Novel method of measurement of in vitro drug uptake in OATP1B3 overexpressing cells in the presence of dextran

Pharmacol Rep. 2024 Apr;76(2):400-415. doi: 10.1007/s43440-024-00583-8. Epub 2024 Mar 26.

Abstract

Background: In predictions about hepatic clearance (CLH), a number of studies explored the role of albumin and transporters in drug uptake by liver cells, challenging the traditional free-drug theory. It was proposed that liver uptake can occur for transporter substrate compounds not only from the drug's unbound form but also directly from the drug-albumin complex, a phenomenon known as uptake facilitated by albumin. In contrast to albumin, dextran does not exhibit binding properties for compounds. However, as a result of its inherent capacity for stabilization, it is widely used to mimic conditions within cells.

Methods: The uptake of eight known substrates of the organic anion-transporting polypeptide 1B3 (OATP1B3) was assessed using a human embryonic kidney cell line (HEK293), which stably overexpresses this transporter. An inert polymer, dextran, was used to simulate cellular conditions, and the results were compared with experiments involving human plasma and human serum albumin (HSA).

Results: This study is the first to demonstrate that dextran increases compound uptake in cells with overexpression of the OATP1B3 transporter. Contrary to the common theory that highly protein-bound ligands interact with hepatocytes to increase drug uptake, the results indicate that dextran's interaction with test compounds does not significantly increase concentrations near the cell membrane surface.

Conclusions: We evaluated the effect of dextran on the uptake of known substrates using OATP1B3 overexpressed in the HEK293 cell line, and we suggest that its impact on drug concentrations in liver cells may differ from the traditional role of plasma proteins and albumin.

Keywords: Dextran; In vitro uptake; OATP1B3.

MeSH terms

  • Albumins
  • Dextrans*
  • HEK293 Cells
  • Hepatocytes / metabolism
  • Humans
  • Liver
  • Liver-Specific Organic Anion Transporter 1 / genetics
  • Liver-Specific Organic Anion Transporter 1 / metabolism
  • Liver-Specific Organic Anion Transporter 1 / pharmacology
  • Membrane Transport Proteins / metabolism
  • Organic Anion Transporters* / genetics
  • Organic Anion Transporters* / metabolism
  • Organic Anion Transporters, Sodium-Independent / genetics
  • Organic Anion Transporters, Sodium-Independent / metabolism
  • Solute Carrier Organic Anion Transporter Family Member 1B3 / genetics
  • Solute Carrier Organic Anion Transporter Family Member 1B3 / metabolism
  • Solute Carrier Organic Anion Transporter Family Member 1B3 / pharmacology

Substances

  • Dextrans
  • Solute Carrier Organic Anion Transporter Family Member 1B3
  • Liver-Specific Organic Anion Transporter 1
  • Organic Anion Transporters
  • Membrane Transport Proteins
  • Albumins
  • Organic Anion Transporters, Sodium-Independent