Quinoxaline-based cyanoacrylate hybrids as multi-target anti-proliferative and anti-inflammatory agents with potent activity against hepatocellular carcinoma: induction of apoptosis and modulation of oxidative stress signaling
Findings identify compound 9 as a promising multi-target quinoxaline-based agent with dual antiproliferative and anti-inflammatory activities, mediated through induction of mitochondrial apoptosis and modulation of oxidative stress and cytokine signaling pathways.
Abstract
A set of quinoxaline-based derivatives incorporating a cyanoacrylate pharmacophore were designed, synthesized, and biologically evaluated for their dual anticancer and anti-inflammatory potential. Cytotoxic activity was assessed against HepG-2, HCT-116, MCF-7, and Panc-1 cancer cell lines using the MTT assay. Among the synthesized compounds, compound 9 exhibited the highest potency, particularly against HepG-2 cells (IC50 = 7.81 ± 1.1 µM), surpassing doxorubicin (IC50 = 15.96 ± 0.61 µM), and demonstrated improved selectivity toward WI-38 normal fibroblasts (IC50 = 67.21 µM; SI ≈ 7.81). Mechanistic investigations in HepG-2 cells revealed that compound 9 induced pronounced G0/G1 cell cycle arrest, increasing the cell population from 54.39% to 86.21% (∼1.6-fold), with a concomitant reduction in S-phase cells (∼3.1-fold decrease). Apoptosis analysis showed a significant increase in total apoptotic cells from 3.12% to 35.14% (∼11.3-fold), predominantly driven by late apoptosis (∼129-fold increase). These findings were supported by gene expression analysis, where compound 9 upregulated p53 (∼5.9-fold), BAX (∼3.7-fold), cytochrome c (∼3.9-fold), and caspase-7 (∼2.6-fold), while downregulating Bcl-2 (∼0.64-fold), indicating activation of the intrinsic mitochondrial apoptotic pathway. In LPS-stimulated RAW264.7 macrophages, compound 9 exhibited potent anti-inflammatory activity without significant cytotoxicity. The compound markedly reduced intracellular reactive oxygen species (ROS) levels in a dose-dependent manner (from 591.52 to 60.62 pg mL−1; ∼9.8-fold reduction) and significantly suppressed nitric oxide production. Furthermore, it downregulated key pro-inflammatory cytokines, including TNF-α (∼0.36-fold), IL-1β (∼0.47-fold), and IL-6 (∼0.51-fold), with effects comparable to celecoxib. Molecular docking studies indicated favorable binding of compound 9 within the active sites of IL-1β, TNF-α, and IL-6Rα, with the highest affinity toward IL-6Rα (−7.93 kcal mol−1). These interactions were further validated by 100 ns molecular dynamics simulations, which supported stable protein–ligand complexes, consistent RMSD profiles, preserved structural compactness, and persistent key interactions throughout the trajectories. Collectively, these findings identify compound 9 as a promising multi-target quinoxaline-based agent with dual antiproliferative and anti-inflammatory activities, mediated through induction of mitochondrial apoptosis and modulation of oxidative stress and cytokine signaling pathways.
Overall, the cytotoxic effects of 7k on MV4-11 cells are attributed to the coordinated induction of ROS-mediated oxidative stress, G0/G1 cell cycle arrest, and caspase-dependent apoptosis, highlighting its promising antiproliferative potential.
G glycine conjugation of a coumarin-quinazoline hybrid has not been reported previously, and the results identify it as a substituent- and cell-line-dependent strategy for improving anticancer potency and selectivity.
N. Çalışkan, M. Emirik, F. Yılmaz et al.· Bioorganic chemistry (Print)· 0 citations
A series of novel pyrimidine-based benzothiazole/phenyl hybrids was rationally designed and synthesized via molecular hybridization to develop promising anticancer agents. The compounds were evaluated against human lung carcinoma (A549) and breast adenocarcinoma (MCF-7) cell lines, while HEK-293 cells were used to assess selectivity toward normal cells. Among the derivatives, K12, K5, and K4 showed potent activity against A549 cells, with IC50 values of 6.18, 7.01, and 7.54 μM, respectively. K5 and K12 also showed significant activity against MCF-7 cells, with IC50 values of 8.33 and 9.30 μM, respectively. All compounds displayed minimal cytotoxicity toward HEK-293 cells, indicating favorable selectivity for cancer cells. UV-Vis DNA-binding studies suggested a probable intercalative binding mode, while DNA nicking assays demonstrated protection against oxidative DNA damage. K12 showed the strongest DNA-binding affinity and DNA-protective activity. Molecular docking against the VEGFR-2 kinase domain (PDB ID: 4ASD), followed by 501 ns MD simulations, revealed stable protein-ligand complexes, supported by RMSD, RMSF, DCCM, PCA, and free energy landscape analyses. MM/GBSA calculations indicated favorable binding free energies, dominated by van der Waals interactions. Overall, K12 emerged as the most promising lead compound, combining potent antiproliferative activity, high selectivity, efficient DNA-binding characteristics, and robust computational performance, highlighting its potential for further development as a novel anticancer agent.
