ACS Bio & Med Chem Au最新文献

{"title":"mSumireF a Monomeric Violet Fluorescent Protein.","authors":"Jacob Kress, Sookyeong Kim, Caitlynn Bryant, Emily Camila Lopez-Lopez, Jiali Zha, Mathew Tantama","doi":"10.1021/acsbiomedchemau.5c00175","DOIUrl":"10.1021/acsbiomedchemau.5c00175","url":null,"abstract":"<p><p>The development of genetically encoded labels for multicolor experiments requires a diverse palette of fluorescent proteins that are well-behaved. Here, we report mSumireF, a monomeric variant of the violet fluorescent protein Sumire. On its own, the canonical monomerizing valine-to-lysine mutation at residue 206 causes a significant loss of brightness for the mSumire variant. We found that brightness is recovered upon the reversion of mSumire's tyrosine at position 165 back to phenylalanine as in its superfolder GFP grandparent. Importantly, this mSumireF variant exhibits significantly less oligomerization tendency in the organized smooth endoplasmic reticulum (OSER) assay in mammalian cells and in protein solution assays. Furthermore, we demonstrate that an mSumireF donor can be effectively paired with the fluorescent protein acceptors mCerulean3, mTurquoise2, and LSSmScarlet to generate FRET-based ATP biosensors. mSumireF thus provides an improved violet fluorescent protein to expand color options with reduced concern of unwanted dimerization.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"6 1","pages":"36-43"},"PeriodicalIF":4.3,"publicationDate":"2026-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921512/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147272103","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Blue Fluorescent Siloxytecans Exhibit Potent Anticancer Activity and Enable Direct Real-Time Quantification of Intracellular Uptake.","authors":"Isabella Lo, Sanika Vaidya, Ashley Mo, Alyssa Yee-Xin Chia, Katelyn Li, Jessica Parvin, Sripathy Sadagopan, Zane D'souza, Olivia Kwok, Jenny Zhang, Isa Baratoff, Shreya Somani, Lekhya Menta, Yining Xie, Thomas Sanchez, Terry Wang, Rebecca Chen, Akira Yamamoto, Joseph Pazzi, Edward Njoo","doi":"10.1021/acsbiomedchemau.5c00155","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.5c00155","url":null,"abstract":"<p><p>Notwithstanding the natural abundance of silicon on Earth, silicon-containing compounds comprise relatively few pharmaceutical drugs, though several silylated natural products have demonstrated greater bioavailability and lipophilicity. Here, we apply this strategy in the preparation and biological evaluation of synthetic camptothecin analogs involving a C-10 silyl ether on SN-38, which not only blocks a site of metabolism but also enhances bright-blue fluorescence properties in such compounds. These siloxytecans exhibit comparable dose- and time-dependent antiproliferative activity in a broad panel of cancer cells. Uniquely, we demonstrate that the enhanced fluorescence of these compounds enables real-time, quantitative visualization of the dynamics and selectivity of intracellular uptake through fluorescence microscopy without the need for extensive sample preparation or installation of auxiliary fluorophores. We further demonstrate that the kinetics of cellular uptake observed by fluorescence microscopy are consistent with time-course washout experiments with subtle differences in anticancer potency in several cell lines. Further cell cycle analysis by flow cytometry and cell-free topoisomerase inhibition studies suggests that these siloxytecans retain the topoisomerase-inhibiting properties of camptothecin and other related topoisomerase I inhibitors. Collectively, these studies highlight the utility of quantitative fluorescence microscopy in investigating mechanisms of biological transport and the anticancer activity of such siloxytecans.