A simple passive equilibration method for loading carboplatin into pre-formed liposomes incubated with ethanol as a temperature dependent permeability enhancer

Abstract A passive equilibration method which relies on addition of candidate drugs to pre-formed liposomes is described as an alternative method for preparing liposome encapsulated drugs. The method is simple, rapid and applicable to liposomes prepared with high (45 mol%) or low (  60 °C. Super-saturated solutions of CBDCA (40 mg/mL) can be prepared at 70 °C and these are stable in the presence of ethanol even when the temperature is reduced to C. maximum CBDCA encapsulation is achieved within 1 h after the CBDCA solution is added to pre-formed DSPC/Chol liposomes in the presence of 30% (v/v) ethanol at 60 °C. When the pre-formed liposomes are mixed with ethanol (30% v/v) at or below 40 °C, the encapsulation efficiency is reduced by an order of magnitude. The method was also applied to liposomes prepared from other compositions include a cholesterol free formulations (containing 1,2-distearoyl-sn-glycero-3-phosphoethanolamine- N -[carboxy(polyethylene glycol)-2000] (DSPE-PEG 2000 )) and a low Chol ( sn -glycero-3-phospho-(1′- rac -glycerol) DSPG)). The cytotoxic activity of CBDCA was unaffected when prepared in this manner and two of the resultant formulations exhibited good stability in vitro and in vivo . The cytotoxic activity of CBDCA was unaffected when prepared in this manner and the resultant formulations exhibited good stability in vitro and in vivo . Pharmacokinetics studies in CD-1 mice indicated that the resulting formulations increased the circulation half life of the associated CBDCA significantly (AUC 0–24 h of CBDCA = 0.016 μg·hr/mL; AUC 0-24h of the DSPC/Chol CBDCA formulation = 1014.0 μg·hr/mL and AUC 0-24h of the DSPC/DSPG/Chol CBDCA formulation = 583.96 μg·hr/mL). Preliminary efficacy studies in Rag-2M mice with established subcutaneous H1975 and U-251 tumors suggest that the therapeutic activity of CBDCA is improved when administered in liposomal formulations. The encapsulation method described here has not been disclosed previously and will have broad applications to drugs that would normally be encapsulated during liposome manufacturing.