Deanna J. Nelson

Preparation and Characterization of Diaspirin Cross-Linked Hemoglobin Solutions for Preclinical Studies

During 1990 and 1991 the capability for repetitive, consecutive production of DCLHb solution to meet a rigorous and complete set of product criteria was demonstrated. In addition, through periodic monitoring of product stored under controlled conditions, the stability of all lots of DCLHb solution during frozen storage was demonstrated for more than a year. In this way, assurance was provided that the DCLHb solution used in preclinical testing met all product criteria throughout the biological testing period.

POLYMERIZATION OF DIASPIRIN CROSSLINKED HEMOGLOBIN (DCLHB) WITH PEG ACTIVATED WITH BENZENESULFONATE BEARING ELECTRON-WITHDRAWING GROUPS

Abstract The hemoglobin-based oxygen carrier solution, DCLHb, was polymerized with novel polyethylene glycol polymerization reagents to provide a solution of monomers to tetramers of hemoglobin molecules. The polyethylene glycol reagents 4 and 5 were designed to be selective toward nucleophilic displacement of the substituted benzenesulfonates by the sulfhydryl groups of β93 Cys of DCLHb.

Diaspirin Crosslinked Hemoglobin (DCLHbTM): Bioanalytical Studies in Swine

These studies were a part of preclinical safety and efficacy studies of DCLHb. Their purpose was to analyze the characteristics of DCLHb during circulation, and the distribution of iron following the administration of DLCHb to swine. Swine were dosed (2 g/kg) with 10 g/dL DCLHb, infused intravenously at a rate of 1 mL/kg/min. Blood samples were collected up to 48 hours post-infusion for analysis. Tissue samples were obtained for iron determination. The data collected showed that the concentration of DCLHb in the plasma gradually decreased, while the concentration of methemoglobin remained low and essentially constant. The oxygen binding characteristics and stability of the crosslink were preserved following infusion, indicating that the DCLHb continued to function as an effective oxygen carrier. Iron concentrations in the liver and kidneys increased as expected, but plasma levels of iron did not saturate the iron binding capacity of transferrin, inferring a controlled process for the release of iron.