Fig. 2: Biophysical characterization of BI-0115-LOX-1 interaction.
a Characterization of small molecule binding to LOX129 by STD NMR. Aromatic region of the off-resonance spectrum showing all expected resonances of BI-0115 (labeled Off-resonance). STD spectrum (subtraction of on- and off-resonance spectrum) displays signals at the respective resonance frequencies of the small molecule indicative for binding to LOX-1 (BI-0115 STD +LOX-1). STD spectrum of BI-0115 in the absence of LOX-1 (BI-0115 STD -LOX-1) shows no signals supporting specific binding of the compound to LOX-1 in the middle row. b Surface plasmon resonance binding analysis of BI-0115 to immobilized LOX129. Binding of BI-0115 was assessed by measuring dose responsive changes (from 0.391 µM to 25 µM) in the refractive index. The Kd of 4.3 µM was determined by a fit to the change of the refractive index at equilibrium (n = 4). The figure shows a representative example of a sensorgramm and affinity fit. c ITC-Titration of 300 µM LOX129 dimer into 40 µM BI-0115 compound to determine the binding affinity (mean Kd = 6.99 µM, n = 3). The left graph presents the raw data. The integrated peak areas with the corresponding fit are shown at the right.
