ACS Infectious Diseases最新文献

{"title":"Deploying Soft Drugs in the Fight against MRSA with Detailed Biological Evaluation of 4-(n-Alkoxy)-phenoxy Betaine Amphiphiles Active against MDR Staphylococcus aureus and Enterococcus sp.","authors":"Dharmendra Katiyar,&nbsp;Abdul Akhir,&nbsp;Grace Kaul,&nbsp;Deepanshi Saxena,&nbsp;Tejender S. Thakur* and Sidharth Chopra*,&nbsp;","doi":"10.1021/acsinfecdis.5c00267","DOIUrl":"https://doi.org/10.1021/acsinfecdis.5c00267","url":null,"abstract":"<p >Antimicrobial resistance is one of the major challenges faced worldwide by the healthcare systems, negatively impinging upon global health and the economy. Methicillin-resistant <i>Staphylococcus aureus</i> (MRSA), which causes skin and mucosal infections, contributes to 64% more death as compared to drug-susceptible infections, which forces researchers to continuously search for new antimicrobials. In this context, we report the synthesis and antimicrobial activity of novel amphiphilic cationic 1,4-alkoxy-phenol derivatives. We synthesized a library of 12 small amphiphilic compounds containing an <span>l</span>-carnitine or betaine as a cationic headgroup and investigated the effect of the hydrocarbon chain length on the antimicrobial activity against a critical and high-priority human pathogen panel. The amphiphilic molecules with a C-10 alkyl chain length (<b>2f</b> and <b>3f</b>) showed substantial antibacterial activity at very low concentrations (1–2 μg/mL). Based on low cytotoxicity (Selectivity Index 20) and MIC of amphiphilic series, we further explored <b>3f</b>, which shows a concentration-dependent bactericidal activity at 10× MIC; it completely wiped out <i>S. aureus</i> ATCC 29213 at 1 h, and when combined with daptomycin, it showed a reduction of ∼6 log₁₀ CFU/mL as compared with untreated at 24 h against <i>S. aureus</i> ATCC 29213 even at 1× MICs. In order to discriminate between nonspecificity in detergent-like action, we further determined the CMC of 3f and found that it is 36-fold higher than MIC, thus further emphasizing its specificity. <b>3f</b> exhibits biofilm eradication properties that are comparable to levofloxacin and better than vancomycin while intracellular killing efficacy of <b>3f</b> was found to be better than vancomycin and similar to levofloxacin. When tested <i>in vivo</i> in the murine skin infection model, it shows activity comparable to fusidic acid as it reduces ∼2 log₁₀ CFU/g compared with that untreated. Considering the potent antimicrobial properties of <b>3f</b>, we hypothesize that it opens up a new avenue for discovery and development of novel antimicrobials active against MDR <i>S. aureus</i> and <i>Enterococcus</i> sp.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2434–2445"},"PeriodicalIF":3.8,"publicationDate":"2025-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient Synthesis of Clovibactin and Its Analogues: Promoting the Discovery of More Potent Antibiotics via Multiple Screenings","authors":"Chengshuo He,&nbsp;Xiaoshu Jing,&nbsp;Biyao Liu,&nbsp;Fei Liu and Kang Jin*,&nbsp;","doi":"10.1021/acsinfecdis.5c00458","DOIUrl":"10.1021/acsinfecdis.5c00458","url":null,"abstract":"<p >Increasingly, antimicrobial resistance has become a major concern for public health. The recently discovered cyclic depsipeptide, clovibactin, is a potential antibiotic candidate with potent activities against multidrug-resistant pathogens. It contains three <span>d</span>-amino acids, including a rare <span>d</span>-hydroxyasparagine residue. Herein, we present the efficient synthesis of the suitably protected <span>d</span>-hydroxyasparagine building block and the natural product clovibactin. Efforts have also been made by us to establish the structure–activity relationship (SAR) of clovibactin and to hunt for more active analogues. After three rounds of screening, the new compounds <b>30</b>, <b>40,</b> and <b>42</b> were found to exhibit higher antibacterial activity than clovibactin and were able to eradicate methicillin-resistant <i>Staphylococcus aureus</i> (MRSA). Their substantial efficacy was also demonstrated in a bactericidal activity assay, an in vitro time-killing assay, a hemolysis assay, and a potential resistance development evaluation. The improved antibacterial activity, low hematotoxicity, and resistance-resistant properties of analogues <b>30</b>, <b>40,</b> and <b>42</b> make them promising for further evaluation of drug-likeness and the development of next-generation antibiotics.