Use of beta-d-thioglucose sodium salt in the preparation of a medicament for alleviating or treating acute kidney injury

Abstract

The application discloses application of beta-D-thioglucose sodium salt (beta-Tg) in preparation of a medicine for relieving or treating acute kidney injury. 11 The molecular formula of the beta-D-thioglucose sodium salt is C6H 11 NaO5S, and a chemical structural formula is as follows: by removing a large amount of active oxygen in a renal tubule, kidney protection is further exerted, and the beta-D-thioglucose sodium salt is especially suitable for being used for the symptoms of drug-induced acute kidney injury and ischemic acute kidney injury, and has a good market prospect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of research and development of medical products, and particularly relates to application of a beta-D-thioglucose sodium salt in preparation of a medicine for relieving or treating acute kidney injury. BACKGROUND

[0002] Acute kidney injury (AKI) is a common critical illness in clinical practice, with more than 50,000 cases per million population, and 1.7 million deaths per year. AKI is a clinical manifestation characterized by rapid decline in kidney excretion, reduced urine output, and increased nitrogen metabolism. Reactive oxygen species (ROS) induced by oxidative stress and inflammation in the body are closely related to AKI in most cases. Since there is no specific treatment method other than kidney dialysis and transplantation, ROS is a key target for preventing AKI. It has been reported that the antioxidant N-acetylcysteine (NAC) has achieved some success in preventing AKI, but the clinical benefits of the drug are uncertain. The kidney plays an important role in maintaining blood glucose balance in the human body, and the glomerulus filters about 180 g of glucose from the plasma every day. As a glucose with antioxidant properties, beta-D-thioglucose sodium salt may open up new prospects for the development of AKI treatment technology in the relief or treatment of AKI. SUMMARY

[0003] In view of the deficiencies of the prior art, the present application provides application of a beta-D-thioglucose sodium salt in preparation of a medicine for relieving or treating acute kidney injury, which effectively relieves or treats acute kidney injury by reducing the level of a large amount of reactive oxygen species in the body, and has good market prospects.

[0004] The application of beta-D-thioglucose sodium salt in preparation of a medicine for relieving or treating acute kidney injury is characterized in that the beta-D-thioglucose sodium salt has a molecular formula of C6H 11 NaO5S, and a chemical structural formula as follows:

[0005]

[0006] As an improvement, the purity of the beta-D-thioglucose sodium salt is 90-97%.

[0007] As an improvement, the beta-D-thioglucose sodium salt is enriched in the mouse kidney and eliminates a large amount of reactive oxygen species in the renal tubule, thereby relieving and treating acute kidney injury.

[0008] As an improvement, the acute kidney injury is drug-induced acute kidney injury and ischemic acute kidney injury.

[0009] As an improvement, the drug inducing acute kidney injury includes cisplatin, aminoglycoside antibiotic, adriamycin, mercuric chloride, or sodium fluoride; the ischemic acute kidney injury is caused by one side kidney ischemia / reperfusion plus contralateral kidney resection method, bilateral kidney pedicle clamping method, glycerol injection method, norepinephrine infusion method, or oleic acid method.

[0010] Beneficial effects:

[0011] Compared with the prior art, the application of the beta-D-thioglucose sodium salt in the preparation of drugs for relieving or treating acute kidney injury is related to the reduction of a large number of active oxygen levels in the body, effectively relieves and treats acute kidney injury, protects kidney function, and has good market prospects. BRIEF DESCRIPTION OF DRAWINGS

[0012] Figure 1 The figure is the clearance rate of beta-D-thioglucose sodium salt on ABTS+ free radicals;

[0013] Figure 2 The figure is the clearance rate of beta-D-thioglucose sodium salt on superoxide anion (·O 2- ) radicals;

[0014] Figure 3 The figure is the acute kidney injury induced by 8ml / kg 50% glycerol, wherein (a) is the determination of urea nitrogen (BUN) in mouse serum, (b) is the determination of creatinine (CRE) in mouse serum, and (c) is the determination of oxidative stress gene superoxide dismutase SOD in mouse kidney tissue;

