2. Academic Validation
  3. Synthesis, biological evaluation and molecular docking studies of N-propylsulfonyl indole-linked hydrazinecarbothioamides as selective ecto-5'-nucleotidase and NTPDase inhibitors

Synthesis, biological evaluation and molecular docking studies of N-propylsulfonyl indole-linked hydrazinecarbothioamides as selective ecto-5'-nucleotidase and NTPDase inhibitors

  • Sci Rep. 2026 May 2;16(1):14419. doi: 10.1038/s41598-026-50728-3.
Zahra Batool 1,  Shireen Mona Dutt 2,  Mariya Al-Rashida 3,  Nicolly Espindola Gelsleichter 4 5,  Julie Pelletier 5,  Jean Sévigny 4 5,  Talha Islam 3,  Mostafa A Ismail 6,  Ajmal Khan 7 8,  Halil Şenol 9,  Rima D Alharthy 10,  Jamshed Iqbal 11,  Zahid Shafiq 12
Affiliations

Affiliations

  • 1 Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan.
  • 2 Centre for Advanced Drug Research, COMSATS University Islamabad, Abbottabad Campus, Abbottabad, 22060, Pakistan.
  • 3 Department of Chemistry, Forman Christian College (A Chartered University), Lahore, Pakistan.
  • 4 Département de microbiologie-infectiologie et d'immunologie, Centres PROTEO et ARThrite, Faculté de Médecine, Université Laval, Quebec City, QC, G1V 0A6, Canada.
  • 5 Axe Maladies infectieuses et immunitaires, Centre de recherche du CHU de Québec - Université Laval, Quebec City, QC, G1V 4G2, Canada.
  • 6 Department of Chemistry, Faculty of Science, Research Center for Advanced Materials Science (RCAMS), King Khalid University, P.O. Box 960, Abha, 61421, Saudi Arabia.
  • 7 Department of Chemical and Biological Engineering, College of Engineering, Korea University, 145 Anam-ro, Seongbuk-gu, Seoul, 02841, Republic of Korea. [email protected].
  • 8 Natural and Medical Sciences Research Centre, University of Nizwa, P.O. Box 33, PC 616, Birkat Al Mauz, Nizwa, Sultanate of Oman. [email protected].
  • 9 Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Bezmialem Vakif University, 34093, Fatih, İstanbul, Türkiye.
  • 10 Department of Chemistry, Rabigh Branch, Science & Arts College, King Abdulaziz University, Rabigh, 21911, Saudi Arabia.
  • 11 Centre for Advanced Drug Research, COMSATS University Islamabad, Abbottabad Campus, Abbottabad, 22060, Pakistan. [email protected].
  • 12 Institute of Chemical Sciences, Bahauddin Zakariya University, Multan, 60800, Pakistan. [email protected].
PMID: 42069907 DOI: 10.1038/s41598-026-50728-3
Abstract

Ectonucleotidases, including NTPDases and ecto-5'-nucleotidase (e-5'NT/CD73), regulate extracellular purinergic signaling by converting ATP to adenosine, a pathway critically involved in immune response, inflammation, and Cancer progression. In this study, a novel library of 22 N-propylsulfonyl-substituted indole-based hydrazinecarbothioamides (5a-5v) was synthesized and structurally characterized. Biological evaluation against human e-5'NT and NTPDase1, -2, -3, and - 8 revealed that several compounds exhibited low micromolar inhibitory activity, with 5n (IC50 = 1.7 µM), 5o (IC50 = 1.7 µM), 5f (IC50 = 1.0 µM), and 5i (IC50 = 1.6 µM) emerging as the most promising derivatives, showing strong potency and isoform selectivity. Structure-activity relationship analysis indicated that both electronic and steric features of substituents significantly influence activity and enzyme preference. Molecular docking studies performed on e-5'NT demonstrated that active compounds adopt consistent binding modes within the catalytic pocket, stabilized by key residues such as Asp-506, Phe-500, Phe-417 and Arg-395. Binding free energy calculations (MM-GBSA) supported strong ligand-protein interactions ( ~ - 70 kcal/mol). The docking protocol was validated by redocking, yielding an RMSD value well below the accepted threshold. Molecular dynamics simulations (500 ns) confirmed stable complex formation, with low RMSD values (~ 1-3 Å), limited residue fluctuations, and persistent interactions with catalytic residues. Surface and compactness parameters (rGyr, SASA) remained stable, indicating consistent ligand accommodation. In silico ADME analysis suggested favorable drug-like properties for most compounds, particularly for the lead candidates. Overall, these findings identify 5n and 5o as the most promising lead compounds, supported by both experimental and computational results, and highlight this scaffold as a valuable platform for the development of selective ectonucleotidase inhibitors.

Keywords

Ecto-5′-nucleotidase; Enzyme inhibition; Molecular docking; NTPDase inhibitors; Thiosemicarbazones.

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