Enhanced plasmid DNA delivery using anionic LPDII by listeriolysin O incorporation

Background A major obstacle to achieving effective DNA-based therapeutics is efficient delivery of the DNA to its site of action in the cell. Upon internalization by endocytosis, the endosomal membrane represents a critical physical barrier preventing access of DNA to the cell cytosol. In order to overcome the membrane barrier and facilitate cytosolic entry, the endosomolytic bacterial protein listeriolysin O (LLO) is a potentially promising agent. Methods LLO was incorporated in an anionic liposome-entrapped polycation-condensed DNA delivery system (LPDII). Plasmid DNA was condensed using protamine sulfate and then complexed to anionic liposomes. LLO was incorporated into the delivery vehicle through encapsulation in anionic, pH-sensitive liposomes. Transfection levels were monitored using a model reporter plasmid encoding luciferase in P388D1 cells, a macrophage-like cell line. Results Transfection using the anionic LPDII delivery platform was enhanced through incorporation of LLO. Additionally, the net charge of the condensate, the lipid composition, and the total amount of LLO-liposomes were all capable of modulating the transfection levels of the vehicle. Importantly, in the presence of serum, transfection levels using the LLO-containing LPDII system were comparable to established cationic lipid delivery systems. Conclusions LLO is capable of facilitating transfection using an anionic LPDII system. This anionic delivery vehicle represents the successful combination of the LPDII system for condensation of the DNA with the unique endosomolytic properties of LLO for improved transfection using plasmid DNA. Copyright © 2005 John Wiley & Sons, Ltd.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/48699/1/750_ftp.pd

Discovery of a Highly Selective JAK2 Inhibitor, BMS-911543, for the Treatment of Myeloproliferative Neoplasms

JAK2 kinase inhibitors are a promising new class of agents for the treatment of myeloproliferative neoplasms and have potential for the treatment of other diseases possessing a deregulated JAK2-STAT pathway. X-ray structure and ADME guided refinement of C-4 heterocycles to address metabolic liability present in dialkylthiazole <b>1</b> led to the discovery of a clinical candidate, BMS-911543 (<b>11</b>), with excellent kinome selectivity, <i>in vivo</i> PD activity, and safety profile