Breast cancer is known as a frequently diagnosed malignancy in women. Over 70% of cases express estrogen receptor α (ERα). The activation of ERα promotes tumor proliferation and progression. Furthermore, mutations in ERα lead to acquired resistance against standard endocrine therapies such as tamoxifen. The induced resistance posed a significant clinical challenge in metastatic breast cancer. In this study, research for identifying novel, naturally derived compounds to inhibit the Y537S-mutated ERα was conducted using in silico methods supported by an in vitro cytotoxicity screen. Molecular docking and molecular dynamics simulations served as the primary in silico screening strategies. The top 2% of candidates were filtered from a docking screen of the IBS natural library composed of over 15,000 chemicals. From this group, 11 compounds were purchased and tested using a cell-based cytotoxicity assay in MCF-7 before advancing to detailed simulations. Five candidates were then advanced to 150 ns MD simulations. A post-MD analysis followed, including MM-PBSA binding free energy calculations and principal component analysis (PCA). The phytochemical ibs-04156 was identified as the most promising overall candidate, predicted to maintain consistent stability across both the wild-type and Y537S-mutated ERα, while ibs-18821 demonstrated potent mutant-specific inhibition via an allosteric mechanism involving spatial deviations in distal helices H3 and H11. This result shows that mutated ERα can be potentially targeted by bioactive phytochemical scaffolds found through molecular modeling, presenting a potential therapeutic strategy for metastatic breast cancer resistant to therapies such as tamoxifen.
PIKfyve inhibitors have evolved from niche probes into powerful tools for studying endolysosomal biology. Although PIKfyve inhibition has revealed important roles in membrane trafficking, autophagy, viral entry, and cancer-cell vulnerability, clinical translation remains limited by a narrow therapeutic window because disruption of PI(3,5)P2 homeostasis causes profound lysosomal dysfunction. Future progress will depend on tuneable or partial inhibitors, improved selectivity, and targeted delivery. PIKfyve therefore provides a useful model for balancing potency, precision, and safety in lipid kinase drug discovery.
Glioblastoma (GBM) is a highly aggressive, poor-prognosis brain tumor classified as WHO grade IV, for which effective treatments remain limited. Although AlkB homolog 3 (ALKBH3), a demethylase for 1-methyladenosine (m1A) and 3-methylcytidine (m3C) in DNA and RNA, has been implicated in cancer cell proliferation, its functional role in GBM remains unclear. We aimed to characterize the role of ALKBH3 in GBM and develop and evaluate a patented ALKBH3 inhibitor, HUHS199, to identify its molecular targets and therapeutic potential. We found that ALKBH3 was highly expressed in clinical GBM specimens; its knockdown inhibited GBM cell proliferation. HUHS199 inhibited GBM cell proliferation in a dose-dependent manner, induced G1 phase cell cycle arrest, and increased m1A levels in RNA. Furthermore, RNA immunoprecipitation-microarray analysis using an anti-m1A antibody identified growth arrest and DNA-damage-inducible protein GADD45 alpha (GADD45A) mRNA as a target of ALKBH3-mediated demethylation. These findings suggest that ALKBH3 promotes GBM cell proliferation via m1A demethylation of GADD45A mRNA. Collectively, this study highlights the therapeutic potential of ALKBH3 inhibition and presents HUHS199 as a first-in-class candidate for GBM treatment.
Emerging evidence indicates that coagulation-related molecular programs are associated with thrombosis, tumor progression, and molecular dysregulation in gastric cancer (GC). However, thrombosis-associated molecular features in GC and their potential links to inherited susceptibility remain insufficiently understood. Integrated analyses of transcriptomic data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets were performed to identify thrombosis-associated genes and establish a machine learning-based prognostic signature. Genome-wide association study (GWAS), expression quantitative trait loci (eQTL), transcriptome-wide association study (TWAS), and Mendelian randomization (MR) analyses were conducted to investigate susceptibility-associated transcriptional programs in GC. Functional assays were used to evaluate candidate genes associated with malignant phenotypes. Single-cell RNA sequencing (scRNA-seq) and cell-cell communication analyses were further performed to characterize cell-type-specific expression patterns and potential intercellular interactions. A total of 22 differentially expressed thrombosis-associated genes were identified, and a prognostic signature comprising 14 genes was established. The signature stratified patients into high- and low-risk groups and showed prognostic performance in both the training and validation cohorts. Integrative GWAS, eQTL, and TWAS analyses identified susceptibility-associated transcriptional programs that were positively correlated with the thrombosis-associated risk score. Silencing ACTN2 and CRYAB significantly reduced GC cell migration and invasion. scRNA-seq analysis revealed relatively high CRYAB expression in neutrophils, and CellChat analysis suggested potential neutrophil-B cell interactions involving COLLAGEN-related signaling. This integrative multi-omics study identified a thrombosis-associated molecular signature linked to prognosis and germline susceptibility-associated transcriptional programs in GC. ACTN2 and CRYAB may represent candidate genes associated with GC cell migration and invasion, while single-cell analysis suggested potential immune-related communication features.
