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Preparation technology of interventional thrombolytic biological product namely urechis unicinctus plasmin

A single ring thorn and biological product technology, applied in the direction of biochemical equipment and methods, enzymes, peptidases, etc., can solve problems such as allergies, intracranial hemorrhage, and high prices

Inactive Publication Date: 2015-03-11
陈良海
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The second-generation thrombolytic drugs mainly refer to tissue plasminogen activator (such as t-PA), etc. t-PA is also called alteplase, which has a good affinity with fibrin and has a strong Thrombus selective, systemic fibrinolytic effect is small, and side effects are small, but its half-life is short (3 minutes), requires a large amount of administration in a short period of time, is expensive, and has the risk of causing intracranial hemorrhage
Lumbrokinase is a kind of substance extracted or molecularly recombined from special earthworms. It has two functions of kinase and plasmin. deafness, nephrotic syndrome and other diseases, but most lumbrokinase products are not a single component, and have side effects such as allergies
Other third-generation thrombolytic drugs (reteplase, defibrase, etc.) still have problems such as short half-life in vivo, poor stability, specificity and safety to be further improved
In addition, the common problem of these three generations of drugs is that they cannot completely dissolve fresh and old thrombus, even if thrombolysis is involved, the thrombus cannot be completely dissolved

Method used

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  • Preparation technology of interventional thrombolytic biological product namely urechis unicinctus plasmin
  • Preparation technology of interventional thrombolytic biological product namely urechis unicinctus plasmin
  • Preparation technology of interventional thrombolytic biological product namely urechis unicinctus plasmin

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Embodiment Construction

[0013] The examples listed below are only to help those skilled in the art to better understand the present invention.

[0014] (1) The preparation process of the crude product in the present invention is as follows: Fresh and alive Echinococcus monoringensis is cultivated overnight in sterile sea water to remove impurities in the body, dissects and collects body cavity fluid, freezes and centrifuges at 2000g for 30 minutes at 4°C, and supernatant passes through 400 Remove lipid insoluble matter by suction filtration with mesh silk cloth and 3-layer filter paper, put in dialysis bag at 4°C overnight for desalination, check the desalination effect, and vacuum freeze-dry to obtain crude product.

[0015] (2) DEAE-Sepahcel anion exchange chromatography: the crude product was dissolved in 50mM ammonium bicarbonate buffer (pH7.8), refrigerated and centrifuged at 10000g at 4°C for 10 minutes, and the supernatant was injected into 50mM ammonium bicarbonate buffer (pH7.8). 8) Equilibr...

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Abstract

The invention relates to a preparation technology of an interventional thrombolytic biological product namely urechis unicinctus plasmin. The technology comprises the following steps: freezing coelomic fluid of urechis unicinctus, then subjecting the fluid to centrifugation and suction filtration, carrying out freeze drying in vacuum to obtain a coarse product, subjecting the coarse product to DEAE (diethyl amino ethanol)-Sepahcel (cellulose glucan gel) anion exchange chromatography, Sephadex (hydroxypropyl glucan gel)G-75 chromatography, benzamidine-Sepharose (agarose gel) chromatography, and freeze drying to further verify that the coarse product is plasmin, and then carrying out an interventional thrombolytic experiment on a rat carotid and cerebral artery embolism model to prove that the urechis unicinctus plasmin can completely dissolve thrombus and does not have any side effect such as allergy, and the like.

Description

Technical field: [0001] The invention relates to a preparation process of a thrombus-dissolving biological product P. monocyticus fibrinolytic enzyme. The biological product can be used for basic research and clinical application of thrombotic diseases. Background technique: [0002] Thrombotic disease is a kind of circulatory system disease that seriously endangers human health because it completely or partially blocks blood vessels after thrombus forms or falls off, causing ischemic necrosis of organs and tissues. Performance of acute myocardial infarction, cerebral infarction, deep vein thrombosis, pulmonary embolism. At present, the main clinical drugs for the treatment of thrombotic diseases include antiplatelet drugs, anticoagulant drugs, and thrombolytic drugs, and the latter is currently the most widely used clinical treatment. Thrombolytic drugs, also known as fibrinolytic drugs, can activate plasmin and promote the dissolution of thrombus. The first generation of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/68A61P9/10
CPCC12N9/6408C12Y304/21007
Inventor 陈良海
Owner 陈良海
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