Application of ligustrazine nitrone derivative in prevention and treatment of diabetic complication diseases

Abstract

The invention relates to an application of a ligustrazine nitrone derivative in prevention and treatment of diabetic complication diseases, and an application of the ligustrazine nitrone derivative or pharmaceutically acceptable salts thereof in preparation of drugs for prevention and / or treatment of diabetic complication diseases. The diabetic complication disease is diabetic nephropathy; the effective dose of the ligustrazine nitrone derivative or the pharmaceutically acceptable salt thereof is 1.0-60 mg / kg, and the ligustrazine nitrone derivative or the pharmaceutically acceptable salt thereof has important clinical use value.

Description

[0001] This application is a divisional application of a Chinese invention patent application filed on May 27, 2017, with application number 201710392539.X and invention name “Application of ligustrazine nitrone derivatives in the prevention and treatment of diabetic complications”. Technical Field

[0002] The invention belongs to the field of pharmaceutical technology, and specifically relates to an application of a ligustrazine nitrone derivative and a pharmaceutical composition thereof in preventing and treating diabetic complications. Background Art

[0003] Diabetes is a metabolic disease characterized by high blood sugar due to insulin secretion defects or insulin action disorders. Continuous high blood sugar and long-term metabolic disorders can lead to damage to tissues and organs throughout the body, especially the eyes, kidneys, cardiovascular and nervous systems, as well as dysfunction and failure. In severe cases, it can cause dehydration, electrolyte disorders and acid-base imbalance, and other acute complications such as ketoacidosis and hyperosmolar coma.

[0004] Diabetic Kidney Disease (DKD) is one of the most important microvascular complications of diabetes. Its clinical features include proteinuria, progressive renal impairment, hypertension, edema, and severe renal failure in the late stage. DKD is currently the leading cause of end-stage renal disease, and about 30% to 40% of diabetic patients suffer from kidney disease. According to the IDF report, the global prevalence of diabetes was 382 million in 2013, and it will increase to 592 million in 25 years. Summary of the invention

[0005] The present invention discloses a novel use of a ligustrazine nitrone derivative and a pharmaceutical composition thereof, namely, the use of the ligustrazine nitrone derivative and the pharmaceutical composition thereof in the preparation of a drug for preventing and treating diabetic complications. The derivative has the following general formula (I):

[0006]

[0007] Wherein, R1 and R3 are each independently C1-C6 alkyl; R2 is C1-C6 alkyl or R4 and R5 are each independently sec-butyl, isobutyl, tert-butyl, cyclopentyl or cyclohexyl.

[0008] Preferably, the C1-C6 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl or n-pentyl.

[0009] Further preferably, the ligustrazine nitrone derivative has the following structural formula:

[0010]

[0011] The present invention provides the application of the ligustrazine nitrone derivative and the pharmaceutical composition thereof in preventing and treating diabetic complications.

[0012] The diabetic complications are preferably diabetic nephropathy and diabetic eye disease. Further preferably, the diabetic eye disease is retinopathy, glaucoma and cataract.

[0013] The present invention also provides the use of the ligustrazine nitrone derivative and its pharmaceutical composition in the preparation of medicines, wherein the medicines can be used to prevent and treat diabetic nephropathy and its complications. The pharmaceutical composition comprises a therapeutically effective dose of the derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0014] The drug containing the ligustrazine nitrone derivative can be used alone or in combination with other drugs to prevent and treat diabetic complications. Other drugs that can be used in combination mainly include commonly used oral antidiabetic drugs in clinical practice, including biguanides, sulfonylureas, glucosidase inhibitors, thiazolidinediones, non-sulfonylurea insulin secretagogues and dipeptidyl peptidase inhibitors, etc.; hypertension drugs, including angiotensin receptor blockers (such as losartan, losartan), angiotensin converting enzyme inhibitors; and drugs for treating diabetic nephropathy (such as folic acid).

