Solutes for the two legs of the thermodynamic cycle were assembled. One leg of the cycle contained the protein (parameterized by the Amber99sb*ILDN force field) from the 4d09 crystal structure , the crystallographic waters, and the hybrid ligands positioned according to their previously determined binding poses. The other branch contained only the hybrid ligands. The structures were then solvated with TIP3P 55 water and 0.15 mM sodium and chloride ions parameterized by Joung and Cheatham 56 in a dodecahedral simulation box with 1.5 Å padding. All subsequent simulations were carried out with Gromacs 2021.6 44 with a 2 fs integration time step.