Central University of Punjab

Thymidylate Synthase (TS) is a validated therapeutic target against cancer. The research examined whether the anticancer effects of lead compound 4d surpassed the action of 5-fluorouracil (5-FU) while monitoring its impact on MCF-7 breast cancer cells under both 2D and 3D experimental conditions. The treatment of MCF-7 cells with 4d at 1 μM and 5 μM concentrations and 5-FU at 1 μM level allowed researchers to measure cytotoxicity, apoptosis, and cell cycle arrest activities. Furthermore, ROS production and TS expression levels, including migration dynamics and 3D spheroid structure integrity, were also examined. The cell viability declined dramatically when cells were exposed to compound 4d, which causes cell cycle blockage at the G₂/M phase and elevated ROS levels, while reducing TS expression. Severe, destructive effects on cell movement and spheroid organization resulted in a dose-dependent death of cells. The compound 4d demonstrated effects that matched the results obtained from using 5-FU. The potential of TS-targeted therapeutic candidate status for breast cancer treatment appears promising, because compound 4d demonstrates strong anticancer effects through multiple pathways.

BRAF is one of the most commonly mutated oncogenes in human cancers, with more than 90 % of mutations occurring at codon 600. Among these, BRAF^V600E is the most prevalent, accounting for nearly 90 % of all BRAF codon 600 mutations, while the less frequent variants are collectively referred to as BRAF^V600WT. BRAF mutations are reported across several cancers, with the highest frequency in malignant melanoma (70-90 %). To target these mutations, a series of phenyl-substituted thioxo-tetrahydro-pyrimidine-benzenesulfonamide hybrids in the [αC-OUT/DFG-IN] conformation were designed. Fourteen derivatives were synthesized through multistep reactions and evaluated for ADME-T properties and in silico binding affinities using molecular docking, further validated by molecular dynamics (MD) simulations of the top-ranked compound. In addition, cytotoxic activity against melanoma cell lines was assessed, followed by kinase inhibition studies on the most potent derivatives against BRAF^V600E/WT. All compounds exhibited favorable ADME-T and drug-like properties. Among them, AV05 demonstrated the highest binding affinity (-8.013 kcal/mol) and potent cytotoxic activity (IC₅₀ = 1.25 ± 0.57 μM), with BRAF^V600E inhibition (0.89 ± 0.78 μM) comparable to sorafenib (-6.189 kcal/mol; IC₅₀ = 0.90 ± 0.21 μM; inhibition = 0.10 ± 0.01 μM). In contrast, AV01 showed the strongest affinity for BRAF^V600WT (-4.954 kcal/mol) and kinase inhibition (0.93 ± 0.28 μM), relative to sorafenib (-12.241 kcal/mol; inhibition = 0.16 ± 0.01 μM). Collectively, computational and biological evaluations revealed that the synthesized hybrids exhibited higher selectivity and potency toward BRAF^V600E than BRAF^V600WT, with AV05 emerging as the most promising compound.

Cardiovascular diseases (CVDs) continue to rise at an alarming rate, contributing to millions of deaths globally. Among them, myocardial infarction (MI), commonly known as a heart attack, remains a leading cause of mortality. Despite extensive research, MI remains incurable, and its complete eradication has yet to be achieved. Mitochondria play a central role in the pathogenesis and potential treatment of MI, and recent studies have identified mitochondrial microRNAs (mito-miRs) as promising molecular regulators. Although the precise mechanisms of mito-miRs remain incompletely understood, emerging evidence suggests their involvement in regulating mitochondrial metabolism, dynamics, ROS production, bioenergetics, and mitochondrial biogenesis. Additionally, mito-miRs influence several forms of programmed cell death, including apoptosis, necrosis, ferroptosis, and pyroptosis. The exact processes governing the translocation of these miRNAs into mitochondria and their intracellular actions remain elusive. Notably, specific miRNAs have been shown to target key cardiac cell types, including cardiomyocytes, endothelial cells, and fibroblasts. Deciphering their mechanistic roles could enable the development of targeted mito-miRNA-based therapeutics. Moreover, their therapeutic efficacy may be enhanced by integrating mito-miRs with stem cell therapies and bioactive compounds, particularly when delivered via nanoparticle-based formulations to ensure targeted delivery within the cardiac microenvironment.