Background: The Epidermal Growth Factor Receptor (EGFR) serves as a core target in tumor-targeted therapy. Mutations in EGFR frequently lead to resistance against clinical Tyrosine Kinase Inhibitors (TKIs), underscoring the critical need to develop novel EGFR-targeting compounds for overcoming drug resistance. Methods: This study performed in silico virtual screening of candidate small-molecule compounds from databases based on the common pharmacophore features of seven clinically used EGFR-TKIs. The candidates were subjected to in silico physicochemical, pharmacochemical, and toxicity evaluations, as well as in silico prediction of kinase inhibitory activity, leading to the identification of ZINC44136419 as a computationally predicted candidate. Computational cross-validation of target proteins was conducted, and EGFR was identified as the putative core target protein through enrichment, interaction network, and mutation frequency analyses. Molecular docking of ZINC44136419 with wild-type and mutant EGFR was performed in silico, using gefitinib as a control to compare docking scores. Results: A computationally predicted kinase inhibitor candidate, ZINC44136419, was obtained. In silico analyses indicated that it exhibited the core pharmacophore features of EGFR-TKIs, was predicted to meet drug-likeness criteria, showed no predicted toxicity risks, was predicted to have high synthetic feasibility, and was identified as possessing a structurally distinct scaffold (low Tanimoto similarity to known inhibitors). In silico analyses revealed predicted selective inhibitory activity against the tyrosine kinase family, with target protein enrichment analysis showing significant association with protein kinase activity, suggesting EGFR as a putative core target protein with high mutation frequency. Notably, ZINC44136419 exhibited a more favorable docking score toward mutant EGFR than toward wild-type EGFR in silico, as compared to gefitinib. discussion: As a novel EGFR inhibitor, ZINC44136419 achieves highly selective inhibition of drug-resistant mutants through its unique structure, providing a lead compound with translational potential for overcoming EGFR-TKI resistance. Discussion: This study relies entirely on computational simulations, which carry inherent methodological limitations. Nevertheless, the favorable druggability and favorable docking scores of this compound toward mutant targets in silico may provide candidate molecules and theoretical foundations for subsequent in vitro experimental validation, delivery system development, and the design of novel agents against EGFR drug resistance. Conclusion: As a computationally predicted EGFR-targeting compound, ZINC44136419 is predicted in silico to show preferential docking interactions with drug-resistant mutants through its structurally distinct scaffold, representing a preliminary candidate with potential translational value for overcoming EGFR-TKI resistance, pending experimental validation.
Background: Chalcones are α,β-unsaturated ketone scaffolds with broad documented bioactivity. No prior study has simultaneously integrated structural validation, PAINS assessment, chemical-space clustering, human-target classification, bioassay hit-rate benchmarking, and patent analysis within a single computational framework. Methods: Seven hundred and twenty-two PubChem chalcone compounds were curated by SMARTS-based E-configured enone validation, ECFP4 k-means clustering, Bemis–Murcko decomposition, QED/Fsp³ profiling, and PAINS filtering. Protein target profiling (476 proteins; 145 human), bioassay benchmarking (12,981 assays; 81,667 data points; ChEMBL 34), and patent analysis (2,148 records) were integrated across PubChem exports. Results: Among 722 compounds, 686 (95.0 %) were E-configured; 50 (6.9 %) were PAINS-flagged; 195 Bemis–Murcko scaffolds (SDI = 0.270); mean QED = 0.444 ± 0.175; k = 2 clusters were statistically optimal. PAINS-flagged compounds showed significantly lower QED (0.392 vs. 0.449; p < 0.001). Among 145 human protein targets across 25 functional classes, key categories included tubulin isoforms (15), ABC transporters, MAO-A/B, and AChE. Bioassay analysis yielded 18,438 active hits (22.6 % overall); hit rates were: MAO 38.6 %, ABC transporters 32.2 %, AChE 31.8 %, and anticancer 31.5 %. China dominated patent filings (38.4 %); A61P35 (anticancer) was the leading IPC category. Discussion: Chalcones demonstrate robust polypharmacological engagement across multiple target classes. The low PAINS prevalence (6.9 %) confirms chemical tractability, while the inverse correlation between hit rate and assay dataset size underscores the need to account for reporting bias. The growing patent portfolio highlights the continued translational relevance of this scaffold. Conclusion: Chalcones represent a tractable, polypharmacological scaffold with target engagement validated across 145 human proteins. Hit-rate patterns provide reliable evidence even after accounting for reporting bias, although translational claims require independent experimental validation.
