Optimization-by-design of hepatotropic lipid nanoparticles targeting the sodium-taurocholate cotransporting polypeptide

Abstract

Active targeting and specific drug delivery to parenchymal liver cells is a promising strategy to treat various liver disorders. Here, we modified synthetic lipid-based nanoparticles with targeting peptides derived from the hepatitis B virus large envelope protein (HBVpreS) to specifically target the sodium-taurocholate cotransporting polypeptide (NTCP; SLC10A1) on the sinusoidal membrane of hepatocytes. Physicochemical properties of targeted nanoparticles were optimized and NTCP-specific, ligand-dependent binding and internalization was confirmed in vitro. The pharmacokinetics and targeting capacity of selected lead formulations was investigated in vivo using the emerging zebrafish screening model. Liposomal nanoparticles modified with 0.25 mol% of a short myristoylated HBV derived peptide, that is Myr-HBVpreS2-31, showed an optimal balance between systemic circulation, avoidance of blood clearance, and targeting capacity. Pronounced liver enrichment, active NTCP-mediated targeting of hepatocytes and efficient cellular internalization were confirmed in mice by 111In gamma scintigraphy and fluorescence microscopy demonstrating the potential use of our hepatotropic, ligand-modified nanoparticles.

Light-triggered switching of liposome surface charge directs delivery of membrane impermeable payloads in vivo

“…To this end, the embryonic zebrafish has provided an invaluable pre-clinical in vivo screening platform, offering unprecedented opportunities to assess, analyse and optimise nanoparticle behaviour over an entire live organism (i.e. visualising total injected doses), at cellular resolution and in real time 42,60,61 . Furthermore, the presence and conserved function of key RES cell types enables predictive assessment of fundamentally important in vivo clearance mechanisms of nanoparticles 25,43 .…”

Optimized Photoactivatable Lipid Nanoparticles Enable Red Light Triggered Drug Release

“…Dox was excited at 488 nm argon laser and CellMask was excited with a 633 nm. [42] Zebrafish Pharmacokinetic Studies: Embryos from Tg(kdrl:eGFP) and Tg(AB/Tübingen) adult zebrafish (Danio rerio) were breaded at 28 °C in zebrafish culture media containing 30.4 ug mL −1 1-phenyl-2-thiourea (PTU) and maintained according to Swiss animal welfare regulations. Embryos were embedded in 0.3% agarose containing tricaine and PTU and injected with a calibrated volume of 1 nL into Duct of Cuvier (48 h post fertilization, hpf) and cardinal vein (72 hpf) using a micromanipulator (Wagner Instrumentenbau KG), a pneumatic Pico Pump PV830 (WPI) and a Leica S8APO microscope (Leica).…”

“…Quantitative image analysis was performed as previously described. [34,42] Dox Release Studies in Zebrafish Embryos: 48 hpf embryos from Tg(abc/ tu) were injected with 2 nL of 3 mg mL −1 Dox containing LNPs into the duct of Cuvier and incubated at 28 °C in PTU containing zebrafish culture media. 24 hpi, one set of zebrafish was irradiated with pulsedlight for additional 24 h. For quantitative analysis, confocal images of five fishes were processed applying a defined threshold, followed by area calculation in Fiji (ImageJ) 2.1.0.…”

Transporter-Targeted Nano-Sized Vehicles for Enhanced and Site-Specific Drug Delivery

“…For decades, the specific target of HBV on the sinusoidal membrane of hepatocytes was unknown until the interaction of the virus with the human sodium-taurocholate cotransporting polypeptide (NTCP/SLC10A1) was identified [ 137 ]. Witzigmann et al modified synthetic lipid-based nanoparticles with targeting peptides derived from the hepatitis B virus large envelope protein (HBVpreS) to specifically target NTCP/SLC10A1 on the sinusoidal membrane of hepatocytes [ 138 ]. NTCP-specific, ligand-dependent binding and internalization were confirmed in vitro.…”