A series of novel bis urea derivatives derived from 1,4-diisocyanatobenzene were successfully designed, synthesized, and evaluated for their anticancer potential. These compounds were systematically screened for in vitro antiproliferative activity against human HeLa cervical cancer cell lines. Structural elucidation was achieved through comprehensive spectroscopic techniques including FT-IR, NMR (¹H and ¹³C), mass spectrometry, and elemental analysis. Among the ten selected derivatives, compounds **3d** (1,1-(1,4-phenylene) bis(3-(2-chlorobenzyl)urea)) and **3e** (1,1-(1,4-phenylene) bis(3-cyclohexylurea)) exhibited the most potent anticancer activity with IC₅₀ values of 5.40 μM and 5.89 μM, respectively, indicating strong growth inhibition.
Further investigation revealed that these two compounds effectively induce apoptosis in HeLa cells. The induction of apoptosis was confirmed by significant upregulation of key executioner caspases—caspase-3 and caspase-9—indicating activation of the intrinsic apoptotic pathway.HMOX1 Antibody Cancer Additionally, both compounds demonstrated notable antifungal activity against *Saccharomyces cerevisiae*, suggesting broader biological interactions.144701-48-4 Molecular Weight The zone of inhibition observed for compound 3d (0.9 ± 0.02 mm) and 3e (0.8 ± 0.05 mm) further supports their bioactive potential.
To understand the molecular mechanism behind this activity, quantum-polarized-ligand docking (QPLD), combined with molecular mechanics (QM/MM) and MM-GBSA calculations, was employed. The results showed that **3d** and **3e** exhibit high binding affinity toward both ERK/MAP kinase (PDB: 4QP2) and CDK2 (PDB: 1AQ1). Notably, **3d** displayed a superior QPLD-MMGBSA binding free energy of -77.924 kcal/mol with CDK2 and -47.800 kcal/mol with ERK, accompanied by multiple strong hydrogen bonds and hydrophobic interactions with critical residues such as Glu12, Lys33, Asp86, and His402.
Long-range molecular dynamics simulations (150 ns) confirmed the stability of the 3d-CDK2 and 3d-ERK complexes.PMID:34061415 Root mean square deviation (RMSD) values remained low throughout the simulation (2.013 Å and 3.704 Å for Cα atoms), while root mean square fluctuation (RMSF) indicated minimal structural variation in key regions. The radius of gyration (rGyr), solvent accessible surface area (SASA), polar surface area (PSA), and intramolecular hydrogen bond count further validated the conformational stability and favorable physicochemical properties of **3d**.
These findings collectively demonstrate that **3d** acts as a dual inhibitor, simultaneously targeting ERK/MAPK and CDK2 signaling pathways. By blocking ERK activation, it disrupts downstream phosphorylation events involving MSK1/2, cdc25, STAT3, and p90RSK, thereby impairing cell proliferation and differentiation. Concurrently, inhibition of CDK2 prevents G1/S transition via hypophosphorylation of Rb protein, leading to cell cycle arrest at the G1/S boundary. This dual mechanism offers a promising strategy for selective anticancer therapy.
In conclusion, the bis urea derivative **3d** emerges as a lead candidate with strong binding affinity, excellent stability, and potent antiproliferative activity. Its ability to modulate two central regulators of cell cycle progression positions it as a highly promising selective agent for future development in targeted cancer therapeutics.MedChemExpress (MCE) offers a wide range of high-quality research chemicals and biochemicals (novel life-science reagents, reference compounds and natural compounds) for scientific use. We have professionally experienced and friendly staff to meet your needs. We are a competent and trustworthy partner for your research and scientific projects.Related websites: https://www.medchemexpress.com
