title: Synthesis of cobalamin dimers using isophthalate cross-linking of corrin ring carboxylates and evaluation of their binding to transcobalamin II.
abstract: Several cobalamin (Cbl) dimers have been prepared for evaluation as potential antiproliferative agents in the treatment of AIDS-related lymphoma. The Cbl dimers were synthesized by cross-linking Cbl carboxylates, produced by acid hydrolysis of the b-, d-, and e-propionamide side chains of cyanocobalamin (CN-Cbl), through an isophthalate molecule. Linking molecules were used between the Cbl carboxylates and the isophthalate moiety. The linkers were incorporated to provide a distance between the two Cbl molecules such that the dimeric Cbls might bind two molecules of transcobalamin II (TCII), the Cbl transport protein in plasma. Initially, the linking moiety used was 1,12-diaminododecane, but the resulting dimers had low aqueous solubility. To improve the solubility of the dimers, 4,7,10-trioxa-1,13-tridecanediamine was employed as the linking moiety. This improved the water solubility of the dimers considerably, while retaining the distance between the Cbl molecules at 41-42 A (fully extended). To introduce additional substitution on Cbl dimers, 5-aminoisophthalic acid was used as the cross-linking reagent. p-Iodobenzoyl and p-(tri-n-butylstannyl)benzoyl conjugates of 5-aminoisophthalate were synthesized and used to prepare Cbl dimers. The stannylbenzoyl-conjugated Cbl dimers were prepared as precursors to be used in radioiodination reactions, and the iodobenzoyl-conjugated Cbl dimers were prepared as HPLC standards for the radioiodinated product. Attempts to iodinate/radioiodinate the stannylbenzoyl Cbl dimers were unsuccessful. Although an explanation for this is not readily apparent, the failure to react may be due to the lipophilicity of the linker used and the steric environment of the two Cbl moieties. A biotinylated derivative of 5-aminoisophthalate was also synthesized and used to prepare biotinylated-Cbl dimers. In a competitive rhTCII binding assay with [57Co]CN-Cbl, Cbl dimers containing the lipophilic diaminododecane linking moiety had decreased binding avidities compared to those of Cbl monomers substituted at the same corrin ring carboxylate. However, Cbl dimers containing the water-solubilizing trioxadiamine linker appeared to have avidities similar to those of the Cbl monomers.
authors.affiliation: Department of Radiation Oncology, University of Washington, Seattle 98195, USA.
authors.forename: P M
authors.initials: PM
authors.lastname: Pathare
authors.forename: D S
authors.initials: DS
authors.lastname: Wilbur
authors.forename: D K
authors.initials: DK
authors.lastname: Hamlin
authors.forename: S
authors.initials: S
authors.lastname: Heusser
authors.forename: E V
authors.initials: EV
authors.lastname: Quadros
authors.forename: P
authors.initials: P
authors.lastname: McLoughlin
authors.forename: A C
authors.initials: AC
authors.lastname: Morgan
mesh_terms: D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D003432:Cross-Linking Reagents; D019281:Dimerization; D006801:Humans; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D010795:Phthalic Acids; D011994:Recombinant Proteins / Q000378:metabolism; D014155:Transcobalamins / Q000378:metabolism*; D014805:Vitamin B 12 / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D003432:Cross-Linking Reagents; D007457:Iodine Radioisotopes; D010795:Phthalic Acids; D011994:Recombinant Proteins; D014155:Transcobalamins; C059768:isophthalate; D014805:Vitamin B 12