Reduced oxygen carriers and their use for the treatment of carboxyhemoglobinemia
A hemoglobin and oxygen carrier technology, applied in the field of carboxyhemoglobinemia, can solve the problem of carbon monoxide scavenging rate in general and unusable
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Embodiment 1
[0171] Example 1: Carbon Monoxide (CO) Scavenging Rapid In Vivo Removal of Carboxyhemoglobin (HbCO) in CO Intoxicated Mice
[0172] It has been previously shown that exposing mice to air containing 1500 ppm CO gas for an average of 50 minutes increases HbCO levels to 64% + / - 1% (PCT Publication No. WO2014 / 150413). Prior to exposure, mice were surgically placed with femoral artery and venous catheters for blood pressure monitoring, blood sampling and infusion with recombinant neuroglobin (rNgb), another type of CO-scavenging globulin, or PBS as a control. Using a Harvard infusion pump, inject 250 μL of 8-12 mM rNgb or PBS into the mice within 4 minutes. Immediately after infusion and every 5 minutes 5 μL of blood was collected for HbCO measurement. Such as figure 1 As shown, rNgb infusion can rapidly reduce HbCO levels compared with PBS control. Specifically, 5 minutes after returning to normal air, HbCO levels in the group that received rNgb decreased by an average of 32.8%...
Embodiment 2
[0173] Example 2: Materials and methods
[0174] This example describes the methods and experimental procedures used for the studies described in Examples 3-8.
[0175] Kinetics of carboxylated RBC mixed with myoglobin
[0176] Red blood cells were obtained by washing 50-100 μL of blood 5-7 times with PBS and centrifuging at 1000 x g for 5-10 min. Dilute washed RBCs in 1-2 ml of PBS. The RBCs were then deoxygenated on ice for 1 h by a stream of argon with slow stirring. For anaerobic experiments, argon was briefly passed through and an excess of sodium dithionite relative to Hb was added to the RBC. Dilute the carboxylated erythrocyte solution at least 4:1 to obtain carboxylated erythrocyte-encapsulated Hb. RBCs were washed twice with degassed PBS (5-10 mM dithionite for anaerobic experiments) and centrifuged at 1000 x g for 5 min in a degassed, septum-covered 15 mL centrifuge tube to remove excess CO. After washing, resuspend the RBCs to a final concentration of 100-200 ...
Embodiment 3
[0186] Example 3: In Vitro Model of CO Intoxication
[0187] Oxygen carriers will scavenge CO from CO-poisoned HbCO complexes. In an in vitro model of this clearance process, 100% HbCO was placed in solution under anaerobic conditions. The concentration of HbCO measured by spectroscopy did not change when PBS was added. When 100% deoxymyoglobin was added to this solution at a ratio of 1:1, more than half of the HbCO was reduced and CO was bound by myoglobin. When insufficient amounts were added to HbCO (110.8 μM HbCO vs 85.3 μM deoxymyoglobin), the HbCO concentration was reduced by one-third (Fig. 4).
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