title: Binding of acellular, native and cross-linked human hemoglobins to haptoglobin: enhanced distribution and clearance in the rat.
abstract: It is well established that hemoglobin resulting from red cell lysis binds to haptoglobin in plasma to form a complex. The increased molecular size precludes its filtration by the kidneys, redirecting it toward hepatocellular entry. Chemically cross-linked hemoglobins are designed to be resistant to renal excretion, even in the absence of haptoglobin. The manner in which binding to haptoglobin influences the pharmacokinetics of acellular cross-linked and native hemoglobins was investigated after intravenous injection of radiolabeled native human hemoglobin and trimesyl-(Lys82)beta-(Lys82)beta cross-linked human hemoglobin, at trace doses, into rats. Under these conditions, there is sufficient plasma haptoglobin for binding with hemoglobin. In vitro binding assayed by size-exclusion chromatography for bound and free hemoglobin revealed that, at <8 muM hemoglobin, native human hemoglobin was completely bound to rat haptoglobin, whereas only approximately 30% of trimesyl-(Lys82)beta-(Lys82)beta cross-linked hemoglobin was bound. Plasma disappearance of low doses (0.31 mumol/kg) of native and cross-linked hemoglobins was monoexponential (half-life = 23 and 33 min, respectively). The volume of distribution (40 vs. 19 ml/kg) and plasma clearance (1.22 vs. 0.4 ml.min(-1).kg(-1)) were higher for native than for cross-linked hemoglobin. Native and cross-linked human hemoglobins were found primarily in the liver, and not in the kidney, heart, lung, or spleen, mostly as degradation products. These pharmacokinetic findings suggest that the binding of hemoglobin to haptoglobin enhances its hepatocellular entry, clearance, and distribution.
authors.affiliation: Davenport Laboratory, Department of Chemistry, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Noam J
authors.initials: NJ
authors.lastname: Ship
authors.forename: Ayça
authors.initials: A
authors.lastname: Toprak
authors.forename: Roseanna P
authors.initials: RP
authors.lastname: Lai
authors.forename: Eric
authors.initials: E
authors.lastname: Tseng
authors.forename: Ronald
authors.initials: R
authors.lastname: Kluger
authors.forename: K Sandy
authors.initials: KS
authors.lastname: Pang
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents; D006242:Haptoglobins / Q000378:metabolism*; D006454:Hemoglobins / Q000493:pharmacokinetics*; D006801:Humans; D008099:Liver / Q000737:chemistry; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D014018:Tissue Distribution
chemical_list: D003432:Cross-Linking Reagents; D006242:Haptoglobins; D006454:Hemoglobins