Biopure (United States)

Establishing an acceptable safety and efficacy profile for hemoglobin based oxygen carriers, HBOCs, is essential. Understanding the data that describes an HBOC "safety profile" requires consideration beyond counting "events" typically associated with intent to treat analysis. The imputation of causation from numerical counts alone, without clinical context, is an incomplete description of a complex situation. Clinical contextualization provides a greater understanding of the clinico-pathological processes involved. Generating alternative hypotheses for the origin of the events, apart from the drug, patient co-morbidities, situational and disease demands, could include protocol design, failure to provide mitigation strategies, patient management issues, or inadequate education of investigators. How clinical contextualization can provide insight for the interpretation of significant safety profile events is discussed in the context of a large phase III clinical trial where an appreciation of factors underlying the differences between intent to treat analysis and other approaches is discussed.

Described in this protocol is a rat preparation that serves as a sensitive, reliable in vivo assay of peripheral vascular resistance. Experiments are conducted on conscious animals 5 days following the surgical implantation of an arterial pressure catheter in the carotid artery and a transit time flow probe on the abdominal aorta. The transit time flow probe measures volume blood flow per unit time, making it possible to calculate a true vascular resistance, a reliable indicator of microvascular tone. After allowing appropriate recovery time between experiments, it is possible to use the animal preparation multiple times. This increases study efficiency by reducing the number of animals required and makes it possible to use paired statistical analysis since multiple treatment interventions, including control conditions, can be evaluated in each animal.

HBOC-201, a hemoglobin-based oxygen carrier, improved physiologic parameters and survival in hemorrhagic shock (HS) animal models. However, resuscitation from HS and the properties of different fluids influence immune responses. The aim of this study was to determine if HBOC-201 significantly alters immune function in traumatic HS. Anesthetized pigs underwent soft tissue injury, controlled hemorrhage of 40% of blood volume, and resuscitation with HBOC-201 or Hextend, or no resuscitation. Sequential whole-blood samples were collected for analyses of leukocyte differential (hematology analyzer), T-lymphocyte subsets (CD3, CD4, and CD8) (FACS), lymphocyte adhesion marker CD49d (alpha4-integrin) expression (FACS), plasma cytokines-tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-10-(ELISA), and lymphocyte apoptosis (annexin-V/propidium iodide staining) (FACS). Statistical analyses were performed by the mixed procedure. Total WBC counts decreased posthemorrhage in both resuscitation groups. Lymphocyte percentages decreased and PMN percentages increased around 4 h posthemorrhage in all groups. CD3 cells decreased in all groups, but CD4 and CD8 cells decreased only in the resuscitation groups. TNF-alpha levels were not detectable in any groups. IL-6 levels were similar across treatment groups (P > 0.05); however, IL-10 levels were higher in the HBOC group, as early as 1 h posthemorrhage (P = 0.04). Increases in lymphocytic CD49d expression levels and apoptosis occurred only in nonresuscitation and Hextend groups, respectively (P < or = 0.01). In comparison with Hextend, HBOC-201 had no significant adverse or beneficial effects on immune function in this model of moderately severe HS in swine, suggesting that it may be safe as a resuscitation fluid in HS patients.

Biopure's hemoglobin-based oxygen carrier, HBOC-201 (Hemopure), enhances oxygen transport by promoting both the convective and diffusive components of transport in the microcirculation. Convective transport is modified by HBOC-201 in three ways; (i) volume expansion promotes organ and tissue perfusion, (ii) the low viscosity of HBOC-201 improves flow to tissues, and (iii) oxygen delivery by HBOC Hb in the plasma is relatively insensitive to mechanisms regulating RBC distribution in the microcirculation. Diffusive oxygen transport is increased by the higher P50 compared with native RBC Hb which increases the off-loading of oxygen to tissues. Oxygen transport is also increased by reducing the diffusional barrier to oxygen transport associated with the plasma, in which oxygen is sparingly soluble. Biopure's HBOC solutions have been shown in vitro and in vivo to take up and off-load oxygen more efficiently than RBC Hb, and when added to blood can increase the efficiency of RBC oxygen transport.

