Anti-CD20 multivalent HPMA copolymer-Fab' conjugates for the direct induction of apoptosis.
A hybrid biomimetic system comprising high-molecular-weight, linear copolymer of N-(2-hydroxypropyl)methacrylamide (HPMA) grafted with multiple Fab' fragments of anti-CD20 monoclonal antibody (mAb) was synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization followed by attachment of Fab' fragments via thioether bonds. Exposure of human non-Hodgkin's lymphoma (NHL) Raji B cells to the multivalent conjugates resulted in crosslinking of CD20 receptors and commencement of apoptosis. Five conjugates with varying molecular weight and valence (amount of Fab' per polymer chain) were synthesized. One of the copolymers contained enzyme degradable peptide sequences (GFLG) in the backbone. The multivalency led to higher avidity and apoptosis induction compared to unconjugated whole mAb. Time-dependent studies showed that the cytotoxicity of conjugates exhibited a slower onset at shorter exposure times than mAb hyper-crosslinked with a secondary Ab; however, at longer time intervals the HPMA copolymer conjugates achieved significantly higher biological efficacies. In addition, study of the relationship between the structure of conjugates and Raji B cell apoptosis revealed that both valency and polymer molecular weight influenced biological activities, while insertion of peptide sequences into the backbone was not a factor in vitro.
Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, Utah 84112, USA.
Te-Wei
TW
Chu
Jiyuan
J
Yang
Jindřich
J
Kopeček
D058846:Antibodies, Monoclonal, Murine-Derived / Q000494:pharmacology; D018951:Antigens, CD20 / Q000737:chemistry* / Q000276:immunology; D017209:Apoptosis; D032701:Biomimetics; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry*; D008228:Lymphoma, Non-Hodgkin / Q000276:immunology* / Q000628:therapy; D008689:Methacrylates / Q000737:chemistry*; D018613:Microscopy, Confocal / Q000379:methods; D008956:Models, Chemical; D011108:Polymers / Q000737:chemistry*; D000069283:Rituximab; D013997:Time Factors
D058846:Antibodies, Monoclonal, Murine-Derived; D018951:Antigens, CD20; D007140:Immunoglobulin Fab Fragments; D008689:Methacrylates; D011108:Polymers; D000069283:Rituximab; D053148:Caspase 3; C032802:hydroxypropyl methacrylate