ACS Chemical Biology最新文献

{"title":"High-Throughput Screening of Potent Drug-like Molecules Targeting 17β-HSD10 for the Treatment of Alzheimer's Disease and Cancer.","authors":"Laura Aitken, Gemma Baillie, Andrew Pannifer, Angus Morrison, Louise L Major, Magnus S Alphey, Ritika Sethi, Martin Timmerman, John Robinson, Jennifer Riley, Yoko Shishikura, Lizbe Koekemoer, Frank Von Delft, Helma Rutjes, Kevin D Read, Philip S Jones, Stuart P McElroy, Terry K Smith, Frank J Gunn-Moore","doi":"10.1021/acschembio.5c00110","DOIUrl":"10.1021/acschembio.5c00110","url":null,"abstract":"<p><p>In this study, the first industrial-scale high-throughput screening of nearly 350,000 drug-like molecules targeting the enzyme 17β-HSD10, a promising therapeutic target for Alzheimer's disease and cancers, is presented. Two novel series of potent 17β-HSD10 inhibitors that demonstrate low nanomolar potency against both the enzyme and <i>in vivo</i> cellular assays with minimal cytotoxicity were identified. These inhibitors were characterized further through a series of assays demonstrating ligand-protein interactions and co-crystallography, revealing un-/non-competitive inhibition with respect to the cofactor NADH, unlike previously published inhibitors. This work significantly advances the development of 17β-HSD10-targeting therapeutics, offering new potential leads for treating Alzheimer's disease and cancers.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":"1544-1559"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12281485/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144323817","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-Throughput Screening Tool to Identify Small Molecule Inhibitors of Telomerase.","authors":"Elisa Aquilanti, Sulyman Barkho, Vincent Bozinov, Lauren Kageler, Max Garrity-Janger, Michael F Mesleh, Steven Horner, Matthew J Ranaghan, Matthew Meyerson","doi":"10.1021/acschembio.5c00244","DOIUrl":"10.1021/acschembio.5c00244","url":null,"abstract":"<p><p>Telomerase reverse transcriptase is a ribonucleoprotein complex that maintains telomere length in rapidly dividing cells, thus enabling cellular immortality. Despite being recognized as an important cancer target for decades, no small molecule telomerase inhibitors have been approved as anticancer therapeutics to date. Several limitations, including the absence of high-throughput screening tools, have posed challenges to the telomerase drug discovery field. Here, we describe a high-throughput, fluorescently coupled screening method employing a chemically modified reporter nucleotide. We utilize the <i>Tribolium castaneum</i> telomerase as a surrogate model as it shares a high degree of active site homology with the human enzyme. We piloted this tool by screening a chemical library of ∼3600 nucleoside mimetics to demonstrate excellent assay quality, and identified 2 compounds with inhibitory activity that were further validated in a direct enzymatic assay. Our work introduces a method that has the potential to uncover novel telomerase inhibitors for further drug discovery efforts.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":"1707-1714"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12281481/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144256626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Three-Enzyme Cascade Catalyzes Conversion of Auramycinone to Resomycin in Chartreusin Biosynthesis.","authors":"Magdalena Niemczura, Aleksi Nuutila, Rongbin Wang, Katariina Rauhanen, S Eric Nybo, Mikko Metsä-Ketelä","doi":"10.1021/acschembio.5c00205","DOIUrl":"10.1021/acschembio.5c00205","url":null,"abstract":"<p><p>Chartreusin is a potent antiproliferative agent that contains a unique aromatic pentacyclic bislactone carbon scaffold. The biosynthesis of type II polyketide aglycone has been extensively investigated and shown to proceed through a tetracyclic anthracycline intermediate. The last remaining unknown steps are the conversion of auramycinone to resomycin C. Here we have discovered three enzymes that play crucial roles in two mechanistically distinct dehydration reactions. We show that ChaX is an NAD(P)H-dependent auramycinone quinone reductase that allows the cyclase-like ChaU to catalyze the formation of 9,10-dehydroauramycinone via a carbanion intermediate. In contrast, the cyclase-like ChaJ, homologous to ChaU, is responsible for subsequent 7,8-dehydration via a canonical carbocation intermediate, yielding resomycin C. The results were confirmed via assembly of the biosynthetic pathway for production of resomycin C in <i>Streptomyces coelicolor</i> M1152Δ<i>matAB</i>. The work expands the catalytic repertoire of the SnoaL protein family, which has previously been associated with anthracycline fourth-ring cyclization and two-component 1-hydroxylation.