In vitro Pharmacokinetic Cell Culture System that Simulates Physiologic Drug and Nanoparticle Exposure to Macrophages

Abstract

Purpose An in vitro dynamic pharmacokinetic (PK) cell culture system was developed to more precisely simulate physiologic nanoparticle/drug exposure. Methods A dynamic PK cell culture system was developed to more closely reflect physiologic nanoparticle/drug concentrations that are changing with time. Macrophages were cultured in standard static and PK cell culture systems with rifampin (RIF; 5 μg/ml) or β-glucan, chitosan coated, poly(lactic-co-glycolic) acid (GLU-CS-PLGA) nanoparticles (RIF equivalent 5 μg/ml) for 6 h. Intracellular RIF concentrations were measured by UPLC/MS. Antimicrobial activity against M. smegmatis was tested in both PK and static systems. Results The dynamic PK cell culture system mimics a one-compartment elimination pharmacokinetic profile to properly mimic in vivo extracellular exposure. GLU-CS-PLGA nanoparticles increased intracellular RIF concentration by 37% compared to free drug in the dynamic cell culture system. GLU-CS-PLGA nanoparticles decreased M. smegmatis colony forming units compared to free drug in the dynamic cell culture system. Conclusions The PK cell culture system developed herein enables more precise simulation of human PK exposure (i.e., drug dosing and drug elimination curves) based on previously obtained PK parameters.

Hybrid Curdlan Poly(γ ‐Glutamic Acid) Nanoassembly for Immune Modulation in Macrophage

“…[ 32 ] Based on these findings our laboratory as well as others have developed nanoparticles with glucan to significantly enhance the immune modulation of macrophage, with the ultimate goal of reducing intracellular M.tb . We have previously shown β‐glucan as a ligand on PLGA‐CS nanoparticles enhanced cytokine production from macrophage [ 6,11 ] and reduced the number of CFU of Mycobacterium smegmatis , a mimetic of TB. While these nanoparticles are efficacious against M. smegmatis , we have found that synthesis of these nanoparticles is a complex, multi‐step synthesis process that is not feasible for scale up processes and the synthesis process ultimately reduces the amount of drug that can be delivered.…”

“…In the current study, we developed a simple curdlan (linear beta‐1,3‐glucan) based nanoparticle that is physically blended with PGA, thus bypassing the need for a PLGA‐CS based nanoparticles ultimately reducing the number of steps in nanoparticle synthesis. Our C‐NPs formed by blending of curdlan and PGA which eliminates the need for chemical modification or conjugation of curdlan that has been done previously [ 5,6,10,12,13,15,35 ] while maintaining the immunostimulatory capabilities of curdlan that is often lost with chemical modification with small molecules. [ 10,15 ] Blending of curdlan and PGA has suitable processability and obtains the multicarboxylates originated from PGA.…”

“…We have previous published studies using curdlan as a targeting ligand to macrophage; [5,6,11] however, synthesis of these nanoparticles is a complex, multi-step synthesis process that is not feasible for scale up processes. Therefore, we sought to develop a nanoformulation of curdlan that had the following properties; i) simple synthesis and ii) retains ability to stimulate macrophage.…”

“…Particulate curdlan binds to Dectin-1 expressed on immune cells such as dendritic cells, monocytes, macrophages, and B-cells, and stimulates immune cells to produce immune modulators such as cytokines. [1,2] It potentially has an inhibitory effect against HIV infection, [3,4] antibacterial therapy against tuberculosis (TB), [5,6] as well as blood anti-coagulant activity. [7] Previous studies have utilized curdlan as structural components of nanoparticles that modulate the immune system.…”

Nanomedicines for Infectious Diseases

“…Kutscher et al report the development of an in vitro dynamic pharmacokinetic cell (macrophage) culture system with the aim of precise simulation of drug elimination profiles in human. Rifampicin loaded PLGA based nanoparticles were used in the model (8). Such a PK model is intended to support translational studies to develop nanomedicines for infectious diseases.…”