title: In vivo distribution and elimination of hemoglobin modified by intramolecular cross-linking with 2-nor-2-formylpyridoxal 5'-phosphate.
abstract: Modified hemoglobin solutions have potential application as plasma expanders with oxygen-transporting capacity. In a previous study it was found that modification of hemoglobin by intramolecular cross-linking with 2-nor-2-formylpyridoxal 5'-phosphate (NFPLP) improves the vascular retention time by a factor of three, and it also improves the oxygen-transporting properties. In the present study we investigated in rats how, after exchange transfusion of a clinically relevant dose, the modified hemoglobin (HbNFPLP) was distributed in the body compared with how the unmodified hemoglobin was distributed. By using a new technetium 99m labeling technique, we found in a scintigraphic study that accumulation of hemoglobin in the kidneys was greatly diminished by the intramolecular cross-linking with NFPLP. These findings were confirmed by light-microscopic observations after diaminobenzidine staining. It was concluded that the impairment of kidney function caused by blockade of the tubuli is not to be expected from HbNFPLP. In the liver and spleen, where the free HbNFPLP is possibly eliminated, some accumulation of 99mTc label was observed, but the major part of the extravascular label was diffusely spread throughout the body. This led to the conclusion that important accumulation of undegraded HbNFPLP does not occur in the liver and spleen. Rapid appearance of both hemoglobin and HbNFPLP in the lymph showed that cross-linking with NFPLP does not prevent the distribution of hemoglobin over the interstitial space in the first hours after administration. However, pharmacokinetic analysis demonstrated that transcapillary transfer contributes only to a limited extent to the disappearance from the circulation. During 24-hour infusions of HbNFPLP, a steady state with a constant plasma concentration was easily reached. The latter experiment indicated that the eliminating system does not become saturated during prolonged administration of large doses of HbNFPLP.
authors.affiliation: Central Laboratory, the Netherlands Red Cross Blood Transfusion Service, Amsterdam.
authors.forename: W K
authors.initials: WK
authors.lastname: Bleeker
authors.forename: J
authors.initials: J
authors.lastname: van der Plas
authors.forename: R I
authors.initials: RI
authors.lastname: Feitsma
authors.forename: J
authors.initials: J
authors.lastname: Agterberg
authors.forename: G
authors.initials: G
authors.lastname: Rigter
authors.forename: A
authors.initials: A
authors.lastname: de Vries-van Rossen
authors.forename: E K
authors.initials: EK
authors.lastname: Pauwels
authors.forename: J C
authors.initials: JC
authors.lastname: Bakker
mesh_terms: D015100:3,3'-Diaminobenzidine / Q000494:pharmacology; D000818:Animals; D003432:Cross-Linking Reagents; D005260:Female; D006454:Hemoglobins / Q000493:pharmacokinetics*; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008196:Lymph / Q000378:metabolism; D008657:Metabolic Clearance Rate; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D051381:Rats; D011919:Rats, Inbred Strains; D013667:Technetium; D014018:Tissue Distribution; D001743:Urinary Bladder / Q000378:metabolism
chemical_list: D003432:Cross-Linking Reagents; D006454:Hemoglobins; D015100:3,3'-Diaminobenzidine; C045124:2-nor-2-formylpyridoxal-5'-phosphate; D011732:Pyridoxal Phosphate; D013667:Technetium