PEGylated PAMAM dendrimers as a potential drug delivery carrier: in vitro and in vivo comparative evaluation of covalently conjugated drug and noncovalent drug inclusion complex.

To understand more about the influence of the types of interaction between drug and PEGylated PAMAM dendrimers on the in vitro and in vivo behavior of drug, methotrexate (MTX) was coupled to PEGylated or non-PEGylated generation 4 PAMAM (G4) through complexing drug within the dendritic architecture and covalently conjugated onto the surface of the dendrimer, respectively. PAMAM was first modified with PEG(5000) chains at three different degrees of substitution. The ability of PEGylated G4 complexing MTX was higher than that of non-PEGylated one. MTX-G4 and MTX-G4-PEG conjugates were synthesized via amide linkages. MTX was readily released from all complexes in isotonic solution, while the conjugates hardly released MTX in the same medium and keep stable in human plasma and the lysosomal medium. There were no obvious differences between complexes and free MTX in cytotoxicity against KB cell line, whereas the conjugates showed the relatively low activity. In vivo study in rodents found that the MTX-G4-PEG conjugate exhibited significantly prolonged blood residence time and the strongest antitumor effects, as compared with MTX-G4, the complexes and MTX. The results indicated that the covalent attachment of drug to PEGylated PAMAM could be more effective for targeted drug delivery.