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2568–2576"},"PeriodicalIF":3.8,"publicationDate":"2025-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144935850","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Common Biological Properties of Mycobacterium tuberculosis MmpL3 Inhibitors","authors":"Lauren Ames,&nbsp;Renee Allen,&nbsp;Helena I. M. Boshoff,&nbsp;Laura A. T. Cleghorn,&nbsp;Curtis A. Engelhart,&nbsp;Dirk Schnappinger and Tanya Parish*,&nbsp;","doi":"10.1021/acsinfecdis.5c00394","DOIUrl":"10.1021/acsinfecdis.5c00394","url":null,"abstract":"<p >MmpL3 is a promising new target for antitubercular drugs, but the microbiological properties of MmpL3 inhibitors are not fully understood. We compared the activity and mode of action of 11 structurally diverse compound series that target MmpL3. We confirmed the activity was via MmpL3 using strains with differential expression of MmpL3. MmpL3 inhibitors had potent activity against replicating <i>M. tuberculosis</i>, with increased activity against intramacrophage bacilli and were rapidly bactericidal. MmpL3 inhibition induced cell wall stress concomitantly with a boost in the ATP levels in <i>M. tuberculosis</i>. Mutation in MmpL3 conferred resistance to all series at different levels. The molecules did not negatively impact membrane potential, pH homeostasis, or induce reactive oxygen species and were inactive against starved bacilli. Our study revealed common features related to the chemical inhibition of MmpL3, enabling the identification of off-target effects and highlighting the potential of such compounds as future drug candidates.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2523–2533"},"PeriodicalIF":3.8,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsinfecdis.5c00394","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144870156","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Outer Membrane Protein Tp92 of Treponema pallidum Induces BeWo Cells Apoptosis and Oxidative Stress via the Nrf2/keap1 Pathway","authors":"Shaobin Huang,&nbsp;Yue Zhao,&nbsp;Shun Xiong,&nbsp;Zhaoping Liu,&nbsp;Xuan Ding,&nbsp;Jiangchen Yao,&nbsp;Ting Lin,&nbsp;Han Yu,&nbsp;Xiaohong Zhang* and Feijun Zhao*,&nbsp;","doi":"10.1021/acsinfecdis.5c00346","DOIUrl":"10.1021/acsinfecdis.5c00346","url":null,"abstract":"<p ><i>Treponema pallidum</i> can be transmitted from the mother to the newborn via the placenta. Pregnancy complications that can arise from this include congenital syphilis, stillbirth, miscarriage, preterm birth, and fetal growth limitation. Tp92 is a protein located in the outer membrane of <i>T.pallidum</i> and has a β-barrel structure. The structural features of this protein are similar to those of other outer membrane proteins found in Gram-negative bacteria. It plays a crucial role in the initial infection of <i>T. pallidum</i>. However, the precise function of Tp92 in placental trophoblast cells remains uncertain. BeWo cells, derived from human choriocarcinoma, are commonly used to model placental trophoblast cells. Their physical structure and chemical composition are similar to undifferentiated trophoblasts. Our study revealed that Tp92 induces apoptosis in BeWo cells through various mechanisms, including reducing cell survival, decreasing mitochondrial membrane potential, increasing the number of TUNEL-positive cells, enhancing caspase-3 and 7 activity, and upregulating Bax and cleaved caspase-3 proteins. Additionally, Tp92 raises reactive oxygen species (ROS) and malondialdehyde levels, decreases superoxide dismutase (SOD) activity, and inhibits Nrf2 translocation to the nucleus, leading to reduced expression of Nrf2-regulated genes HO-1, NQO1, and SOD-1. Activation of the Nrf2/Keap1 pathway with the Nrf2 activator TBHQ can restore its function and inhibit apoptosis. This reveals that Tp92 induces oxidative stress and cell death by increasing ROS and inhibiting the Nrf2 pathway, offering a potential therapeutic target for pregnancy and congenital syphilis.