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Oxygen carrier based on hemoglobin-haptoglobin compound and preparation method of oxygen carrier

A technology of haptoglobin and hemoglobin, which is applied in the field of hemoglobin oxygen carrier-hemoglobin complex oxygen carrier and its preparation, can solve the problems of increasing the mortality rate of subjects, achieve good clinical application prospects, overcome Deficiency, no hypertensive response effect

Inactive Publication Date: 2015-01-28
CHENGDU MILITARY GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the results of clinical trials show that HBOCs may cause side effects such as cardiotoxicity, hypertension, and gastrointestinal reactions, and increase the mortality of subjects

Method used

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  • Oxygen carrier based on hemoglobin-haptoglobin compound and preparation method of oxygen carrier
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  • Oxygen carrier based on hemoglobin-haptoglobin compound and preparation method of oxygen carrier

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Embodiment 1

[0036] Embodiment 1 The present invention is based on the preparation of hemoglobin-haptoglobin complex oxygen carrier

[0037] 1. Preparation method

[0038]Take 0.1 μmol of purified human umbilical cord blood hemoglobin (provided by the Institute of Blood Transfusion, Chinese Academy of Medical Sciences), dissolve it in 10 ml of phosphate buffer (pH 7.4), fill it with pure oxygen for 5 minutes, and add 0.1 μmol of haptoglobin (binding The molar ratio of globin to hemoglobin is about 1:1), then, stir and react in the dark at 0-4°C for 30 minutes, and dialyze the reaction solution with a dialysis bag with a molecular weight cut-off of 100kD until the dialysate outside the dialysis bag is in the 400-600nm region until no absorption. After the dialysis is completed, add 1 μmol of ascorbic acid under stirring, stir and react in the dark at 0-4°C for 5 minutes, adjust the pH value to 7.4 with 0.1M bicarbonate solution, freeze-dry, and obtain the oxygen-carrying carrier of the hem...

Embodiment 2

[0043] Example 2 The present invention is based on the preparation of hemoglobin-haptoglobin complex oxygen carrier

[0044] 1. Preparation method

[0045] Take 0.2 μmol of purified human umbilical cord blood hemoglobin (provided by the Institute of Blood Transfusion, Chinese Academy of Medical Sciences), dissolve it in 10ml of phosphate buffer (pH7.4), fill it with mixed gas (95% oxygen + 5% carbon dioxide) for 5 minutes, and stir Add 0.1 μmol of haptoglobin (the molar ratio of haptoglobin to hemoglobin is about 1:2), then stir and react in the dark at 0-4°C for 5 minutes, and the reaction solution is dialyzed with a dialysis bag with a molecular weight cut-off of 100kD until it is out of the dialysis bag The dialysate has no absorption in the 400-600nm region. After the dialysis is completed, add 0.5 μmol of superoxide dismutase and glutathione under stirring, and add 1.2 μmol of pyridoxal phosphate (the molar ratio of pyridoxal phosphate: hemoglobin is about 6:1), 0~4℃ St...

Embodiment 3

[0050] Example 3 The present invention is based on the preparation of hemoglobin-haptoglobin complex oxygen carrier

[0051] 1. Preparation method

[0052] Take 0.5 μmol of polymerized human cord blood hemoglobin (provided by the Institute of Blood Transfusion, Chinese Academy of Medical Sciences), dissolve it in 10 ml of phosphate buffer (pH 7.4), fill it with a mixed gas (95% oxygen + 5% carbon dioxide) for 10 minutes, and stir Add 0.1 μmol of haptoglobin (the molar ratio of haptoglobin to hemoglobin is about 1:5), then stir and react in the dark at 0-4°C for 15 minutes, and the reaction solution is dialyzed with a dialysis bag with a molecular weight cut-off of 100kD until it is out of the dialysis bag The dialysate has no absorption in the 400-600nm region. After the dialysis is completed, add 2.5 μmol of superoxide dismutase and glutathione under stirring, and add 2 μmol of pyridoxal phosphate (the molar ratio of pyridoxal phosphate: hemoglobin is about 4:1), and keep aw...

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Abstract

The invention discloses an oxygen carrier based on a hemoglobin-haptoglobin compound and a preparation method of the oxygen carrier. The oxygen carrier comprises haptoglobin, hemoglobin, antioxidant and pyridoxal phosphate. The hemoglobin-haptoglobin compound is formed as a unit for bringing the oxygen carrying activity into play by virtue of special combination ability of the haptoglobin and the hemoglobin. The oxygen carrier based on the hemoglobin-haptoglobin compound disclosed by the invention is free of hypertension reaction after being infused, and capable of protecting an isolated heart. And therefore, the oxygen carrier can be used for clinical blood transfusion, protecting vital organs including the heart and the like from ischemia reperfusion injuries, and has good application prospects and economical benefits.

Description

technical field [0001] The invention relates to the field of blood substitutes, in particular to an oxygen carrier of a hemoglobin oxygen carrier-haptoglobin complex and a preparation method thereof. Background technique [0002] Blood transfusion has played an important role in modern medicine and is an indispensable medical method for clinical operations, disaster relief and battlefield treatment. However, traditional blood transfusion still has many disadvantages. For example, its blood source is single, mainly relying on blood donation, and due to the complexity of human blood types, it needs to be used after strict matching, which limits its application in the treatment of acute and severe patients and in emergencies. What's more serious is that blood is easily polluted by viruses such as hepatitis and AIDS, and the problem of old and new blood is becoming more and more concerned, which threatens the safety of blood transfusion. With the development of medical technol...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/42A61P7/08
Inventor 吴畏刘英海刘合年巩固殷亮万勇孙小琴李涛杨倩
Owner CHENGDU MILITARY GENERAL HOSPITAL OF PLA
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