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Pillworm fibrinolysis active protein and application thereof

A kind of active protein, the technology of murine, applied in the field of murine fibrinolytic active protein and its application, to achieve the effect of strong antithrombotic activity in vivo, simple process and mild thrombolytic effect

Active Publication Date: 2017-05-10
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few oral drugs currently on the market that have both thrombolytic and anticoagulant effects. Therefore, it is imperative to find a thrombolytic anticoagulant that has fewer side effects and avoids reocclusion and reinfarction.

Method used

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  • Pillworm fibrinolysis active protein and application thereof
  • Pillworm fibrinolysis active protein and application thereof
  • Pillworm fibrinolysis active protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Grinding and enzymatic hydrolyzate of muscaria

[0031] 1. Preparation of superfine pulverized material by wet method

[0032] Weigh 100.0 g of dried muffin, add distilled water 8 times the mass of muffin, soak in a water bath at 25°C for 1 hour, add it into an ultrafine pulverizer, and perform wet ultrafine pulverization at 25°C for 10 minutes. Centrifuge the pulverized slurry at 10,000 r / min for 10 min at 4°C, take the supernatant, and freeze-dry to obtain 9.6 g of ultrafine pulverized material by wet method.

[0033] 1. Preparation of the enzymatic hydrolyzate of the pulverized mousewort

[0034] Weigh 500 mg of the ultra-fine pulverized material by the wet method, and resuspend it in 50 mL of artificial gastric juice (see the preparation of artificial gastric juice in the 2010 edition of Chinese Pharmacopoeia, the concentration of pepsin is 10 mg / mL), to ensure that the pepsin in the artificial gastric juice (batch number 313A0313, Solarbio company) is e...

Embodiment 2

[0037] Example 2 The preparation of the extract of chrysanthemum with thrombolytic and anticoagulant activity

[0038] (1) HiTrap Capto Q ion exchange chromatography

[0039] HiTrap Capto Q ion-exchange chromatography column (100ml, purchased from GE Healthcare, USA) was equilibrated with 5 column volumes of equilibration buffer I (pH8.0, concentration 20mmol / L Tris-HCl buffer solution), and the flow rate was 1.0mL / min. Use 20mmol / L Tris-HCl buffer (pH8.0) to prepare the freeze-dried powder of the enzymatic hydrolyzate of the squirrel powder (pepsin and trypsin) into a 10mg / mL solution, and load it on the equilibrated The HiTrap Capto Q ion-exchange chromatography column, first elute the unadsorbed protein with equilibration buffer Ⅰ, and then use elution buffer Ⅰ (pH8.0 containing 1mol / L NaCl, 20mmol / LTris-HCl Buffer) and equilibration buffer I for elution, elution is 2 column volumes, the volume ratio of elution buffer I and equilibration buffer I is 1:4, and each 2 mL is...

Embodiment 3

[0072] Embodiment 3 fibrinolytic active protein PSLTro02 thrombolytic anticoagulant activity

[0073] 1. Antithrombotic activity in vivo

[0074]Get 40 SD rats, half male and half male, randomly divided into 5 groups: normal saline group (i.e. blank control group), urokinase group and lumbrokinase group (positive control group), fibrinolytic active protein PSLTro02 high-dose group and low-dose group. In the dosage group, each group was administered intragastrically for 5 consecutive days, once a day, and the dosage is shown in Table 3. Anesthetize the above-mentioned 5 groups of SD rats with 2% pentobarbital sodium solution (40 mg / kg) to make an arteriovenous loop thrombosis model. The rats are fixed on the operating board in the supine position, and the right common carotid artery and the left common carotid artery are separated. For the lateral external jugular vein, place a weighed surgical thread about 8 cm long in a polyethylene tube filled with heparin sodium saline (50...

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Abstract

The invention provides a pillworm fibrinolysis active protein and an application thereof, and relates to the pharmaceutical field. The pillworm fibrinolysis active protein is obtained by carrying out the enzymolysis to smashed pillworm by successively using pepsin and trypsin, the molecular weight of the pillworm fibrinolysis active protein is 8-10 k Da, and the pillworm fibrinolysis active protein has the thrombolysis and anticoagulant activity. Tested by experiments, a rat has the stronger in-vivo antithrombotic activity after oral administration, and the thrombolysis effect is moderate relatively. The pillworm fibrinolysis active protein can be applied for preparing oral drugs for treating thrombotic diseases.

Description

technical field [0001] The invention relates to the field of pharmacy, in particular to a murine fibrinolytic active protein and its application. Background technique [0002] Thrombosis is the root cause of three deadly cardiovascular diseases (heart attack, stroke and venous thromboembolism), with a high incidence rate, showing a trend of increasing worldwide. According to the data provided by the International Thrombosis and Hemostasis Society, in the United States, 100,000-300,000 people die from venous thrombosis every year; in Europe, 500,000 people die from venous thrombosis every year, exceeding AIDS, breast cancer, prostate cancer and highway The sum of traffic accident deaths; in China, the number of deaths due to thrombotic diseases and their complications is also large. At present, although antithrombotic drugs in clinical use can greatly reduce the mortality and disability rates, these thrombolytic drugs still have disadvantages, such as large bleeding side eff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12P21/06C07K1/36C07K1/34C07K1/18C07K1/16A61K38/17A61P7/02
CPCA61K38/00C07K14/43509C12P21/06
Inventor 李博吴勉华郭立玮朱华旭王正俊
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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