The anticancer efficacy of 4-aryl-2-hydrazinothiazole derivatives, which combine a thiazole known for its wide range of medicinal applications and a hydrazide/hydrazone structure known for its unique biological properties, has been determined in numerous studies. In this study, novel 2-[4-[(2-(4-substituted thiazol-2-yl)hydrazono)methyl]phenoxy]acetic acid derivatives (2a-l) were synthesized and evaluated for their anticancer activity in MCF-7 breast cancer, A549 lung cancer, and L929 normal cell lines using cytotoxicity, apoptosis, and caspase-3 activation assays. The compounds were found to exhibit high cytotoxicity, particularly showing more selective and high-potential antiproliferative activity on the MCF-7 cell line. In apoptosis studies, compounds 2c, 2d, and 2h caused programmed cell death in MCF-7 cells exceeding 24%, close to cisplatin. Among these compounds, the derivative 2c containing 4-methoxyphenyl caused caspase-3 activation at a level similar to cisplatin. To conduct in silico studies of the compounds, molecular docking studies with the (4QTX) caspase-3 enzyme, molecular dynamics simulation for three compounds (2d, 2g and 2l), and Density Functional Theory (DFT) studies were performed. These studies have shown that the carboxylate groups in the compounds form strong and stable interactions with Arg64 and Arg207.
Human African trypanosomiasis (HAT), caused by Trypanosoma brucei, remains a neglected tropical disease with a critical shortage of therapeutic options, underscoring the need for new chemotypes. Cell division cycle-2-related kinase 12 (CRK12) has emerged as a genetically essential and chemically validated target in kinetoplastids, yet experimentally supported CRK12-targeting chemotypes against T. brucei remain limited, and no experimental CRK12 structure is available. Here, we developed an integrated computational-experimental screening workflow to accelerate CRK12-guided hit discovery, combining ligand-based prescreening, structure-based virtual screening, molecular dynamics (MD) refinement, and experimental validation. An 8.6-million-compound library was prescreened using MACCS fingerprint screening and complex-based pharmacophore screening, followed by staged docking and MD simulations to prioritize candidates for experimental testing. Notably, four of ten purchased compounds showed strong growth inhibition against T. brucei at 10 μM, with IC50 values of 6.09, 1.47, 0.81, and 1.33 μM, demonstrating a promising hit rate for this data-limited target. Binding-mode analysis revealed a conserved hinge-anchoring interaction pattern in the ATP-binding pocket, providing a structural rationale for follow-up analogue design. Overall, this study identifies new chemical starting points for anti-T. brucei drug discovery and demonstrates the utility of an AlphaFold2-enabled CRK12-guided screening strategy for targets with limited structural and ligand data.
Nucleic Acid Therapeutics (NATs), including Antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), and messenger RNAs (mRNAs), are a rapidly developing class of therapeutics capable of specifically regulating previously considered undruggable and inaccessible genes and pathways for modification by small molecules and antibodies. Despite promising results, the utilization of NATs in clinical practice is complicated by the potential off-target effects, such as activation of the immune response and organ-specific toxicity, which cannot be effectively predicted based solely on primary structure, chemotype descriptors, or off-target effects predictors developed in silico. A combination of multiple omics technologies, including proteomics/metabolomics/single-cell transcriptomics, helps researchers elucidate the interaction of drug compounds with biological targets. This allows for the detection of changes not only at the pathway and cellular level but also early signs of toxicity in parallel. Thus, in this context, this review offers a mechanistic view on the use of multi-omics strategies for the investigation of NATs-induced biological effects to analyze the mechanism of action of chemically modified ASOs, siRNAs and mRNA conjugates. The review also discusses case studies in which multi-omics data have been used to improve therapeutic development. By examining individual layers of molecules separately, a more holistic understanding of treatment mechanisms can be achieved, which is helpful for the discovery of biomarkers and the development of next-generation nucleic acid drugs.