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"6 1","pages":"1-14"},"PeriodicalIF":4.3,"publicationDate":"2025-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921511/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147272094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Is Thiourea the Weak Link? An Investigation of the In Vitro and In Vivo Destabilization of [²⁰³Pb]- and [²¹²Pb]Pb²⁺ Thiourea-Based Bioconjugates.","authors":"Brooke L McNeil, Luke Wharton, Chao-Cheng Chen, Helen Merkens, Milena Čolović, Cristina Rodríguez-Rodríguez, John Wilson, Stefan Mair, Chengcheng Zhang, Anthony W McDonagh, Kuo-Shyan Lin, François Bénard, Frank Wuest, Paul Schaffer, Caterina F Ramogida","doi":"10.1021/acsbiomedchemau.5c00224","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.5c00224","url":null,"abstract":"<p><p>Fibroblast activation protein (FAP) is overexpressed in a variety of cancers, making it an attractive target for bifunctional chelator-based radiopharmaceuticals. This study initially aimed to assess the effect of chelator structure on the biodistribution of ²⁰³Pb/²¹²Pb-labeled FAP inhibitor (FAPI) bioconjugates. However, suboptimal in vivo biodistribution and imaging results suggested the bioconjugate was unstable. RadioHPLC analysis of urine samples suggest the thiourea bond, formed during conjugation between an amine on the biomolecule, and an isothiocyanate-functionalized chelator, is unstable in vivo, resulting in detachment of the radiometal-chelator complex from the targeting vector, resulting in poor tumor accumulation. To determine whether this instability was specific to the FAPI system, a peptide-based (Cyclic melanocyte stimulating hormone, CycMSH) bioconjugate targeting the melanocortin-1 receptor was synthesized using the same thiourea linkage. Identical metabolites were observed, supporting the hypothesis that thiourea bonds are unstable in vivo with this theranostic isotope pair. Subsequently, the effect of bioconjugation chemistry, specifically thiourea and amide bonds, on the stability and biodistribution of ²⁰³Pb/²¹²Pb-labeled bioconjugates was assessed. Modifying the bioconjugation linker to be an amide bond, formed by utilizing a chelate containing an active ester instead of an isothiocyanate, led to significantly improved in vitro and in vivo stability, as demonstrated by radioHPLC and biodistribution and imaging studies in both models. These findings highlight the importance of the choice of bioconjugation chemistry in the development of lead-based radiopharmaceuticals and emphasize the importance of selecting stable linkages to ensure optimal radiometal retention and tumor targeting.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"6 1","pages":"56-77"},"PeriodicalIF":4.3,"publicationDate":"2025-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921517/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147272077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Tutorial Review on Surface Plasmon Resonance Biosensors: Applications in Biomedicine.","authors":"Antony Chirco, Elisabetta Meacci, Giancarlo Margheri","doi":"10.1021/acsbiomedchemau.5c00182","DOIUrl":"10.1021/acsbiomedchemau.5c00182","url":null,"abstract":"<p><p>Surface Plasmon Resonance (SPR) has proven to be one of the most effective technologies in terms of specificity, affinity, and determination of kinetic parameters for evaluating interactions between macromolecules. The focus of this tutorial is to give an overview of the recent advances and applications of SPR biosensors in biomedicine that are presented emphasizing the potentiality for the detection of very low abundant compounds, which, in recent years, have assumed great importance for prevention and early diagnosis of various diseases in biomedicine. The real-time detection of important biomarkers such as tumor markers, viruses, and toxins but also of compounds of interest such as drugs and hormones allows point-of-care analysis and monitoring of disease progression quickly and in a less invasive manner. Over the past years, several technical innovations have been introduced to SPR devices, which have gone through a process of miniaturization, portability, flexibility, and cost reduction. These characteristics are in line with the advantages of SPR biosensors over other biosensing techniques, i.e., to be label-free detection systems and their capacity to observe in real-time the interactions between a variety of molecules of interest at the metal surface. Recent advances in SPR sensor technology, such as LSPR, SPRi, and SPRM, attempted to improve the sensitivity and performance of molecule detection.