ACS Infectious Diseases最新文献

{"title":"High-Throughput Repurposing Screen Reveals Compounds with Activity against <i>Toxoplasma gondii</i> Bradyzoites.","authors":"Taher Uddin, Jing Xia, Yong Fu, Case W McNamara, Arnab K Chatterjee, L David Sibley","doi":"10.1021/acsinfecdis.4c00689","DOIUrl":"10.1021/acsinfecdis.4c00689","url":null,"abstract":"<p><p><i>Toxoplasma gondii</i> causes widespread chronic infections that are not cured by current treatments due to the inability to affect semidormant bradyzoite stages within tissue cysts. To identify compounds to eliminate chronic infection, we developed an HTS using a recently characterized strain of <i>T. gondii</i> that undergoes efficient conversion to bradyzoites in vitro. Stage-specific expression of luciferase was used to selectively monitor the growth inhibition of bradyzoites by the Library of Pharmacological Active Compounds, consisting of 1280 drug-like compounds. We identified 44 compounds with >50% inhibitory effects against bradyzoites, including new highly potent compounds, several of which have precedent for antimicrobial activity. Subsequent characterization of the compound sanguinarine sulfate revealed potent and rapid killing against in vitro-produced bradyzoites and bradyzoites harvested from chronically infected mice, including potent activity against intact cysts. These findings provide a platform for expanded screening and identify promising compounds for further preclinical development against <i>T. gondii</i> bradyzoites that are responsible for chronic infection.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"600-609"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11915370/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143397485","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":"C10-Benzoate Esters of Anhydrotetracycline Inhibit Tetracycline Destructases and Recover Tetracycline Antibacterial Activity.","authors":"Emily E Williford, Yao-Peng Xue, Wai Kwan Tang, Ruihao Li, Katherine V Jones, Kevin S Blake, Helen C Blaine, Xiang Lian, Christina L Stallings, Niraj H Tolia, Gautam Dantas, Timothy A Wencewicz","doi":"10.1021/acsinfecdis.4c00912","DOIUrl":"10.1021/acsinfecdis.4c00912","url":null,"abstract":"<p><p>Tetracyclines (TCs) are an important class of antibiotics threatened by enzymatic inactivation. These tetracycline-inactivating enzymes, also known as tetracycline destructases (TDases), are a subfamily of class A flavin monooxygenases (FMOs) that catalyze hydroxyl group transfer and oxygen insertion (Baeyer-Villiger type) reactions on TC substrate scaffolds. Semisynthetic modification of TCs (e.g., tigecycline, omadacycline, eravacycline, and sarecycline) has proven effective in evading certain resistance mechanisms, such as ribosomal protection and efflux, but does not protect against TDase-mediated resistance. Here, we report the design, synthesis, and evaluation of a new series of 22 semisynthetic TDase inhibitors that explore D-ring substitution of anhydrotetracycline (aTC) including 14 C10-benzoate ester and eight C9-benzamides. Overall, the C10-benzoate esters displayed enhanced bioactivity and water solubility compared to the corresponding C9-benzamides featuring the same heterocyclic aryl side chains. The C10-benzoate ester derivatives of aTC were prepared in a high-yield one-step synthesis without the need for protecting groups. The C10-esters are water-soluble, stable toward hydrolysis, and display dose-dependent rescue of tetracycline antibiotic activity in <i>E. coli</i> expressing two types of tetracycline destructases, represented by TetX7 (Type 1) and Tet50 (Type 2). The best inhibitors recovered tetracycline antibiotic activity at concentrations as low as 2 μM, producing synergistic scores <0.5 in the fractional inhibitory concentration index (FICI) against TDase-expressing strains of <i>E. coli</i> and clinical <i>P. aeruginosa</i>. The C10-benzoate ester derivatives of aTC reported here are promising new leads for the development of tetracycline drug combination therapies to overcome TDase-mediated antibiotic resistance.