title: Scale up and pharmacokinetic study of a novel mutated chimeric tissue plasminogen activator (mt-PA) in rats.
abstract: Because of high mortality caused by cardiovascular diseases, various fibrinolytic agents with diverse pharmacokinetic and pharmacodynamic properties have been developed. A novel mutated chimeric tissue plasminogen activator (mt-PA) was developed by the removal of first three domains of t-PA, insertion of GHRP sequence and mutation towards resistance to plasminogen activator inhibitor-1 (PAI-1). Mt-PA protein was expressed in Expi293F cells. The expression level of mt-PA was found to be 5000 IU/mL. Following purification, the pharmacokinetic properties of mt-PA were evaluated in three doses in rats. Data related to mt-PA were best fitted to two compartment model. With the increase in dose, the Area Under the plasma concentration-time Curve (AUC0→∞) increased. The elimination half-life (t1/2) of mt-PA was in the range of 19.1-26.1 min in three doses while that of Alteplase was 8.3 min. The plasma clearance (CLp) of mt-PA ranged from 3.8 to 5.9 mL/min in three doses, which was several times lower than that of Alteplase (142.6 mL/min). The mean residence time (MRT) of mt-PA ranged from 23.3-31.8 min in three doses, which was 4-5 times greater than that of Alteplase (6 min). Mt-PA showed extended half-life and mean residence time and is a good candidate for further clinical studies.
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
authors.forename: Mozhgan
authors.initials: M
authors.lastname: Raigani
authors.affiliation: Biopharmaceutics Division, Department of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran 14155/6451 Iran.
authors.forename: Mohammad-Reza
authors.initials: MR
authors.lastname: Rouini
authors.affiliation: Department of human vaccines and sera, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran.
authors.forename: Ali-Akbar
authors.initials: AA
authors.lastname: Golabchifar
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
authors.forename: Esmat
authors.initials: E
authors.lastname: Mirabzadeh
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
authors.forename: Behrouz
authors.initials: B
authors.lastname: Vaziri
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
authors.forename: Farzaneh
authors.initials: F
authors.lastname: Barkhordari
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Davami
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
authors.forename: Fereidoun
authors.initials: F
authors.lastname: Mahboudi
mesh_terms: D000818:Animals; D019540:Area Under Curve; D005260:Female; D057809:HEK293 Cells; D006207:Half-Life; D006801:Humans; D016296:Mutagenesis; D009842:Oligopeptides / Q000235:genetics; D012372:ROC Curve; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000493:pharmacokinetics; D010959:Tissue Plasminogen Activator / Q000235:genetics / Q000378:metabolism*
chemical_list: D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C048770:glycyl-histidyl-arginyl-proline; D010959:Tissue Plasminogen Activator