SLAS Discovery最新文献

{"title":"A high-throughput approach to evaluating NCp7 RNA binding activity for HIV-1 drug discovery","authors":"Joanna K. Winstone ,&nbsp;Rikki Uhrich ,&nbsp;Thibault Alle ,&nbsp;Brian C. Kraemer","doi":"10.1016/j.slasd.2025.100260","DOIUrl":"10.1016/j.slasd.2025.100260","url":null,"abstract":"<div><div>The HIV-1 epidemic broadly impacts healthcare. There remains a continued need for improved anti-viral therapies resilient to the development of drug resistance. HIV-1 nucleocapsid protein 7 (NCp7) seems a prime drug target due to its unique nucleic acid chaperone activity required for multiple viral processes. NCp7 RNA binding activity has been shown to increase viral production and infectivity within the host. Here we introduce a high-throughput AlphaScreen assay to evaluate NCp7 RNA binding activity and validate its specificity and sensitivity using a known inhibitor. We also demonstrate the utility of this assay by performing a drug-repurposing screen, which identified seven confirmed inhibitors of NCp7 RNA binding and two confirmed enhancers of NCp7 RNA binding. This tool will aid in future NCp7-targeted drug discovery initiatives for the treatment of HIV-1 infection.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100260"},"PeriodicalIF":2.7,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144850062","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of novel inflammasome inhibitors via cellular NLRP3 target engagement assays","authors":"Francisco Castillo ,&nbsp;Thomas A. Mackenzie ,&nbsp;Elisabeth Domingo ,&nbsp;Inmaculada Iañez ,&nbsp;Matthew B. Robers ,&nbsp;Jennifer Wilkinson ,&nbsp;Erika Kay-Tsumagari ,&nbsp;Martha O’Brien ,&nbsp;Olga Genilloud ,&nbsp;Rosario Fernandez-Godino ,&nbsp;Maria C. Ramos","doi":"10.1016/j.slasd.2025.100258","DOIUrl":"10.1016/j.slasd.2025.100258","url":null,"abstract":"<div><div>The NLRP3 (NOD-like receptor family, pyrin domain-containing protein 3) inflammasome, a multiprotein complex, plays a crucial role in triggering the release of pro-inflammatory cytokines like interleukin-1 beta. Abnormal activation of NLRP3 can mediate an aberrant immune response to viral infections and is associated with inflammatory diseases. In this study, the goal was to identify bioactive, potent, and specific inhibitors of NLRP3 that could modulate the inflammasome pathway and assess their potential therapeutic relevance. An innovative workflow was assembled by setting up a robust cellular-based high throughput screening (HTS) target engagement (TE) tool to identify potent NLRP3 inhibitors and validate their functional effect on the inflammasome downstream signaling cascade. A subset of 2,500 compounds from the European Chemical Biology Library (ECBL) was screened and validated inhibitors were subjected to a similarity study by state-of-the-art computational tools to comprehend their specific impact on inflammasomal signaling nodes upstream of NLRP3 and to propose feasible anti-inflammatory drugs. Ultimately, ten compounds were selected and validated in functional checkpoints of the NLRP3 inflammasome pathway, such as caspase-1 activity and IL-1β release, proving the validity of this HTS TE method for identifying NLRP3 inhibitors. Subsequent similarity-based clustering suggested the organization of the active compounds into three primary groups, linked to NF-κB signaling, ROS-induced NLRP3 activation, and NLRP3 induction pathway in response to microbial and related insults. Overall, these findings demonstrate the robustness and efficiency of the target-engagement methodology to capture bioactive inflammasome inhibitors with diverse mechanisms of action. Seven of the identified NLRP3 inhibitors were characterized as novel inflammasome inhibitors with therapeutic potential.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100258"},"PeriodicalIF":2.7,"publicationDate":"2025-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144823308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An overview of drugging the bacterial cytoskeleton, rod, and divisome systems","authors":"Elvis Awuni","doi":"10.1016/j.slasd.2025.100261","DOIUrl":"10.1016/j.slasd.2025.100261","url":null,"abstract":"<div><div>Bacterial infections and antibiotic resistance remain significant threats to global health, with millions of related deaths recorded annually. Projections that antibacterial resistance-related deaths could reach alarming proportions in the coming years, along with the shortcomings of current interventions, highlight the need for new drug targets, novel antibiotics, and revised strategies and policy actions. The bacterial cytoskeleton, rod, and divisome systems (BCRDs) perform vital cellular roles and serve as a reserve of numerous potential