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"738-749"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11915366/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143254157","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":"Activity-Based Protein Profiling Identifies an α-Amylase Family Protein Contributing to the Virulence of Methicillin-Resistant <i>Staphylococcus aureus</i>.","authors":"Md Jalal Uddin, Kjersti Julin, Herman S Overkleeft, Mona Johannessen, Christian S Lentz","doi":"10.1021/acsinfecdis.4c00638","DOIUrl":"10.1021/acsinfecdis.4c00638","url":null,"abstract":"<p><p>In search of new putative antimicrobial drug targets in methicillin-resistant <i>Staphylococcus aureus</i>, we aimed to identify and characterize retaining glycosidase activities in this bacterial pathogen. Using activity-based protein profiling (ABPP), a panel of 7 fluorescent probes was screened to detect activities of diverse retaining glycosidase families. Based on this, a cocktail of 3 biotinylated probes (targeting α-glucosidases, β-galactosidases and α-fucosidases) was used for target enrichment and three glycoside hydrolase family proteins were identified by mass-spectrometry: 6-phospho-β-glucosidase (BglA), α-amylase family protein trehalase C (TreC), and autolysin (Atl). The physiological relevance of previously uncharacterized BglA and TreC was addressed in CRISPRi and inhibitor studies with the putative TreC inhibitor α-cyclophellitol-aziridine. Silencing of <i>tre</i>C did not affect bacterial growth in rich media, but reduced biofilm formation <i>in vitro</i>, and attenuated virulence during <i>Galleria mellonella</i> infection, warranting future investigations into the biochemical function of this enzyme.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"573-583"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11915364/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143363253","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":"<i>Acinetobacter baumannii</i> Employs a Rare Fatty Acid Desaturase for Niche-Specific Host Adaptation.","authors":"Felise G Adams, Saleh Alquethamy, Jack K Waters, Brynley J Davies, Ella Haracic, Jeffrey D Nanson, James C Paton, Jade K Forwood, Karl A Hassan, Erin B Brazel, Claudia Trappetti, Bart A Eijkelkamp","doi":"10.1021/acsinfecdis.4c00765","DOIUrl":"10.1021/acsinfecdis.4c00765","url":null,"abstract":"<p><p><i>Acinetobacter baumannii</i> is a hospital-associated pathogen with unique fatty acid homeostasis features. This includes a reliance on desaturases for proliferation, due to an inability to generate unsaturated fatty acids during the synthesis cycles. However, there are various unexplained gaps in <i>A. baumannii</i> fatty acid homeostasis, such as the desaturation of <i>de novo</i> synthesized fatty acids. We identified a conserved desaturase (DesC) with a rare structural feature that may have roles in coordinating fatty acids with acyl carrier protein conjugants. We showed that DesC can generate fatty acids with <i>cis</i> double bonds in the delta-9 position. Profiling of <i>A. baumannii</i> fatty acids and mRNA transcripts emphasized its significance during fatty acid synthesis. DesC was found to be most critical in mouse niches where <i>A. baumannii</i> relies on fatty acid synthesis. This work has contributed to our understanding of core metabolic features that are key to the disease potential of <i>A. baumannii</i>.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"550-557"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143447317","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":"Expanding the Landscape of Dual Action Antifolate Antibacterials through 2,4-Diamino-1,6-dihydro-1,3,5-triazines.","authors":"John D Georgiades, Daniel A Berkovich, Samuel R McKee, Angela R Smith, Banumathi Sankaran, Kelly N Flentie, Carlos H Castañeda, Daniel G Grohmann, Ram Rohatgi, Carrie Lasky, Twila A Mason, James E Champine, Patricia A Miller, Ute Möllmann, Garrett C Moraski, Scott G Franzblau, Marvin J Miller, Christina L Stallings, Joseph M Jez, Bruce A Hathaway, Timothy A Wencewicz","doi":"10.1021/acsinfecdis.4c00768","DOIUrl":"10.1021/acsinfecdis.4c00768","url":null,"abstract":"<p><p>Antibiotics that operate <i>via</i> multiple mechanisms of action are a promising strategy to combat growing resistance. Previous studies have shown that dual action antifolates