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This project reproduces a minimal free energy perturbation (FEP) workflow for two PIM kinase inhibitors using OpenFE / OpenMM

System Overview

  • Protein: Human PIM1 kinase
  • Ligands:
    • Compound 8 (PIM447 / LGH447) — PDB 5DWR
    • Compound 3 / 5c — PDB 4N70 (same compound, renumbered between 2013–2015 papers)
  • Experimental Ki (PIM1):
    • 5c/3: 1 pM
    • 8: 6 pM
      → ~6× tighter binding for 5c/3
      → ΔΔG (exp, 8→3) ≈ −1.06 kcal mol⁻¹

Simulation Summary

  • Software: OpenFE 1.7 + OpenMM 8.2
  • Protocol: Minimal RBFE (12 λ per leg, 250 ps prod / 50 ps eq per λ)
  • Legs: complex (Protein + Ligand) and solvent (water box)
  • Runs: 6 replicates per leg
  • Temperature: 300 K Pressure: 1 bar Solvent: explicit TIP3P

Results

Leg ⟨ΔG⟩ (kcal mol⁻¹) SEM
Complex −24.25 0.08
Solvent −22.77 0.07
ΔΔG(8 → 3) −1.48 ± 0.11

Predicted fold-change: e^(ΔΔG / RT) ≈ 12× tighter for 3/5c vs 8
Experimental fold-change: ≈ 6× tighter
Agreement: same direction, slightly larger magnitude (+0.4 kcal mol⁻¹ deviation)

Interpretation

  • The FEP reproduced the correct trend with reasonable magnitude.
  • Sign flip checks confirm proper ligand ordering (negative ΔΔG = j tighter).
  • Minor overestimation likely stems from limited sampling (250 ps per λ).

Files

  • rbfe_python_tutorial_two_ligands.ipynb — notebook performing setup, run, and analysis
  • — ligand coordinates
  • — receptor coordinates
  • README.md — this summary

References

  1. J. Med. Chem. 2015, 58, 6599–6616 — Identification of N-(4-((1R,3S,5S)-3-amino-5-methylcyclohexyl)… (LGH447) — reports 6 pM Ki for compound 8.
  2. ACS Med. Chem. Lett. 2013, 4, 436–440 — Structure-Guided Optimization of Pan-PIM Inhibitors — reports 1 pM Ki for compound 5c (same as 3). """

readme_path = Path("/mnt/data/README.md") readme_path.write_text(readme_text) readme_path

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