Entrapment of small molecules and nucleic acid-based drugs in liposomes.
In the past two decades there have been major advances in the development of liposomal drug delivery systems suitable for applications ranging from cancer chemotherapy to gene therapy. In general, an optimized system consists of liposomes with a diameter of approximately 100 nm that possess a long circulation lifetime (half-life >5 h). Such liposomes will circulate sufficiently long to take advantage of a phenomenon known as disease site targeting, wherein liposomes accumulate at sites of disease, such as tumors, as a result of the leaky vasculature and reduced blood flow exhibited by the diseased tissue. The extended circulation lifetime is achieved by the use of saturated lipids and cholesterol or by the presence of PEG-containing lipids. This chapter will focus on the methodology required for the generation of two very different classes of liposomal carrier systems: those containing conventional small molecular weight (usually anticancer) drugs and those containing larger genetic (oligonucleotide and plasmid DNA) drugs. Initially, we will examine the encapsulation of small, weakly basic drugs within liposomes in response to transmembrane pH and ion gradients. Procedures will be described for the formation of large unilamellar vesicles (LUVs) by extrusion methods and for loading anticancer drugs into LUVs in response to transmembrane pH gradients. Three methods for generating transmembrane pH gradients will be discussed: (1) the use of intravesicular citrate buffer, (2) the use of transmembrane ammonia gradients, and (3) ionophore-mediated generation of pH gradients via transmembrane ion gradients. We will also discuss the loading of doxorubicin into LUVs by formation of drug-metal ion complexes. Different approaches are required for encapsulating macromolecules within LUVs. Plasmid DNA can be encapsulated by a detergent-dialysis approach, giving rise to stabilized plasmid-lipid particles, vectors with potential for systemic gene delivery. Antisense oligonucleotides can be spontaneously entrapped upon electrostatic interaction with ethanol-destabilized cationic liposomes, giving rise to small multilamellar systems known as stabilized antisense-lipid particles (SALP). These vectors have the potential to regulate gene expression.
Fenske
David B
DB
Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada.
Cullis
Pieter R
PR
D000890:Anti-Infective Agents / Q000378:metabolism; D000903:Antibiotics, Antineoplastic / Q000378:metabolism; D002021:Buffers; D002939:Ciprofloxacin / Q000378:metabolism; D004317:Doxorubicin / Q000378:metabolism; D016503:Drug Delivery Systems; D018014:Gene Transfer Techniques; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007476:Ionophores / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry; D009696:Nucleic Acids / Q000008:administration & dosage*; D016376:Oligonucleotides, Antisense / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000008:administration & dosage*; D010743:Phospholipids / Q000737:chemistry; D010957:Plasmids / Q000235:genetics / Q000378:metabolism
D000890:Anti-Infective Agents; D000903:Antibiotics, Antineoplastic; D002021:Buffers; D007476:Ionophores; D008081:Liposomes; D009696:Nucleic Acids; D016376:Oligonucleotides, Antisense; D004364:Pharmaceutical Preparations; D010743:Phospholipids; D002939:Ciprofloxacin; D004317:Doxorubicin