Karan Kamle, Shreyansh R. Mevada, Reena Hirani et al.· Bioorganic chemistry (Print)· 0 citations
The high mortality rate of prostate cancer remains a major threat to men's health worldwide, highlighting an urgent need for novel therapeutic agents with improved efficacy and safety profiles. In this study, we employed a rational structural modification strategy to synthesize 14 novel 1,2,3-triazole-hybridized derivatives of piperlongumine (PL), a natural alkaloid with proven anticancer activity. The antiproliferative activity of these derivatives was evaluated in vitro against multiple cancer cell lines (PC-3, A549, T47D, and SMMC-7721), with normal HEK293 cells used to assess their selectivity. All synthesized derivatives (1 M-14 M) exhibited significant antitumor efficacy, with lead compound 11 M showing pronounced inhibition of PC-3 cell proliferation and migration. Mechanistic studies revealed that 11 M induced G2/M phase arrest and triggered mitochondrial-dependent apoptosis, while concurrently activating the ROS-MAPK signaling pathway and autophagic flux. Notably, 11 M attenuated DNMT3A-mediated global DNA hypomethylation. In vivo validation in PC-3 xenograft mice confirmed that 11 M exerted robust tumor growth suppression with low systemic toxicity, accompanied by DNMT3A downregulation and MAPK pathway activation, substantiating its multi-target therapeutic mechanism.
It is demonstrated that benzofuran-furan-pyrano[2,3-c]pyrazole hybrids possess promising multifaceted in vitro anticancer activity by inhibiting cancer cell proliferation, migration, invasion, and clonogenicity while inducing ROS-mediated mitochondrial apoptosis, highlighting these hybrid scaffolds as attractive candidates for further anticancer drug development.
E. Hutanu, Bassam A. Najri, A. Abdelsalam et al.· RSC Advances· 0 citations
The development of dual-directed anticancer agents has emerged as an effective strategy to simultaneously modulate tumor proliferation and angiogenesis while overcoming resistance associated with single-target therapies. In the present study, a novel series of benzofuran-based aryl urea derivatives incorporating a 1,3,4-thiadiazole linker were designed as dual VEGFR-2/BRAFWT inhibitors using sorafenib as a pharmacophoric template. The in vitro cytotoxic activity of the synthesized compounds against cervix HeLa, prostate PC-3, colon HCT-116, and breast MCF-7 cancer cell lines was evaluated. Most derivatives showed variable activity, with 5-bromobenzofuran analogues displaying superior potency. Compound 7j emerged as the most active analogue, with IC50 values of 7.83-13.27 μM and reduced toxicity toward normal lung fibroblast WI-38 cells. Enzymatic assays revealed potent dual inhibition of VEGFR-2 and BRAFWT by 7j (IC50 = 0.044 and 0.071 μM, respectively), outperforming sorafenib and vemurafenib. Mechanistic studies showed that 7j induced G2/M cell cycle arrest and promoted apoptosis in HeLa cells. This effect was associated with upregulation of BAX, p53, and caspase-9, downregulation of Bcl-2, and activation of the intrinsic apoptotic pathway. Additionally, in silico studies including molecular docking and molecular dynamics simulations demonstrated stable ligand-target interactions and favorable binding modes of 7j across both VEGFR-2 and BRAF, supporting the proposed hybrid design strategy. Overall, compound 7j represents a promising dual VEGFR-2/BRAFWT inhibitor and highlights benzofuran-based scaffolds as valuable platforms for anticancer drug development.
Marwa I Serag, Mohamed R. Elnagar, Wafaa A. Ewes et al.· Bioorganic & Medicinal Chemi...· 0 citations
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