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":"1457-1463"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12281474/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144537387","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Photoaffinity Ligand of Cystic Fibrosis Corrector VX-445 Identifies SCCPDH as an Off-Target.","authors":"Minsoo Kim, Kwangho Kim, Jesun Lee, Lea A Barny, Toya D Scaggs, Ian M Romaine, KyuOk Jeon, Simona G Codreanu, Stacy D Sherrod, John A McLean, Anjaparavanda P Naren, Gary A Sulikowski, Lars Plate","doi":"10.1021/acschembio.5c00157","DOIUrl":"10.1021/acschembio.5c00157","url":null,"abstract":"<p><p>Cystic fibrosis (CF) pharmacological correctors improve the cystic fibrosis transmembrane conductance regulator (CFTR) protein trafficking and function. Several side effects of these correctors and adverse drug interactions have been reported, emphasizing the need to understand off-targets. We synthesized VU439, a functionalized photoaffinity ligand (PAL) of VX-445. Chemoproteomics analysis by mass spectrometry (MS) was used to identify cross-linked proteins in CF bronchial epithelial cells expressing F508del CFTR. We identified saccharopine dehydrogenase-like oxidoreductase (SCCPDH), an uncharacterized putative oxidoreductase, as a VX-445-specific off-target. We also characterized changes in the metabolomic profiles of cells overexpressing SCCPDH to determine the consequence of binding of VX-445 to SCCPDH. These data show dysregulation of amino acid metabolism and a potential inhibitory activity of VX-445 on SCCPDH. The identified off-target may explain the exacerbation of psychological symptoms observed in the clinic, thus emphasizing the need for further optimization of correctors.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":"1560-1573"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12281486/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144332045","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Target-Responsive DNA Hydrogels with Encapsulation and Release Properties Using Programmable CRISPR-Cas12a.","authors":"Ram J Tharu, Emmett Hanson, Mehmet V Yigit","doi":"10.1021/acschembio.5c00355","DOIUrl":"10.1021/acschembio.5c00355","url":null,"abstract":"<p><p>We report the development of a DNA hydrogel that disassembles and releases its payload in response to a target of interest. The DNA hydrogel is assembled from Y-shaped DNA motifs with polyA domains and cross-linked <i>via</i> the small molecule cyanuric acid through hydrogen bonding. The hydrogel's structural integrity was rapidly assessed using a simple, instrumentation-free capillary migration assay that provides results within seconds. To evaluate its responsiveness to enzymatic degradation, the hydrogel was exposed to nonspecific nuclease activity using <i>DNase I</i>, resulting in increased mobility and decrease in fluorescence. Later, CRISPR-Cas12a was incorporated to enable programmable, target-specific hydrogel disassembly using a conserved genomic region from <i>Salmonella typhimurium</i>. Guided by crRNA sequences, the target sequences activated Cas12a to selectively degrade hydrogels. This process enabled the controlled release of various payloads, including a small-molecule drug, a fluorescent dye, a nanoparticle-based MRI contrast agent conjugated to a chemotherapeutic agent, and a model protein. To evaluate whether the hydrogel disassembly can be selectively programmed to an intended target, we tested its responsiveness against two serotypes of <i>Salmonella</i>, <i>i.e.