[0015] Figure 4 The figure is the acute kidney injury induced by 12ml / kg 50% glycerol, wherein (a) is the determination of urea nitrogen (BUN) in mouse serum, (b) is the determination of creatinine (CRE) in mouse serum, and (c) is the determination of oxidative stress gene superoxide dismutase SOD in mouse kidney tissue;

[0016] Figure 5 The figure is the evaluation of the treatment effect of beta-D-thioglucose sodium salt injected once every 24 hours, wherein (a) is the determination of urea nitrogen (BUN) in mouse serum, (b) is the determination of creatinine (CRE) in mouse serum, and (c) is the determination of oxidative stress gene superoxide dismutase SOD in mouse kidney tissue;

[0017] Figure 6The pathological results of HE staining of kidney tissues of mice for evaluating the therapeutic effect of β-D-thioglucose sodium salt in the specific embodiment of the present application by injection once every 24 hours, wherein (a) is the HE staining result of healthy mice, (b) is the HE staining result of mice treated once with β-D-thioglucose sodium salt, (c) is the HE staining result of mice treated twice with β-D-thioglucose sodium salt, and (d) is the HE staining result of mice treated three times with β-D-thioglucose sodium salt;

[0018] Figure 7 The pharmacokinetics of β-D-thioglucose sodium salt in mice in the present application, wherein (a) is the calibration curve of β-D-thioglucose sodium salt in plasma by liquid chromatograph (HPLC), and (b) is the pharmacokinetics of β-D-thioglucose sodium salt in mice for 3 hours. DETAILED DESCRIPTION

[0019] In order to make the purpose, technical scheme and effect of the present application more clear, the present application will be further described below in combination with specific embodiments and drawings. It should be understood that the specific embodiments described herein are only used to explain the present application and do not limit the present application.

[0020] In the experimental methods described in the embodiments, if not specifically stated, they are all conventional methods; and the reagents and materials, if not specifically stated, can be obtained from commercial channels.

[0021] β-D-thioglucose sodium salt purchased from J&K Chemical was selected.

[0022] Embodiment 1

[0023] Effect of β-D-thioglucose sodium salt on scavenging ·ABTS+ free radicals

[0024] The above β-D-thioglucose sodium salt was used to prepare a solution of 0-40 mg / ml for determining the scavenging effect on ·ABTS+ free radicals, and the scavenging efficiency of ·ABTS+ free radicals was determined by total antioxidant capacity detection kit (ABTS method) (Shanghai Biyun Tian Biological Co., Ltd.), and the specific test method was carried out according to the scheme provided by the manufacturer.

[0025] As shown in Figure 1 , β-D-thioglucose sodium salt can effectively scavenge ·ABTS+ free radicals, and has concentration-dependent characteristics in the measured concentration range.

[0026] Embodiment 2:

[0027] Effect of β-D-thioglucose sodium salt on scavenging superoxide anion (·O 2- ) radicals

[0028] The above β-D-thioglucose sodium salt was used to prepare a solution of 0-40 mg / ml to determine the scavenging superoxide anion (·O 2- ) and the efficiency of scavenging superoxide anion (·O 2- ) was determined by total SOD activity detection kit (WST-8 method) (Shanghai Biyun Tian Biological Co., Ltd.) according to the manufacturer's protocol. The results are shown in Figure 2 From the figure, it can be seen that β-D-thioglucose sodium salt can effectively scavenge superoxide anion (·O 2- ) and has concentration-dependent characteristics in the measured concentration range.

[0029] Example 3:

[0030] Urea nitrogen (BUN) and creatinine (CRE) level test

[0031] The whole blood sample was placed at room temperature for 2 hours, centrifuged at 3000 rpm for 15 minutes, and the supernatant was transported on dry ice for determination by automatic biochemical analyzer (determined by Wuhan Seville Biological Technology Co., Ltd.).