[0015] The ligustrazine derivatives TBN and TN-2 provided by the present invention can be used in combination with existing clinical drugs for treating diabetes and diabetic nephropathy to improve the efficacy through synergistic effects, reduce the side effects of existing clinical drugs, and improve the benefit / risk ratio of clinical drug use.

[0016] The ligustrazine nitrone derivative can be prepared into various dosage forms with available drug carriers, including tablets, granules, injections, powders, capsules and suspensions.

[0017] Preferably, the therapeutically effective dose of the ligustrazine nitrone derivative is 0.001-2 g / kg. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Effect of ligustrazine derivatives on body weight ( Figure 1 a) Water intake ( Figure 1 b) and dietary intake ( Figure 1 c) impact. ### P<0.001, ## P<0.01 compared with the control group (Control); *** P<0.001,

[0019] *P<0.05 compared with the model group (Model).

[0020] Figure 2 Effect of ligustrazine derivatives on STZ-induced DKD urinary protein ( Figure 2 a) and rat blood glucose ( Figure 2 b) impact. ### P<0.001 compared with the control group (Control); *** P<0.001, ** P<0.05, * P<0.01 compared with the model group (Model).

[0021] Figure 3 Effect of ligustrazine derivatives on serum creatinine ( Figure 3 a) Urea nitrogen ( Figure 3 b) Cholesterol ( Figure 3 c) and triglycerides ( Figure 3 d) Horizontal influence. ### P<0.001, ## P<0.01 compared with the control group (Control); ** P<0.05, * P<0.01 compared with the model group (Model).

[0022] Figure 4 The effect of ligustrazine derivatives on renal indexes of STZ-induced DKD rats. ### P<0.001 compared with the control group (Control); ** P<0.05, * P<0.01 compared with the model group (Model).

[0023] Figure 5 The number of cases and time of occurrence of retinopathy in diabetic nephropathy model rats were listed. DETAILED DESCRIPTION

[0024] The technical solution of the present invention will be clearly and completely described below in conjunction with the specific embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Those skilled in the art should understand that modifications to the specific embodiments of the present invention or equivalent replacement of some technical features without departing from the spirit of the technical solution of the present invention should be included in the scope of protection of the present invention.

[0025] Example 1. Preparation and grouping of STZ-induced diabetic nephropathy rat model

[0026] 1. Model making

[0027] SD rats (200±10g) were selected as modeling animals. They were fasted for 12h before injection. STZ was dissolved in citric acid buffer at a concentration of 1% during injection. After weighing the fasted rats, 55mg / kg STZ was injected intraperitoneally and placed in a cage to ensure that the rats had sufficient drinking water for 24h. STZ injection needs to be fast and completed within 10min. The normal control group was injected with an equal volume of citric acid-sodium citrate buffer at pH 4.5. The animal status was observed (polydipsia and polyuria occurred), and blood was collected from the tail vein 3 weeks after STZ injection. Fasting blood glucose ≥16.7mmol / L was measured as the modeling standard for diabetic nephropathy in rats.

[0028] 2. Grouping

[0029] DKD rats were randomly divided into 6 groups and given different drugs. The experiment was terminated after 6 weeks of drug administration to observe the protective effects of the drugs on DKD rats.

[0030] Example 2. Effects of ligustrazine derivatives on body weight, diet and drinking water in STZ-induced DKD rats

[0031] The general condition and weight changes of the rats after modeling were observed weekly, including the rats' activities, mental state, coat color, diet, water intake, and urine volume, etc. The water intake and feed intake were recorded weekly.

[0032] Effects of ligustrazine derivatives on body weight in rats with STZ-induced diabetic nephropathy Figure 1 As shown in a, TBN and TN-2 had little effect on the body weight of diabetic nephropathy rats; the changes in water intake and feed intake of diabetic nephropathy rats were shown in Figure 1 b and Figure 1 As shown in c, the water intake and diet of DKD rats in the TBN and TN-2 treatment groups were significantly lower than those in the model group, indicating that TBN and TN-2 can slow down the progression of diabetes in DKD rats, thereby improving the diabetic symptoms of polydipsia and polyphagia in DKD rats.

