[0062] The groups were divided as follows: normal control (NC) group, diabetes mellitus (DM) group, diabetes mellitus + saline (DM+Saline) group, diabetes mellitus + low-dose stachyose (DM+ST+L, ST concentration 0.5 g·kg–1·d–1) group, diabetes mellitus + medium-dose stachyose (DM+ST-M, ST concentration 1 g·kg–1·d–1) group, and diabetes mellitus + high-dose stachyose (DM+ST-H, ST concentration 2 g·kg–1·d–1) group. The mice in each group were weighed and their blood glucose levels were measured every 2 weeks, and urine specimens were collected every 4 weeks. This continued for 8 weeks, and plasma and kidney tissue specimens were collected according to the experimental requirements.

[0063] 2. Biochemical analysis of urine samples

[0064] Twenty-four-hour urine samples from the mice in each group were collected. After centrifugation (3000 rpm, 10 min), the supernatant was taken for detection; uALB was determined by immunoturbidimetry (wavelength 600 nm), and uCr was determined by an enzymatic method (wavelength 500 - 550 nm). The instrument parameters were set, calibrated, and quality controlled strictly in accordance with the reagent instructions. The ACR calculation formula is the ratio of uALB (mg / L) to uCr (g / L), and the normal reference range is <30 mg / g.

[0065] 3. Biochemical analysis of plasma samples

[0066] Whole blood from the orbital venous plexus of the mice was collected into an EDTA anticoagulant tube. Plasma was separated by centrifugation at 3000×g for 10 min at 4℃ and stored at -80℃ for later use. For the detection of melibiose, a HILIC chromatographic column (2.1×100 mm, 1.7 μm) was used, and gradient elution was performed with 10 mM ammonium formate aqueous solution (containing 0.1% formic acid)-acetonitrile as the mobile phase. The mass spectrometry was performed in the electrospray negative ion mode, and multiple reaction monitoring (MRM) was carried out by selecting m / z341.1→89.0 (melibiose) and m / z347.1→95.0 ([13C6]-melibiose internal standard). The samples were analyzed by injection after protein precipitation with methanol. The detection of inflammatory factors was strictly carried out according to the instructions of the commercial mouse TNF-α and IL-6 ELISA kits. The absorbance value at 450 nm was measured by an enzyme-labeled instrument, and the concentration of each index was calculated according to the standard curve.

[0067] 4. Data analysis

[0068] GraphPad Prism 9.5 software was used for statistical analysis and image construction. The data were expressed as mean ± standard deviation (Mean±SDs), and one-way ANOVA two-way variance analysis was used for comparison between three or more groups.

[0069] To sum up, Figure 1It is a schematic diagram showing the changes in ACR, TNF-α, IL-6, kidney inflammatory factor levels, and ultrastructure of glomerular endothelial cells among each group of mice treated with MEL in the embodiments of the present invention (n = 8).

[0070] Compared with the normal control group, the levels of urinary ACR, plasma inflammatory factors (TNF-α, IL-6), and kidney inflammatory factors (TNF-α, IL-1β, IL-6) in diabetic mice were significantly increased, and the ultrastructure of glomerular endothelial cells was severely damaged; after treatment with melibiose for 8 weeks, urinary ACR decreased in a dose-dependent manner, and plasma and kidney inflammatory factors also decreased in a dose-dependent manner, and the ultrastructure of glomerular endothelial cells was significantly improved (see Figure 1 ), indicating that melibiose can improve inflammation in diabetic mice and reduce proteinuria.

[0071] Figure 2 It is a schematic diagram showing the levels of TNF-α and IL-6 secreted by glomerular endothelial cells in each group treated with MEL in the embodiments of the present invention, as well as the protein expression levels and quantification of ET-1, OCLN, ICAM-1, and ZO-1 in each group of cells.

[0072] Compared with the normal control group, the inflammatory factors (TNF-α, IL-6) secreted by glomerular endothelial cells treated with high glucose were significantly increased, the expression levels of endothelial injury-related proteins (ET-1, ICAM-1) in the cells were significantly increased, and the expression levels of tight junction-related proteins (OCLN, ZO-1) were significantly decreased; after treatment with melibiose for 48 hours, plasma inflammatory factors decreased significantly, the expression levels of endothelial injury-related proteins in the cells decreased significantly, and the expression levels of tight junction-related proteins increased significantly (see Figure 2 ), indicating that melibiose can improve glomerular endothelial cell injury and dysfunction.

[0073] Figure 3 It is a schematic diagram showing the metabolic pathway of stachyose and melibiose in the embodiments of the present invention (see Figure 3 A), and at the same time, it is a schematic diagram showing the changes in plasma melibiose, TNF-α, IL-6 levels, and ACR levels among each group of mice treated with stachyose in the embodiments of the present invention (n = 6).

[0074] From the metabolic pathway of melibiose, it can be known that stachyose is the precursor of melibiose, and its bioavailability is extremely low (about 3%), but stachyose can be metabolized by intestinal flora to produce melibiose. One molecule of stachyose can decompose into one molecule of melibiose, and melibiose is absorbed into the blood to play a role. Supplementing stachyose can also avoid the influence of gastric acid on melibiose and ensure the concentration of melibiose in the plasma.

[0075] After treating diabetic mice with stachyose for 8 weeks, it was found that the level of melibiose in plasma increased, and the urinary ACR of the mice decreased in a dose-dependent manner (see Figure 3 ), indicating that stachyose, the precursor of melibiose, can reduce proteinuria in diabetic mice by increasing the concentration of melibiose in plasma. Therefore, the present invention can actually also provide an application of stachyose, the precursor of melibiose, in the preparation of a drug for improving diabetic nephropathy.

[0076] In the present invention, in a type 2 diabetic mouse model, by intervening with different concentrations of melibiose, the changes in urinary protein-related indexes and plasma inflammatory factors of mice in different groups were detected, and it was verified that melibiose can significantly reduce proteinuria in diabetic mice, improve inflammation, and play a role in protecting the kidneys.

[0077] Finally, it should be noted that the above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, for those skilled in the art, they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. Use of a pharmaceutical composition in the preparation of a drug for treating diabetic nephropathy, the pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable excipient, characterized in that: The active ingredient is selected from one or more of melibiose or its pharmaceutically acceptable salts, isomers and precursors.

2. The use according to claim 1, characterized in that The pharmaceutical composition achieves the treatment of diabetic nephropathy by inhibiting the levels of TNF-α and IL-6 in the renal inflammatory microenvironment, repairing the damage of glomerular endothelial cells in diabetic nephropathy, and reducing the urine protein / creatinine ratio (ACR).

3. The use according to claim 1, characterized in that The melibiose precursor is stachyose, which is metabolized by intestinal flora to produce melibiose, and the plasma melibiose concentration is positively correlated with the administered dose.

4. The use according to claim 1, characterized in that The dosage of the pharmaceutical composition administered to diabetic mice is 0.5-2 g·kg -1 ·d -1 , and the ACR decreased by ≥30% after continuous administration for at least 8 weeks.

5. The use according to claim 1, characterized in that: The pharmaceutical composition is selected from oral preparations, and the dosage form is solid powder or oral liquid.

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