WO2013138964A1

WO2013138964A1 - Application of iso-daphnetin compound in preparation of anti-diabetic medicines

Info

Publication number: WO2013138964A1
Application number: PCT/CN2012/001297
Authority: WO
Inventors: 张卫东; 单磊; 苏娟; 李慧梁; 柳润辉; 沈云亨; 徐希科; 张寿德
Original assignee: 中国人民解放军第二军医大学
Priority date: 2012-03-19
Filing date: 2012-09-24
Publication date: 2013-09-26
Also published as: CN102626407A

Abstract

The present invention provides an application of an iso-daphnetin compound in preparation of anti-diabetic medicines. The structural formula of the iso-daphnetin is described in the following. In the present invention, high-sugar and high-fat forage combined with a small dosage of streptozotocin (STZ) is applied to establish a type 2 diabetes rat model. Male SD adult rats are fed with high-sugar and high-fat forage for one month to induce insulin resistance and injected with a small dosage of STZ (25 mg/kg) into the abdominal cavity to establish the type 2 diabetes rat model. Then, a given medicine is injected into the abdominal cavity. The blood sugar, serum insulin, glucagon, metal state, weights, water intake, food intake, and so on of the rats are used as test indexes. An experimental result shows that after treatment with the iso-daphnetin, the blood sugar values and the serum insulin levels of the type 2 diabetes rats are significantly decreased (p < 0.01), and when the insulin is significantly increased, the blood sugar levels of the dosage groups are not notably different (p > 0.05). Therefore, the iso-daphnetin can be used for preparation of anti-diabetic medicines. According to the present invention, the medicine is a pharmaceutical composition formed by the iso-daphnetin as an active ingredient and a medicinal carrier.

Description

Application of isospermidine compound in preparing antidiabetic drugs

The invention relates to drugs. Specifically, it relates to the use of iso-rhimelin compounds in the preparation of anti-diabetic drugs. Background technique

In recent years, the incidence of type 2 diabetes has risen year by year, seriously threatening human health. Current research shows that patients with type 2 diabetes have three major defects: insulin resistance, beta cell dysfunction, and hepatic glucose excess. Oral anti-hyperglycemic drugs commonly used in clinical practice include: PPAR Y agonists, sulfonylureas that promote insulin secretion, Lenin, and biguanides. The above drugs are only effective within a certain range, can not effectively control the development of diabetes, and have limited tolerance or varying degrees of adverse reactions, such as: glucose intolerance, hypoglycemia, weight gain, lactic acidosis and edema.

Isoflavin is from the Daphne plant of Phylaceae (Daphne), Phnom Penh (Daphne)

Isoflavin structure:

Source s Phnom Penh Daphne odora Thunb. var. marginata There are about 44 species of Chinese genus, which are rich in resources. Its roots, stems, leaves and flowers can be used as medicines. It is sweet and non-toxic. It has the functions of clearing heat and reducing fire, reducing inflammation, swelling, promoting blood circulation, and relieving pain. According to the "Chinese Medicine Dictionary", Ruixiang leaves can cure sore and wind pain; flowers can cure sore and swollen teeth; roots can cure snake bites, bruises and other diseases. In recent years, there have been many studies on the pharmacological activities of the extracts of Rhizoma rugulosa. However, there have been no reports of chemical studies, such as analgesic effect, anti-hypoxia, anti-lipid peroxidation and anti-aging effects. In addition, Jinbian Ruixiang is small. The cellular immunity, non-specific immunity and erythrocyte immune function of the mouse are promoted. The inventors conducted a systematic chemical composition study on Jinbian Ruixiang, and found that the chemical component of ruthenium has a good anti-diabetic effect. So far, there has been no report on the anti-diabetic effect of Phnom Penh. Summary of the invention

The technical problem to be solved by the present invention is to overcome the above-mentioned deficiencies, to study the new use of the sirolimus, and to design an effective anti-diabetic drug. The present invention provides the use of isolisin in the preparation of an antidiabetic drug.

The ruthenium sulphate provided by the invention is obtained by the following method - the medicinal material of Phnom Penh is extracted by 75% ethanol, and the extract is concentrated under reduced pressure to obtain an extract, and the obtained crude extract is extracted with petroleum ether, chloroform and ethyl acetate respectively. , the ethyl acetate fraction was subjected to repeated silica gel H (200 to 300 mesh) column chromatography, chloroform: acetone (10: 1-1: 1), chloroform: methanol (50: 1-1: 1) gradient elution, Sephadex LH -20 column chromatography and reverse phase silica gel (Merck), the compound was obtained.

