Development of a Rapid Mass Spectrometric Determination of AMP and Cyclic AMP for PDE3 Activity Study: Application and Computational Analysis for Evaluating the Effect of a Novel 2-oxo-1,2-dihydropyridine-3-carbonitrile Derivative as PDE-3 Inhibitor.
Ilaria CicaliniBarbara De FilippisNicola GambacortaAntonio Di MicheleSilvia ValentinuzziAlessandra AmmazzalorsoAlice Della ValleRosa AmorosoNicolotti OrazioPiero Del BoccioLetizia GiampietroPublished in: Molecules (Basel, Switzerland) (2020)
A simple, quick, easy and cheap tandem mass spectrometry (MS/MS) method for the determination of adenosine monophosphate (AMP) and cyclic adenosine monophosphate (cAMP) has been newly developed. This novel MS/MS method was applied for the evaluation of the inhibitory effect of a novel 2-oxo-1,2-dihydropyridine-3-carbonitrile derivative, also named DF492, on PDE3 enzyme activity in comparison to its parent drug milrinone. Molecule DF492, with an IC50 of 409.5 nM, showed an inhibition of PDE3 greater than milrinone (IC50 = 703.1 nM). To explain the inhibitory potential of DF492, molecular docking studies toward the human PDE3A were carried out with the aim of predicting the binding mode of DF492. The presence of different bulkier decorating fragments in DF492 was pursued to shift affinity of this novel molecule toward PDE3A compared to milrinone in accordance with both the theoretical and experimental results. The described mass spectrometric approach could have a wider potential use in kinetic and biomedical studies and could be applied for the determination of other phosphodiesterase inhibitor molecules.
Keyphrases
- protein kinase
- solid phase extraction
- molecular docking
- tandem mass spectrometry
- high performance liquid chromatography
- ultra high performance liquid chromatography
- ms ms
- molecularly imprinted
- liquid chromatography tandem mass spectrometry
- simultaneous determination
- endothelial cells
- photodynamic therapy
- gas chromatography
- molecular dynamics simulations
- risk assessment
- human health
- mass spectrometry
- water soluble