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.
Abstract
The development of anticancer agents capable of affecting multiple cancer-related cellular processes remains an important strategy in cancer drug discovery. In this study, a novel series of benzofuran-furan-pyrano[2,3-c]pyrazole hybrids was synthesized and comprehensively investigated through in vitro biological assays and in silico analyses. The synthesized compounds exhibited concentration-dependent cytotoxicity against LNCaP, MDA-MB-231, A549, and Caco-2 cancer cells, with EC50 values ranging from 20.49 to 274.37 µM while displaying lower toxicity toward non-malignant HUVEC cells. Among the synthesized derivatives, compounds 4j, 4h, and 4d showed the strongest antiproliferative activities and significantly suppressed cancer cell migration, invasion, and colony formation. Mechanistic studies demonstrated that the active hybrids induced G1/S phase cell cycle arrest, increased intracellular reactive oxygen species (ROS) production, disrupted mitochondrial membrane potential, and promoted apoptotic cell death. These findings were further supported by quantitative real-time PCR, which revealed downregulation of BCL2 together with upregulation of BAX, CASP3, CASP8, CASP9, and TP53, indicating activation of both intrinsic and extrinsic apoptotic pathways. In silico analysis of absorption, distribution, metabolism, excretion, and toxicity (ADMET) predicted favorable drug-likeness and pharmacokinetic properties for the synthesized hybrids. Molecular docking suggested potential interactions with CDK2, the androgen receptor, VEGFR-2, and Bcl-2, with compound 4f exhibiting the most favorable overall docking profile. Collectively, these findings demonstrate 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.
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
Cancer remains a leading cause of mortality worldwide, necessitating the continuous development of more effective and selective therapeutic agents. Among heterocyclic scaffolds, 1,3,4-thiadiazole derivatives have attracted considerable attention due to their diverse pharmacological properties and favorable physicochemical characteristics. In this study, a novel thiadiazole-based compound namely 2-((5-acetamido-1,3,4-thiadiazol-2-yl)thio)-N-(naphthalen-2-yl)acetamide, was successfully synthesized in excellent yield (99%) and high thermal stability and was fully structurally characterized using FT-IR, NMR, and mass spectrometry. The anticancer potential of the synthesized compound was evaluated in vitro against three human cancer cell lines, namely breast adenocarcinoma (MCF-7), colorectal carcinoma (HCT-116), and hepatocellular carcinoma (HepG-2), using the MTT assay. The compound exhibited pronounced cytotoxic activity against MCF-7 and HCT-116 cell lines, with IC50 values of 1.07 ± 0.03 µM and 2.09 ± 0.55 µM, respectively, demonstrating superior potency compared to the reference drug doxorubicin. In contrast, no significant activity was observed against HepG-2 cells, indicating a degree of selectivity. These findings highlight the potential of the 1,3,4-thiadiazole scaffold as a promising platform for the development of novel anticancer agents with enhanced efficacy and selectivity. In silico investigations, including molecular docking, molecular dynamics simulations, and MM/GBSA analysis, revealed stable binding of the synthesized compound within the VEGFR2 active site, supported by favorable binding free energy and key ligand–residue interactions.
Mostafa Sayed, Ehdaa Mohammed, Doha H. Aboubaker et al.· RSC Advances· 0 citations
It is suggested that 4c exerts anticancer activity through ROS-mediated mitochondrial apoptosis and cell-cycle disruption, highlighting its promise as an oncotherapeutic candidate for TNBC and Dalton lymphoma.
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
Cancer continues to be a leading cause of global mortality, highlighting the ongoing need for novel anticancer compounds that offer high efficacy with improved side effect profiles. In the present study, a series of 3H-1,2-dithiole-3-thione derivatives (DTT-S1-18) were synthesized as promising anticancer agents, and the structures of products were confirmed by spectral techniques. H2S-releasing experiments showed that most of the compounds released higher amounts of H2S slowly over time compared to standard ADT-OH. All compounds were tested for antiproliferative activity on HT-29, PC-3, MCF-7, and HUVEC cell lines. Compounds DTT-S6 (3-nitrophenyl derivative) and DTT-S8 (methionine derivative) have the lowest IC50 values of 41.6 and 38.9 µM on the MCF-7 cell line, respectively. Based on the wound healing and colony formation assays performed in MCF-7 cells, the wound areas were not significantly changed after treatment with compounds DTT-S6 and DTT-S8, whereas compound DTT-S8 at double IC50 dose inhibited colony formation by 81.82%. In addition, molecular docking, MD simulations, MM/GBSA binding free energy calculations, and binary QSAR analyses were performed to explore the potential target interactions and predicted activity profiles of the synthesized compounds toward inflammation-related proteins, including COX-1, COX-2, 5-LOX, and iNOS, thereby supporting the development of mechanistic hypotheses for future validation. Furthermore, structure-activity relationship (SAR) analyses were conducted to correlate the structural characteristics of the synthesized compounds with their H2S releasing potential and biological profiles. Overall, this work integrates experimental anticancer evaluation with computational pathway and structure-based cancer/inflammation analyses to characterize novel DTT-based H2S donors. The findings identify particularly compound DTT-S8, as a promising in vitro anticancer candidate, while the computational results suggest a putative involvement of inflammation-related targets, particularly the COX-2/5-LOX axis, which requires direct biochemical and cellular validation.
Semra Altunsoy, Y. Yilmaz, Tuğba Güngör et al.· Molecular diversity· 0 citations
Cancer remains a leading cause of mortality worldwide, necessitating the development of new, selective, and potent therapeutic agents. In this study, a novel, metal-free, one-pot sequential synthetic approach was developed for the synthesis of carbazolyl-thiazolidinone (CTZD) derivatives via the reaction of N-octylcarbazole-3-carbaldehyde with variety of aromatic and aliphatic primary and secondary amines and thioglycolic acid. This strategy efficiently yielded a diverse range of CTZD derivatives (4a-p) in moderate to high yields (20-95%). The synthesized compounds were characterized by FTIR, NMR (1H, 13C, DEPT, APT), and HRMS. Their in vitro cytotoxicity was tested on human leukemia cell lines NB4, K562 and U937 using MTT assays, where four derivatives (4e, 4i, 4j, and 4o) exhibited potent, concentration-dependent antiproliferative activity over the tested concentration range (1.25-10 μM). As c-MYC is a key regulator of cell proliferation, qRT-PCR analysis demonstrated that these four derivatives significantly downregulated c-MYC mRNA expression, with compound 4j producing the greatest reduction, suggesting a potential association with modulation of the Wnt/β-catenin pathway. DNA fragmentation analysis showed no detectable late-stage apoptosis, indicating that the observed c-MYC downregulation and antiproliferative effects were not associated with late-stage apoptotic cell death. The ADME/T analysis of all compounds showed favorable pharmacokinetic profiles with prediction of good oral absorption (HIA >92%) and no hERG I liability. Molecular docking studies demonstrated strong binding affinities of these compounds to β-catenin protein (PDB ID: 7ZRB) with compound 4i showing strongest affinity with ΔG = -8.10 kcal/mol via H-bonds with Ser473, Asn430, Arg469 and His470 amino acid residues. The developed metal-free synthesis provided a sustainable route to bioactive carbazolyl-thiazolidinones, and derivatives 4e, 4i, 4j, 4o could be promising leads for targeting Wnt/β-catenin/c-MYC signaling in leukemia.
S. Shehzadi, Sana Naveed, D. Khan et al.· Bioorganic chemistry (Print)· 0 citations