Drug repurposing strategy to identify the putative leads against the Epidermal growth factor receptor (EGFR) from the USFDA-approved drug pool: Investigating the utility as an anticancer agent.

Coumarin as a privileged natural product scaffold towards the development of antibacterial agents targeting ESKAPE pathogens.

Antimicrobial resistance (AMR) has emerged as a global healthcare crisis, necessitating the discovery of potent antibacterial agents. In the present investigation, we report the discovery, in vitro and ex vivo antibacterial efficacy studies, and mechanism of action of an unexplored benzimidazole derivative (BI-10), a promising broad-spectrum antibacterial agent with significant efficacy against Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus (MRSA). BI-10 demonstrated a minimum Inhibitory Concentration (MIC) of 2.4 μg/ml (6.25 µM) against these priority pathogens. In vitro assessments revealed that BI-10 possessed rapid bactericidal activity, anti-biofilm potential and showed synergistic interactions with conventional antibiotics. Ex vivo efficacy studies using various mammalian cell lines demonstrated strong intracellular killing. Moreover, BI-10 showed the ability to prevent both adhesion and invasion of pathogens in different mammalian cell infection models. The membrane-disrupting nature of BI-10 against both pathogens was established using scanning electron microscopy, membrane permeability, depolarization, and integrity assays. Multiple in silico analyses further demonstrated drug-likeness and the biocompatibility profile of BI-10. Altogether, our findings establish BI-10 as a potent, broad-spectrum antimicrobial agent with robust in vitro and ex vivo efficacy and bacterial cell membrane-disrupting activity. This study highlights BI-10 as a valuable antibacterial lead molecule for the development of a new antimicrobial agent against multidrug-resistant pathogens.

Tauopathies arise when normal functions of the tau protein in axonal transport and neuronal maintenance are disrupted by an imbalance between kinases and phosphatases. Dysregulation of key kinases such as dual-specificity Tyrosine-Regulated Kinase 1A (DYRK1A), Tau Tubulin Kinase 1 (TTBK1), and ABL Proto-Oncogene 1, and Non-Receptor Tyrosine Kinase (ABL1) drives excessive tau phosphorylation and neurofibrillary tangle accumulation. DYRK1A regulates MAPT exon 10 splicing and phosphorylates tau at multiple Ser/Thr residues, priming it for further phosphorylation by other kinases. TTBK1 phosphorylates tau at disease-associated epitopes within the microtubule-binding domain, promoting detachment from microtubules and aggregation. ABL1 phosphorylates tau at tyrosine residues, linking tau modification with Aβ-induced synaptic dysfunction. These events collectively drive tau hyperphosphorylation, misfolding, and neurofibrillary pathology characteristic of tauopathies. To identify natural product-derived multitarget inhibitors for these kinases, we developed a comprehensive machine learning (ML) workflow trained on bioactivity data from ChEMBL and BindingDB. We implemented five distinct classifiers: CatBoost, Support Vector Machine (SVM), k-Nearest Neighbors (KNN), Naive Bayes, and XGBoost. Stratified sampling and SMOTE were employed to address class imbalance for DYRK1A and ABL1, while Bemis-Murcko scaffold splitting was used to ensure rigorous evaluation of the data-scarce TTBK1 data set. A soft-voting ensemble model, integrating optimized CatBoost, XGBoost, and SVM, demonstrated superior performance. This robust ensemble was deployed to screen ∼695,000 natural compounds from the COCONUT 2.0 database. The resulting hits were refined through consensus molecular docking and deep learning-based rescoring (GNINA), leading to the identification of two high-potential lead molecules, CNP0591834.1 and CNP0484145.0. Validation using 1 μs molecular dynamics simulations confirmed their conformational stability and strong binding affinities. Steered MD further demonstrated their superior mechanical resistance to unbinding, particularly in DYRK1A and ABL1 complexes. Overall, this integrative computational framework highlights these two natural compounds as potent multitarget leads with strong potential to mitigate tau-hyperphosphorylation-driven neurodegeneration.

Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid (5 F), an ent-kaurane diterpenoid from the Chinese folk medicine Pteris semipinnata L., has been reported to be a potential anticancer agent. However, its moderate activity and toxicity have hindered its drug development. In the present study, a novel series of 5 F derivatives were successfully synthesised through hetero-Michael addition reactions, and their antiproliferative potential was systematically evaluated against a panel of human cancer cell lines. Among them, compound 29 was approximately 7-fold more potent than the parent 5 F in HCT116 cells, with an IC50 value of 0.957 μM. Furthermore, 29 exhibited comparable anti-tumor activity to 5-fluorouracil (5FU) in HCT116 xenograft nude mice, with reduced intrinsic toxicity. These results suggest that 29 could be considered a promising lead molecule for the treatment of cancer.

