The purpose of the present study is to evaluate the ability of ESMACS and TIES to estimate binding affinities of a set of 22 ligands (Table ) to the protein target. For the SMYD3 systems studied here, the binding site of the protein is well structured (Figure ), and the binding mode for the scaffold of the congeneric compounds (Table ) is well-defined in the crystal structure. It is thus likely, based on our foregoing criteria, that a reasonable prediction can be achieved, although the relatively large size of the binding site, the presence of multiple components in the structure including several crystallographic water molecules involved in binding, and the rotatable bonds at the R2 site of the compounds (Table ) still pose a challenge for the conformational sampling and hence the accuracy and precision of the predictions.