Currently, there is a lack of effective disease-modifying drugs for osteoarthritis (OA), and existing treatment methods often accompany significant adverse reactions. Schisandra chinensis (SC), as a traditional Chinese medicine with anti-inflammatory activity, may have therapeutic potential for OA, but its specific molecular mechanism is not yet clear. This study adopts network pharmacology strategy to explore the potential mechanism of SC intervention in OA. Screen SC active ingredients through TCMSP database and obtain OA-related targets by combining OMIM, TTD, and GeneCards databases. Venn analysis shows that there are only 5 common targets between SC and OA; Further analysis revealed that PTGS2 is the only candidate target with diagnostic value (AUC = 0.702, p = 0.015), while AR, ESR1, DPP4, and CHRM2 did not show clinical diagnostic efficacy. Functional enrichment analysis suggests that common targets mainly involve steroid hormone responses and extracellular matrix (ECM) related pathways, with a significant enrichment in "response to steroid hormones" (adjusted p = 1.2 × 10 -5), and GSEA also shows activation of the ECM-receptor interaction pathway (NES = 1.87, p = 0.002). The molecular docking results indicate that the SC active ingredients MOL008957 and MOL008978 can form stable binding with PTGS2. The qRT-PCR results further confirmed that the expression of PTGS2 was significantly downregulated in the OA model after SC intervention. Single cell transcriptome analysis showed that PTGS2 was mainly enriched in monocytes and gradually increased with the pseudo temporal progression; Cell communication analysis revealed that monocytes can interact with NK cells and T cells through the MIF-CD74 + CXCR4 and MIF-CD74 + CD44 signaling axes. After virtual knockout of PTGS2, downstream regulatory networks suggest that GSN may be a key target gene, mainly involving collagen containing extracellular matrix and ECM structural components. In summary, SC may exert its anti OA effect mainly by selectively inhibiting PTGS2 and maintaining the homeostasis of ECM related pathways. PTGS2 is expected to become a candidate biomarker for OA and a potential target for SC intervention in OA, but further experimental verification of the specific efficacy and mechanism of SC active ingredients is still needed.
Esophageal squamous cell carcinoma (ESCC) features epithelial heterogeneity and an immunosuppressive microenvironment, yet clinically relevant malignant epithelial states remain poorly defined. We integrated four single-cell RNA-sequencing datasets and 10 bulk transcriptomic cohorts totaling 1318 samples, and applied cross-cohort differential expression, survival analysis, and a machine-learning framework of 113 algorithm combinations to screen for malignant epithelial cell-associated biomarkers. GNGT1 was prioritized for its consistent upregulation, prognostic association, and limited prior characterization in ESCC. Across independent cohorts, GNGT1 exhibited favorable diagnostic performance, while high expression correlated with poorer overall survival and more advanced local tumor status. Immune deconvolution consistently linked GNGT1-high tumors to reduced CD8+ T-cell infiltration, lower immune scores, and decreased cytotoxic T-cell markers. Single-cell analyses further associated GNGT1-high epithelial cells with altered epithelial-immune communication involving MIF, prostaglandin, and CXCL signaling, alongside enrichment of epithelial-mesenchymal transition, mTORC1, and proliferative programs; qPCR confirmed elevated GNGT1 expression in ESCC cell lines. This study defines GNGT1 as a marker of an immunosuppressive malignant epithelial state, thereby bridging epithelial heterogeneity with immune remodeling in ESCC. These findings support GNGT1 as a candidate diagnostic, prognostic, and biologically informative biomarker warranting mechanistic and clinical validation.
Colorectal cancer (CRC) is the second most common cause of cancer-related deaths in the United States. The incidence of early-onset CRC (EOCRC) has been rising in the past few decades. Given the multifaceted nature of EOCRC, the genetic and environmental factors contributing to this malignancy have remained elusive. Systems genetics is a multi-omics tool that analyzes multiple genes as a collective network. Transcriptional correlation, protein-protein interaction network, functional annotation, and drug-gene interaction analyzes enable investigators to explore the functional roles of target genes and aid with discovery of novel diagnostic biomarkers and potential therapeutics. To clarify the objective of this work, this manuscript serves three purposes: a narrative review of recent systems genetics applications in EOCRC research, a step-by-step protocol for conducting these analyses, and a discussion of current advances in systems genetics to address limitations in generalizability and context dependence. Through this manuscript, we envision a future in which emerging oncology researchers adopt these tools into their investigations of EOCRC and other malignancies.
