Identification of Novel Small-Molecule Janus Kinase 1 and 2 Inhibitors Through Structure-Based Virtual Screening, In Vitro Assays, and Molecular Dynamics Simulations

Avcı A., Taşci H., Sağlık Özkan B. N., Tozkoparan Köprücü B., Gökhan Kelekçi N.

ACS OMEGA, cilt.1, ss.1-17, 2026 (SCI-Expanded, Scopus)

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 1
  • Basım Tarihi: 2026
  • Doi Numarası: 10.1021/acsomega.6c02611
  • Dergi Adı: ACS OMEGA
  • Derginin Tarandığı İndeksler: Scopus, Science Citation Index Expanded (SCI-EXPANDED), Chemical Abstracts Core, Directory of Open Access Journals
  • Sayfa Sayıları: ss.1-17
  • Hacettepe Üniversitesi Adresli: Evet

Özet

Janus kinases (JAKs) are central mediators of cytokine-driven signal transducer and activator of transcription (STAT) signaling, and dysregulation of the JAK–STAT axis is implicated in inflammatory, autoimmune, and neoplastic diseases. To identify new small-molecule inhibitors of Janus kinase 1 (JAK1) and Janus kinase 2 (JAK2), we implemented an integrated virtual screening workflow combining structure-based docking and ligand-based prioritization using the InterBioScreen (IBS) compound library. A drug-likeness filter reduced 521,627 IBS molecules to 116,064 candidates, which were then docked into the ATP-binding sites of JAK1 (PDB ID: 4EI4) and JAK2 (PDB ID: 6VGL) using Glide SP; compounds were prioritized using docking score thresholds of <− 8.5 kcal/mol (JAK1) and <− 9.0 kcal/mol (JAK2), yielding 407 JAK1- and 298 JAK2-focused hits. Subsequent analysis shortlisted 42 candidates for enzymatic evaluation. In vitro kinase assays identified multiple nanomolar inhibitors, including compound 1–3 (IC50 = 0.032 μM) among the most potent for JAK1 and compound 2–8 (IC50 = 0.026 μM) for JAK2. Finally, 100 ns molecular dynamics (MD) simulations supported stable binding for representative complexes, with persistent hinge-region interactions for compound 1–3 in JAK1 (e.g., Glu957/Leu959) and compound 2–8 in JAK2 (e.g., Glu930/Leu932), consistent with a stable interaction network in the active site. Collectively, these results define validated IBS-derived hit scaffolds for further optimization and selectivity profiling toward JAK-targeted therapeutics.