title: Characterization of a long-acting recombinant human serum albumin-atrial natriuretic factor (ANF) expressed in Pichia pastoris.
abstract: The cardiac hormone atrial natriuretic factor (ANF) combines pharmacological properties of drugs used to treat essential hypertension (EH), congestive heart failure (CHF) and acute myocardial infarction (AMI). Treatment of CHF or AMI patients with an intravenous (iv) infusion of the circulating form of ANF (ANF(99-126)) produces significant clinical improvement. The short half-life (5 min) and peptide nature of ANF impose logistic restrictions for chronic administration. To increase its half-life, we fused ANF and human serum albumin (HSA) mini-genes by recombination in Pichia pastoris. The activity of three configurations of the fusion protein was tested in vitro and in vivo. The fusion protein that comprised of C-terminus HSA connected to N-terminus ANF via a hexaglycine linker showed the best outcome; it increased cGMP production in vitro. In vivo an iv bolus of HSA-ANF into mice increased significantly plasma cGMP levels and lowered blood pressure (BP) for up to 6 h hence successfully extended ANF half-life in plasma while retaining its biological activity. HSA-ANF represents the basis for development in the chronic therapeutic use of ANF.
authors.affiliation: University of Ottawa Heart Institute, Ottawa, ON, Canada K1Y 4E7. mdebold@ottawaheart.ca
authors.forename: Mercedes Kuroski
authors.initials: MK
authors.lastname: de Bold
authors.forename: William P
authors.initials: WP
authors.lastname: Sheffield
authors.forename: Amy
authors.initials: A
authors.lastname: Martinuk
authors.forename: Varsha
authors.initials: V
authors.lastname: Bhakta
authors.forename: Louise
authors.initials: L
authors.lastname: Eltringham-Smith
authors.forename: Adolfo J
authors.initials: AJ
authors.lastname: de Bold
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000627:therapeutic use*; D001794:Blood Pressure / Q000187:drug effects*; D006152:Cyclic GMP / Q000378:metabolism*; D006207:Half-Life; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000627:therapeutic use; D010843:Pichia / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000097:blood / Q000627:therapeutic use*; D012709:Serum Albumin / Q000235:genetics*
chemical_list: D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; C054547:atrial natriuretic factor prohormone (1-30), human; D009320:Atrial Natriuretic Factor; D006152:Cyclic GMP