therapeutic targets. The components of the BCRDs play different roles but share some relationships, suggesting the possibility of exploiting synergistic, polytherapeutic, and polypharmacological effects with antibiotics to mitigate bacterial resistance. Unfortunately, few drug targets within the BCRDs have been validated, and bacterial resistance to the inhibitors and approved antibiotics poses a challenge to the health and pharmaceutical industries. This review provides a concise but comprehensive overview of drugging the BCRDs, emphasizing the relationships and druggable potentials, validated targets, inhibitors, challenges, interventions, prospects, perspectives, and future directions geared toward reinvigorating research and overcoming bottlenecks in the sector. Overall, the material presented and discussed could facilitate the identification and validation of new therapeutic targets, the discovery and development of novel clinical drugs, and the revision of strategies and policy interventions to augment the fight against antibiotic resistance.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100261"},"PeriodicalIF":2.7,"publicationDate":"2025-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144818588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aurora 2.0: A fluorogenic dye library for expanding the capability of protein-adaptive differential scanning fluorimetry (paDSF)","authors":"Annemarie F. Charvat ,&nbsp;Kayleigh Mason-Chalmers ,&nbsp;Aneta Grabinska-Rogala ,&nbsp;Shloka Shivakumar ,&nbsp;Zachary Gale-Day ,&nbsp;Taiasean Wu ,&nbsp;Zoe Millbern ,&nbsp;Jonathan B. Grimm ,&nbsp;Emma C. Carroll ,&nbsp;K․ Peter R․ Nilsson ,&nbsp;Luke D. Lavis ,&nbsp;Nelson R. Vinueza ,&nbsp;Jason E. Gestwicki","doi":"10.1016/j.slasd.2025.100259","DOIUrl":"10.1016/j.slasd.2025.100259","url":null,"abstract":"<div><div>Differential Scanning Fluorimetry (DSF) is a biophysical assay that is used to estimate protein stability <em>in vitro</em>. In a DSF experiment, the increased fluorescence of a solvatochromatic dye, such as Sypro Orange, is used to detect the unfolding of a protein during heating. However, Sypro Orange is only compatible with a minority of proteins (&lt; 30 %), limiting the scope of this method. We recently reported that protein-adaptive DSF (paDSF) can partially solve this problem, wherein the protein is initially pre-screened against ∼300 chemically diverse dyes, termed the Aurora collection. While this approach significantly improves the number of targets amenable to DSF, it still fails to produce protein-dye pairs for some proteins. Here, we report the expansion of the dye collection to Aurora 2.0, which includes a total of 517 structurally diverse molecules and multiple new chemotypes. To assess performance, these dyes were screened against a panel of ∼100 proteins, which were selected, in part, to represent the most challenging targets (<em>e.g.</em> small size). From this effort, Aurora 2.0 achieved an impressive success rate of 94 %, including producing dyes for some targets that were not matched in the original collection. These findings support the idea that larger, more chemically diverse libraries improve the likelihood of detecting melting transitions across a wider range of proteins. We propose that Aurora 2.0 makes paDSF an increasingly powerful method for studying protein stability, ligand binding and other biophysical properties in high throughput.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100259"},"PeriodicalIF":2.7,"publicationDate":"2025-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144818589","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TREM2 hit discovery using temperature-related intensity change (TRIC) technology: A proof-of-concept high-throughput screening approach","authors":"Natalie Fuchs ,&nbsp;Katarzyna Kuncewicz ,&nbsp;Farida El Gaamouch ,&nbsp;Moustafa T. Gabr","doi":"10.1016/j.slasd.2025.100255","DOIUrl":"10.1016/j.slasd.2025.100255","url":null,"abstract":"<div><div>Triggering receptor expressed on myeloid cells 2 (TREM2) is an immunomodulatory receptor implicated in both neurodegenerative diseases and cancer. Depending on the context, TREM2 agonists or inhibitors hold therapeutic potential. To date, the majority of TREM2-targeted strategies have centered on monoclonal antibodies (mAbs), which face limitations such as poor tissue penetration and potential immunogenic side effects. To overcome these challenges and expand the chemical space for TREM2-targeting agents, we developed a high-throughput screening (HTS) platform to identify novel small molecule TREM2 binders. Using temperature-related intensity change (TRIC) technology in a 384-well plate format (NanoTemper Dianthus), we screened two focused