S-palmitoylation is a crucial post-translational modification that regulates diverse cellular processes, particularly signalling pathways. The ZDHHC family of enzymes catalyzes this modification, however, the role of ZDHHC16 in cancer warrants in-depth investigation. This study presents a comprehensive pan-cancer analysis of ZDHHC16, examining its expression patterns, clinical relevance, associations with immune responses, genomic characteristics, and potential as a therapeutic target. We further analysed single-cell and spatial transcriptomic data to identify specific cell populations associated with ZDHHC16 dysregulation. Our findings reveal that ZDHHC16 dysregulation is tissue-specific and promotes tumor progression and modulates immune responses, highlighting its potential as a therapeutic target in several cancers, including ACC and BRCA. In ACC and BRCA, high ZDHHC16 expression correlates with shorter overall survival (p < 0.001) and regulates PD-L1 expression.
Hyperuricemia (HUA) is a common metabolic disorder with limited safe and effective therapeutic options. This study integrated GEO dataset mining and network pharmacology to explore the anti-HUA efficacy and mechanism of pomegranate peel polyphenol extract (PPE). In a mouse model of HUA induced by potassium oxonate and 5% fructose water, PPE significantly reduced serum, urinary, and fecal uric acid levels, attenuated the increases in creatinine and blood urea nitrogen, improved estimated glomerular filtration rate, and ameliorated renal pathological damage, inflammation, and xanthine oxidase activity. Integration of GEO-derived HUA-related genes and computational target prediction for PPE identified 44 common targets, and protein-protein interaction network analysis revealed core targets including Akt1. KEGG enrichment highlighted the PI3K-AKT signaling pathway as a key mediator. Western blotting in vivo and in vitro confirmed that PPE suppressed PI3K-AKT phosphorylation and downregulated the urate reabsorption transporters URAT1 and GLUT9. Furthermore, experiments in UA-induced HK-2 cells demonstrated that ellagic acid, a major bioactive component of PPE, acted through the same pathway. These findings indicate that PPE reduces uric acid levels and protects the kidney via modulation of the PI3K-AKT pathway, providing an integrative data-driven rationale for its potential as a functional food or pharmaceutical agent against HUA.
The tetrahedral DNA nanostructure (TDN) has emerged as a premier, highly programmable framework for targeted drug delivery and diagnostics. This review provides a definitive, chemistry-first design guide for TDN engineering, systematically structured across three translational tiers. First, we examine foundational derivatives, detailing how precise modifications across the three components of the nucleotide scaffold confer essential nuclease resistance and structural longevity. Second, a diverse array of conjugation strategies is compiled, mapping the chemical mechanics of stable and responsive covalent linkages alongside distinct non-covalent loading modalities, including intercalation, metallo-coordination, and groove binding. Third, we explore advanced derivatives for materials science, detailing how merging TDNs with traditional nanotechnology or smart polymeric matrices yields hybrid architectures engineered into sophisticated analytical tools and biomaterials. Advancing this platform will entail integration of AI-driven computational models and navigating critical physiological barriers, establishing the functionalized TDN as a definitive cornerstone of next-generation nanomedicine.
A series of new substituted 1,6-dihydropyrimidinones was designed, synthesized, and biologically evaluated as potential antibacterial candidates and β-lactamase enzyme inhibitors. All the synthesized compounds were tested for their antibacterial activity against Staphylococcus aureus, Bacillus subtilis as a gram-positive species whereas Pseudomonas aeruginosa as a gram-negative species was utilized. Most of the compounds exhibited moderate antibacterial activity compared to the reference drug amoxicillin. Furthermore, clinical antibacterial tests were conducted on β-lactamase resistant strains, including Acinetobacter baumannii, Bacillus subtilis, and Staphylococcus aureus. Results indicated that the candidate compounds possess potentiation effect, indicating their role as effective β-lactamase enzyme inhibitors. Whereas in-vitro assay performed on β-lactamase enzymes revealed that compounds 3d, 3i, and 7b (IC50 = 0.758, 0.400, and 0.524 nM) have lower IC50 values compared to clavulanic acid (IC50 = 0.934 nM). These findings suggested that compounds 3d, 3i, and 7b are considered as promising hits for further exploration of potent and selective β-lactamase inhibitors.