Piper betle, or betel vine, is a common chewing plant used for recreational purposes. Recently, there has been an increased interest in the cancer chemotherapeutic potential of dietary plants due to their antioxidant, antiinflammatory, and anti-mutagenic actions. While previous studies have demonstrated that P. betle leave extract inhibits the proliferation of breast cancer cells in vitro, the underlying mechanisms of its anti-cancer activity remain poorly understood. In this study, we investigated the effect of P. betle leaf extract on breast cancer cell lines and identified the possible mechanisms involved. Cell viability was assessed using the MTT assay. Annexin V and caspase-3/7 assays were used to determine apoptosis (programmed cell death). Major bioactive constituents in the extract were identified by using LC-MS and GC-MS techniques. We found that P. betle leaves' extract inhibited the proliferation of all the breast cancer cell lines. Mechanistic analyses revealed that P. betle mediated inhibition of cell growth is a result of cell cycle arrest and DNA fragmentation. Moreover, the expression of apoptotic genes increased, ultimately leading to the activation of caspase-3/7 and cell death. The major compounds identified were hydroxychavicol, methyl palmitate, (E)- methyl octadec-9-enoate, and coniferaldehyde. We therefore speculate that the anti-cancer effect of P. betle is largely dependent on these phytochemicals. In line with previously reported studies, our results demonstrate, for the first time, the anti-cancer effect of P. betle extract on the AU565 a HER2+ breast cancer cell line. Together with in silico and mechanistic approaches, we established that major compounds identified in the P. betle extract could be further evaluated as potential lead molecules against breast cancer. In summary, we report here for the first time that P. betle extract inhibits the proliferation of the AU565 (HER2+) breast cancer cell line, which may aid in advancing treatment options, particularly for HER2+ breast cancer.</p>.

Mechanistic insights into marine-derived PDE6D inhibitors disrupting prenyl-binding to modulate leukemia-associated RAS trafficking.

Targeting p38α, p38γ, and ERα with bioactive compounds from royal jelly for anti-breast cancer activity: A comprehensive in-silico study using fingerprint analysis, molecular dynamics, MEP, and PCA.

ABSTRACT Cancer's global prevalence demands novel therapeutic agents with improved efficacy and selectivity. In this study, six azo‐containing derivatives featuring trifluoromethyl (─CF 3 ) groups and a coumarin scaffold were comprehensively examined, five of which were newly synthesized. Their chemical structures were confirmed using FTIR, UV–Vis, and NMR ( 1 H and 13 C) spectroscopy. DFT calculations (B3LYP/6‐311++G(d,p)) supported the experimental data, showing strong agreement between theoretical and observed spectra. All compounds fulfilled Lipinski's rule of five, according to ADMEt studies; however, compounds 4b and 4c showed the lowest projected toxicity (LD 50 : 6480 mg/kg). High binding affinities toward cancer‐related VEGFR2 protein targets (PDB IDs: 3VO3, 6GQO, 3VHE, and 3WZD) were found via molecular docking. The compound that interacted with 3VHE the most was compound 5a (–11.2 kcal/mol). The 6GQO–4a complex was shown to be the most stable (–120.099 kJ/mol) by MM/PBSA analysis. This finding was confirmed by 100 ns molecular dynamics simulations that showed constant hydrogen bonding and stable RMSD values. These results point to compound 4a as a promising lead molecule for the development of anticancer drugs, which calls for additional in vitro and in vivo research.

Polyphenol oxidase (PPO) has been the subject of many inhibition studies due to its presence in many species, causing enzymatic browning in fruits and vegetables, and its pigmentation role in mammalian species. In the present study, the inhibitory properties of twelve 1-alkyl-1H-benzimidazolium sulfonates and two 1-alkyl-1H-benzimidazolium iodides on PPO activity have been investigated. The PPO enzyme was purified from banana using Sepharose 4B-tyrosine-p-aminobenzoic acid affinity gel chromatography. The effects of 14 different synthesized benzimidazole derivatives on the PPO enzyme were investigated, and they were found to significantly inhibit the enzyme in question. When ADME predictions were examined, it was seen that all compounds that include methylbenzenesulfonate structure showed good pharmacokinetic properties; only compounds 3a and 3b, which contain iodide, violated the Ghose rules. These compounds, which were effective at low concentrations, may serve as promising lead molecules for the development of PPO inhibitors with potential applications in food preservation, cosmetic formulations, and pharmaceutical research, pending further in vivo and formulation-based studies.