[0033] Example 3. Effects of ligustrazine derivatives on STZ-induced retinopathy in DKD rats

[0034] During the experiment, the retinopathy of rats was observed during daily administration, and the dates when retinopathy occurred in rats of different groups were recorded.

[0035] Retinopathy is one of the common complications of diabetic nephropathy and is highly correlated with diabetic nephropathy. Effects of ligustrazine derivatives on STZ-induced retinopathy in DKD rats Figure 5 As shown, after treatment with TBN and TN-2, the number of DKD rats with retinopathy decreased and the time point of retinopathy onset was relatively delayed. The results showed that ligustrazine derivatives TBN and TN-2 can reduce and delay the occurrence of retinopathy.

[0036] Example 4. Effects of ligustrazine derivatives on blood glucose and urine protein in STZ-induced DKD rats

[0037] During the experiment, the blood glucose of rats was monitored after modeling and administration, and urine was collected once for 24 hours to detect urine protein.

[0038] Effects of ligustrazine derivatives on blood glucose in STZ-induced DKD rats Figure 2 As shown in a, the blood glucose of rats increased significantly after 3 weeks of STZ induction, and decreased significantly after 6 weeks of TBN and TN-2 treatment. Figure 2 As shown in b, the urine protein content in the urine of DKD rats treated with TBN and TN-2 was significantly reduced, and the combination of TBN and losartan was superior to TBN treatment alone or losartan treatment alone.

[0039] Example 5. Effects of ligustrazine derivatives on serum biochemical parameters in STZ-induced DKD rats

[0040] After 6 weeks of administration, blood was collected from the abdominal aorta of the rats after anesthesia, and after standing for 1 hour, it was centrifuged at 3000 rpm for 10 minutes and stored at -70°C. The levels of creatinine, urea nitrogen, cholesterol and triglyceride in serum were detected using an automatic biochemical analyzer.

[0041] Serum creatinine, urea nitrogen, cholesterol and triglyceride levels can reflect the body's lipid metabolism, carbohydrate metabolism and kidney function. Effects of ligustrazine derivatives on serum creatinine, urea nitrogen, cholesterol and triglyceride in STZ-induced DKD rats Figure 3 As shown, TBN and TN-2 can significantly reduce serum creatinine ( Figure 3 a) Urea nitrogen ( Figure 3 b) Cholesterol ( Figure 3 c) and triglycerides ( Figure 3 d) in a dose-dependent manner, showing an improving effect on lipid metabolism, carbohydrate metabolism and renal function in rats with STZ-induced diabetic nephropathy.

[0042] Example 6. Effects of ligustrazine derivatives on renal indexes in STZ-induced DKD rats

[0043] After 6 weeks of administration, the abdominal cavity was opened with autoclaved surgical instruments to uniformly separate the kidney tissue, washed with physiological saline, dried with filter paper, weighed with a fine balance, and then stored in a -80°C refrigerator for later use. The kidney index (relative kidney weight) is kidney weight (mg) / body weight (g) = BW / KW.

[0044] The renal index can reflect the pathological condition of diabetic nephropathy to a certain extent. Figure 4 As shown, TBN and TN-2 could significantly reduce the renal index of DKD rats, indicating that TBN and TN-2 could delay the disease progression of diabetic nephropathy.

Claims

1. Use of ligustrazine nitrone derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for preventing and / or treating diabetic complications; the diabetic complications are diabetic nephropathy; the effective dose of ligustrazine nitrone derivatives or pharmaceutically acceptable salts thereof is 1.0-60 mg / kg; the ligustrazine nitrone derivatives have the following structural formula:

2. The use according to claim 1, characterized in that: The effective dose is 10-60 mg / kg.

3. The use according to claim 1, characterized in that: The ligustrazine nitrone derivative is TBN.

4. The use according to any one of claims 1 to 3, wherein the dosage form of the drug is tablets, granules, injections, powders, capsules or suspensions.

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