formed from a pyrroloquinazolinediamine core can inhibit the growth of bacterial pathogens without developing resistance. In this work, we expand the scope of dual action antifolates by repurposing the 2,4-diamino-1,6-dihydro-1,3,5-triazine (DADHT) cycloguanil scaffold to a variety of derivatives designed to inhibit dihydrofolate reductase (DHFR) and disrupt bacterial membranes. Dual mechanism DADHTs have activity against a variety of target pathogens, including <i>Mycobacterium tuberculosis</i>, <i>Mycobacterium abscessus</i>, and <i>Pseudomonas aeruginosa</i>, among other <i>ESKAPEE</i> organisms. Through X-ray crystallography, we confirmed engagement of the <i>Escherichia coli</i> DHFR target and found that some DADHTs stabilize a previously unobserved conformation of the enzyme but, broadly, bind in the occluded conformation. Using <i>in vitro</i> inhibition of purified <i>E. coli</i> and <i>Staphylococcus aureus</i> DHFR and disruption of <i>E. coli</i> membranes, we determined that alkyl substitution of dihydrotriazine at the 6-position best optimizes the DADHT's two mechanisms of action. By employing both mechanisms, the DADHT spectrum of activity was extended beyond the scope of traditional antifolates. We are optimistic that the dual mechanism approach, particularly through the action of antifolates, offers a unique means of combating hard-to-treat bacterial infections.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"689-702"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11908892/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143412304","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":"Pharmacokinetic and Pharmacodynamics of Clofazimine Nano-in-Microparticles: Enhanced Brain Delivery and CNS Tuberculosis Amelioration via Intranasal Administration.","authors":"Krishna Jadhav, Agrim Jhilta, Raghuraj Singh, Swarnima Negi, Shweta Sharma, Rahul Shukla, Amit Kumar Singh, Rahul Kumar Verma","doi":"10.1021/acsinfecdis.4c00767","DOIUrl":"10.1021/acsinfecdis.4c00767","url":null,"abstract":"<p><p><i>Mycobacterium tuberculosis</i> (Mtb) demonstrates a proclivity for infecting extrapulmonary sites, notably the brain. Treating these extrapulmonary tuberculosis (TB) manifestations is challenging due to the difficulty of drug delivery across the blood-brain barrier. Clofazimine (CLF) has exhibited promising activity against Mtb, including multidrug-resistant variants, in vitro and in preclinical animal models. However, its clinical implication is restricted owing to poor physicochemical and pharmacokinetic properties. This study aims to develop CLF nano-in-microparticles (CLF-NIMs) for brain drug delivery for central nervous system TB (CNS-TB) treatment via the intranasal route. Simultaneously, the potential dissemination of TB bacilli to the brain was investigated. Following treatment, colony-forming unit (CFU) enumeration was conducted in both the brain and lung tissues to assess mycobacterial burden. Concurrently, drug concentrations were quantified in serum, brain, and lung tissue, enabling a comprehensive evaluation of pharmacokinetics and tissue-specific drug distribution. In pharmacokinetic investigations of CLF-NIMs, significant accumulation of CLF was observed in brain tissue compared to orally administered CLF, surpassing the minimum inhibitory concentration of CLF. In a murine CNS-TB model, intranasal insufflation of CLF-NIMs for 4 weeks led to a substantial reduction (∼0.99 ± 0.57 Log10CFU/gram) in CFU count in the brain compared to oral administration of CLF (2.45 ± 0.47 Log10CFU/gram). These promising preclinical results indicate that CLF-NIMs are well-tolerated and exhibit significant anti-TB activity in a murine CNS-TB model.