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 6","pages":"922-946"},"PeriodicalIF":4.3,"publicationDate":"2025-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12715632/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145805727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structural and Biochemical Characterization of <i>Fusobacterium nucleatum</i> Enoyl-ACP Reductase II (FabK) Reveals the Basis for Bacterial Species-Specific Inhibition.","authors":"Kristiana Avad, Osama Alaidi, Destiny Okpomo, Fahad Bin Aziz Pavel, Darcy Doran, Madeline Matheson, Dianqing Sun, Julian Hurdle, Kirk E Hevener","doi":"10.1021/acsbiomedchemau.5c00199","DOIUrl":"10.1021/acsbiomedchemau.5c00199","url":null,"abstract":"<p><p><i>Fusobacterium nucleatum</i> is a Gram-negative anaerobic bacterium ubiquitous in the oral cavity and increasingly recognized for its involvement in diverse clinical conditions, including periodontal disease, inflammatory bowel disease, premature birth, and several forms of cancer. These associations highlight the need for narrow-spectrum antibacterial agents directed against <i>F. nucleatum</i> to avoid disruption of beneficial microflora and limit the rise of antibiotic resistance. Recent studies have identified the fusobacterial fatty acid synthesis pathway (FAS-II) enzyme, enoyl-acyl carrier protein (ACP) reductase, <i>Fn</i>FabK, as an essential and promising target for selective antibacterial intervention. However, there is a lack of detailed structural information, which has hindered the validation of <i>Fn</i>FabK's druggability and the discovery of new inhibitors. Here, we present a comprehensive characterization of <i>Fn</i>FabK, including its cocrystal structure solved at 2.25 Å resolution and its biochemical and biophysical interactions with a series of potent small-molecule inhibitors. Our analyses revealed that these inhibitors display low to submicromolar activity against <i>Fn</i>FabK, with notable selectivity and differential activity when tested against FabK homologues from other bacterial pathogens. Importantly, the unique structural features of the <i>Fn</i>FabK active site, elucidated through these crystallographic studies, provide a mechanistic basis for species-specific inhibition. These findings not only validate <i>Fn</i>FabK as a druggable target but also furnish critical insights into the design of next-generation narrow-spectrum antibacterial agents.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"6 1","pages":"44-55"},"PeriodicalIF":4.3,"publicationDate":"2025-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921513/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147272129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of Indole- and Pyrazole-fused Glycyrrhetinic Acid Derivatives as Potent PTP1B Inhibitors: In Silico, In Vitro, In Vivo, and Metabolomic Studies.","authors":"Mitzi López-Sánchez, Hannya Mendoza-Mota, Ledy De-la-Cruz-Martínez, Félix Matadamas-Martínez, Diana Laura Torres-Chacón, Rosendo Martínez-Arellano, Juan Francisco Palacios-Espinosa, Jaime Pérez-Villanueva, Martin González-Andrade, José Carlos Páez-Franco, Julio César Almanza-Pérez, Francisco Cortés-Benítez","doi":"10.1021/acsbiomedchemau.5c00164","DOIUrl":"https://doi.org/10.1021/acsbiomedchemau.5c00164","url":null,"abstract":"<p><p>Protein tyrosine phosphatase 1B (PTP1B) is a crucial enzyme involved in regulating insulin and leptin signaling pathways, making it a promising target for treating type 2 diabetes. In this study, we synthesized 14 derivatives of indole- and pyrazole-fused glycyrrhetinic acid (GA) and evaluated their effects on PTP1B, using both its long (<i>h</i>PTP1B_1-400) and short (<i>h</i>PTP1B_1-285) forms. We analyzed enzymatic kinetics and selectivity over T-cell protein tyrosine phosphatase (TCPTP). Molecular docking and molecular dynamics simulations were performed to understand the binding mode of the compounds within PTP1B. Untargeted metabolomics, using gas chromatography-mass spectrometry, assessed metabolic changes caused by the most potent PTP1B inhibitors in HepG2 cells. We also evaluated these inhibitors <i>in vivo</i> to determine their effects on insulin sensitivity through the insulin tolerance test (ITT) in streptozotocin (STZ)-induced diabetic mice. Two compounds, <b>4b</b> (indole-fused) and <b>5g</b> (pyrazole-fused), showed greater potency against the long form of PTP1B compared to the short form, indicating that both compounds preferentially bind to the disordered C-terminal region of PTP1B. Molecular docking and molecular dynamics studies supported this finding. Furthermore, enzymatic kinetics revealed that compounds <b>4b</b> and <b>5g</b> function as uncompetitive inhibitors of PTP1B, with <i>K</i> _i values of 0.32 and 0.72 μM, respectively. Notably, both GA derivatives exhibited more pronounced inhibition of PTP1B compared to the well-established inhibitors ursolic acid and Ertiprotafib, while also demonstrating selectivity over TCPTP. Metabolomic analysis revealed that these compounds increased pantothenic acid and glycine levels, while decreasing glucose and fatty acid levels in HepG2 cells, suggesting enhanced glycolysis and reduced lipogenesis. Both compounds exhibited low cytotoxicity in HFF-1 cells and significantly reduced glucose levels in the ITT in STZ-induced diabetic mice, outperforming the insulin-sensitizing drug Pioglitazone.