Monitoring antiviral active metabolite (N-hydroxycytidine) levels in plasma in presence of carboxylesterase-2 inhibitor verapamil using copper tetracyanoquinodimethane enhanced sensor

This study focuses on the determination of the active metabolite of molnupiravir (MOL), N-hydroxycytidine (NHC), using a square wave voltammetric (SWV) method. Carboxylesterase-2 enzymes catalyze the conversion of MOL prodrug into NHC. However, co-administration of verapamil (VER), a carboxylesterase-2 inhibitor, may reduce the levels of NHC, leading to decreasing its antiviral activity. In this context, the levels of NHC and VER were simultaneously monitored using a carbon paste electrode modified with phase I of copper tetracyanoquinodimethane (CuTCNQ) which is highly conductive charge transfer complex. The as-designed sensor was characterized successfully using various spectroscopic techniques and Scanning Electron Microscopy (SEM). The electrochemical behavior of the newly fabricated probe was examined using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). This method demonstrated its efficacy in measuring NHC and VER levels in rabbit plasma samples, showing high sensitivity and selectivity. The calibration plots for the simultaneous quantitation of NHC and VER displayed excellent linearity over the concentration ranges of 50-1600 nmol/L for NHC and 10-250 nmol/L for VER. The limits of detection (LOD) and quantitation (LOQ) in rabbit plasma were found to be 15.2 and 50.8 nmol/L for NHC and 2.9 and 9.9 nmol/L for VER, respectively. Moreover, fundamental pharmacokinetic parameters were calculated for NHC before and after co-administration of VER. The results suggest that the SWV method using CuTCNQ-modified CPE can be a useful tool for the determination of NHC and VER levels in plasma samples, with potential applications in the monitoring of drugdrug interactions involving carboxylesterase-2 inhibitors.

Automated summary

This study developed a new method for determining the active metabolite of molnupiravir, N-hydroxycytidine, and simultaneously monitoring the levels of the metabolite and the inhibitor verapamil in samples. The method showed high sensitivity, selectivity, and was able to quantify N-hydroxycytidine and verapamil levels in plasma samples. This method can be a useful tool for monitoring drug-drug interactions involving carboxylesterase-2 inhibitors.