Quantitative study on a simple electrochemical dsDNA-pregabalin biosensor; multi-spectroscopic, molecular docking and...

Area of Science:

• Biochemistry
• Molecular Biology
• Analytical Chemistry

Background:

• Pregabalin (PGB), a GABA analog, is widely used for neuropathic pain, fibromyalgia, and neuralgia.
• Understanding drug-target interactions at the molecular level is crucial for drug development and efficacy.
• Deoxyribonucleic acid (DNA) is a potential target for various therapeutic agents.

Purpose of the Study:

• To investigate the binding mechanism and interactions between Pregabalin (PGB) and double-stranded fish sperm deoxyribonucleic acid (dsDNA).
• To develop a sensitive electrochemical biosensor for the detection of PGB using its interaction with dsDNA.

Main Methods:

• Spectroscopic techniques (UV-Vis absorption spectroscopy, fluorescence spectroscopy)
• Electrochemical methods (differential pulse voltammetry)
• Molecular docking and molecular dynamics simulations
• Binding studies using ethidium bromide and Hoechst-33258 assays

Main Results:

• Experimental and computational data confirmed that PGB binds to dsDNA primarily through minor groove binding.
• The binding constant (Kb) was determined to be 2.41×10^4 ± 0.30 at 298 K, with a binding stoichiometry (n) of 1.21 ± 0.09.
• A sensitive PGB-dsDNA biosensor was developed, enabling detection in the μM range with LOD of 0.57 μM and LOQ of 1.91 μM.

Conclusions:

• Pregabalin interacts with dsDNA via minor groove binding, elucidated by a combination of experimental and theoretical approaches.
• The established PGB-dsDNA interaction provides a foundation for developing novel electrochemical biosensing platforms for PGB quantification.
• This study offers insights into the molecular interactions of PGB with DNA, potentially relevant for understanding its pharmacological effects and developing new therapeutic strategies.