The KRASG12C mutation is a critical therapeutic target in the management of solid tumors, owing to its role in oncogenic signaling. Recent advances in covalent inhibitors that target mutant KRAS cysteine-12 have demonstrated the potential to halt aberrant signaling associated with this historically "undruggable" target. Here, we report the identification of 6-cyanoquinazoline covalent irreversible KRASG12C inhibitors. Lead optimization used structure-based design to identify novel switch-II pocket-binding motifs and in silico models to forecast in vitro metabolic stability and permeability. Human dose was improved by maximizing the rate of covalent modification (kobs/[I]) of KRASG12C-GDP, along with optimizing ADME parameters, to identify potent, orally bioavailable lead molecule 13de which demonstrated significant antitumor efficacy in the NCI-H1373 human lung adenocarcinoma xenograft model. Studies evaluating KRASG12C-GDP covalent target engagement, pharmacokinetics, and tumor growth inhibition estimated the efficacious human dose of 13de to be 192 mg administered once daily (QD), using allometric scaling.

Anti-Theileria equi activity of methanolic extract of Artemisia scoparia: In vitro efficacy, in vivo safety, and identification of lead molecules.

Discovery of novel indole derivatives as LRH-1 antagonists for the treatment of castration resistant prostate cancer.

Endophytic bacteria are an untapped source of bioactive compounds with potential antimicrobial applications. This study focused on isolating and evaluating endophytic bacterial strains from marigold (Tagetes spp.) for their antimicrobial activity and optimizing the production of bioactive compounds under various conditions. Twenty bacterial isolates were obtained from roots, stems, leaves, and flowers of marigold, with isolate F2 exhibiting the highest antimicrobial activity. Screening against E. coli, S. aureus, P. aeruginosa, and C. albicans revealed significant zones of inhibition, particularly for F2 (25 mm against E. coli and 23 mm against C. albicans), demonstrating its potential as a promising antimicrobial producer. The effects of temperature, carbon, and nitrogen sources, pH, and incubation time on antimicrobial production were investigated. Optimal production was observed at 35°C, pH 7.0, and 72 hours of incubation, with lactose and yeast extract identified as the most effective carbon and nitrogen sources, respectively. Purification through column chromatography and HPLC identified a bioactive compound at 12.254 minutes retention time, with mass spectrometry confirming a molecular weight of ~575 Da. Comparative analysis revealed the novelty and enhanced efficacy of this compound, highlighting its potential as a lead molecule for novel antibiotic development. This study uniquely explores endophytic bacteria from marigold as a source of bioactive compounds and identifies F2 as Bacillus kochii with potent antimicrobial activity. The structural analysis of a novel compound further underscores its pharmaceutical significance, contributing to the search for new antibiotics to combat antimicrobial resistance.

India's rich biodiversity is complemented by its vast knowledge of medicinal plants and traditional practices, with plant-based compounds having been harnessed as therapeutic agents for centuries. Cancer, a leading cause of global mortality, is fundamentally driven by DNA abnormalities that disrupt the function of key metabolic proteins. Ras proteins show a vital role in cell signalling, regulating cell proliferation and apoptosis, but mutations in these proteins may results in uncontrolled cell growth. This study investigated the anticancer potential of phytochemicals against mutated H-RAS, N-RAS, and K-RAS (RAS proteins) using molecular docking analysis. Aegle marmelos, rich in over a hundred phytochemicals with reported anticancer properties, was the source of 89 molecules screened as ligands. Seventy-eight molecules exhibited strong binding affinity (∆Gbind ≤ -5 kcal/mol) for H-Ras, 74 for K-Ras, and 77 for N-Ras, with α-amyrin and lupeol emerging as top leads for H-Ras, betulinic acid, β-amyrin, and lupeol for K-Ras, and βand α-amyrins for N-Ras. α-Amyrin emerged as a promising lead molecule against mutated Ras proteins, showing strong potential against H-Ras and N-Ras, and comparable efficacy to top leads against K-Ras. To overcome the challenges observed in drug likeness prediction connected with alpha amyrin, nano emulsion formulations strategies can improve its bioavailability and efficacy. Further validation of the lead molecules' anticancer efficacy requires comprehensive preclinical and clinical studies to confirm their biological activity.