compound libraries comprising over 1,200 molecules. From this screen, 18 preliminary hits (1.44 % hit rate) were identified and subsequently validated by dose-response binding studies using microscale thermophoresis (MST), yielding four validated hits (0.32 % hit rate) with binding affinities in the high to medium micromolar range (e.g., <strong>T2337</strong>, <em>K</em>_D = 22.4 µM). The binding of the top hit, <strong>T2337</strong>, was further validated using surface plasmon resonance (SPR). Additionally, we assessed the functional activity of all four validated hits in a cellular assay measuring TREM2-mediated Syk phosphorylation in HEK293 cells co-expressing human TREM2 and its adaptor protein DAP12. These findings establish a robust and scalable platform for the discovery of small molecule TREM2 modulators and serve as a proof-of-concept for broader HTS campaigns targeting TREM2.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100255"},"PeriodicalIF":2.7,"publicationDate":"2025-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144763963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Off-target effects of oligonucleotides and approaches of preclinical assessments","authors":"Haiwen Ruan ,&nbsp;Dehu Dou ,&nbsp;Jing Lu ,&nbsp;Xia Xiao ,&nbsp;Xinjiang Gong ,&nbsp;Xuefeng Zhang","doi":"10.1016/j.slasd.2025.100254","DOIUrl":"10.1016/j.slasd.2025.100254","url":null,"abstract":"<div><div>Oligonucleotide-based therapies, such as antisense oligonucleotides (ASOs), small interfering RNAs (siRNAs), represent a class of therapeutic agents that specifically target gene transcription or translation mechanisms through sequence specificity. These pharmaceuticals exhibit significant promise in the treatment of genetic disorders, including spinal muscular atrophy, as well as malignancies, viral infections, and metabolic diseases. Nonetheless, unintended toxicity continues to pose a considerable challenge and remain a critical safety concern in the development of oligonucleotide therapeutics (ONTs). Off-target toxicity may be caused by hybridization to sequences that are similar but not identical to the target, hybridization-independent sequence related, or sequence- and hybridization-independent effects. The effects may result in diminished transcript levels, decreased translation rates, or anomalous splicing, employing same molecular pathways and protein machinery as the desired on-target effects. Currently, there exists no established methodology for the systematic identification and evaluation of off-target toxicity, which may hinder the optimization of safety approaches. This review delineates significant nonclinical toxicities and clinical adverse effects by summarizing and analyzing approved oligonucleotides with their off-target assays, encompassing the limitations of nonclinical off-target effects and the potential off-target mechanisms. Plus, it discusses and emphasizes the factors that lead to the off target of ONTs, systematically offers approaches and workflows of preclinical assessments to enhance the transfer value of oligonucleotide therapies from nonclinical to clinical trials by managing unavoidable off-target effects.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100254"},"PeriodicalIF":2.7,"publicationDate":"2025-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144721301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bonferroni’s method, not Tukey’s, should be used to control the total number of false positives when making multiple pairwise comparisons in experiments with few replicates","authors":"Adam Zweifach","doi":"10.1016/j.slasd.2025.100253","DOIUrl":"10.1016/j.slasd.2025.100253","url":null,"abstract":"<div><div>Statistical tests can be used to help determine whether experimental manipulations produce effects. In tests of means, when more than two groups are compared the total number of Type 1 errors (false positive results) increases unless a correction is used. Tukey’s method is thought to offer good control of the number of false positives and high statistical power when all pairwise comparisons are made. However, the number of replicates in laboratory experiments is often quite low, and small sample sizes can undermine assumptions underlying statistical methods. I used simulations to investigate how well ANOVA followed by different post-hoc tests controls the total number of false positives when there are 3- 6 experimental groups and 2- 6 experimental replicates, conditions that span the range of typical values. Tukey’s method, one of the most common, allows too many. I investigated 11 other approaches to controlling false positives and found that none is as effective as the simple Bonferroni correction or offers much more power. I conclude that researchers should not make all pairwise comparisons using ANOVA followed by Tukey’s method but instead use Bonferroni’s method on a limited number of pre-selected comparisons.