Synthesis, Acoustic Stability, and Pharmacologic Activities of Papaverine-Loaded Echogenic Liposomes for Ultrasound Controlled Drug Delivery

Abstract

Background: development of encapsulated therapeutics that could be released upon ultrasound exposure has strong implications for enhancing drug effects at the target site. We have developed echogenic liposomes (ELIP) suitable for ultrasound imaging of blood flow and ultrasound-mediated intravascular drug release. Papaverine was chosen as the test drug because its clinical application requires high concentration in the target vascular bed but low concentration in the systemic circulation. Methods: the procedure for preparation of standard ELIP was modified by including Papaverine hydrochloride in the lipid hydration solution, followed by three freeze-thaw cycles to increase encapsulation of the drug. Sizing and encapsulation pharmacokinetics were performed using a Coulter counter and a phosphodiesterase activity assay. Stability of Papaverine-loaded ELIP (PELIP) was monitored with a clinical diagnostic ultrasound scanner equipped with a linear array transducer at a center frequency of 4.5 MHz by assessing the mean digital intensity within a region of interest over time. The stability of PELIP was compared to those of standard ELIP and Optison™. Results: relative to standard ELIP, PELIP were larger (median diameter = 1.88 ± 0.10 μm for PELIP vs 1.08 ± 0.15 μm for ELIP) and had lower Mean Gray Scale Values (MGSV) (92 ± 24.8 for PELIP compared to 142.3 ± 10.7 for ELIP at lipid concentrations of 50 μg/ml). The maximum loading efficiency and mean encapsulated concentration were 24% ± 7% and 2.1 ± 0.7 mg/ml, respectively. Papaverine retained its phosphodiesterase inhibitory activity when associated with PELIP. Furthermore, a fraction of this activity remained latent until released by dissolution of liposomal membranes with detergent. The stability of both PELIP and standard ELIP were similar, but both are greater than that of Optison™. Conclusions: our results suggest that PELIP have desirable physical, biochemical, biological, and acoustic characteristics for potential in vivo administration and ultrasound-controlled drug delivery.

Keywords:

• Liposomes
• papaverine
• drug encapsulation
• ultrasound contrast agents

This study was presented in part at the American College of Cardiology Scientific Sessions, New Orleans, Louisiana, March 24–27, 2007. The work was supported in part by grant no. HL-074002 from the National Heart, Lung, and Blood Institute, National Institute of Health, Bethesda, Maryland, and a grant from the American Institute of Ultrasound in Medicine.