Figure 2.
Synthesized CypD inhibitor C-9 binds to immobilized CypD in a dose-dependent manner. Surface plasmon resonance (SPR) analysis of CypD–sulfonamide derivative binding interaction, where the globally fit data (black lines) were overlaid with experimental data (red lines). Human recombinant CypD protein (10 μg/mL) was immobilized directly on the hydrophilic carboxy methylated dextran matrix of the CM5 sensor chip (Biacore-3000) using the standard primary amine coupling reaction according to standard procedures. Compound C-9 (2.5, 5, 10, 15, and 20 μM) was injected at a flow rate of 40 μL/min for the determination of the kinetic and equilibrium constants. All data analyses were carried out using BIA evaluation software, and sensor grams were processed by automatic correction for nonspecific bulk refractive index effects. The dissociation constant (KD) was determined as indicated at 25 °C.