title: Development of a carbon-14 labeling approach to support disposition studies with a pegylated biologic.
abstract: Although it is widely accepted that one can extend the pharmacokinetic half-life of a therapeutic protein by covalent conjugation with polyethylene glycol (PEG), the disposition properties of such biologics have not yet been fully evaluated. Therefore, a novel [¹⁴C]-labeling method was developed that can be applied to a biologic conjugated with PEG through a maleimide-cysteine reaction. The method was used to study the tissue and tumor distribution of a PEGylated Adnectin, a recombinant protein derived from the 10th type III domain of fibronectin, in nude mice bearing human xenograft tumors. The PEGylated Adnectin contained a 40-kDa branched PEG (P40B) that was labeled with [¹⁴C] at the linker region between the PEG and Adnectin, without compromising cellular activity and plasma half-life in mice. After a single intravenous or intraperitoneal dose (33 mg/kg, 1.7 μCi per mouse) of [¹⁴C]-P40B-Adnectin, quantitative whole-body autoradiography analysis revealed that the liver had the highest uptake of the radioactivity among nontumor tissues, followed by the kidneys and lung. The muscle and brain showed the least penetration of the radioactivity among all tissues examined. In addition, the [¹⁴C]-P40B-EI-tandem penetrated into the tumor tissue, although the extent of accumulation was largely dependent on tumor type. Therefore, it was possible to assess the tissue distribution of a PEGylated biologic after it had been [¹⁴C] labeled using the novel method described herein.
authors.affiliation: Metabolism and Pharmacokinetics, Pharmaceutical Candidate Optimization, Bristol-Myers Squibb Research and Development, Route 206 and Province Line Rd., Princeton, NJ 08543, USA. haiqing.wang@bms.com
authors.forename: Haiqing
authors.initials: H
authors.lastname: Wang
authors.forename: Lifei
authors.initials: L
authors.lastname: Wang
authors.forename: Kai
authors.initials: K
authors.lastname: Cao
authors.forename: Stuart L
authors.initials: SL
authors.lastname: Emanuel
authors.forename: Paul
authors.initials: P
authors.lastname: Morin
authors.forename: Zheng
authors.initials: Z
authors.lastname: Lin
authors.forename: Guoxiang
authors.initials: G
authors.lastname: Shen
authors.forename: Jennifer
authors.initials: J
authors.lastname: Hosbach
authors.forename: Donglu
authors.initials: D
authors.lastname: Zhang
authors.forename: Samuel
authors.initials: S
authors.lastname: Bonacorsi
authors.forename: A David
authors.initials: AD
authors.lastname: Rodrigues
authors.forename: Zheng
authors.initials: Z
authors.lastname: Yang
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D001345:Autoradiography; D001688:Biological Products / Q000008:administration & dosage / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D002250:Carbon Radioisotopes / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics*; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000378:metabolism; D005260:Female; D005353:Fibronectins / Q000008:administration & dosage / Q000138:chemical synthesis / Q000235:genetics / Q000493:pharmacokinetics*; D006801:Humans; D007274:Injections, Intraperitoneal; D007275:Injections, Intravenous; D007553:Isotope Labeling / Q000379:methods*; D051379:Mice; D008819:Mice, Nude; D009154:Mutation; D009369:Neoplasms / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation; D011092:Polyethylene Glycols / Q000008:administration & dosage / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D015202:Protein Engineering; D017526:Receptor, IGF Type 1 / Q000378:metabolism; D011994:Recombinant Proteins / Q000493:pharmacokinetics; D014018:Tissue Distribution; D047368:Tumor Burden; D051598:Whole Body Imaging
chemical_list: D000970:Antineoplastic Agents; D001688:Biological Products; D002250:Carbon Radioisotopes; D005353:Fibronectins; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; C512478:EGFR protein, human; D066246:ErbB Receptors; D017526:Receptor, IGF Type 1