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2500–2514"},"PeriodicalIF":3.8,"publicationDate":"2025-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144881642","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comprehensive Identification of β-Lactam Antibiotic Polypharmacology in Mycobacterium tuberculosis","authors":"Kaylyn L. Devlin,&nbsp;Emily Hutchinson,&nbsp;Hailey N. Dearing,&nbsp;Samantha R. Levine,&nbsp;Deseree J. Reid,&nbsp;Damon T. Leach,&nbsp;Lydia H. Griggs,&nbsp;Gerard X. Lomas,&nbsp;Leo J. Gorham,&nbsp;Aaron T. Wright,&nbsp;Gyanu Lamichhane,&nbsp;Vivian S. Lin and Kimberly E. Beatty*,&nbsp;","doi":"10.1021/acsinfecdis.5c00233","DOIUrl":"10.1021/acsinfecdis.5c00233","url":null,"abstract":"<p >Infections with <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>) cause tuberculosis (TB), which requires at least 6 months of treatment with multiple antibiotics. There is emergent interest in using β-lactam antibiotics to improve treatment outcomes for patients. These drugs target cell wall biosynthesis, but a comprehensive list of enzymes inhibited by β-lactams in <i>Mtb</i> is lacking. In the current study, we sought to identify and characterize <i>Mtb</i> enzymes inhibited by β-lactam antibiotics using physiological conditions representative of both acute and chronic TB disease. We used new activity-based probes based on the β-lactam antibiotic meropenem due to its approval by the World Health Organization for TB treatment. Activity-based probes label enzymes based on both substrate specificity and catalytic mechanism, enabling precise identification of drug targets. We identified previously undiscovered targets of meropenem in addition to known cell wall biosynthetic enzymes. We validated β-lactam binding and hydrolysis for six newly identified targets: Rv1723, Rv2257c, Rv0309, DapE (Rv1202), MurI (Rv1338), and LipD (Rv1923). Our results demonstrate that there are at least 30 enzymes in <i>Mtb</i> vulnerable to inhibition by meropenem. This is many more β-lactam targets than historically described, suggesting that efficacy in <i>Mtb</i> is a direct result of polypharmacology.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2422–2433"},"PeriodicalIF":3.8,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144870157","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Long-Term Neuroimaging Findings in a Murine Model of Human Extraparenchymal Neurocysticercosis","authors":"Alejandro Méndez,&nbsp;Agnes Fleury,&nbsp;Roger Carrillo-Mezo,&nbsp;Juan A. Hernández-Aceves,&nbsp;Montserrat Mejía-Hernández,&nbsp;Nelly Villalobos,&nbsp;Marisela Hernández,&nbsp;Raúl Bobes,&nbsp;Luis Concha,&nbsp;Juan J. Ortiz-Retana,&nbsp;Marta Romano,&nbsp;Pedro Tadao Hamamoto Filho,&nbsp;Gladis Fragoso,&nbsp;José Alejandro Espinosa-Cerón* and Edda Sciutto*,&nbsp;","doi":"10.1021/acsinfecdis.5c00431","DOIUrl":"10.1021/acsinfecdis.5c00431","url":null,"abstract":"<p >Neurocysticercosis is caused by the establishment of <i>Taenia solium</i> cysticerci in the central nervous system. The extraparenchymal form (ExP-NCC) is the most severe clinical presentation that may remain asymptomatic for years. Current treatment involves cysticidal drugs (albendazole and/or praziquantel) combined with glucocorticoids to manage the associated neuroinflammation; however, only ∼30% of patients respond effectively. This highlights the need to improve therapeutic strategies. Herein, the experimental murine model of human ExP-NCC was further characterized to improve its usefulness in testing new therapies. In humans, cysts grow slowly in the basal cisterns of the subarachnoid space, and patients become symptomatic years after the infection. Thus, a long-term follow-up was performed by using magnetic resonance imaging (MRI) with sequences allowing volumetric analysis. MRI confirmed NCC in 77% of infected rats, all exhibiting extraparenchymal localization and persistently elevated levels of HP10, a marker of viable cysticerci. Imaging also enabled precise cyst localization and estimation of the parasite-occupied volume.