Computer-aided evaluation of two fluorescent hexanoic acid piperazineamides as potential ligands for insect cytochromes P450

Insects represent a large variable class of animals on Earth, playing important roles as pests and as model organisms for scientific research devoted to, for instance, pesticide development, brain functions, and pathologies like Alzheimer's and Parkinson's disease. Insects have cytochrome P450, a unique class of monooxygenases, which are responsible for reactions of fatty acid conversion into hormone-like substances or cuticular hydrocarbons, as well as for destroying pesticides.

Previously, we reported the synthesis of two fluorescent hexanoic acid piperazineamides, namely, N-hexanoyl-N’-(7-nitrobenzofurazan-4-yl)-piperazine (NpipHex) and N-hexanoyl-ciprofloxacin (CPFHex), with the ability to fluoresce. Because there are no available experimentally solved structures of insect P450s, we have created a small library (120 structures) of the enzymes from Lucilia cuprina and L. sericata (a polinator and a cause of cattle myiasis) and Locusta migratoria (a pest) using the AI-based platforms AlphaFold and AlphaFold2/GoogleColab. Autodock Vina (AV) together with the authored helper software for high-throughput virtual screening (HTVS) FYTdock was used for both hem residue positioning and the docking of the derivatives. It was found that affine binding with AV scores of -9 and lower was found, for instance, for NpipHex and CYP6a8 of L. cuprina (Uniprot id A0A0L0CMB2, score -9.9), of which the Drosophila homologue has been recently described as a fatty acid hydroxylase; CYP12A7 of L. cuprina (Uniprot id A9X930, score -9.8); and CYP307A2 of D.melanogaster (A8Y592, score -9.9).

Taking these results together, we have created a virtual library of some insect P450s models suitable for HTVS and found potential new affine interactions with our fluorescent hexanoic acid derivatives, providing a method for the in silico estimation of their new biological properties with respect to the development of new pest control tools.

This study was supported by GSPSR (Belarus) № 20210560.