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Oxygen carrier system, artificial oxygen carrier, and reducing agent
| Details |
Inventors: Tsuchida, Eishun; Takeoka, Shinji; Teramura, Yuji; Atoji, Tomoyasu;
Assignee: Oxygenix Co., Ltd (Tokyo, JP)
Primary Examiner: Carlson; Karen Cochrane
Assistant Examiner: Tsay; Marsha
Attorney, Agent or Firm: Oblon, Spivak, McClelland, Maier & Neustadt, P.C.
An oxygen carrier system includes an artificial oxygen carrier including hemoglobin vesicles encapsulating hemoglobin and an intermediate electron mediator having a standard electrode potential of 0.05 V to 0.56 V (vs. the standard hydrogen electrode) in the vesicles, and a reducing agent which is added to an external aqueous phase of the hemoglobin vesicle to reduce methemoglobin when the hemoglobin encapsulated in the hemoglobin vesicle is oxidized to the methemoglobin. The reducing agent has, as an active component, at least one compound selected from the group consisting of a thiol compound and a reducing sugar. |
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DETAILED DESCRIPTION OF THE INVENTION The present invention will be explained in detail below.
The oxygen carrier system of the present invention comprises an artificial oxygen carrier containing a given hemoglobin vesicle, and a reducing agent.
The hemoglobin vesicle contains hemoglobin and an intermediate electron mediator therein.
Hemoglobin encapsulated in the vesicle exerts central function of an artificial oxygen carrier, and binds to or releases oxygen due to a difference in oxygen partial pressure.
Hemoglobin can be obtained from erythrocyte derived from a living body, human erythrocyte derived from donation, or erythrocyte derived from livestock such as pig, sheep and cow.
For example, according to the known procedure, an erythrocyte membrane (stroma) is removed from erythrocyte by a hypotonic hemolysis method, this is heated at 60° C.
for 1 hour or longer to inactivate virus, and is purified.
The thus obtained high purity stroma-free hemoglobin can be suitably used.
By the heat treatment noted above, a methemoglobin reductase system present in erythrocyte is denatured, and is inactivated.
Alternatively, recombinant hemoglobin derived from a recombinant gene of cells of Escherichia coli and yeast, or animal or plant cells is purified and concentrated, which can be used.
A component constituting a membrane structure of the vesicle is not limited as far as it is an amphiphilic molecule or a surfactant which forms a bilayer membrane and has high biocompatibility, but lipid, preferably, phospholipid is used.
Phospholipids used in the present invention is not particularly limited, but glycerophospholipid such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, and diphosphatidylglycerol, and sphingophospholipid such as sphingomyelin can be used.
A mixture of these phospholipids may be used.
It is preferable to add cholesterol to a vesicle membrane.
The intermediate electron mediator used in the present invention is encapsulated as an oxidized form in the hemoglobin vesicle
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