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"665-675"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143412305","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":"Rv2741 Promotes Mycobacterium Survival by Modulating Macrophage Function via the IL-1α-MAPK Axis.","authors":"Xintong He, Yonglin He, Xichuan Deng, Nan Lu, Anlong Li, Sijia Gao, Shiyan He, Yuran Wang, Nanzhe Fu, Zijie Wang, Yuxin Nie, Lei Xu","doi":"10.1021/acsinfecdis.4c00790","DOIUrl":"10.1021/acsinfecdis.4c00790","url":null,"abstract":"<p><p>One of the primary healthcare problems in the world today is tuberculosis (TB), a chronic infectious illness brought on by <i>Mycobacterium tuberculosis</i> (<i>M. tuberculosis</i>). A distinct family of PE_PGRS proteins, encoded by the <i>M. tuberculosis</i> genome, has attracted more attention because of their involvement in immune evasion and bacterial pathogenicity. Nevertheless, the specific functions and mechanisms of action for the majority of PE_PGRS proteins remain largely unexplored. This study focuses on the Rv2741 (PE_PGRS47) gene, which is exclusively present in pathogenic mycobacteria. To examine the function of Rv2741 in host-pathogen interactions, we created recombinant strains of <i>Mycobacterium smegmatis</i> (<i>M. smegmatis</i>) that expressed the <i>M. tuberculosis</i> Rv2741 gene. IL-1α was found to be a key mediator of host response modulation by Rv2741. Rv2741 downregulates the secretion of IL-1α and inhibits the MAPK signaling pathway, particularly the p38 and ERK1/2 pathways, thereby cooperatively inhibiting macrophage autophagy and apoptosis. Meanwhile, the decrease in IL-1α secretion directly leads to changes in the cytokine secretion pattern and a reduction in nitric oxide (NO) production. This multifaceted regulatory mechanism ultimately favors the survival of <i>M. smegmatis</i> in macrophages. This research significantly expands our understanding of Rv2741 function, revealing its crucial role as a multifunctional virulence factor in the immune evasion of <i>M. tuberculosis</i>.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"676-688"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143513989","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":"<i>Treponema pallidum</i> Flagellin FlaB3 Activates Inflammation and Inhibits Autophagy in HMC3 Cells via the TLR4 Pathway.","authors":"Fangzhi Du, Zhiyu Lu, Qingyun Wu, Xu Zhang, Xiaoli Zheng, Ruili Zhang, Qianqiu Wang","doi":"10.1021/acsinfecdis.4c01064","DOIUrl":"10.1021/acsinfecdis.4c01064","url":null,"abstract":"<p><p>Neurosyphilis, a neurological manifestation of syphilis, is closely related to neuroinflammation. Autophagy, a fundamental cellular mechanism that mediates the degradation of intracellular components, plays a crucial role in immune regulation and inflammation. Microglia, resident immune cells in the brain, are central to these processes. However, the interplay between autophagy and neuroinflammation in the context of neurosyphilis remains poorly understood. In this research, the recombinant <i>Treponema pallidum</i> flagellin, FlaB3, was constructed to treat human microglia clone 3 (HMC3) cells and HMC3 cells in which TLR4 (Toll-like receptor 4) had been knocked down. We discovered that FlaB3 promotes IL-6 and IL-8 secretion through the TLR4 pathway. We also observed that FlaB3 regulates the expression of autophagy-related proteins Beclin1, LC3B, and P62 via the TLR4/PI3K/AKT/mTOR pathway, thereby inhibiting autophagy and autophagic flux in HMC3 cells. Subsequently, we discovered that the concentration of soluble amyloid β_1-42 (Aβ_1-42) was decreased in the cerebrospinal fluid of neurosyphilis patients. Immunofluorescence analysis further revealed that FlaB3 suppresses the degradation of Aβ by autophagosomes in HMC3 cells. Additionally, treatment with the autophagy activators Rapamycin and LY294002 decreased the levels of IL-6 and IL-8 secretion, indicating that autophagy modulates inflammation in HMC3 cells. In summary, our study demonstrates that FlaB3 promotes inflammation in HMC3 cells by inhibiting autophagy. This inhibition also impedes Aβ degradation, providing new insights into the pathogenesis of neurosyphilis.