Benzenesulfonylpiperazines have been previously identified as a promising class against Chagas disease and leishmaniasis, two parasitic neglected tropical diseases. Thus, the pharmacokinetic profile of two potential leads against visceral leishmaniasis was assessed in vitro. Both lead candidates 1 and 2 exhibited a satisfactory ADME profile, with 2 proving slightly superior in terms of metabolic stability. Therefore, after complementary toxicity assessment which revealed that 2 did not exert hepatotoxicity in vitro or acute toxicity in vivo, the evaluation of its efficacy in a mouse model highlighted that 2 indeed displays antileishmanial efficacy in vivo. Albeit moderate at a dose of 50 mg/kg/day, its efficacy increased at a dose of 100 mg/kg/day, reducing the parasite burden by 90% in the spleen of infected mice, though safety concerns were raised at this dose. In vitro investigation of its mode of action revealed that 2 exerts a cytostatic effect on Leishmania infantum promastigotes, promoting cell cycle arrest during the G0/G1 phase which may potentially be linked to the production of reactive oxygen species.
Ischemic stroke (IS) is a cerebrovascular disease with high mortality and disability rates, currently lacking effective therapeutic targets. The STING inhibitor SN-011 shows potential in IS treatment, but its mechanism of action remains unclear. This study aims to explore the key molecular mechanisms of SN-011 in treating IS through bioinformatics approaches. IS transcriptome datasets were analyzed to identify differentially expressed genes. Mendelian randomization using brain eQTL and IS-GWAS data identified genes with causal relationships to IS. Single-cell transcriptome, pseudo-time trajectory, intercellular communication, and transcription factor regulatory network analyzes were performed. Molecular docking and DARTS-WB assay validated SN-011 binding to transcription factors. Transcriptomic analysis identified 77 intersecting genes. Mendelian randomization revealed ADGRE5 as a protective gene for IS (OR < 1), significantly downregulated in venous endothelial cells (vECs) during disease progression. Cell communication analysis showed ADGRE5-high vECs interact with immune, glial, and stromal cells via LAMININ (Lamb2-CD44, Lamb2-Itga6+Itgb1, Lamb2-Dag1 pair) and JAM signaling pathways. Transcription factor analysis identified JUNB as a negative regulator of ADGRE5. Molecular docking (-6.8 kcal/mol) combined with an in vitro DARTS-WB assay confirmed the interaction between SN-011 and JUNB. In OGD-induced endothelial cell injury models, SN-011 suppressed JUNB expression, restored ADGRE5 expression inhibited by JUNB overexpression, reversed the downregulation of LAMB2 and CD44, and reduced the expression of the pro-inflammatory cytokines IL-6 and IL-1β. Notably, blockade of LAMB2 largely abolished these protective effects, indicating that the anti-inflammatory and endothelial-protective activities of SN-011 are mediated, at least in part, through restoration of the LAMB2-CD44 signaling axis. ADGRE5 downregulation in vECs may impair vascular repair by disrupting LAMININ-mediated intercellular communication. SN-011 may exert neuroprotective effects by targeting JUNB to upregulate ADGRE5 expression and restore the vEC-centered cellular communication network, providing a theoretical basis for SN-011 as a potential IS therapeutic.
Osteoarthritis (OA) progression is driven by inflammatory mediators and immune dysregulation within the joint. Curcumin (Cur) possesses multi-target therapeutic potential. However, its clinical application is limited by poor solubility and a short half-life. In this study, we developed injectable Cur-loaded GelMA microspheres (Cur-MS) and evaluated their effects in IL-1β stimulated chondrocytes, cartilage organoids, and a monosodium iodoacetate (MIA) induced rat OA model, complemented by public transcriptomic analysis (GSE114007). The Cur-MS demonstrated uniform size distribution, favorable biocompatibility, and sustained curcumin release. Treatment with Cur-MS significantly reduced chondrocyte apoptosis, reactive oxygen species levels, and hypertrophic markers; restored COL-II and ACAN synthesis; and downregulated MMP13 and inflammatory gene expression. In vivo, Cur-MS improved joint space, alleviated pain, decreased synovial CD68 positive macrophage infiltration and levels of IL-6 and TNF-α, and enhanced chondrogenic gene expression. Public transcriptomic data corroborated these findings, revealing upregulation of MMP13 and downregulation of SOX9 in OA cartilage, consistent with our experimental targets. Collectively, this study provides the first multi-model evidence combined with transcriptomic validation, demonstrating that Cur-MS not only directly protects chondrocytes and restores extracellular matrix homeostasis but also modulates the joint inflammatory immune microenvironment. These findings suggest that Cur-MS represents a promising locally sustained-release therapeutic strategy for OA.