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"6 1","pages":"15-35"},"PeriodicalIF":4.3,"publicationDate":"2025-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921516/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147272143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Photoactivatable, Biomimetic Ligand photoMultiTASQ Traps DNA/RNA G‑Quadruplexes and Their Protein Binding Partners.","authors":"Sandy Raevens, Yehuda M Danino, Cécile Desingle, Francesco Rota Sperti, Marc Pirrotta, Lucie Mazzucotelli, Léa Letissier, Eran Hornstein, Sarah Cianferani, Oscar Hernandez-Alba, Ibai E Valverde, David Monchaud","doi":"10.1021/acsbiomedchemau.5c00162","DOIUrl":"10.1021/acsbiomedchemau.5c00162","url":null,"abstract":"<p><p>The biological role of DNA and RNA G-quadruplexes (G4s) is relayed to the cellular circuitry by a plethora of proteins known as G4-binding proteins (G4PBs). It is thus critical to decipher the formation of the G4/G4BP complexes to accurately understand their involvement in G4-mediated regulatory mechanisms. While hundreds of G4-interacting compounds (G4 ligands) have been used to uncover G4 biology in a rather indirect manner, only a handful of multivalent ligands allowing for identifying associated proteins have been designed, mostly relying on the covalent capture of G4BPs upon photoactivation. We report herein on such a multivalent ligand named photoMultiTASQ, which belongs to a family of biomimetic and smart G4 ligands known as template-assembled synthetic G-quartets (TASQs). We show how photoMultiTASQ interacts with and photo-cross-links both DNA/RNA G4s and related G4BPs, and develop a new mass spectrometry (MS)-based technique to characterize the covalent adducts. Collectively, these results make photoMultiTASQ a new molecular tool in the arsenal of chemical cross-linking and isolation by pull-down (Chem-CLIP) technologies that uniquely identify targets and validate target engagement in cells.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 6","pages":"994-1006"},"PeriodicalIF":4.3,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12715623/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145805782","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Manipulation of 7‑Finger Zinc Finger Nuclease Increases the Efficiency of Genome Editing in Human Cells.","authors":"Shota Katayama, Masahiro Watanabe, Wataru Nomura, Takashi Yamamoto","doi":"10.1021/acsbiomedchemau.5c00174","DOIUrl":"10.1021/acsbiomedchemau.5c00174","url":null,"abstract":"<p><p>Genome editing tools have great potential for medicinal use. Among them, zinc finger nucleases (ZFNs) are smaller in size than transcriptional activator-like effector nucleases and CRISPR-Cas9. Therefore, ZFNs are easily packed into a viral vector with limited cargo space, including adeno-associated viral vectors. Furthermore, because ZFN patents expired in 2020, high patent royalties are not required for application. Although functional 6-finger ZFNs can be easily prepared by modular assembly, it has been extremely difficult to produce functional 7-finger ZFNs, which are expected to have higher target specificity than 6-finger ZFNs in some cases. Herein we describe the construction of 7-finger ZFNs and the improvement in genome editing efficiency, which is generally lower in 7-finger ZFNs than in 6-finger ZFNs. Modular assembly of 7-finger ZFNs was achieved using a specific mutation, and the original genome editing efficiency was increased by up to 19%. Furthermore, 7-finger ZFNs showed reduced off-target effects, exhibiting higher target specificity than the corresponding 6-finger ZFNs. Our study provides critical insights for safer and more specific genome editing.