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"773-783"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143555302","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":"C7-Substituted Quinolines as Potent Inhibitors of AdeG Efflux Pumps in <i><i>Acinetobacter baumannii</i></i>.","authors":"Yiling Zhu, Charlotte K Hind, Taha Al-Adhami, Matthew E Wand, Melanie Clifford, J Mark Sutton, Khondaker Miraz Rahman","doi":"10.1021/acsinfecdis.4c00705","DOIUrl":"10.1021/acsinfecdis.4c00705","url":null,"abstract":"<p><p>Efflux, mediated by a series of multidrug efflux pumps, is a major contributor to antibiotic resistance in Gram-negative bacteria. Efflux pump inhibitors (EPIs), which can block efflux, have the potential to be used as adjuvant therapies to resensitize bacteria to existing antibiotics. In this study, 36 quinoline-based compounds were synthesized as potential EPIs targeting resistance nodulation division (RND) family pumps in the multidrug-resistant pathogen <i><i>Acinetobacter baumannii</i></i>. In <i><i>A. baumannii</i></i> strains with overexpressed AdeFGH (chloramphenicol-adapted) and AdeABC (AYE, Ab5075-UW), these compounds enhanced Hoechst dye accumulation, indicating general efflux inhibition, and potentiated chloramphenicol, which is an AdeG substrate. The research focused on two generations of quinoline compounds, with modifications at the C-7 position of first-generation compounds to improve hydrophobic interactions with the Phe loop in the AdeG efflux pump, to generate second-generation compounds. The modified quinolines showed strong pump inhibition and significant chloramphenicol potentiation, with MIC reductions of 4- to 64-fold. Notably, compounds <b>1.8</b> and <b>3.8</b> exhibited the highest inhibitory activity, while compounds <b>1.3</b> and <b>3.3</b> showed up to 64-fold potentiation, highlighting the importance of specific structural features at the C-7 position for efflux pump inhibition. The study also revealed selective inhibition of AdeFGH over AdeABC, with no potentiation observed for gentamicin, showing the specificity of these quinoline-based inhibitors. Importantly, the compounds showed no toxicity in a <i>Galleria mellonella</i> model at a dose level of 20 mg/kg, highlighting their suitability as potential antibiotic adjuvants for combating bacterial resistance.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"626-638"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11915368/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143522188","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":"Targeting Two-Tetrad RNA G-Quadruplex in the SARS-CoV-2 RNA Genome Using Tetraphenylethene Derivatives for Antiviral Therapy.","authors":"Payal Gupta, Rushikesh M Khadake, Oinam Ningthemmani Singh, Harshad A Mirgane, Dharmender Gupta, Sheshanath V Bhosale, Sudhanshu Vrati, Milan Surjit, Ambadas B Rode","doi":"10.1021/acsinfecdis.5c00058","DOIUrl":"10.1021/acsinfecdis.5c00058","url":null,"abstract":"<p><p>Targeting the specific RNA conformations that are crucial for SARS-CoV-2 replication is a viable antiviral approach. The SARS-CoV-2 genome contains GG repeats capable of forming unstable two-tetrad G-quadruplex (GQ) structures, which exist as a mix of conformations, including hairpin (Hp), intra-, and intermolecular GQs. RGQ-1, originating from the nucleocapsid gene's ORF, adopts a dynamic equilibrium of conformations, including intramolecular hairpin and G-quadruplex (Hp-GQ) structures, as confirmed by CD analysis. In this study, tetraphenylethene (TPE) derivatives were developed to target the Hp-GQ conformational equilibrium of RGQ-1. EMSA, fluorescence spectroscopy, and ITC assays confirmed that two TPE derivatives, TPE-MePy and TPE-Allyl Py, bind to RGQ-1. CD thermal melting experiments indicate that RGQ-1 is stabilized by 8.56 and 12.54 °C in the presence of TPE-MePy and TPE-Allyl Py, respectively. Additionally, luciferase assays demonstrated that TPE derivatives suppressed luciferase activity by 2.2-fold and 3.6-fold, respectively, shifting the HpGQ equilibrium toward the GQ conformation, as suggested by CD spectroscopy. Treatment of SARS-CoV-2-infected A549 cells with TPE derivatives reduced the levels of viral RNA, spikes, and nucleocapsid proteins. To explore their antiviral mechanism, preinfection and postinfection treatments were tested, revealing that the TPE derivatives specifically suppressed the postentry stages of viral replication without affecting viral entry. These findings highlight the therapeutic potential of TPE derivatives in inhibiting key gene expressions critical for SARS-CoV-2 replication.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":"784-795"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143522190","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}