title: Application of accelerator mass spectrometry to macromolecules: preclinical pharmacokinetic studies on a polybisphosphonate.
abstract: Data on the use of accelerator mass spectrometry (AMS) in conjunction with in vivo studies of macromolecular drugs are scarce. The present study shows the versatility of this technique when investigating the pharmacokinetics (PK) of a macromolecular drug candidate, a polybisphosphonate conjugate (ODX). The aforementioned is a polymer (molecular weight ~30 kDa) constituting a carbohydrate backbone with covalently linked ligands (aldendronate and aminoguanidine) and is intended for treatment of osteoporosis and the therapy of bone metastasis from prostate cancer. The conjugate is prepared through partial oxidation of the carbohydrate and sequential coupling of the ligands by reductive amination. (14)C was incorporated in the conjugate by means of coupling a commercially available (14)C-lysine in the conjugation sequence. Fifteen rats were injected intravenously with (14)C-labelled ODX (150 µg, 14 Bq/rat) and blood samples were collected at 1, 2, 4, 6, and 24 h post-injection (3 rats/time point). Liver, spleen and kidney samples were collected at 4 and 24 h post-injection. Blood from each time point (triplicate) were collected for AMS measurement determining the isotopic ratio ((14)C/(12)C) and consequently the drug concentration in blood. ODX showed a transient presence in blood circulation; 93% of the total dose was cleared from the circulation within 1 h. The half-life after 1 h was estimated to be about 3 h; 0.7% of the administered (14)C dose of ODX remained in circulation after 24 h. The major (14)C accumulation was in the liver, the spleen and the kidneys indicating the probable route of metabolism and excretion. This study demonstrates the versatility of AMS for pharmacological in vivo studies of macromolecules. Labelling with (14)C is relatively simple, inexpensive and the method requires minimal radioactivity, eliminating the need for radioprotection precautions in contrast to methods using scintillation counting.
authors.affiliation: Ion Physics, Department of Physics and Astronomy, Uppsala University, Uppsala, Sweden. mehran.salehpour@angstrom.uu.se
authors.forename: Mehran
authors.initials: M
authors.lastname: Salehpour
authors.forename: Karl
authors.initials: K
authors.lastname: Håkansson
authors.forename: Urban
authors.initials: U
authors.lastname: Höglund
authors.forename: Annika
authors.initials: A
authors.lastname: Grahn-Westin
authors.forename: Sten
authors.initials: S
authors.lastname: Nilsson
authors.forename: Marcela
authors.initials: M
authors.lastname: Márquez
authors.forename: Göran
authors.initials: G
authors.lastname: Possnert
authors.forename: Anders R
authors.initials: AR
authors.lastname: Holmberg
mesh_terms: D019386:Alendronate / Q000737:chemistry; D000818:Animals; D002247:Carbon Isotopes / Q000737:chemistry; D004164:Diphosphonates / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006146:Guanidines / Q000737:chemistry; D046911:Macromolecular Substances / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D011108:Polymers / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014018:Tissue Distribution
chemical_list: D002247:Carbon Isotopes; D004164:Diphosphonates; D006146:Guanidines; D046911:Macromolecular Substances; D011108:Polymers; C004479:pimagedine; D019386:Alendronate