Introduction: Chemotherapy-Induced Alopecia (CIA) is a stressful effect associated with cancer therapy, mainly affecting hair follicles. Epigallocatechin Gallate (EGCG), a bioactive polyphenol, can be used for its antioxidant and hair growth-promoting effects. Hence, in this study, an EGCG- and olive oil-loaded emulgel was formulated to treat CIA. Method: EGCG-loaded emulgel was formulated using a factorial design with a QbD approach. Characterisation included rheological studies, transmission electron microscopy, particle size analysis, spreadability, and swelling index analysis. In vitro release and ex vivo studies were also performed. An in vivo study was conducted in rats, and the presence of EGCG in serum was detected by HPLC. Results: The optimised emulgel showed optimum rheological behaviour, and the particle size was 0.470 ± 0.14 μm with a zeta potential of −32.9 ± 2.3 mV. The in vitro release study showed maximum release of 59.44 ± 3.10% at the 7th hour. Ex vivo studies showed measurable skin deposition (212.08 ± 7.22μg/cm²) and limited transdermal permeation (23.50 ± 2.14% at 12 h). In vivo studies in a rat model demonstrated significant hair growth in the group treated with the optimised formulation. Furthermore, HPLC analysis of serum samples showed no detectable EGCG peak above the limit of detection of the method, suggesting minimal or no systemic absorption under the current analytical conditions. Discussion: The CIA adversely affects quality of life, and current treatments have limited efficacy. EGCG and olive oil show hair growth–promoting and antioxidant effects but suffer from poor dermal delivery and stability. Emulgel enhanced the stability and also provided significant sustained release. An in vivo study demonstrated that EGCG–olive oil–loaded emulgel can be a promising treatment strategy for CIA. Conclusion: Emulgel can be used to deliver EGCG, which addresses the limitations of formulations available and potentially can be adopted as a therapeutic approach in healthcare.
Introduction/Objective: Hypoxia is a critical component of the tumor microenvironment contributing to metabolic reprogramming and drug resistance. Subsequently, elucidating the molecular mechanisms involved in hypoxia regulation in tumor cell models is essential. Methods: This study used CRISPR/Cas9 to knock out hepatocyte nuclear factor 4-alpha (HNF4-α) and hypoxia-inducible factor 1-alpha (HIF1-α) in HEPG2 cells. This study then assessed the effects of both knockouts (KO) on proliferation, migration, drug sensitivity, mitochondrial membrane stability, and gene expression in HEPG2 cells under both hypoxic and normoxic conditions. Results: Our data demonstrate that HIF1-α KO, but not HNF4-α KO, sensitizes HEPG2 cells to doxorubicin under hypoxic conditions. Additionally, the KO of both HIF1-α and HNF4-α enhances cell migration, irrespective of oxygen levels. Importantly, both HIF1-α and HNF4-α are required for cellular response to hypoxia and the maintenance of mitochondrial function, as evidenced by the loss of mitochondrial membrane potential increase under hypoxia in KO cells. Microarray-based transcriptomic profiling of HEPG2 cells revealed separation between HNF4-α and HIF1-α KO cells and controls under hypoxic versus normoxic conditions, with a markedly higher number of differentially expressed genes in hypoxia (up to 5,595 genes; adjusted p < 0.05, log2FC ±1.5). Both HNF4-α and HIF1-α contribute to hypoxia-responsive pathways, with impaired induction of key genes such as VEGFA and AK4 in KO cells, while GSEA indicated minimal effects of HNF4-α loss under hypoxia but pronounced metabolic pathway disruption in HIF1-α KO cells. Moreover, key regulators of mitochondrial function and mitophagy, including FUNDC1, MIEF1, and MIEF2, were upregulated in control hypoxic cells but not in KO cells. Discussion: These findings suggest that, in this single HEPG2 cell line model, HNF4-α contributes to the regulation of cell migration and mitochondrial stability under hypoxic conditions. Conclusion: Further investigation into the role of HNF4-α in tumor biology is encouraged.
Introduction: Impaired mitophagy in cancer cells leads to the accumulation of dysfunctional mitochondria, elevated ROS, and activation of pro-survival pathways, promoting tumour growth and metabolic adaptation. ROCK2, a serine/threonine kinase overexpressed in cancer, negatively regulates Parkin-mediated mitophagy, making it a rational therapeutic target. Methods: This study used an integrated in silico pipeline to identify phytocompounds from Indian medicinal plants predicted to inhibit ROCK2 and potentially modulate mitophagy. This study filtered a library of 31,496 natural product compounds from 325 medicinal plants (IMPPAT 2.0 database) using SwissADME based on Lipinski's, Veber's, Ghose's, and Pfizer's rules, yielding 598 drug-like compounds. Bioactivity scoring identified 49 NPCs with kinase inhibitory potential, which underwent Glide SP docking against ROCK2 (PDB: 7JNT), followed by ProTox-II toxicity profiling and XP docking. Results: Six compounds (Quercetin, Hydnocarpin, Annomontine, Camptothecin, 9- Methoxycamptothecin, and Methoxycamptothecin) showed binding affinities comparable to or exceeding reference inhibitors Fasudil and Y27632, with key interactions at conserved catalytic residues (Met172, Asp218, Asn219, Lys216). MD simulations showed that Hydnocarpin formed the most stable protein-ligand complex, with a consistent RMSD, persistent hydrogen bonding with Glu170, Met172, and Lys216, and minimal residue fluctuations, resembling Fasudil's dynamic profile. Quercetin, despite favourable docking scores, showed structural instability. Discussion: Integrated docking and MD analyses indicate Hydnocarpin exhibits superior binding stability compared to other screened phytocompounds, suggesting more favourable predicted binding under physiologically relevant simulation conditions. Conclusion: Hydnocarpin emerges as the most promising ROCK2 inhibitor among screened phytocompounds, supporting its potential as a mitophagy-modulating anticancer agent warranting further in vitro validation, as all findings reported here are computational predictions.