The invention adopts a high-sugar and high-fat diet combined with a low-dose streptozotocin (STZ) type 2 diabetic rat model, which is in line with the pathogenesis of human common type 2 diabetes, and is beneficial for the discovery of therapeutic drugs for type 2 diabetes. We fed a male SD adult rat with high-sugar and high-fat diet for 1 month, induced insulin resistance, and administered a small dose of streptozotocin (STZ) 25 mg/kg intraperitoneally to establish a rat model of type 2 diabetes. Then, the drug is administered intraperitoneally, and the blood glucose, serum insulin, glucagon, mental state, body weight, water intake, food intake, and urine are used as detection indexes. The results of the experiment showed that the blood glucose level and glucagon level of type 2 diabetic rats decreased significantly after treatment with iso-rhimexin (P<0.01), while insulin increased significantly, and there was no significant difference in blood glucose between the groups. (P>0.05). Therefore, isorubicin can be used to prepare an anti-diabetic drug. In the experiment of the present invention, the oral hypoglycemic agent metformin was used as a positive control, and the results showed that the isoxanthin and the positive drug metformin have considerable activity and have the functions of protecting and repairing islet cells.

The pharmaceutical composition of the present invention comprises a pharmaceutical composition comprising isosericin as an active ingredient and a pharmaceutically acceptable carrier. The invention has good anti-diabetic effect, and has the functions of protecting and repairing islet cells. In addition, the active ingredient isorubicin is derived from an extract of natural products, and is superior to chemical drugs and hypoglycemic agents in side effects and the like. Large clinical application value. detailed description

Example 1

Preparation of ruthenium

7. 5kg Phnom Penh fragrant herbs (commercially available) were pulverized and extracted with 150L 75% ethanol. The extract was concentrated under reduced pressure to obtain 8L extract. After the extract was diluted with 10L of water, 100L petroleum ether and 100L chloroform were used. 100 L of ethyl acetate was extracted, and the ethyl acetate fraction was collected and weighed to 125 g. The ethyl acetate fraction was subjected to two times of silica gel H (200 to 300 mesh) column chromatography, chloroform:acetone (10 _: 1 to 1 _: 1), chloroform. : Methanol (50: 1~1: 1) gradient elution, Sephadex LH-20 column chromatography and reversed-phase silica gel (Merck), to obtain 108 mg of the compound isotretin.

The isoericin prepared in Example 1 was used in the following experiment. Example 2

Experimental study on the effects of sirolimus on serum insulin and glucagon in rats. In this experiment, high-sugar and high-fat diet combined with low-dose streptozotocin (STZ) type 2 diabetic rat model was used. The pathogenesis of type 2 diabetes is conducive to the discovery of therapeutic drugs for type 2 diabetes. Male SD adult rats were fed with high-sugar and high-fat diet for 1 month to induce insulin resistance. Low-dose streptozotocin (STZ) 25 mg/kg was administered intraperitoneally to establish a rat model of type 2 diabetes. Then, the drug was administered intraperitoneally, and the blood glucose, serum insulin, glucagon, mental state, body weight, water intake, food intake, and urine were measured as indicators (Pharmacological Research. 2005, 52: 313-320). Experimental steps -

1. Material Irradix is prepared by the preparation of Example 1.

1. 1 Experimental animals and main reagents: 60 healthy male Wistar rats, weighing (180±20) g, provided by Shanghai Slac Laboratory Animal Co., Ltd. Streptozotocin (Sigma, USA); Metformin Hydrochloride (Beijing Yongkang Pharmaceutical Co., Ltd.); normal saline, sodium citrate, citric acid, picric acid, urethane, distilled water are all commercially available products.

1. 2 Main instruments: TP-5000A electronic balance (division value 0. 5 g, Xiangyi Tianping Instrument Factory); GT-1640 Kyoto blood glucose meter (SUPER GLUCOCARD II) and test paper (Japan ARKRAY, Inct); visible ~ UV Spectrophotometer (Shimadzu, Japan); SHA-B-Electrical thermostatic water bath (Changzhou Guohua Electric Co., Ltd.).