A Hb-based oxygen carrier, HBOC-301, is licensed by the FDA for use as a single-dose product to treat anemia in canines. The objective of this study was to investigate the immunopathologic consequences of multiple doses of HBOC-301. HBOC-301 was administered intravenously at 1.3 g per kg (10 mL/kg) nine times over 50 weeks to each of eight splenectomized beagles. During interim weeks, HBOC-301-specific IgG antibody was quantified in serum by immunoassay. Immunofluorescence studies were performed on thin sections of control and test dog livers and kidneys to detect any IgG, IgM, IgA, or C3 deposition. Physiology (p50) studies evaluated the capacity of circulating HBOC-301-specific IgG antibody to competitively block the binding of oxygen to HBOC-301. HBOC-301-specific IgG antibody was detected in seven of eight HBOC-301-treated dogs by Week 6. Peak antibody levels occurred by Week 10 (after the third dose). Immunofluorescence studies detected comparable IgG, IgA, IgM, or C3 deposition patterns in the kidneys and livers of both control and test dogs, which indicated no selective antibody-mediated deposition from multiple HBOC-301 administrations. Moreover, HBOC-301-specific IgG antibody in serum was unable to inhibit oxygen binding to the HBOC-301 in vitro, which indicated that circulating antibody did not diminish the oxygen-binding capacity of HBOC-301. The immunologic, histologic, and physiologic data support the safety of multidose administration of HBOC-301 in canines.

We investigated the effect of bovine polymerized hemoglobin-based oxygen carrying (HBOC) solution on myocardial oxygen consumption (MVO2) and substrate use. At 15 min after the end of HBOC infusion (20% blood volume, i.v.) in nine permanently instrumented conscious dogs, mean arterial pressure and coronary blood flow were both increased by 41+/-5% and 93+/-20% (p<0.01) without affecting late diastolic coronary resistance and left ventricular dP/dtmax. Administration of HBOC did not affect arterial PO2 or O2 content, but significantly decreased coronary sinus PO2 and O2 content by 21+/-3% and 36+/-3%, respectively. MVO2 was increased from 7.2+/-0.8 to 15+/-1.8 ml O2/min (p<0.01). Despite an increase in triple product from 44+/-2 to 56+/-3 (p<0.01) 15 min after HBOC, the ratio of MVO2 and triple product was markedly elevated by 62+/-19%. Myocardial free fatty acid consumption was decreased from 14+/-1 to 4.5+/-2.2 microEq/min, whereas consumption of lactate increased from 19+/-6 to 69+/-10 micromol/ min and that of glucose increased from 1.0+/-0.5 to 10+/-3 mg/min (all p values, <0.05). These metabolic changes were not observed in dogs that received angiotensin II at a dose used (20-40 ng/kg/min, i.v.) to match those hemodynamic effects of HBOC. These results suggest that administration of HBOC increases coronary blood flow and MVO2 and shifts cardiac metabolism from using free fatty acid to using lactate and glucose in conscious dogs at rest. These metabolic changes are independent of the HBOC-induced change in hemodynamics.

Red blood cell (RBC) replacement solutions are being developed as alternatives to allogeneic RBC use in blood transfusions in the treatment of massive trauma, to achieve hemodynamic stability during elective surgery, and to increase oxygen-carrying capacity in anemia. Hemoglobin-based oxygen carrier (HBOC)-201 (Biopure Corp.) is a purified, sterile, isosmotic glutaraldehyde-polymerized bovine hemoglobin. Because this product is acellular, blood components containing this substance appear hemolyzed. This study reports on the interferences produced by the presence of HBOC-201 in a variety of clinical assays. This product was added in vitro at concentrations up to 60 g/L (6.0 g/dL) to normal human serum, plasma, or whole blood before testing for serum chemistries, coagulation profiles, and hematology and blood bank assays. In addition, a set of normal human sera containing HBOC-201 was supplemented with various therapeutic drugs and assayed for these agents. The results of these studies demonstrate that the presence of HBOC-201 in blood components does not result in significant analytical interference that would be of concern with many clinical assays at HBOC-201 concentrations encountered during routine clinical use of this RBC replacement solution in patients.

Hemoglobin-based oxygen carrier-201 (HBOC) contains a solution of glutaraldehyde-polymerized bovine hemoglobin. The hemoglobin molecule absorbs light at several wavelengths. The presence of HBOC increases plasma and serum hemoglobin, producing the appearance of hemolysis. Young lists >80 analytes for which clinical laboratory tests are affected by hemolysis [1] . In preparation for a multicenter study to evaluate the effect of the use and safety of HBOC (Biopure Corp.), we studied the effect of this drug on most of our instruments and analytes that might be used in management of our patients during the planned clinical trial. The instruments studied were: …