Synthesis, biophysical properties and pharmacokinetics of ultrahigh molecular weight tense and relaxed state polymerized bovine hemoglobins.
Hemoglobin-based oxygen carriers (HBOC) are currently being developed as red blood cell (RBC) substitutes for use in transfusion medicine. Despite significant commercial development, late stage clinical results of polymerized hemoglobin (PolyHb) solutions hamper development. We synthesized two types of PolyHbs with ultrahigh molecular weights: tense (T) state PolyHb (M(W)=16.59 MDa and P(50)=41 mmHg) and relaxed (R) state PolyHb (M(W)=26.33 MDa and P(50)=0.66 mmHg). By maintaining Hb in either the T- or R-state during the polymerization reaction, we were able to synthesize ultrahigh molecular weight PolyHbs in distinct quaternary states with no tetrameric Hb, high viscosity, low colloid osmotic pressure and the ability to maintain O(2) dissociation, CO association and NO dioxygenation reactions. The PolyHbs elicited some in vitro RBC aggregation that was less than 6% dextran (500 kDa) but more than 5% human serum albumin. In vitro, T-state PolybHb autoxidized faster than R-state PolybHb as expected from previously reported studies, conversely, when administered to guinea pigs as a 20% exchange transfusion, R-state PolybHb oxidized faster and to a greater extent than T-state PolybHb, suggesting a more complex oxidative processes in vivo. Our findings also demonstrate that T-state PolybHb exhibited a longer circulating half-life, slower clearance and longer systemic exposure time compared to R-state PolybHb.
Laboratory of Biochemistry and Vascular Biology, Division of Hematology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, USA.
Paul W
PW
Buehler
Yipin
Y
Zhou
Pedro
P
Cabrales
Yiping
Y
Jia
Guoyong
G
Sun
David R
DR
Harris
Amy G
AG
Tsai
Marcos
M
Intaglietta
Andre F
AF
Palmer
D000818:Animals; D001703:Biophysics; D001704:Biopolymers / Q000737:chemistry / Q000493:pharmacokinetics*; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D004903:Erythrocyte Aggregation; D006454:Hemoglobins / Q000138:chemical synthesis* / Q000737:chemistry / Q000493:pharmacokinetics*; D008970:Molecular Weight; D012996:Solutions
D001704:Biopolymers; D006454:Hemoglobins; D012996:Solutions