2. Model establishment: 60 healthy Wi star rats, 10 randomly selected as normal control group (I), fed with conventional diet. 50 rats of the model were replicated. After 8 weeks of high-calorie diet, a small dose of 35 mg I kg STZ was intraperitoneally injected (STZ was diluted to 1 with 0.1 mol of sodium citrate-sodium citrate solution (pH 4.0). % of the solution), 72 h after the determination of fasting blood glucose stable at 11 mmol I blood glucose 29 ramol IL for the type 2 diabetes model required for this trial. The diabetic model rats were randomly divided into 5 groups according to the blood glucose level, and 10 rats in each group were: normal control group (1), diabetes model group (11), low-dose group of different components (111), medium dose Group (IV), high dose group (V) (dissolved in a solution of 0.5% sodium carboxymethylcellulose (CMC-Na)) and metformin positive control (VI). 5%的carboxymethylcellulose was administered to groups I and II, respectively. Sodium solution; Groups III, IV, and V were given veratrosin 20, 40, 60 mg/kg per day (isoruthenin dissolved in 0.5% sodium carboxymethylcellulose (CMC-Na) solution) The content of the substance was 5m _g /ml); the group VI was administered with m ^ mww Π Π VVII 100 mg / kg of metformin. The time for intragastric administration was fixed at 9:00 am every day for a period of 4 weeks.

3. Collection and treatment of samples: The rats in each group were fasted for 16 h at the same time before the test and at the weekend of the 3rd, 6th, 9th and 12th week of the test. On the second day, blood was taken from the tail and blood glucose was determined. The fasting blood glucose level was measured by the instrument; after the end of the 4-week gavage test, blood was taken from the abdominal aorta, serum was separated, and serum insulin and glucagon were detected by radioimmunoassay.

4. Data processing: The data was statistically analyzed using one-wayANOVA in the SPSS16.0 software package. The measurement data was expressed as the mean standard deviation (X soil s ;), and the DUNCAN multiple comparison between groups was a =0. 05.

5. Results and analysis: As can be seen from the table below, blood glucose levels and serum insulin levels were significantly decreased in type 2 diabetic rats after treatment with iso-rhimelin compared with pre-replication model and before gavage treatment (P <0.01), and there was no significant difference in blood glucose between the dose groups (P>0.05).

Table 1 Effect of different ruthenium on fasting blood glucose in rats

Blood sugar / (mmol/L)

Before copying the model SI after the end of the stomach

6.12±0.5 6.65±1.3 6.68±0.9 6.44±0.6 16. 33 soil 0. 9 15·21±0.6 6.52 soil 0.7 16. 54±1. 1 9.12±0.6 6.66±0.6 16. 62 ±0. 8 8.99±0.7 6.34±0.9 16. 50±1. 2 9.03±0.9 6.47±1.0 16.59±1. 3 8.89±1.1 Note: The data in the table are x±5, n=10, (P<0.01). Of serum on serum insulin and glucagon in rats

Treatment of insulin (pmol/L) glucagon (pg/mL)

48.66 ±10.55 301.12±51.44

24.56 ±9.88 438.22±61.77

42.55±9.63 341.22±55.23

43.12±11.23 332.45±40.23

43.56 ±12.33 312.22±30.69

24.12±16.54 446.23 ±54, 63

Note: The data in the table is x±5, n=10, (P<0.01). Conclusion

The above experiments show that the isoforms of the present invention are related to the type 2 sugar established by streptozotocin (STZ) combined with high fat diet. The urinary rat model has better glycemic inhibitory activity and is equivalent to the positive drug metformin. At the same time, the changes of serum insulin and glucagon levels in this experiment indicate that isorubicin has the function of protecting and repairing islet cells, and can reduce the level of glucagon in serum and inhibit the possible pathogenesis of diabetes. . Therefore, isorubicin can be developed as a drug for treating diabetes.

Claims

Rights request

1. The use of isoprenein for the preparation of an antidiabetic drug, characterized in that the isoeugenol has the following structural formula:

The application according to claim 1, characterized in that the isoxanthin is obtained by the following method: the medicinal material of Phnom Penh is extracted with 75% ethanol, and the extract is concentrated under reduced pressure to obtain a crude extract. The extract was extracted with petroleum ether, chloroform and ethyl acetate respectively, and the ethyl acetate fraction was subjected to repeated silica gel H 200 to 300 column chromatography, chloroform: acetone 10: 1-1: 1, chloroform: methanol 50: 1-1 : 1 Gradient elution, Sephadex LH-20 column chromatography and reversed phase silica gel were used to separate the compound isorubicin.

3. Use according to claim 1, characterized in that the medicament is a pharmaceutical composition consisting of isospermidine as an active ingredient and a pharmaceutically acceptable carrier.