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2534–2541"},"PeriodicalIF":3.8,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsinfecdis.5c00431","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144870158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Periplasmic Binding Protein YiuA Enables Yersinia pestis to Scavenge Fe(III)-Catechol Siderophores","authors":"Emil Thomsen,&nbsp;Parker Stow,&nbsp;Daniel Roth and Alison Butler*,&nbsp;","doi":"10.1021/acsinfecdis.5c00524","DOIUrl":"10.1021/acsinfecdis.5c00524","url":null,"abstract":"<p ><i>Yersinia pestis</i>, the pathogen causing plague, requires iron to grow. <i>Y. pestis</i> employs several uptake pathways for iron, including the siderophore yersiniabactin, as well as hemin and inorganic iron. The <i>Y. pestis</i> iron assimilation repertoire further harbors the uncharacterized YiuRABC pathway, presumed to transport an unidentified Fe(III)-siderophore(s). Through intrinsic fluorescence quenching of the periplasmic binding protein YiuA, we discovered that YiuA displays high affinity toward Fe(III) complexes of the hydrolysis products of enterobactin, Fe(III)-[di(DHB-^LSer)] and Fe(III)-[DHB-^LSer]₂, with K_d values of 27.6 ± 4.2 nM and 28.2 ± 6.9 nM, respectively, as well as the bis-catechol siderophore butanochelin, with K_d 0.76 ± 0.17 nM. By comparison, YiuA has a much weaker affinity for intact Fe(III)-enterobactin, K_d 444.7 ± 20.6 nM. Electronic circular dichroism spectroscopy reveals YiuA has a strong preference for binding Λ configured Fe(III)-siderophores, which can be achieved with the Fe(III) bis-catechol complexes but not Fe(III)-enterobactin.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2391–2397"},"PeriodicalIF":3.8,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144853938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ultrasensitive Aptamer-Based Metal–Organic Framework-on-Metal–Organic Framework Platform for Clinical Detection of KPC-2 Klebsiella pneumoniae and Multidrug-Resistant Acinetobacter baumannii","authors":"Dandan Shi,&nbsp;Ruiwen Li,&nbsp;Shaoning Yu and Guoqing Qian*,&nbsp;","doi":"10.1021/acsinfecdis.5c00349","DOIUrl":"10.1021/acsinfecdis.5c00349","url":null,"abstract":"<p >The escalating crisis of hospital-acquired multidrug-resistant (MDR) infections, particularly <i><i>Klebsiella pneumoniae</i></i> carbapenemase 2-expressing <i><i>K. pneumoniae</i></i> (KPC-2 KP) and MDR-<i><i>Acinetobacter baumannii</i></i> (AB), demands rapid diagnostic solutions. We developed a dual nanozyme-powered colorimetric aptasensor leveraging a cascade amplification mechanism, a metal–organic framework (MOF)-on-MOF nanostructure with peroxidase-like activity. Cu-MOF@PMOF(Fe) integrates catechol oxidase-like activity, with the Cu-MOF core oxidizing catechol to generate H₂O₂, and the PMOF(Fe) shell utilizes H₂O₂ to oxidize the TMB substrate, producing dual-wavelength signals at 370 and 652 nm for ultrasensitive detection. Functionalized with pathogen-specific aptamers, the system achieves selective bacterial capture within 40 min, quantifying 10–10⁸ CFU/mL with detection limits of 7 CFU/mL for KPC-2 KP and 6 CFU/mL for MDR-AB. Clinical validation using cerebrospinal fluid, peritoneal fluid, serum, and bile samples demonstrated robust performance in complex matrices (91.2–112.2% recovery rates). Therefore, this platform provides a rapid (&lt;1 h), sensitive, and clinically adaptable solution for combating MDR bacterial infections, bridging advanced materials with diagnostic microbiology.