Nanoformulated Antiretroviral Drug Combinations Extend Drug Release and Antiretroviral Responses in HIV-1-Infected Macrophages: Implications for NeuroAIDS Therapeutics

Abstract

We posit that improvements in pharmacokinetics and biodistributions of antiretroviral therapies (ART) for human immunodeficiency virus-type one infected people can be achieved through developments in nanoformulations. To this end, we manufactured nanoparticles of atazanavir, efavirenz, and ritonavir (termed nanoART) and treated human monocyte-derived macrophages (MDM) in combination therapies. This resulted in improved drug uptake, release and antiretroviral efficacy over monotherapy. MDM rapidly, within minutes, ingested nanoART combinations, at equal or similar rates, as individual formulations. Combination nanoART ingested by MDM facilitated drug release from 15 to > 20 days. These findings are noteworthy as a nanoART cell-mediated drug delivery provides a means to deliver therapeutics to viral sanctuaries, such as the central nervous system during progressive human immunodeficiency virus-type one infection. The work brings us yet another step closer to realizing the utility of nanoART for virus-infected people.

Endosomal Trafficking of Nanoformulated Antiretroviral Therapy Facilitates Drug Particle Carriage and HIV Clearance

“…Cells were exposed to native ATV or nanoATV for 24 h at 30 and 70 g/ml for each treatment. Previous studies in our laboratory had shown that these concentrations of native and nanoATV are not toxic to MDM (5,6). Substantial (about 3 orders of magnitude) differences were observed in the rate and extent of drug uptake between the native and nanoATV treatment groups (Fig.…”

“…This is seen in nanoART's abilities to maintain consistent plasma and tissue drug levels, as demonstrated in our previous studies (4). Nonetheless, to facilitate clearance of human immunodeficiency virus type 1 (HIV-1), antiretroviral drugs need to be effectively delivered to viral sanctuaries (5). This can target persistent or restricted infection (6)(7)(8).…”

Comparative manufacture and cell-based delivery of antiretroviral nanoformulations

“…12 Cell supernatant samples (10 µL) were mixed with 10 µL of 100 mM Tris-HCl (pH 7.9), 300 mM KCl, 10 mM DTT, and 0.1% nonyl phenoxylpolyethoxylethanol-40 (NP-40) in a 96-well plate. The samples were incubated at 37°C for 15 minutes.…”

“…The samples were incubated at 37°C for 15 minutes. Twenty-five microliters of a solution containing 50 mM Tris-HCl (pH 7.9), 150 mM KCl, 5 mM DTT, 15 mM MgCl 2 , 0.05% NP-40, 10 µg/mL poly(A), 0.250 U/mL oligo d(T), [12][13][14][15][16][17][18] and 10 µCi/mL 3 H-TTP was then added to each well and incubated at 37°C for 18 hours. Following incubation, 50 µL of ice-cold 10% trichloroacetic acid was added to each well, and the well contents were harvested onto glass fiber filters and assessed for 3 H-TTP incorporation by β-scintillation spectroscopy.…”

“…To this end, improvements in drug pharmacodynamics were described for ART nanoformulations (nanoART). 4,11,12 Prior works demonstrated that such nanoART could be carried in monocyte-macrophages and reach viral sanctuaries. [11][12][13][14][15] These studies also showed that size, shape, and charge of crystalline indinavir (IDV), ritonavir (RTV), atazanavir (ATV), and efavirenz (EFV) affect uptake, release, cytotoxicities, and antiretroviral responses.…”