Biochemical determinants of clearance of tissue-type plasminogen activator from the circulation.
Biochemical modification of tissue-type plasminogen activator (t-PA) designed to alter pharmacokinetics and pharmacodynamics offers promise for development of pharmaceuticals particularly suitable for treatment of specific disorders and for induction of coronary thrombolysis by intramuscular as well as intravenous administration. Accordingly, to identify biochemical determinants of clearance of t-PA from the circulation, we injected rabbits intravenously with three different preparations of t-PA synthesized from the same human gene and expressed in Chinese hamster ovary cells cultured under disparate conditions. Influences of glycosylation on clearance were defined by experiments with enzymatically treated t-PA in which clearance was assessed with concomitant administration of selected neoglycoproteins that compete with t-PA for specific glycoprotein receptors. The role of an intact active catalytic site, as reflected by differences in clearance with and without prior treatment of t-PA with the protease inhibitor PPACK, was defined also. Results indicate that clearance is altered by inhibition of the active site and that the nature and extent of glycosylation--not evident simply by analysis of peptide structure--influence clearance as well. These findings suggest that mannose/N-acetylglucosamine-specific glycoprotein receptors expressed on hepatic reticuloendothelial cells participate in clearance of t-PA from the circulation but that galactose-specific glycoprotein receptors probably do not. The observations may explain differences in clearance seen with different preparations of t-PA that have been seen in clinical pilot studies and may identify biochemical determinants of clearance amenable to modification for development of agents with potentially desirable, specific biological properties.
D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D006207:Half-Life; D006801:Humans; D008099:Liver / Q000378:metabolism; D008358:Mannose / Q000378:metabolism; D037241:Mannose-Binding Lectins; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D011817:Rabbits; D011971:Receptors, Immunologic / Q000378:metabolism; D013329:Structure-Activity Relationship; D010959:Tissue Plasminogen Activator / Q000493:pharmacokinetics*
D006023:Glycoproteins; D037241:Mannose-Binding Lectins; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D011971:Receptors, Immunologic; C032193:glycoprotein receptor GPIb-IX; C035313:mannose-N-acetylglucosamine receptor; D010959:Tissue Plasminogen Activator; D008358:Mannose; D000117:Acetylglucosamine