The global spread of drug-resistant Plasmodium falciparum, particularly artemisinin-resistant strains, underscores the urgent need for novel antimalarial agents with distinct mechanisms of action and improved therapeutic potential. In this study, we employed an integrated in silico and in vitro strategy to identify compounds with inhibitory activity against P. falciparum apicoplast Gyrase B (PfGyrB). High-throughput virtual screening identified hit compounds with favorable predicted interactions against the target protein, which were subsequently evaluated using biochemical ATPase inhibition and parasite growth inhibition assays. UNC8153 and Fexofenadine hydrochloride demonstrated time-dependent antiplasmodial activity, with lower IC50 values at 96 h than at 48 h under prolonged exposure conditions. UNC8153 exhibited greater antiplasmodial activity (~25-fold) than the reference compound novobiocin during the second intraerythrocytic cycle. Morphological analysis indicated impaired parasite development during the second intraerythrocytic cycle under prolonged exposure conditions. Importantly, UNC8153 retained inhibitory activity against the artemisinin-resistant C580Y strain and exhibited minimal cytotoxicity toward HEK-293 cells under the tested experimental conditions. Structural similarity analysis indicated that the identified compounds are chemically distinct from currently used antimalarial drugs, supporting their structural novelty. Collectively, these findings identify UNC8153 as a structurally distinct new antiplasmodial scaffold warranting further mechanistic, pharmacological, and in vivo evaluation.
Replication is started by DNA primase, which synthesizes an oligoribonucleotide primer that DNA polymerases extend. Effective chemotherapy is still needed to manage and treat leishmaniasis, which continues to pose a threat to public health around the world. Repurposing drugs offers alternative uses for drugs with established pharmacological effects, saving money and increasing the pool of human resources available to create novel anti-leishmanials. Here, we used a non-radioactive primase-pyrophosphatase assay to biochemically characterize the Leishmania donovani nuclear DNA primase (LdPri) subunits, followed by in silico evaluation of inhibitors targeting LdPri. The best-evaluated inhibitors showed LdPri inhibition in vitro under conditions similar to those of the primase-pyrophosphatase experiment. The MTT assay also confirmed the inhibitors' anti-leishmanial properties. Parasite growth and morphological analysis were performed by culturing cells in the presence of inhibitors. Biochemical characterization revealed that LdPriS activity was unstable, but was highly stabilized upon association with LdPriL in LdPri. LdPri was found to be thermostable and possesses 3'-terminal nucleotidyltransferase activity. Additionally, in silico and in vitro studies evaluated Pritelivir (BAY 57-1293) as the most efficient LdPri complex inhibitor, followed by Epigallocatechin Gallate (EGCG). Moreover, kinetic studies showed that Pritelivir and EGCG exhibit competitive and uncompetitive inhibition, respectively, for both NTP and DNA substrates. Pritelivir effectively disrupted the L. donovani cell cycle, and the replication mechanism in treated promastigotes showed irregular shapes and short flagella.
The COVID-19 pandemic highlighted the role of rapid viral mutation and global connectivity in accelerating viral emergence and spread, emphasising the necessity for expedited and adaptable antiviral drug discovery and development. Despite ongoing efforts to develop effective, low-toxicity therapeutics, the number of antivirals that have achieved clinical approval remains limited. The shortfall is especially significant in developing countries, where access to new antivirals is limited by high prices, few options, import dependence, unstable supply chains and weak purchasing systems. The challenge is further heightened by the emergence of increasingly drug-resistant variants while vaccines often provide inadequate protection against newly mutated or novel viruses. Consequently, the identification of novel antiviral agents that are both effective and cost-efficient via innovative strategies for antiviral drug discovery is essential to manage and control viral infections. Therefore, this review examines the different challenges associated with conventional antiviral drugs alongside recent strategies in antiviral drug discovery and development, such as the exploitation of plant secondary metabolites with antiviral properties, advanced microscopy technologies, computer-aided drug design, artificial intelligence and machine learning, gene-editing technologies, drug combination therapy and nanotechnology-enhanced drug delivery systems. Additionally, this study proposes a simple decision-focused pathway integrating natural products, computation and targeted delivery to guide candidate prioritisation, optimisation and translation from discovery to implementation. While these emerging strategies offer considerable promise, challenges related to validation, toxicity, scalability and equitable access remain important considerations for successful clinical translation. Future research should therefore integrate complementary technologies to accelerate the development of effective antiviral agents against current and emerging viral threats.