[0032] Example 4:

[0033] Evaluation of β-D-thioglucose sodium salt for alleviating acute kidney injury

[0034] All experimental operations were in accordance with the animal use and care system approved by the clinical center animal care and use committee. After 15 hours of water deprivation without food restriction, Balb / c female mice (6-8 weeks) were injected with 8 ml / kg 50% glycerol solution in the hind leg muscle to establish a mouse model of acute kidney injury, and were injected with phosphate buffer solution or β-D-thioglucose sodium salt at 2 hours and 5 hours, respectively.

[0035] The mice were randomly divided into two groups: (1) acute kidney injury mice injected with phosphate buffer solution; (2) acute kidney injury mice injected with β-D-thioglucose sodium salt, and the mice were sacrificed after 24 hours, the mouse blood was centrifuged to obtain serum, and the urea nitrogen and creatinine content were tested. The kidneys (left: stored in a refrigerator at minus 80°C; right: immersed in formalin) were taken for testing kidney SOD level and HE staining pathological detection, respectively.

[0036] Among them, the injection used phosphate buffer solution was 100 μl, and the β-D-thioglucose sodium salt was 40 mg / ml, 100 μl.

[0037] Example 5:

[0038] Evaluation of β-D-thioglucose sodium salt for alleviating acute kidney injury

[0039] All experimental procedures were performed in accordance with the institutionally approved animal use and care guidelines of the Clinical Center Animal Care and Use Committee. Acute kidney injury model was established by intramuscular injection of 12 ml / kg 50% glycerol solution in the hind leg of Balb / c female mice (6-8 weeks) after 15 hours of water deprivation without food restriction. Phosphate buffered solution or β-D-sodium thio-dextrose was injected at 2 hours and 5 hours, respectively.

[0040] Mice were randomly divided into two groups: (1) acute kidney injury mice injected with phosphate buffered solution; (2) acute kidney injury mice injected with β-D-sodium thio-dextrose. Mice were sacrificed 24 hours later, and serum was obtained by centrifugation of mouse blood to test urea nitrogen and creatinine content. Kidneys (left: stored in -80°C freezer; right: immersed in formalin) were taken to test kidney SOD levels and HE staining for pathological detection, respectively.

[0041] Injection of phosphate buffered solution was 100 μl, and β-D-sodium thio-dextrose was 40 mg / ml, 100 μl.

[0042] Example 6:

[0043] Evaluation of mouse kidney tissue oxidative stress gene superoxide dismutase SOD index

[0044] Mouse kidney tissue oxidative stress gene superoxide dismutase SOD was determined by total SOD activity detection kit (WST-8 method) (Shanghai Biyun Tian Biological Co., Ltd.). The test was performed according to the manufacturer's provided protocol.

[0045] As shown in Figures 3-4 , the urea nitrogen and creatinine levels of glycerol-induced acute kidney injury mice increased significantly. The urea nitrogen and creatinine content of acute kidney injury mice injected with β-D-sodium thio-dextrose was significantly lower than that of mice injected only with phosphate buffered solution, and was close to the level of healthy mice. The urea nitrogen and creatinine content of more severe acute kidney injury mice also decreased significantly. At the same time, the mouse kidney oxidative stress gene superoxide dismutase SOD was significantly restored, while the SOD level of acute kidney injury mice was decreased, further proving that β-D-sodium thio-dextrose can act as a reducing agent, remove ROS, restore SOD level, further alleviate acute kidney injury, and thus protect the kidney.

[0046] Example 7:

[0047] Evaluation of acute kidney injury after multiple injections of β-D-sodium thio-dextrose

[0048] All experimental procedures were performed in accordance with the animal use and health protocols approved by the Clinical Center Animal Health and Use Committee. Balb / c female mice (6-8 weeks old) were deprived of water but not restricted to food for 15 hours before an acute kidney injury model was established by intramuscular injection of 8 ml / kg of 50% glycerol solution in the hind leg. β-D-thioglucosinolate was injected at 2, 24, and 48 hours.

