5.1. pH-Sensitive PEG Release While normal tissues and blood have a physiological pH near 7.4, human tumors have lower pH values (~6.0/6.5) because of an elevated rate of glycolysis [275, 276]. pH-sensitive bonds have been developed for the coupling of PEG to liposomes [277] (Figure 1). pH-sensitive liposomes achieved

a higher concentration Inhibitors,research,lifescience,medical of cargo in the cytoplasm and nucleus than non-pH-sensitive PEGylated liposomes in vitro and allowed faster intratumoral content release in vivo [278, 279]. In addition to tumor sensitivity, pH sensitive BIBF 1120 manufacturer groups can potentiate the efficacy of targeted drug-loaded liposomes. Folate-targeting of daunorubicin-loaded liposomes by incorporation of a pH-sensitive folate-PEG-cholesterol hemisuccinate (CHEMS) conjugate combined tumor targeting and increased Inhibitors,research,lifescience,medical drug release at the tumor site with improved chemotherapeutic activity over untargeted liposomes [280]. Similarly, untargeted cisplatin-loaded liposomes or EGFR-targeted gemcitabine-loaded liposomes incorporating CHEMS had superior antitumor activity over untargeted drug-loaded liposomes or free drugs [281, 282]. Obata et al. used a

glutamic acid-based zwitterionic lipid (1,5-dihexadecyl N,N-diglutamyl-lysyl-L-glutamate) as titratable lipid for doxorubicin delivery [283]. These liposomes showed a charge inversion from negative to positive at acidic Inhibitors,research,lifescience,medical pH with endosomal escape leading to higher doxorubicin delivery in the cytoplasm and higher toxicity in vitro over conventional liposomes. This resulted in superior antitumor activity in vivo. Biswas et al. developed a new pH-sensitive DSPE-PEG-hydrazone-PEG2000 conjugate for attachment of ligands to the liposome surface Inhibitors,research,lifescience,medical [284]. In their work, the cell penetrating peptide (TATp) was unmasked after PEG release at acidic pH allowing efficient cellular uptake. Recently, three Inhibitors,research,lifescience,medical new approaches for generation of pH sensitivity have been reported. First, by electrostatic adsorption

of negatively charged carboxyl-modified gold nanoparticles to the surface of cationic liposomes (egg dipalmitoylphosphatidylcholine/DOTAP 9:1 weight ratio) at pH 7 (pKa of 5 for the carboxylic below group) [285]. Authors reported detachment of gold nanoparticles at acidic pH due to protonation of the carboxyl groups and speculated that a similar strategy could be applied with negative charged liposomes and amine-modified gold nanoparticles. Second, a platform for finely tuned pH-induced PEG release was introduced using phenyl-substituted-vinyl-ether-(PIVE)-PEG lipid conjugates [286]. Liposomes containing PIVE showed pH-induced dePEGylation and content release at acidic pH whereas they were stable at physiological pH. Third, ligand unmasking by acidic pH-induced membrane reorganization has been introduced as a reversible ligand-masking strategy.