2. Academic Validation
  3. Multi-target pyrazolopyrimidine-coumarin derivatives as potent CA IX/XII and tubulin polymerization inhibitors: Design, synthesis, and biological evaluation

Multi-target pyrazolopyrimidine-coumarin derivatives as potent CA IX/XII and tubulin polymerization inhibitors: Design, synthesis, and biological evaluation

  • Eur J Med Chem. 2026 Jun 5:310:118789. doi: 10.1016/j.ejmech.2026.118789.
Mahmoud S Elkotamy 1,  Mohamed A Abdelrahman 2,  Simone Giovannuzzi 3,  Mahmoud Abdelrahman Alkabbani 4,  Alessio Nocentini 3,  Claudiu T Supuran 3,  Wagdy M Eldehna 5,  Hatem A Abdel-Aziz 6,  Sahar M Abou-Seri 7
Affiliations

Affiliations

  • 1 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Alsalam University, Algharbia, Kafr Alzayat, 31611, Egypt.
  • 2 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Egyptian Russian University, Badr City, Cairo, 11829, Egypt; Department of Pharmaceutical Chemistry, College of Pharmacy, University of Kut, Wasit, 52001, Iraq.
  • 3 Department of NEUROFARBA, Section of Pharmaceutical and Nutraceutical Sciences, University of Florence, Polo Scientifico, Via U. Schiff 6, Firenze, 50019, Italy.
  • 4 Pharmacology and Toxicology Department, Faculty of Pharmacy, Egyptian Russian University, Cairo, Badr City, 11829, Egypt.
  • 5 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Kafrelsheikh University, P.O. Box 33516, Kafrelsheikh, Egypt. Electronic address: [email protected].
  • 6 Department of Applied Organic Chemistry, National Research Center, Dokki, Cairo, 12622, Egypt. Electronic address: [email protected].
  • 7 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, Cairo, 11562, Egypt. Electronic address: [email protected].
PMID: 41905101 DOI: 10.1016/j.ejmech.2026.118789
Abstract

In the current medical era, developing multi-target Anticancer agents that simultaneously engage complementary cellular vulnerabilities represents a promising strategy to overcome the inherent limitations of single-target therapeutics. Building on insights from our previous investigation, we synthesized a novel series of coumarin-pyrazolo [1,5-a]pyrimidine hybrids (13a-n) designed to target tumor-associated carbonic anhydrases (CA IX/XII) and tubulin polymerization through rational molecular hybridization. Structure-activity relationship analysis revealed that a zinc-binding sulfonamide group was essential for CA inhibitory activity; coumarin-only analogs failed (Ki > 100 μM). Compounds 13g and 13n, bearing sulfonamide functionality, demonstrated nanomolar potency against hCA IX and hCA XII. Compound 13n emerged as the superior lead, achieving balanced dual-target inhibition: hCA IX (Ki = 27.1 nM), hCA XII (Ki = 20.9 nM), and tubulin polymerization (IC50 = 6.35 μM). Broad-spectrum NCI-60 screening revealed 13n's potent antiproliferative activity across nine Cancer types (GI50: 2.48-31.00 μM). Treatment of MCF-7 breast Cancer cells with 13n resulted in significant G2/M phase arrest (13.81% to 31.97%) and robust induction of Apoptosis (37-fold relative to control), mediated by p53-dependent signaling. Molecular analysis revealed elevated p53 expression (3.43-fold), increased Bax level (12.34-fold), reduced Bcl-2 expression (4.37-fold), and enhanced caspase-7 activation (7.35-fold). Molecular docking studies confirmed zinc coordination within the CA active sites and optimal positioning within the tubulin colchicine-binding pocket. In summary, compound 13n validates the design strategies employed to develop an efficient multi-target Anticancer candidate, positioning this lead compound for further preclinical development.

Keywords

Carbonic anhydrase; Coumarin; MCF7; Pyrazolopyrimidine; Tubulin polymerization.

Figures
Products