title: Using small-angle neutron scattering to study the solution conformation of N-(2-hydroxypropyl)methacrylamide copolymer-doxorubicin conjugates.
abstract: Our past research developed two N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-doxorubicin (Dox) conjugates that became the first synthetic polymer-anticancer conjugates to be evaluated clinically. The first, FCE28068, contained Dox bound to the polymeric carrier via a tetrapeptidic linker (glycine-phenylalanine-leucine-glycine (GFLG)) (Mw approximately 30,000 g/mol; approximately 8 wt % drug), and the second, FCE28069, contained additionally galactosamine (Gal) (Mw approximately 30,000 g/mol; approximately 7.5 wt % Dox) again bound by a GFLG linker. Galactosamine was included to promote hepatocyte/hepatoma targeting via the asialoglycoprotein receptor. Both conjugates showed antitumor activity and were clinically less toxic than free Dox (2-5 fold). However, despite their similar chemical characteristics, the conjugates displayed a significantly different maximum-tolerated dose (MTD) in patients. The aim of this study, therefore, was to use small-angle neutron scattering (SANS) to explore the solution behavior of a small library of HPMA polymer conjugates including FCE28068, FCE28069, and their pharmaceutical formulations, plus as reference compounds HPMA copolymer-GFLG conjugates containing aminopropanol (Ap) or galactosamine (Gal) alone (i.e., without Dox). The SANS data obtained showed that HPMA copolymer-GFLG-Ap conjugates (containing 5 and 10 mol % side chains) showed evidence of polymer aggregation, however, no indication of aggregation was observed for FCE28068 and FCE28069 over the concentration range studied (2.5-50 mg/mL). Clear differences in the scattering behavior for the two conjugates were observed at equivalent concentration. Data were best fitted by a model for polydisperse Gaussian coils, and the HPMA copolymer-Dox conjugate with Gal (FCE28069) exhibited a larger radius of gyration (Rg) (by approximately 2.5 nm) compared to FCE28068. In conclusion, we have shown that SANS will be a valuable tool to elucidate conformation-performance relationships for polymer-drug conjugates.
authors.affiliation: School of Chemistry, Cardiff University, Main Building, Park Place, Cardiff CF10 3AT, United Kingdom. paula3@cardiff.ac.uk
authors.forename: Alison
authors.initials: A
authors.lastname: Paul
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Vicent
authors.forename: Ruth
authors.initials: R
authors.lastname: Duncan
mesh_terms: D000178:Acrylamides / Q000737:chemistry*; D004317:Doxorubicin / Q000737:chemistry*; D005688:Galactosamine / Q000737:chemistry*; D008968:Molecular Conformation; D033363:Neutron Diffraction; D053838:Scattering, Small Angle; D012996:Solutions / Q000737:chemistry
chemical_list: D000178:Acrylamides; C438845:HPMA coploymer-doxorubicin-galactosamine; D012996:Solutions; C482378:N-(2-hydroxypropyl)methacrylamide co-polymer-doxorubicin conjugate; D005688:Galactosamine; D004317:Doxorubicin