TARGETABILITY OF NOVEL IMMUNOLIPOSOMES MODIFIED WITH AMPHIPATHIC POLY(ETHYLENE GLYCOL)S CONJUGATED AT THEIR DISTAL TERMINALS TO MONOCLONAL-ANTIBODIES

Distearoyl-N-(3-carboxypropionoyl poly(ethylene glycol) succinyl)phosp hatidylethanolamine (DSPE-PEG-COOH) was newly synthesized and used to prepare novel immunoliposomes carrying monoclonal antibodies at the di stal ends of the PEG chains (Type C). Liposomes were prepared from egg phosphatidylcholine (ePC) and cholesterol (CH) (2:1, m/m) containing 6 mol% of DSPE-PEG-COOH, and a monoclonal IgG antibody, 34A, which is highly specific to pulmonary endothelial cells, was conjugated to the carboxyl groups of DSPE-PEG-COOH to give various amounts of antibody m olecules per liposome. Other immunoliposomes with PEG coating (Type B) or without PEG coating (an earlier type of immunoliposome, Type A) we re prepared for comparison. The average molecular weight of PEG in Typ e B or C immunoliposomes was 2000. Type B and Type C liposomes without antibodies showed prolonged circulation time and reduced reticulo-end othelial system (RES) uptake owing to the presence of PEG. These three different types of 34A-immunoliposomes with 30-35 antibody molecules per vesicle were injected into mice to test the immunotargetability to the lung. The efficiency of lung binding of 34A-Type B was one-half o f that of 34A-Type A, though a large amount of 34A-Type B remained in the blood circulation for a long time, suggesting that the steric hind rance of PEG chains reduced not only the immunospecific antibody-antig en binding, but also the RES uptake. The degree of lung binding of 34A -Type C was about 1.3-fold higher than that of 34A-Type A, indicating that recognition by the antibodies attached to the PEG terminal was no t sterically hindered and that the free PEG (i.e., that not carrying a ntibody) was effective in increasing the blood concentration of immuno liposomes by enabling them to evade RES uptake. The latter phenomenon was confirmed by using nonspecific antibody-Type C immunoliposomes (14 -Type C), which showed a high blood level for a long time. Our approac h provides a simple means of conjugating antibodies directly to the di stal end of PEG which is already bound to the liposome membrane, and s hould contribute to the development of superior targetable drug delive ry vehicles for use in diagnostics and therapy.