Liposome-mediated delivery of antiviral agents to human immunodeficiency virus-infected cells.
Intracellular delivery of novel macromolecular drugs against human immunodeficiency virus type-1 (HIV-1), including antisense oligodeoxynucleotides, ribozymes and therapeutic genes, may be achieved by encapsulation in or association with certain types of liposomes. Liposomes may also protect these drugs against nucleases. Low-molecular-weight, charged antiviral drugs may also be delivered more efficiently via liposomes. Liposomes were targeted to HIV-1-infected cells via covalently coupled soluble CD4. An HIV-1 protease inhibitor encapsulated in conventional negatively charged multilamellar liposomes was about 10-fold more effective and had a lower EC90 than the free drug in inhibiting HIV-1 production in human monocyte-derived macrophages. The drug encapsulated in sterically stabilized liposomes was as effective as the free drug. The EC50 of the reverse transcriptase inhibitor 9-(2-phosphonylmethoxyethyl)adenine (PMEA) was reduced by an order of magnitude when delivered to HIV-1-infected macrophages in pH-sensitive liposomes. A 15-mer antisense oligodeoxynucleotide against the Rev response element was ineffective in free form against HIV-1 replication in macrophages, while delivery of the oligonucleotide in pH-sensitive liposomes inhibited virus replication. The oligodeoxynucleotide encapsulated in sterically stabilized pH-sensitive liposomes with prolonged circulation in vivo, which were recently developed in the laboratories of the authors, was also highly effective. A ribozyme complementary to HIV-1 5'-LTR delivered in pH-sensitive liposomes inhibited virus production by 90%, while the free ribozyme caused only a slight inhibition. Cationic liposome-mediated co-transfection of the HIV-regulated diphtheria toxin A fragment gene and a proviral HIV clone into HeLa cells completely inhibited virus production, while the frame-shifted mutant gene was ineffective. Co-transfection of the proviral genome and a gene encoding a Rev-binding aptamer into HeLa cells via transferrin-associated cationic liposomes inhibited virus production. These studies indicate that liposomes can be used to facilitate the intracellular delivery of certain anti-HIV agents and to enhance their therapeutic effects. These properties may be particularly advantageous in the development of novel macromolecular drugs, which may be necessary because of the emergence of virus strains resistant to the currently available drugs.
Düzgüneş
N
N
Department of Microbiology, School of Dentistry, University of the Pacific, San Francisco, CA 94115, USA. nduzgune@uop.edu
Pretzer
E
E
Simões
S
S
Slepushkin
V
V
Konopka
K
K
Flasher
D
D
de Lima
M C
MC
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D019380:Anti-HIV Agents / Q000008:administration & dosage*; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers; D015316:Genetic Therapy / Q000379:methods*; D006367:HeLa Cells / Q000382:microbiology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000008:administration & dosage*; D008264:Macrophages / Q000382:microbiology; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage; D063065:Organophosphonates; D011480:Protease Inhibitors / Q000008:administration & dosage; D013997:Time Factors
D019380:Anti-HIV Agents; D004337:Drug Carriers; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D063065:Organophosphonates; D011480:Protease Inhibitors; C053001:adefovir; D000225:Adenine