In order to better understand the molecular behavior and to pave the way for potential future applications, the current work aims to synthesize the compound of ethyl 3-oxobutanoate, and use computational techniques to investigate its properties. It is well known that medicinal plants may contain components that can be used to treat illnesses. Hybanthus enneaspermus (L.) F. Muell, as it is commonly known, is a herb or shrub that grows in tropical and subtropical regions all over the world. It belongs to the family Violaceae. Because of its many pharmacological activities, H. enneaspermus (L), which contains a wide range of phytoconstituents, is a powerful medicine for treating a variety of acute or chronic illnesses. The potential to extract the material properties that could be used for particular applications is a major motivator for the thorough investigation of H. enneaspermus (L) and its spectroscopic implications. Ethyl 3-oxobutanoate is frequently employed as an intermediate in the synthesis of different pharmaceutical compounds as well as in the creation of fragrances and flavorings. FT-Raman and NMR spectra were recorded to analyze the structure of the material. The permitted regions, identification of secondary structures, confirmation of protein structure, and understanding of protein folding are all provided by the Ramachandran plot analysis. The Fukui function, the molecular electrostatic potential (MEP), and quantum chemical methods were used to analyze and visualize the electron density distribution within various molecular systems. Taking a sufficient quantity of other substances would not cause any problems because this substance conforms to Lipinski’s fifth rule. The findings offer crucial new details regarding the compound’s interactions, stability, and structural alterations in biological systems. The obtained thermodynamic properties fall between 100 and 700 K. In contrast to ΔH0 m, which decreases with increasing temperature, the gradients of C0 p,m and S0 m increase. The temperature-dependent thermodynamic analysis implemented in this work improves knowledge of the compound’s physiological behavior and provides important information for pharmaceutical and biotechnological applications that are sensitive to temperature. MESP provided additional reactivity information by identifying the electrophilic and nucleophilic regions of the molecule. The drug-likeness factors were computed taking into account the biological characteristics of the lead molecule.

Ebola virus disease remains one of the most serious viral infections with no approved small-molecule treatments. The Ebola virus glycoprotein (EBOV-GP), which enables the virus's entry to host cells, is a promising target for drug discovery. In this study, a multistage computer-aided drug discovery approach was used to identify new specific EBOV-GP inhibitors. A reliable QSAR model was built using 55 terpenoid derivatives. This model was able to predict the activity of newly designed compounds with good accuracy and validated statistical metrics (Rtr2 = 0.70; Rext2 = 0.73). It was subsequently applied to screen over 15,500 newly generated compounds from three lead molecules by fragment-based design tools. Predicted activity, binding affinity toward EBOV-GP, and good ADMET drug-like properties prioritized the eleven most promising hits. Through 150 ns molecular dynamics simulations, these compounds remained stable in the EBOV-GP binding site. Further binding free energy analysis (MM/PBSA) showed strong binding affinities, especially for the compounds L-60, L-832, M-1618, and L-1366. This study showed how combining QSAR, fragment-based design, docking, ADMET, and molecular dynamics could help in identifying potent and safe small molecules against the EBOV-GP. The top compounds are ready for further experimental and in vitro biological testing.

