Once the starting phase-points have been generated in Step 4, we are ready to launch the second major computational and massively parallel stage in the vDSSB approach, namely the swarm of the bound state annihilation trajectories and the swarm of the unbound state growth trajectories.
The annihilation trajectories are generated in a sub-directory 
fsdam to be created by the end-user of the bound state
directory containing the HREM data (USER_SCRATCH/bound) and by
the application script  maketprQ.sh  provided in the
SI.zip archive attached to the JCIM note. In the specific example of
      the the complex PF-07321332-3CLpro, the script generates 384 folders
(fsdam/bi) containing the GROMACS tpr
file to switch off the ligand atomic charges in 0.375 ns, starting
from the configurations generated in Step 4.
The fast switching simulations are submitted on the HPC using the
batch submission
script submit.slr (also
  provided in the SI.zip archive attached to the JCIM note) which allows:
maketprQ.sh;
fsdam/bi folders using as starting point the
      confout.gro configuration file of previous run;
 maketprQ.sh  and submit.slr use
the mdp files 
transitionQ_mdp and 
transitionvdw.mdp also provided in SI.zip attached to the JCIM
note. The bound state ligand annihilation job requires about two wall
clock time hours on the Marconi100 HPC platform.
Using the bash script 
maketprvdw.sh (also provided in the SI.zip attached to JCIM
note), the tpr GROMACS files to perform unbound fast
switching alchemical simulations are generated in
the fsdam user-created subdirectory of the main HREM
directory for the unbound state 
  USER_SCRATCH/unbound.
The script creates for 192 folders (fsdam/ui)
containing the tpr files to switch on the LJ ligand-water
interactions in 0.360 ns, starting from the configurations generated
in Step 4.
The simulations are submitted
      using submit.slr which allows:
tpr files generated with
  maketprvdw.sh;
  tpr files using as starting point the confout.gro
    configuration file of previous run;
maketprvdw.sh and submit.slr use the mdp
files transitionvdw.mdp and transitionQ.mdp
provided in SI.zip attached to the JCIM note. The unbound ligand growth required few wall clock minutes on the Marconi100
  HPC cluster.