title: Aggregation properties of a HPMA-camptothecin copolymer in isotonic solutions.
abstract: Copolymers of camptothecin (CPT) and [N-(2-hydroxypropyl) methacrylamide] (HPMA) are novel anticancer drugs that show improved pharmacological profile in animal models as compared to the free drug CPT. We investigate here the aggregation properties of a HPMA-glycyl-6-aminohexanoyl-glycyl-CPT copolymer ( approximately 20,000 Da). The molecular size of HPMA-copolymer CPT is followed over 5 orders of magnitudes of concentration in isotonic buffer by measuring either the time resolved fluorescence anisotropy (FA) of CPT or the autocorrelation function of the light scattered by the copolymer. A detailed analysis of these data suggests the presence of elongated structures with axial ratio approximately 3 in the range 0.1-0.5 microg/ml and aggregates with association number higher than 2 in more concentrated solutions (up to 10 mg/ml). The binding affinity of HPMA-copolymer CPT for serum albumin is inversely dependent on the degree of aggregation of the copolymer. We also show that the copolymer concentration in plasma from mice treated with an active, non-toxic, dose of HPMA-copolymer CPT, decreases from 3 to 0.01 mg/ml in 72 h. In the same range of concentrations in vitro, we do not detect hydrophobic aggregates of polymers with high (>3) association number. Our study indicates that the circulating HPMA-copolymer CPT in mice should not undergo extensive aggregation and should interact with serum albumin more weakly than free CPT.
authors.affiliation: Department of Physics, Università di Milano-Bicocca, and Istituto Nazionale per la Fisica della Materia, Piazza della Scienza 3, 20126 Milan, Italy. giuseppe.chirico@unimib.it
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Chirico
authors.forename: Maddalena
authors.initials: M
authors.lastname: Collini
authors.forename: Francesca
authors.initials: F
authors.lastname: Olivini
authors.forename: Moreno
authors.initials: M
authors.lastname: Zamai
authors.forename: Enrico
authors.initials: E
authors.lastname: Frigerio
authors.forename: Valeria R
authors.initials: VR
authors.lastname: Caiolfa
mesh_terms: D016880:Anisotropy; D002166:Camptothecin / Q000737:chemistry*; D007201:Indicator Dilution Techniques; D007552:Isotonic Solutions / Q000737:chemistry; D008689:Methacrylates / Q000737:chemistry*; D015394:Molecular Structure; D011108:Polymers / Q000737:chemistry*; D012709:Serum Albumin / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013997:Time Factors
chemical_list: D007552:Isotonic Solutions; D008689:Methacrylates; D011108:Polymers; D012709:Serum Albumin; C032802:hydroxypropyl methacrylate; D002166:Camptothecin