</i>, conserved genomic regions from <i>Salmonella enteritidis</i> and <i>S. typhimurium</i>. To test the disassembly of this novel DNA hydrogel in the presence of a full genome, we tested the hydrogel with the <i>S. typhimurium</i> genome. The target genome induced an increase in the hydrogel's mobility and loss in fluorescence with as few as 50 copies of full genome. The results demonstrate the potential of these CRISPR-responsive DNA hydrogels as intelligent platforms for target-induced imaging and therapeutic agent release, and biosensing applications.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":"1805-1812"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12281480/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tryptamine Metabolism and Functionalization in Gut Commensal Bacteria Expand Human Tryptamine Signaling Responses.","authors":"Hyun Bong Park, Deguang Song, Mytien Nguyen, Noah W Palm, Jason M Crawford","doi":"10.1021/acschembio.5c00313","DOIUrl":"10.1021/acschembio.5c00313","url":null,"abstract":"<p><p>Gut microbes secrete specialized small molecules that broadly influence human physiology. Despite their potential significance, the variety of functional small molecules known in the gut is relatively limited. Here, we screened the supernatants from human fecal-derived bacterial cultures to explore their agonist effects on the human G protein-coupled receptors (GPCRs), melatonin receptor types 1A and 1B (MTNR1A, MTNR1B). Chemical analysis of the supernatant-soluble molecules of <i>Clostridium sporogenes</i>, a prominent gut commensal identified in the screen, led to the characterization of agonists for these two melatonin receptors. Specifically, through bioactivity-assisted isolation and characterization, we identified three small molecules, <b>1</b>-<b>3</b>, including two previously uncharacterized metabolites, which were synthesized to confirm their structures. While the structure of <b>1</b> features a urea core symmetrically disubstituted with tryptamine moieties, <b>2</b> and <b>3</b> harbor a monomeric tryptamine functionalized with methyl carbamate and <i>N</i>-acetyl groups, respectively. These structural characterization efforts illuminated downstream functional consequences of tryptamine metabolism in <i>C. sporogenes</i>. Additional GPCR screening analyses revealed that <b>2</b> activates melatonin receptors and the purinergic P2RY11 receptor, whereas <b>1</b> serves as an agonist for the semiorphan receptor GPR55. Interestingly, <b>1</b> also exhibits significant inhibitory activity against inflammatory soluble epoxide hydrolase with a half-maximal inhibitory concentration of 420 nM. Single-cell RNA sequencing analysis of the gut tissue from mice orally treated with <b>1</b> relative to the solvent vehicle control revealed that <b>1</b> specifically decreased the frequency of GPR55- and granzyme K-expressing effector-like CD8 T cells in the intraepithelial lymphocyte population. Overall, this study broadens our understanding of tryptamine-derived signaling at the human-microbe interface.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":"1775-1782"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144511248","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phospholipase PAFAH2 Mediates Ferroptosis Surveillance and Lipid Remodeling to Promote Resistance in KEAP1 Mutant Cancers.","authors":"Stephen B Ruiz, Daniel E Tylawsky, Janki Shah, Michelle Saoi, Brandon Cuevas, Shanay Desai, Boglarka Racz, Ana Marie Perea, Arianna R Izawa-Ishiguro, Justin Cross, Daniel A Heller","doi":"10.1021/acschembio.5c00273","DOIUrl":"10.1021/acschembio.5c00273","url":null,"abstract":"<p><p>Although ferroptosis resistance is prevalent among many cancer cell types, precisely how ferroptosis surveillance mechanisms are induced remains elusive due to the heterogeneity of the cellular mutational status and metabolic states. Here, we find that phospholipase PAFAH2 regulates ferroptosis through its unique ability to specifically detoxify membrane-bound oxidized phospholipids in KEAP1 mutant and NRF2-active cancer cells. We show that the genetic or chemical perturbation of PAFAH2 is sufficient to sensitize KEAP1 mutant lung adenocarcinoma cells to ferroptosis. Lipidomic analyses reveal that PAFAH2 inhibition shifts the cellular lipidome to a distinctly ferroptosis state characterized by the enrichment of key phospholipids previously identified to be important in ferroptosis, like ether-linked phosphatidylethanolamines. Finally, we comparatively assessed the antitumor efficacy of PAFAH2 inhibitor monotherapy versus cotreatment with a nanoparticle-stabilized GPX4 inhibitor formulation. Our findings support that the broad applicability of PAFAH2 inhibition can be used in ferroptosis induction and abrogation of ferroptosis resistance across cancer types.