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2491–2499"},"PeriodicalIF":3.8,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144833457","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dynamic Proteomic and PTMomic Characterization of Mycobacteria after Clinical Pharmaceutical Intervention","authors":"Jun Yin,&nbsp;Liming Wang,&nbsp;Hailong Jin,&nbsp;Mingya Zhang,&nbsp;Tian Jiang,&nbsp;Yunbo Kan,&nbsp;Tianxian Liu,&nbsp;Feng Su,&nbsp;Lei Zhao,&nbsp;Yi Li,&nbsp;Shiyang Shen,&nbsp;Lu Zhou,&nbsp;Minjia Tan,&nbsp;Yuanlin Song*,&nbsp;Lijie Tan* and Jun-Yu Xu*,&nbsp;","doi":"10.1021/acsinfecdis.4c00891","DOIUrl":"10.1021/acsinfecdis.4c00891","url":null,"abstract":"<p >Tuberculosis, a major global health threat, necessitates understanding the pharmacological mechanisms of current drugs to combat multidrug resistance. In addition to alterations at the proteome level, the dynamic changes occurring at various levels of post-translational modifications (PTMs) following pharmacological intervention remain unclear. In our current study, we employed a quantitative proteomic approach to systematically analyze the dynamic molecular alterations at both the proteome and PTM levels in response to clinical drugs, including ethambutol, bedaquiline, moxifloxacin, and streptomycin. Our findings revealed enriched bioprocesses beyond known functions, phosphorylation-level changes in kinases and phosphatases, and increased acetylation levels with all four drugs. Overexpression of CobB in <i>Mycobacterium smegmatis</i> significantly increased its susceptibility to ethambutol, indicating enhanced drug sensitivity. Our study provides integrated multiomics resources for understanding the dynamic molecular characteristics and drug resistance associated with clinical drug interventions and proposes novel therapeutic strategies targeting the PTM levels.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2398–2410"},"PeriodicalIF":3.8,"publicationDate":"2025-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144820112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and Characterization of a Metal Binding-Independent Rift Valley Fever Virus Endonuclease Inhibitor","authors":"Cara D. Kirby,&nbsp;Gai Liu,&nbsp;Farheen Fatma,&nbsp;Maris R. Pedlow,&nbsp;Narendran G-Dayanandan,&nbsp;Nitin Sharma,&nbsp;Zachary D. Frey,&nbsp;Rachael E. Rush,&nbsp;Steven C. Cardinale,&nbsp;Andrzej M. Krezel,&nbsp;Amy L. Hartman,&nbsp;Terry L. Bowlin,&nbsp;Daisy W. Leung and Gaya K. Amarasinghe*,&nbsp;","doi":"10.1021/acsinfecdis.5c00088","DOIUrl":"10.1021/acsinfecdis.5c00088","url":null,"abstract":"<p >The viral cap-snatching mechanism, facilitated by the endonuclease (EndoN) domain of viral polymerases, is critical for viral gene transcription and translation and is therefore an attractive target for antiviral development. The successful development of EndoN inhibitor XOFLUZA (Baloxavir marboxil) has opened the possibility that the EndoN domain of negative-sense RNA viruses (NSVs) can be targeted in a similar fashion. Rift Valley Fever Virus (RVFV) belongs to the <i>Bunyaviricetes</i> class, which includes other pathogens with pandemic potential, such as severe fever with thrombocytopenia syndrome virus (SFTSV) and Heartland virus (HRTV). Here, we identify MBXC-4522 as a RVFV EndoN inhibitor using a high-throughput FRET-based assay. We screened a library of &gt;20,000 compounds and identified those that target the RVFV EndoN domain. MBXC-4522, a spiro piperidine pyrido pyridine, directly binds the RVFV EndoN domain, increases protein stability, and inhibits EndoN activity. MBXC-4522 acts in a metal binding-independent mechanism, while XOFLUZA’s mode of action is metal binding-dependent. Infectious assays also support the ability of MBXC-4522 to inhibit pathogenic RVFV (ZH501), suggesting that hit-to-lead optimization and future <i>in vivo</i> validation are warranted.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":"11 9","pages":"2373–2382"},"PeriodicalIF":3.8,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acsinfecdis.5c00088","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144815265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}