A two-step targeting approach for delivery of doxorubicin-loaded liposomes to tumour cells in vivo.
A two-step targeting approach was used to deliver doxorubicin-loaded liposomes to a murine tumour cell (P388 leukaemia) grown in culture and, more importantly, in vivo. Targeting was mediated through the use of an antibody specific for the Thy 1.2 antigen that is highly expressed on P388 cells. Briefly, the approach consists of prelabeling target cells with biotinylated anti-Thy 1.2 antibody prior to administration of drug-loaded liposomes that have streptavidin covalently attached to their surface. Results from in vitro studies demonstrate that a 30-fold increase in cell-associated lipid and a 20-fold increase in cell-associated doxorubicin can be achieved over control liposomes using this two-step procedure. Flow-cytometry and fluorescent-microscopy data were used to confirm that P388 cells can be stably labeled with the biotinylated anti-Thy 1.2 antibody in vivo. Subsequently, liposome-targeting studies were initiated in vivo, where target cell binding was assessed following i.p. or i.v. injection of doxorubicin-loaded liposomes into animals bearing P388 tumours prelabeled with biotinylated antibody. A streptavidin-mediated 3.7-fold increase in cell-associated lipid and drug was achieved when the liposomes were given i.p. When doxorubicin-loaded streptavidin liposomes were injected i.v., P388 cells located in the peritoneal cavity were specifically labeled, although the efficiency of this targeting reaction was low. Less than a 2-fold increase in cell-associated lipid was achieved through the use of target-specific (streptavidin-coated) liposomes. These studies demonstrate that the presence of a well-labeled target cell population within the peritoneal cavity will not promote accumulation of an i.v. injected, targeted liposomal drug. Furthermore, the importance of separating target-cell-specific binding from non-specific uptake by tumour-associated macrophages is discussed.
Longman
S A
SA
University of British Columbia, Department of Biochemistry, Faculty of Medicine, Vancouver, Canada.
Cullis
P R
PR
Choi
L
L
de Jong
G
G
Bally
M B
MB
D000818:Animals; D001426:Bacterial Proteins; D004317:Doxorubicin / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000627:therapeutic use; D004337:Drug Carriers; D005260:Female; D005434:Flow Cytometry; D007941:Leukemia P388 / Q000188:drug therapy*; D008081:Liposomes; D008657:Metabolic Clearance Rate; D051379:Mice; D008815:Mice, Inbred Strains; D008856:Microscopy, Fluorescence; D010533:Peritoneal Lavage; D010713:Phosphatidylcholines; D019809:Streptavidin
D001426:Bacterial Proteins; D004337:Drug Carriers; D008081:Liposomes; D010713:Phosphatidylcholines; D004317:Doxorubicin; D019809:Streptavidin; C010942:1,2-distearoyllecithin