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Process for preparing hirudin peptide for reducing harmful bleeding reaction of hirudo

A technology of adverse reactions and dried leeches, applied in the fields of medicine and biology, can solve the problems of bleeding reactions and bleeding adverse reactions that leech preparations cannot be used for a long time

Inactive Publication Date: 2017-10-24
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When leech powder is used clinically for the treatment of diseases, taking too much leech powder or taking it for a long time may cause bleeding adverse reactions. The main reason why leech preparations cannot be used for a long time is also because it is easy to cause bleeding reactions (internal bleeding)

Method used

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  • Process for preparing hirudin peptide for reducing harmful bleeding reaction of hirudo
  • Process for preparing hirudin peptide for reducing harmful bleeding reaction of hirudo
  • Process for preparing hirudin peptide for reducing harmful bleeding reaction of hirudo

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The preparation of leech extract under the different conditions of embodiment 1

[0043] Based on different purposes, the preparation of leech extract has different combinations. In addition, the current research based on leeches is mainly the separation and purification of active substances, and the enzymatic hydrolysis of leeches is simulated in vitro by simulating the environment of the human gastrointestinal tract. The metabolic transformation of substances also plays a role that cannot be ignored. During the oral administration of most Chinese herbal medicines, it is unavoidable to come into contact with intestinal microorganisms in the intestines and be metabolized by them, and then absorbed into the blood by the gastrointestinal tract. Microorganisms have excellent enzyme systems that allow them to participate in catalysis and hydrolysis processes.

[0044] In the present invention, the whole dry leech is used as a raw material, and four kinds of leech extracts a...

Embodiment 2

[0095] Example 2 Effects of different leech extracts on coagulation activity of endothelial cells

[0096] VECs are a continuous monolayer of flat cells on the surface of the lumen of blood vessels. In addition to mediating the exchange of substances between blood and tissues, VECs play a major role in inflammation and coagulation activation. VEC can play a two-way regulation role in the dynamic balance of hemostasis and anticoagulation in the body by synthesizing and releasing a variety of coagulation-related factors. The procoagulation-related factors expressed by it mainly include: TF, thromboxane A2 (ThromboxaneA2, TXA2), vWF, platelet activating factor (Platelet activating factor, PAF), PAI, etc.; the anti-coagulation related factors mainly include: endothelium-derived vasodilator factor NO , eNOs, TM, antithrombin-III (AntithrombinIII, AT-III), PS, tPA, TFPI, PGI2, heparan sulfate (heparinsulfate, HS) and so on. The present invention explores the influence of different ...

Embodiment 3

[0141] Example 3 Effect of PHL on molecules related to endothelial cell dysfunction

[0142] 3.1 Establishment of endothelial dysfunction model

[0143] Select the well-growing EA.hy926 cells to digest and centrifuge, resuspend with DMEM medium containing 10% FBS, count the cells, and divide them into 2×10 5 Each well was inoculated on a 6-well plate, cultivated in an incubator until the confluence was about 80%, and TNF-α was added at a concentration of 0, 1, 5, 10 and 100 ng / ml, and placed in an incubator to continue culturing, respectively, at 6 After 12 and 24 hours, the corresponding total cellular RNA was extracted, and the relevant inflammatory factors IL-6 and MCP-1 were subjected to PCR, electrophoresis and mapping, so as to screen out the appropriate concentration and time of TNF-α treatment.

[0144] RT-PCR method was used to detect the expression levels of IL-6 and MCP-1 mRNA in EA.hy926 endothelial cells after 0, 1, 5, 10 and 100ng / ml TNF-α acted on EA.hy926 endo...

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Abstract

The invention discloses a process for preparing hirudin peptide for reducing harmful bleeding reaction of hirudo. The process comprises the following steps: grinding and homogenating dried hirudo, adding 10000NFU / g of pepsin at 36-38 DEG C and pH of 2.0, and performing enzymolysis for 8 hours and above; performing alcohol precipitation on enzymatic hydrolysate, and concentrating and lyophilizing the filtrate to obtain hirudo extract. The hirudo extract PHL prepared by using the method can be used for inhibiting the process of atherosclerosis on the premise of not influencing the blood coagulation factor of endothelial cells. A reliable preparation process is provided to a safe and reliable novel oral preparation which can be clinically used for preventing atherosclerosis for a long time and has weak harmful bleeding reaction.

Description

technical field [0001] The invention belongs to the field of medicine and biotechnology, and specifically relates to a preparation process of leech peptides for alleviating the adverse reactions of leech bleeding, in particular to a preparation process of the leech peptides for alleviating the adverse reactions of leech bleeding and having the effect of preventing and treating atherosclerosis. Background technique [0002] Atherosclerosis (AS) is a lipid metabolism defect disease characterized by the accumulation of lipid and fibrous substances in large and medium-sized elastic blood vessels, which may lead to ischemia in the heart, brain or limbs due to blood vessel blockage, resulting in infarction. Therefore, AS is also considered to be the main cause of many cardiovascular and cerebrovascular diseases, including myocardial infarction, coronary heart disease and cerebral thrombosis. To be precise, AS lesions represent a series of highly specific cellular and molecular re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/30A61K38/58A61P9/10A61P7/02A61P9/08
CPCC12P21/06A61K38/00C07K14/815
Inventor 宋淑亮吉爱国李雪
Owner SHANDONG UNIV
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