Neutralization of Daboxin P activities by rationally designed aptamers

Area of Science:

• Biochemistry
• Molecular Biology
• Pharmacology

Background:

• Current snakebite antivenoms face challenges with efficacy and safety, necessitating novel therapeutic strategies.
• Aptamers, highly specific nucleic acid molecules, offer potential for both diagnostics and therapeutics in snakebite management.

Purpose of the Study:

• To design and validate nucleic acid aptamers targeting snake venom phospholipase A2 (PLA2).
• To assess the inhibitory potential of these aptamers against PLA2 enzymatic and biological activities.

Main Methods:

• In silico docking and molecular dynamics simulations to predict aptamer binding to PLA2 enzymes.
• Electrophoretic mobility shift assays (EMSA) to confirm aptamer-PLA2 binding.
• In vitro assays to measure inhibition of PLA2 enzymatic and anticoagulant activities.

Main Results:

• Designed aptamers demonstrated stable binding to the active and Ca2+ binding sites of various snake venom PLA2 enzymes.
• Aptamers effectively inhibited soluble PLA2 (sPLA2) activity (IC50: 0.52–0.77 μM) and anticoagulant effects of specific snake venoms.
• Inhibition was specific to PLA2 and anticoagulant activities, with no effect on proteolytic or hemolytic functions.

Conclusions:

• Nucleic acid aptamers can be rationally designed to specifically inhibit the biochemical and biological activities of snake venom PLA2.
• These findings support the development of aptamer-based therapeutics and diagnostics for snakebite envenomation.