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100253"},"PeriodicalIF":2.7,"publicationDate":"2025-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144700594","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Overcoming a false-positive mechanism in RapidFire MRM-based high throughput screening","authors":"De Lin ,&nbsp;Lesley-Anne Pearson ,&nbsp;Shamshad Ahmad,&nbsp;Sandra O’Neill,&nbsp;John Post,&nbsp;Colin Robinson,&nbsp;Duncan E. Scott,&nbsp;Ian H. Gilbert","doi":"10.1016/j.slasd.2025.100252","DOIUrl":"10.1016/j.slasd.2025.100252","url":null,"abstract":"<div><div>False-positives plague high-throughput screening in general and are costly as they consume resource and time to resolve. Methods that can rapidly identify such compounds at the initial screen are therefore of great value. Advances in mass spectrometry have led to the ability to screen inhibitors in drug discovery applications by direct detection of an enzyme reaction product. The technique is free from some of the artefacts that trouble classical assays such as fluorescence interference. Its direct nature negates the need for coupling enzymes and hence is simpler with fewer opportunities for artefacts. Despite its myriad advantages, we report here a mechanism for false-positive hits which has not been reported in the literature. Further we have developed a pipeline for detecting these false-positive hits and suggest a method to mitigate against them.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100252"},"PeriodicalIF":2.7,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144676734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An enzyme activity-based workflow for the identification and characterization of covalent inhibitors","authors":"Buyun Tang,&nbsp;Tatiana Gladysheva,&nbsp;Paul Lang","doi":"10.1016/j.slasd.2025.100250","DOIUrl":"10.1016/j.slasd.2025.100250","url":null,"abstract":"<div><div>The field of covalent drug development has advanced rapidly, offering promising therapeutic potential due to the ability of these drugs to form slowly reversible or irreversible bonds with target proteins, resulting in prolonged pharmacodynamic effects. This distinctive mechanism of action has sparked resurging interest in covalent inhibitors across various disease areas, including oncology, neurological disorders, and infectious diseases. However, characterization of covalent inhibitors poses unique challenges, highlighting the need for simplified and robust assay methods. This protocol describes an enzyme activity-based workflow designed to identify and characterize covalent inhibitors efficiently. By streamlining the evaluation process, this approach enhances the reliability and reproducibility of covalent inhibitor assessment, ultimately accelerating the discovery and optimization of novel covalent therapeutics.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100250"},"PeriodicalIF":2.7,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144676733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nanobodies: A new frontier in antiviral therapies","authors":"Ahmed Mohammed ,&nbsp;Mujahed I. Muustafa","doi":"10.1016/j.slasd.2025.100251","DOIUrl":"10.1016/j.slasd.2025.100251","url":null,"abstract":"<div><div>Nanobodies, derived from the immune systems of camelids such as alpacas and llamas, represent a novel class of therapeutics with significant potential in fighting respiratory viral infections, such as SARS-CoV-2 and influenza. Nanobodies are small biomolecules that are highly stable and have unique binding features that allow for the effective neutralization of viral particles and inhibition of viral replication. This review highlights the advantages of nanobodies over traditional antibodies, including cost-effective production and enhanced specificity for target antigens. We discuss the mechanisms through which nanobodies block viral entry, their applications in diagnostics, and the methodologies for their development, such as phage display technology. Furthermore, we explore the efficacy of nanobodies in preclinical studies and their potential in clinical settings. As research progresses, structural optimization and the exploration of combination therapies may enhance their therapeutic efficacy, providing a promising approach for addressing global health challenges caused by emerging viral pathogens.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100251"},"PeriodicalIF":2.7,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144669122","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}