Abstract We have developed an antibody-enzyme fusion protein designed to overcome cancer treatment limitations due to immunosuppressive tumor microenvironments (TMEs) through a unique three-pronged mechanism of action. Our approach consists of a tumor-targeting nanobody fused to a xanthine oxidase (XO) enzyme domain modified to maximize superoxide production activity. The mechanism of action consists of: 1) the tumor-targeting nanobody domain binds to target-expressing cells, inhibiting growth with selectivity; 2) the XO enzyme domain produces extracellular superoxide, triggering cell death of the target and surrounding cells; and 3) the dying cells undergo immunogenic cell death (ICD), releasing damage-associated molecular patterns (DAMPs) into the TME to recruit immune cells. This plug-and-play approach delivers therapeutic enzymes specifically to the tumor microenvironment, bypassing conventional limitations and establishing a novel platform for oncology treatment. In this study, we investigated the feasibility of our approach in models of glioblastoma (GBM) using an EGFR-targeted fusion protein. A key molecular feature of GBM is EGFR amplification and mutation, however current EGFR-targeted therapies like cetuximab or kinase inhibitors have limited efficacy due to heterogeneous tumors and adaptive resistance. Due to an immunosuppressive TME, GBM tumors are frequently resistant to immunotherapies such as checkpoint blockade and CAR-T therapy. Approaches designed to induce an inflammatory response show promise in GBM, such as oncolytic viruses that trigger ICD. To validate the target binding and enzyme activity of our lead molecule, VISK-103, we have performed enzyme activity assays, ELISA binding assays, and flow cytometric analyses. To demonstrate proof of concept, we have measured efficacy both in vitro and in vivo, using cell viability assays and a mouse GBM xenograft model. In addition, we have employed several in vitro techniques and have demonstrated production of ICD markers (e.g. eATP, HMGB1) and measured immune activation via co-culture experiments. Our results indicate that VISK-103 treatment can bind to EGFR-positive GBM cells with a KD in the nanomolar range. VISK-103 treatment induces cell death in EGFR-positive cells with a sub uM IC50 in vitro for GBM cell lines. In a xenograft model of GBM, VISK-103 treatment led to a ∼50% reduction in tumor burden and prolonged animal survival. Additionally, VISK-103 treatment causes release of DAMPs and activation of dendritic cells in co-culture models with an over 3-fold increase in IL-1b release using VISK-103-treated cells compared to a chemotherapy control. Taken together, we are developing a first-in-class, plug-and-play platform using an antibody-enzyme scaffold that couples cell-selective killing with immune system engagement to provide a multi-pronged approach to GBM treatment. Citation Format: Camille H. Cushman, Bushra Dabbagh, Karistan Swan, Aakanksha Singh Parihar, Daniel T. Dransfield, Eric Fossel. Targeting glioblastoma using an antibody-enzyme fusion protein with a multi-pronged mechanism of action [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Brain Cancer; 2026 Mar 23-25; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2026;86(6_Suppl):Abstract nr A019.

Identifying potent lead molecules for specific targets remains a major bottleneck in drug discovery. As structural information about proteins becomes increasingly available, ultra-large virtual screenings (ULVSs) which computationally evaluate billions of molecules offer a powerful way to accelerate early-stage drug discovery. Here, we introduce AdaptiveFlow, an open-source platform designed to make ULVSs more accessible, scalable, and efficient. AdaptiveFlow provides free access to a screening-ready version of the Enamine REAL Space, the largest library of ready-to-dock, drug-like molecules, containing 69 billion compounds that we prepared using the ligand preparation module of the platform. A key innovation of the platform is its use of a multi-dimensional grid of molecular properties, which helps researchers explore and prioritize chemical space more effectively and reduce the computational costs by a factor of approximately 1000. This grid forms the basis of a new method for identifying promising regions of chemical space, enabling systematic exploration and prioritization of compound libraries. An optional active learning component can further accelerate this process by adaptively steering the search toward molecules most likely to bind a given target. To support a broad range of applications, AdaptiveFlow is compatible with over 1,500 docking methods. The platform achieves near-linear scaling on up to 5.6 million CPUs in the AWS Cloud, setting a new benchmark for large-scale cloud computing in drug discovery. Using this approach, we identified nanomolar inhibitors of two disease-relevant targets: ferroptosis suppressor protein 1 (FSP1) and poly(ADP-ribose) polymerase 1 (PARP-1). By leveraging newly solved crystal structures of FSP1 in complex with NAD+, FAD, and coenzyme Q1, we validated these hits experimentally and determined the first co-crystal structures of FSP1 bound to small-molecule inhibitors, enabling insights into inhibitor binding mechanisms previously unknown. With its high scalability, flexibility, and open accessibility, AdaptiveFlow offers a powerful new resource for discovering and optimizing drug candidates at an unprecedented scale and speed.

In this work, a synthetic strategy based on the reaction of 5-bromo-2-(4-fluorophenyl)-4-methyl-2H-1,2,3-triazole with various thioaromatic derivatives were applied to afford a series of thioether derivatives of 1,2,3-triazole-N-oxides in yields of 52–80%. The obtained compounds were shown to be characterized by low cytotoxicity towards human embryonic kidney (HEK293) cells with IC50 values ranging from 128 to 76 µM and high toxicity towards malignant rhabdomyosarcoma cancer (Rd) cells with IC50 values ranging from 93 to 22 µM. These compounds were screened for the ability to induce nitric oxide production in cells via DAF-FM fluorescence. Several compounds demonstrated a twofold increase in nitric oxide levels relative to the control. Two leader molecules demonstrating the lowest cytotoxicity values and the effect on nitric oxide production in rhabdomyosarcoma cells were identified. Thus, the method could be considered as a convenient one for qualitative or semi-quantitative determination of nitric oxide in a screening process of large number of compounds to define lead molecules capable of influencing nitric oxide production.