title: Intravenous application of an anticalin dramatically lowers plasma digoxin levels and reduces its toxic effects in rats.
abstract: Lipocalins tailored with high affinity for prescribed ligands, so-called anticalins, constitute promising candidates as antidotes. Here, we present an animal study to investigate both pharmacokinetic and clinical effects of an anticalin specific for the digitalis compound digoxin. Intravenous digoxin (2.5-50 μg/kg/min) was administered to rats until first changes in the ECG occurred (dose finding study) or a priori for 30 min (kinetic study). The anticalin DigA16(H86N), dubbed DigiCal, was administered intravenously at absolute doses of 1, 5, 10 and 20 mg, while the control group received isotonic saline. Hemodynamic changes, several ECG parameters and digoxin concentration in plasma were monitored at given time intervals. After DigiCal administration free digoxin concentration in plasma ultrafiltrate declined dramatically within 1 min to the presumably non-toxic range. There was also a significant and DigiCal dose-dependent effect on longer survival, less ECG alterations, arrhythmia, and improved hemodynamics. Infusion of a lower digoxin dose (2.5 μg/kg/min) resulted in a more sustained reduction of free digoxin in plasma after DigiCal administration compared to a higher digoxin dose (25 μg/kg/min), whereas ECG and hemodynamic parameters did not markedly differ, reflecting the known relative insensitivity of rats towards digoxin toxicity. Notably, we observed a re-increase of free digoxin in plasma some time after bolus administration of DigiCal, which was presumably due to toxin redistribution from tissue in combination with the relatively fast renal clearance of the rather small protein antidote. We conclude that anticalins with appropriately engineered drug-binding activities and, possibly, prolonged plasma half-life offer prospects for next-generation antidotal therapy.
authors: Department of Toxicology, Klinikum rechts der Isar, Munich, Germany. Florian.Eyer@mac.com
authors: Florian
authors: F
authors: Eyer
authors: Werner
authors: W
authors: Steimer
authors: Thomas
authors: T
authors: Nitzsche
authors: Nicole
authors: N
authors: Jung
authors: Heidi
authors: H
authors: Neuberger
authors: Christine
authors: C
authors: Müller
authors: Martin
authors: M
authors: Schlapschy
authors: Thomas
authors: T
authors: Zilker
authors: Arne
authors: A
authors: Skerra
mesh_terms: D000818:Animals; D000931:Antidotes / Q000008:administration & dosage / Q000494:pharmacology*; D002316:Cardiotonic Agents / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000633:toxicity*; D004077:Digoxin / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004562:Electrocardiography; D006207:Half-Life; D007262:Infusions, Intravenous; D054834:Lipocalins / Q000008:administration & dosage / Q000494:pharmacology*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015996:Survival Rate; D013997:Time Factors
chemical_list: D000931:Antidotes; D002316:Cardiotonic Agents; C575984:DigA16; D054834:Lipocalins; D004077:Digoxin