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 6","pages":"1016-1026"},"PeriodicalIF":4.3,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12715528/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145805737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Leader-Independent C‑Terminal Modification by a Radical <i>S</i>‑Adenosyl‑l‑methionine Maturase Enables Macrocyclic GLP-1-Like Peptides.","authors":"Jacob K Pedigo, Karsten A S Eastman, Vahe Bandarian","doi":"10.1021/acsbiomedchemau.5c00152","DOIUrl":"10.1021/acsbiomedchemau.5c00152","url":null,"abstract":"<p><p>Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a rapidly expanding family of natural products in which biosynthetic maturase enzymes tailor ribosomal precursors into bioactive products. Classical RiPP maturation relies on an N-terminal leader sequence in the precursor peptide and a complementary RiPP-recognition element in the enzyme to guide substrate binding. Herein, we interrogated PapB, a radical <i>S</i>-adenosyl-l-methionine RiPP maturase known to introduce thio-(seleno)-ether cross-links and discovered that its catalytic reach extends well beyond this paradigm. PapB efficiently processes substrates that lack any canonical leader sequence, demonstrating <i>bona fide</i> leader-independent activity. To highlight the practical value of this capability, we applied PapB to three therapeutically relevant analogues of glucagon-like peptide pathway agonists to generate C-terminal cyclic structures. In every case, the enzyme achieved complete conversion of the linear to the thioether macrocyclized peptide. These results establish PapB as a versatile, plug-and-play biocatalyst for late-stage macrocyclization of structurally diverse peptides, opening a direct route to conformationally constrained therapeutic candidates without the need for leader tags.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 6","pages":"1007-1015"},"PeriodicalIF":4.3,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12715619/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145805813","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"1,2,4-Triazole-Based First-in-Class Non-Nucleoside Inhibitors of the Bacterial Enzyme MraY.","authors":"Tomayo Berida, Tzu-Yu Huang, Stefanie C Weck, Marcel Lutz, Samuel R McKee, Nathalie Kagerah, Destinee L Manning, Mohamed E Jahan, Sushil Mishra, Jennifer Herrmann, Rolf Müller, Robert J Doerksen, Christina L Stallings, Christian Ducho, Sudeshna Roy","doi":"10.1021/acsbiomedchemau.5c00158","DOIUrl":"10.1021/acsbiomedchemau.5c00158","url":null,"abstract":"<p><p>MraY is an essential bacterial enzyme for peptidoglycan synthesis in cell walls and serves as a promising but unrealized target for developing effective antibacterial drugs. Nature has provided a remarkable array of nucleoside inhibitors of MraY, and researchers have skillfully refined these structures to develop inhibitors that effectively mimic natural products. Yet, both natural products and their synthetic variants often face challenges regarding inadequate <i>in vivo</i> efficacy, and the intricate nature of these structures complicates their synthesis and exploration of structure-activity relationships (SAR). Here, we present our findings on the discovery of first-in-class small molecule MraY inhibitors that are non-nucleoside-derived, based on 1,2,4-triazoles, using a structure-based drug design strategy. By leveraging the structural roadmap of the MraY binding site, we discovered the initial hit compound <b>1</b> with an IC₅₀ value of 171 μM <i>in vitro</i> against MraY from <i>Staphylococcus aureus</i> (MraY _<i>SA</i> ) that was refined to compound <b>12a</b>, exhibiting an IC₅₀ value of 25 μM. Molecular docking studies against MraY _<i>SA</i> provided critical insights into how the binding interactions of compounds directly influence their activity. Furthermore, we report that these compounds show broad-spectrum antibacterial activity against critical pathogens such as <i>Enterococcus</i> spp., methicillin-resistant <i>S. aureus</i> (MRSA), vancomycin-resistant <i>Enterococci</i> (VRE) strains, <i>Acinetobacter baumannii</i>, and <i>Mycobacterium tuberculosis</i>. This study showcases novel non-nucleoside inhibitors as a compelling proof-of-concept for crafting the next generation of antibacterial agents targeting MraY.</p>","PeriodicalId":29802,"journal":{"name":"ACS Bio & Med Chem Au","volume":"5 6","pages":"966-981"},"PeriodicalIF":4.3,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12715639/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145805729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}