Polymeric Nanoparticles Containing Combination Antiretroviral Drugs for HIV Type 1 Treatment

Abstract

The use of combination antiretroviral nanoparticles (cART NPs) was investigated as a novel treatment approach for the inhibition of HIV-1 replication. We developed nanoparticles of biodegradable polymer, poly-(dl-lactide-coglycolic acid; PLGA) containing efavirenz (EFV) and boosted lopinavir (lopinavir/ritonavir; LPV/r) by a highpressure homogenization method. The method resulted in > 79% drug entrapment efficiency for each of the three drugs. The average size of cART NPs was 138.3 -55.4 nm as measured by dynamic light scanning, confirmed by scanning electron microscopy (SEM) with an average surface charge of -13.7 -4.5. Lissamine-rhodamine-labeled fluorescent PLGA NPs exhibited efficient uptake in nonimmune (HeLa cells) and immune (H9 T cells) cells as measured by confocal microscopy. Cells treated with cART NPs resulted in minimal loss of cell viability over 28 days. Subcellular fractionation studies demonstrated that HIV-1-infected H9 monocytic cells treated with cART NPs contained significantly ( p < 0.05) higher nuclear, cytoskeleton, and membrane antiretroviral drug levels compared to cells treated with drug solutions alone. Finally, cART NPs efficiently inhibited HIV-1 infection and transduction. The IC 50 for each of the three drugs in the cART NPs was < 31 nM. These experiments demonstrate the efficacy of a novel PLGA NPs formulation for the delivery of cART to inhibit HIV-1 replication.

Preparation, characterization, and safety evaluation of poly(lactide-co-glycolide) nanoparticles for protein delivery into macrophages

“…In 2013, Shibata et al reported no significant cytotoxicity up to 28 days of treatment with 500 µg/mL of PLGA NPs using three cell models including the human monocyte U937 cell model. 39 Therefore, we and others have demonstrated that PLGA NPs do not affect cell activity even at concentrations well above therapeutically relevant doses (100 µg/mL and below), confirming their potential as delivery nanocarriers for therapeutic applications. Interestingly, in this study, we observed that cells treated with PLGA-BSA NPs showed an increase in cell proliferation.…”

“…The results of our cytotoxicity assays support previous studies showing that PLGA NPs do not significantly affect human MDM cell viability. 38,39,[56][57][58] In the present study, PLGA NPs were not found to be cytotoxic for neither TZM-bl nor human MDMs when cells were exposed for 48 hours at concentrations up to 4,000 or 500 µg/mL, respectively. In 2013, Shibata et al reported no significant cytotoxicity up to 28 days of treatment with 500 µg/mL of PLGA NPs using three cell models including the human monocyte U937 cell model.…”

“…20,24 However, despite major breakthroughs in the development of nanopolymeric particles for vaccine and drug delivery into macrophages, there are still many limitations to overcome before their use in clinical applications. 33,34 Although PLGA co-polymer has been used in several studies as micro/nanoparticles (NPs) delivery systems with a certain degree of success 20,30,32,[35][36][37] including delivery of antimicrobial drugs into monocytes/macrophages, 29,[38][39][40][41][42] their safety for therapeutic application is still unknown. Indeed, few studies have focused on the inflammatory profile of PLGA nanopolymeric carriers and, to our knowledge, no study has yet investigated the effect of PLGA-based NPs on polymorphonuclear neutrophil (PMN) apoptosis and recruitment.…”

“…Therefore, as shown in this study and previous studies, the use of polymeric-based nanocarriers is a promising strategy to deliver therapeutic agents in human MDMs and to target pathogens that proliferate inside macrophages including visceral leishmaniasis, malaria, and tuberculosis. 29,[38][39][40] It is well known that PLGA-based material is suitable for therapeutic application since it is biodegradable and show low toxic effects. The results of our cytotoxicity assays support previous studies showing that PLGA NPs do not significantly affect human MDM cell viability.…”

Anti-HIV Drug-Combination Nanoparticles Enhance Plasma Drug Exposure Duration as Well as Triple-Drug Combination Levels in Cells Within Lymph Nodes and Blood in Primates

“…While a number of investigators have developed single-drug nanoparticles to be combined at the time of administration, 22,23 this approach may result in heterogeneity of intracellular drug concentrations and potentially harbor drug-resistant virus. Solid polymeric particles 100-600 nm in diameter containing multiple crystalline drugs have been synthesized, 24 but these are much larger than anti-HIV LNPs and may not be suitable for lymphatic uptake. 25 While the mechanisms of drug action in anti-HIV LNPs remain to be elucidated, cellular uptake of intact nanoparticles followed by pH-or phospholipase-dependent release of active drug likely leads to enhanced intracellular levels.…”