title: Reassessment of diethylenetriaminepentaacetic acid (DTPA) as a chelating agent for indium-111 labeling of polypeptides using a newly synthesized monoreactive DTPA derivative.
abstract: Previous studies on indium-111 (111In) labeling of polypeptides and peptides using cyclic diethylenetriaminepentaacetic dianhydride (cDTPA) as a bifunctional chelating agent (BCA) have indicated that DTPA might be a useful BCA for 111In labeling of polypeptides at high specific activities when DTPA can be incorporated without inducing intra- or intermolecular cross-linking. To investigate this hypothesis, a monoreactive DTPA derivative with a maleimide group as the peptide binding site (MDTPA) was designed and synthesized. A monoclonal antibody (OST7, IgG1) was used as a model polypeptide, and conjugation of MDTPA with OST7, 111In radiolabeling of MDTPA-OST7, and the stability of 111In-MDTPA-OST7 were investigated using cDTPA and benzyl-EDTA derivatives as references. SDS-PAGE analysis demonstrated that while cDTPA induced intramolecular cross-linking, no such undesirable side reactions were observed with MDTPA. MDTPA generated 111In-labeled OST7 with high radiochemical yields at higher specific activities than those produced using cDTPA and benzyl-EDTA derivatives as the BCAs. Incubation of each 111In-labeled OST7 in human serum indicated that MDTPA generated 111In-labeled OST7 of much higher and a little lower stability than those derived from cDTPA and benzyl-EDTA derivatives, respectively. These findings indicated that the low in vivo stability of cDTPA-conjugated antibody reported previously is not attributable to low stability of 111In-DTPA but to formation of intramolecular cross-linking during cDTPA conjugation reactions. The present study also indicated that MDTPA and its precursor, the tetra-tert-butyl derivative of DTPA, would be useful BCAs for 111In radiolabeling of polypeptides that have rapid blood clearance with high specific activities.
authors.affiliation: Department of Radiopharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan. arano@pharm.kyoto-u.ac.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Arano
authors.forename: T
authors.initials: T
authors.lastname: Uezono
authors.forename: H
authors.initials: H
authors.lastname: Akizawa
authors.forename: M
authors.initials: M
authors.lastname: Ono
authors.forename: K
authors.initials: K
authors.lastname: Wakisaka
authors.forename: M
authors.initials: M
authors.lastname: Nakayama
authors.forename: H
authors.initials: H
authors.lastname: Sakahara
authors.forename: J
authors.initials: J
authors.lastname: Konishi
authors.forename: A
authors.initials: A
authors.lastname: Yokoyama
mesh_terms: D002614:Chelating Agents; D004355:Drug Stability; D006801:Humans; D007205:Indium Radioisotopes; D007553:Isotope Labeling; D004369:Pentetic Acid
chemical_list: D002614:Chelating Agents; D007205:Indium Radioisotopes; D004369:Pentetic Acid