Preparation of myocardium peptide

A technology of cardiopeptin and copeptin, which is applied in the field of biochemistry, can solve the problems of ingredients, uses and effects that are not specified in detail, and achieve the effects of promoting damage repair, preventing allergic reactions, and promoting protein synthesis

Inactive Publication Date: 2004-12-08
大连珍奥药业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] However, the above-mentioned patented technology only roughly separates, purifies, and conducts a simple activity test on the bioactive peptide, without specifying its specific components, uses, and effects.

Method used

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  • Preparation of myocardium peptide
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  • Preparation of myocardium peptide

Examples

Experimental program
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Effect test

Embodiment 1

[0059] This experimental example involves the physical and chemical properties, purity, content and activity of cardiopeptin solution

[0060] 1. Physicochemical properties, purity and content of cardiopeptin

[0061] The cardiopeptin obtained by the preparation method of the present invention is a small molecule active polypeptide, which has stable biological activity in the range of pH 3-8, is sensitive to proteinase K, does not change in biological activity at 85°C for 10 minutes, and is stable under freezing or freeze-drying conditions ; Through HPLC analysis, the weight average molecular weight is less than 10000Da ( figure 1 ), preferably weight average molecular weight 2000-8000Da.

[0062] Table 1 The effect of different pH conditions and different time on the activity of myocardin (MTT method) (x±s, n=8)

[0063] Cardiopeptin activity OD value (x±s)

[0064] Grouping 30μg / ml 5μg / ml

[0065] Normal control group 0.691±0.032 **

[0066] Adriam...

Embodiment 2

[0227] The present invention has carried out the main pharmacodynamics test to described cardiopeptin, observed cardiopeptin on cardiac muscle morphological index, physiological index and biochemical index on whole body and isolated myocardial ischemia and ischemia-reperfusion model and its effect on myocardial oxygen consumption. The research results are as follows:

[0228] 1. Effect of cardiopeptin on myocardial ultrastructural damage caused by ischemia-reperfusion

[0229] According to the method of reference literature, after ligation of coronary artery LAD in rats for 5 minutes, cardiopeptin or control drug was injected sublingually, the ligation thread was loosened after 10 minutes of myocardial ischemia, reperfusion was performed for 30 minutes, and lead II ECG was recorded at the same time. After reperfusion, blood was collected from the abdominal aorta, the heart was taken out, rinsed with normal saline through the aorta perfusion, then perfused with 6% glutaraldehy...

Embodiment 3

[0388] Take 1 kg of healthy suckling pig ventricular muscle, wash and chop, add 1 kg of sterilized distilled water to homogenize at a speed of 3000rpm / min, freeze the homogenate at -20°C for 24 hours, repeat 3 times after thawing, and heat to 75°C , filter and remove residue with a plate and frame filter (purchased from Guangzhou Institute of Medical Devices) of model XAS03-172 / 8, select 10u medium speed filter paper for use, obtain the crude filtrate, and the hollow fiber column (the specification is F60, purchased from Switzerland) for the crude filtrate Jinbao Company) ultrafiltration, obtaining molecular weight is 12Kd fine filtrate, with ultrafiltration membrane (the specification is 10Kd, Millipore Company) ultrafiltration, molecular weight cut-off is 150ml of cardiopeptin solution of 9500Da, concentrates product with the reverse osmosis concentration column of Millipore Company.

[0389] The solution of the cardiopeptin was checked to meet the quality standard, then filt...

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Abstract

A process for preparing the myocardial peptide from the ventricular myocardium of health mammal other than man through washing, cutting, adding distilled water, homogenizing, repeated freezing and thawing 3-4 times, heating to 65-95 deg.C, filtering by plate-and-frame filter, ultrafiltering by hollow fibre column, ultrafiltering by membrane ultrafilter, reverse osmosing for concentrating, filtering for removing bacteria, and freeze drying. Its advantages are high purity and high stability.

Description

technical field [0001] The invention relates to a preparation method of cardiopeptin. Specifically, the invention relates to a preparation method for extracting cardiopeptin from the heart of healthy mammals other than humans, and belongs to the field of biochemical technology. Background technique [0002] "Myocardial protection" has been a hot spot in the research of heart surgery and heart surgery in recent years. Recent data show that many changes occur in ischemic and hypoxic cardiomyocytes, including intracellular calcium overload, free radical production, membrane damage, ATP (adenosine triphosphate, ATP) level decline, oxygen depletion and so on. [0003] The improvement and popularization of cardiac surgery has relieved the pain of many patients and improved people's quality of life. With the continuous improvement of people's requirements for myocardial protection, basic and clinical research has become more and more in-depth. Myocardial protection in clinical c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/34A61K38/17A61P9/06A61P9/10C07K1/34C07K14/435
Inventor 孔祥平邹清雁万华印杨联萍张海松李恕王日升梁强阮忠生
Owner 大连珍奥药业股份有限公司
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