Potential Transmembrane Proteins-transporters of Chelidonic Acid for its Intracellular Uptake: In Silico Simulation

Area of Science:

• Pharmacology
• Biochemistry
• Computational Biology

Background:

• Small molecules, defined as biologically active organic compounds under 1 kDa, are crucial in drug development due to their cell membrane permeability and ability to modulate intracellular signaling.
• Chelidonic acid (ChA), a small molecule (184 Da), exhibits diverse biological activities, yet its cellular uptake mechanisms and molecular targets are not well-understood.
• Understanding ChA's transport is essential for its therapeutic applications.

Purpose of the Study:

• To investigate the uptake of Cheildonic acid (ChA) by human cells.
• To identify potential transporter proteins involved in the intracellular transport of ChA.
• To utilize integrated in silico and in vitro methodologies for comprehensive analysis.

Main Methods:

• In vitro co-culturing of human MCF-7 cells with ChA, followed by High-Performance Liquid Chromatography (HPLC) to quantify residual ChA.
• In silico screening of candidate transporter proteins from databases, molecular docking using Autodock Vina, and molecular dynamics simulations (50 ns) with GROMACS.
• Analysis of protein-ligand interactions, channel visualization using HOLE and VMD, and statistical analysis with R (Newey-West estimator, Welch's t-test).

Main Results:

• Human MCF-7 cancer cells demonstrate the capacity to internalize Cheildonic acid (ChA) through common cellular transport mechanisms.
• Initial screening identified six transmembrane proteins as potential transporters for ChA.
• Further analysis highlighted GLUT3, SVCT1, and URAT1 as the most promising candidates, exhibiting structural and functional compatibility for ChA transport.

Conclusions:

• This research enhances the understanding of Cheildonic acid's (ChA) pharmacokinetic and pharmacodynamic properties.
• The identified transporters provide a foundation for the rational design of novel pharmaceutical agents based on ChA.
• The findings pave the way for optimizing ChA-based drug development strategies.