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":"1739-1755"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144537385","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a HiBiT Peptide-Based NanoBRET Ligand Binding Assay for Galanin Receptor 1 in Live Cells.","authors":"Hu Zhu, Harrison C Daly, Tae Gyun Yang, Josh Born, Gisela Andrea Camacho-Hernandez, Mingyang Yuan, Mari Inglese, Vinoth Kumar Chenniappan, Kris Zimmerman, Robin Hurst, Xin Hu, Amy Hauck Newman, Sergi Ferré, Rachel Friedman Ohana, Matthew D Hall, Samarjit Patnaik","doi":"10.1021/acschembio.5c00166","DOIUrl":"10.1021/acschembio.5c00166","url":null,"abstract":"<p><p>Galanin is a neuroendocrine peptide that regulates a wide range of physiological functions, including feeding and energy homeostasis, mood and anxiety, and modulation of pain. The function of the galanin peptide is mediated through its three galanin receptors, namely, GALR1, GALR2, and GALR3, which belong to the G protein-coupled receptor family. To measure the interaction of ligands with galanin receptor 1 (GALR1) in living cells, we developed a novel HiBiT peptide-based NanoBRET ligand binding assay. We generated six bioluminescence resonance energy transfer (BRET) tracers composed of modified and truncated galanin peptide derivatives tagged with a 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY) acceptor fluorophore. The fluorophore-tagged peptide tracers were evaluated in cells expressing GALR1 tagged with HiBiT, an 11-amino acid subunit of NanoLuc, which, upon high affinity complementation with the cell-impermeable subunit LgBiT, reconstituted a functional NanoLuc luciferase. Addition of the furimazine substrate induced BRET to the BODIPY fluorophore acceptor component of the galanin-derived peptide tracers and produced a fluorescence signal output. Using this BRET assay, we characterized the binding affinity and binding kinetics of tracers with GALR1 in both equilibrium and real time. To validate our assay, we evaluated the binding affinity and function of a panel of unmodified galanin-derived peptide ligands through the competitive displacement of bound fluorescent galanin tracers. Our data showed that the binding affinity of these galanin peptide ligands correlated well with their rank order in β-arrestin recruitment and internalization functional assays. This study demonstrates that the HiBiT peptide-based NanoBRET ligand binding assay is a valuable system for studying the ligand engagement of GALR1 in living cells, offering an alternative to neuropeptide radioligand binding assays.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":"1594-1608"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144564147","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"","authors":"Mathilde Pucher,&nbsp;Kirrthana Makenthirathasan,&nbsp;Hadrien Jalaber,&nbsp;Thomas LeSaux,&nbsp;Oliver Nüsse,&nbsp;Gilles Doisneau,&nbsp;Yann Bourdreux,&nbsp;Blaise Gatin-Fraudet,&nbsp;Ludovic Jullien,&nbsp;Boris Vauzeilles,&nbsp;Dominique Guianvarc’h,&nbsp;Marie Erard and Dominique Urban*,&nbsp;","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 7","pages":"XXX-XXX XXX-XXX"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acschembio.5c00156","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144646167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"","authors":"Laura Aitken,&nbsp;Gemma Baillie,&nbsp;Andrew Pannifer,&nbsp;Angus Morrison,&nbsp;Louise L. Major,&nbsp;Magnus S. Alphey,&nbsp;Ritika Sethi,&nbsp;Martin Timmerman,&nbsp;John Robinson,&nbsp;Jennifer Riley,&nbsp;Yoko Shishikura,&nbsp;Lizbe Koekemoer,&nbsp;Frank Von Delft,&nbsp;Helma Rutjes,&nbsp;Kevin D. Read,&nbsp;Philip S. Jones,&nbsp;Stuart P. McElroy,&nbsp;Terry K. Smith and Frank J. Gunn-Moore*,&nbsp;","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 7","pages":"XXX-XXX XXX-XXX"},"PeriodicalIF":3.5,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acschembio.5c00110","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144646171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}