Pyrimidine‐Derived Scaffolds Targeting VEGFR‐2, EGFR, and HER‐2: Synthesis, Anticancer/Immunomodulatory Evaluation, and In Silico Analysis

Abstract

ABSTRACT Following the common structural features of VEGFR‐2, EGFR, and HER‐2 kinase inhibitors, two newly synthesized series of 3,4‐dihydropyrimidine 2(1 H )‐(thio)ones‐5‐carboxamide ( 2a–l ) and thiouracil‐5‐sulfonyl derivatives ( 4a–e ) were evaluated for anti‐proliferative activity against colorectal (HCT‐116) and breast ( MCF‐7) cancer cell lines. Among the designed series, Compounds 2h and 2l were the most potent derivatives across both tumor models, exhibiting IC 50 values of 23.3 and 30.9 μM (HCT‐116), respectively, and 31.5 and 39.2 μM (MCF‐7), respectively, compared to sorafenib (IC 50 = 8.8 and 11.6 μM, respectively). Interestingly, 2h and 2l displayed a favorable safety profile toward the normal human cell line WI‐38, with IC 50 > 200 μM comparable to sorafenib (IC 50 = 192 μM). In vitro enzymatic assays, 2h and 2l derivatives emerged as potent multi‐kinase inhibitors targeting VEGFR‐2, EGFR, and HER‐2 with IC 50 = 0.20, 0.21, and 0.19 μM, respectively, and IC 50 = 0.67, 0.53, and 0.40 μM, respectively, hence circumventing cancer resistance mechanisms. The most potent Compound 2h prompted apoptosis and necrosis at % G0‐G1 phase in HCT‐116 via the activation of caspase‐3 and 8, with significant downregulation of anti‐apoptotic protein Bcl‐2. Additionally, Compound 2h markedly suppressed immunomodulatory proteins TNF‐α and IL‐6 levels by 80.9% and 88.2%, respectively, in comparison to dexamethasone (82.7% and 93.2%, respectively), which attenuates tumor‐promoting inflammation, disrupting pro‐survival signaling pathways and metastasis. In silico ADMET, toxicity, and molecular docking studies were performed; notably, the docking simulations generated an interesting hypothesis for potential direct binding to Bcl‐2, an effect distinct from the experimentally observed downstream downregulation of its expression. The present results pave the way for further developing of 2h candidate as a multi‐tyrosine kinases inhibitor toward colon cancer cells.