[0049] Mice were randomly divided into three groups: (1) mice with acute kidney injury were treated once with β-D-thioglucose sodium; (2) mice with acute kidney injury were treated twice with β-D-thioglucose sodium; and (3) mice with acute kidney injury were treated three times with β-D-thioglucose sodium. Mice were sacrificed 24 hours later, and serum was obtained by centrifugation of mouse blood. Urea nitrogen and creatinine levels were tested. Kidneys were also collected (left: stored at -80°C; right: easily soaked in formalin) to test kidney SOD levels and for HE staining pathological examination.

[0050] The treatment involved injecting β-D-thioglucosinolate sodium salt at a dose of 40 mg / ml (100 μl) over a period of 24 hours.

[0051] like Figure 5 As shown, with increasing treatment frequency, the serum creatinine level in mice decreased to the level of healthy mice, while the SOD level gradually recovered and approached the level of healthy mice. Although the serum urea nitrogen level in mice did not return to normal, the effect of alleviating acute kidney injury was significant. Based on the combined effects of serum urea nitrogen and creatinine levels, SOD content, and β-D-thioglucosinolate sodium salt, this treatment has therapeutic and alleviating effects on acute kidney injury in mice.

[0052] Example 8:

[0053] HE staining pathological examination

[0054] Kidney tissue was soaked in formalin solution and fixed with 4% paraformaldehyde. The pathological samples were then trimmed, dehydrated, embedded, sectioned, stained, and mounted for microscopic examination (measured by Wuhan Sewell Biotechnology Co., Ltd.).

[0055] like Figure 6 As shown, after multiple treatments with β-D-thioglucose sodium salt, the glomeruli in the renal cortex were evenly distributed, with only a small amount of eosinophilic droplets visible in the renal capsule and tubules, and a small amount of protein casts formed. No obvious inflammatory cell infiltration was observed, and the treatment effect was significant.

[0056] Example 9:

[0057] Pharmacokinetics of β-D-thioglucose sodium salt in mice

[0058] Chromatographic conditions: Agilent 5TC-C18 column (250 x 4.6 mm); mobile phase: methanol: water = 4:1; absorbance: 240 nm; column temperature: 30 °C; flow rate: 0.5 ml / min; injection volume: 10 μl.

[0059] Blood was collected from the retro-orbital venous plexus of mice into EDTA-treated centrifuge tubes, centrifuged at 6000 rpm for 10 minutes, and the supernatant plasma was added to 400 μl of chromatographic methanol to precipitate the protein, vortexed for about 3 minutes, centrifuged at 12000 rpm for 10 minutes, and the supernatant was tested by HPLC.

[0060] The pharmacokinetics of β-D-thioglucose sodium salt in mice is shown in Table 1, and the half-life thereof in mice is 0.264 hours. Figure 7

[0061] In summary, the β-D-thioglucose sodium salt of the present application can effectively scavenge free radicals and superoxide anions, and has the ability to scavenge reactive oxygen species in vivo. In addition, the β-D-thioglucose sodium salt shows good effects in mice with acute kidney injury induced by glycerol.

[0062] The above merely describes the preferred embodiments of the present application, and the protection scope of the present application is not limited thereto, and any simple changes or equivalent replacements of the technical solutions within the technical scope disclosed by the present application are within the protection scope of the present application.​

Claims

1. The use of β-D-thioglucose sodium salt in the preparation of drugs for relieving or treating acute kidney injury, characterized in that, The molecular formula of the β-D-thioglucose sodium salt is C6H. 11 NaO5S has the following chemical structural formula: 。 2. The application according to claim 1, characterized in that, The purity of the β-D-thioglucose sodium salt is 90-97%.

3. The application according to claim 1, characterized in that, The acute kidney injury mentioned refers to drug-induced acute kidney injury and ischemic acute kidney injury.

4. The application according to claim 3, characterized in that, The drugs used in the drug-induced acute kidney injury are cisplatin, aminoglycoside antibiotics, doxorubicin, mercuric chloride, or sodium fluoride; the ischemic acute kidney injury is caused by unilateral renal ischemia / reperfusion combined with contralateral nephrectomy, bilateral renal pedicle clamping, glycerol injection, norepinephrine infusion, or oleic acid method.

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