Cross-linked hemoglobin preparation method and organ perfusate comprising hemoglobin

A technology of hemoglobin and organ perfusion, which is applied in the field of organ preservation in medical technology, can solve the problems of non-specific cross-linking reaction, poor molecular weight stability, increased blood viscosity, etc., and solve the adverse consequences of acidosis and lysosomal enzyme activation, The effect of reducing the content and increasing the adding speed

Inactive Publication Date: 2019-12-13
REDPHARM BEIJING BIOPHARMACEUTICAL INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But glutaraldehyde crosslinking also has its obvious disadvantages. First, the crosslinking reaction is non-specific, because there are about 40 lysine side chain amino groups and four chain terminal amino groups on the surface of hemoglobin that can participate in the reaction, and the reaction is difficult to control. The molecular weight distribution of the product ranges from 32kD to over 1000kD, resulting in a heterogeneo...

Method used

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  • Cross-linked hemoglobin preparation method and organ perfusate comprising hemoglobin
  • Cross-linked hemoglobin preparation method and organ perfusate comprising hemoglobin
  • Cross-linked hemoglobin preparation method and organ perfusate comprising hemoglobin

Examples

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Embodiment 1

[0040] Such as figure 1 As shown, dilute the hemoglobin of deoxygenation to 2g / dl, join in the anaerobic reactor 12, its liquid level 10 height is no more than half of the height of the reactor 12, fix the atomizer 1 at the reactor 12 top, in the atomizer 1 is connected to the liquid inlet pipe 2 and the inlet pipe 3, and the balance gas pipe 4 is installed on the top of the reactor 12 to adjust the pressure in the reactor 12, the reactor 12 is closed, the stirring rotor 11 at the bottom is opened, and the reaction is heated through the water bath jacket 13 When the liquid reaches 42°C, open the valve 5 of the liquid inlet pipe, add the cross-linking agent glutaraldehyde, adjust the liquid inlet speed to 260ml / min, open the valve 8 of the inlet pipe, and feed an inert gas, preferably high-purity nitrogen, and adjust the gas flow to make the exchange Joint agent mist and reaction liquid surface 10 fully contact, improve reaction efficiency, regulate and control the exhaust valv...

Embodiment 2

[0043] The preparation method is the same as in Example 1, except that before adding the cross-linking agent, N-acetyl-L-cysteine ​​(NAC) is added to the diluted deoxygenated hemoglobin solution, and the final concentration of the added NAC is 3 mg / ml, the mass ratio of hemoglobin to NAC is 20:3, and the molecular weight distribution of the obtained cross-linked hemoglobin is shown in Table 1.

Embodiment 3

[0045] The preparation method is the same as in Example 1, except that before adding the cross-linking agent, glutathione is added to the diluted deoxygenated hemoglobin solution, and the final concentration of the added glutathione is 3 mg / ml, and the prepared cross-linking agent The molecular weight distribution of linked hemoglobin is shown in Table 1.

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Abstract

The invention discloses a cross-linked hemoglobin preparation method, wherein the preparation method comprises the following steps: a, diluting deoxidized hemoglobin, and adding the diluted deoxidizedhemoglobin into an anaerobic reactor; b, arranging an atomizer at the top of the anaerobic reactor, adding a cross-linking agent into the reactor through the atomizer, and carrying out a cross-linking reaction; and c, adding a reducing agent to terminate the cross-linking reaction, and removing the unreacted cross-linking agent and the terminating reducing agent in an ultrafiltration liquid changing manner to obtain the cross-linked hemoglobin. The cross-linked hemoglobin prepared by the preparation method contains low contents of macromolecules (greater than 500 kD) and micromolecules (lessthan 64 kD); by adding the cross-linked hemoglobin into organ perfusate, the phenomena of organ acidosis and lysosomal enzyme activation adverse consequences caused by lack of an oxygen carrying agentin conventional perfusate and cell damage caused by excessive free radicals generated during reperfusion can be solved, and the perfusate is quite simple and convenient to prepare, store, transport and use.

Description

technical field [0001] The invention relates to the technical field of organ preservation in medical technology, in particular to a preparation method of cross-linked hemoglobin, and in particular to an organ perfusate containing the cross-linked hemoglobin. Background technique [0002] Many human diseases come from the necrosis of cells, tissues, and organs, resulting in loss of function, disability, and even death. With the development of modern medicine, organ transplantation technology is becoming more and more mature. From the perspective of surgery: organ transplantation technology is perfect , head replacement surgery is not a big problem. In fact, organ transplantation still faces various difficulties, and the preservation of isolated organs is one of them. Organ transplantation requires the transplantation of living organs. Therefore, from the time when the organ for transplantation is cut off from the donor to the time when its main blood vessels are connected to...

Claims

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Application Information

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IPC IPC(8): C07K14/805C07K14/415C07K19/00A01N1/02
CPCC07K14/805C07K14/415C07K5/0215A01N1/0247C07K2319/00
Inventor 游可为史国营孙新宇陈浩源
Owner REDPHARM BEIJING BIOPHARMACEUTICAL INST CO LTD
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