SLAS Discovery最新文献

{"title":"Recapitulation of NOD/RIPK2 signaling in iPSC-derived macrophages","authors":"Mozhgan Dehghan Harati ,&nbsp;Jim King ,&nbsp;Simon Langer ,&nbsp;Florian Binder ,&nbsp;Ralf Heilker","doi":"10.1016/j.slasd.2024.100185","DOIUrl":"10.1016/j.slasd.2024.100185","url":null,"abstract":"<div><div>Human induced pluripotent stem cell (iPSC)-derived macrophages (IDMs) present a valuable substitute for monocyte-derived macrophages (MDMs) in order to study inflammation pathways in vitro. Through optimization of an IDM differentiation protocol, a six-fold increase in the production yield of myeloid progenitors was achieved. The derived IDMs were further characterized with respect to nucleotide-binding oligomerization domain (NOD) and receptor-interacting serine/threonine-protein kinase 2 (RIPK2) signaling, a key regulatory pathway for autoimmune diseases. The IDM cells recapitulated MDM biology with respect to the proinflammatory chemokine and inflammatory cytokine fingerprint more closely than THP-1 cells. When assessing RIPK2 modulation effect on tumor necrosis factor α (TNF-α), a cardinal mediator of inflammation, a similar pharmacological effect of RIPK2 inhibitors was observed in IDMs and MDMs. Additionally, IDMs and MDMs displayed a similar transcription and pathway profile in response to NOD1/2 stimulation and pharmacological inhibition of RIPK2. In summary, the enhanced myeloid production yield in the improved IDM differentiation protocol offers new opportunities for utilizing physiologically relevant macrophage models in the context of inflammatory diseases.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 7","pages":"Article 100185"},"PeriodicalIF":2.7,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142334048","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combination screen in multi-cell type tumor spheroids reveals interaction between aryl hydrocarbon receptor antagonists and E1 ubiquitin-activating enzyme inhibitor","authors":"Thomas S. Dexheimer ,&nbsp;Nathan P. Coussens ,&nbsp;Thomas Silvers ,&nbsp;Eric M. Jones ,&nbsp;Li Chen ,&nbsp;Jianwen Fang ,&nbsp;Joel Morris ,&nbsp;Jeffrey A. Moscow ,&nbsp;James H. Doroshow ,&nbsp;Beverly A. Teicher","doi":"10.1016/j.slasd.2024.100186","DOIUrl":"10.1016/j.slasd.2024.100186","url":null,"abstract":"<div><div>The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates genes of drug transporters and metabolic enzymes to detoxify small molecule xenobiotics. It has a complex role in cancer biology, influencing both the progression and suppression of tumors by modulating malignant properties of tumor cells and anti-tumor immunity, depending on the specific tumor type and developmental stage. This has led to the discovery and development of selective AhR modulators, including BAY 2416964 which is currently in clinical trials. To identify small molecule anticancer agents that might be combined with AhR antagonists for cancer therapy, a high-throughput combination screen was performed using multi-cell type tumor spheroids grown from malignant cells, endothelial cells, and mesenchymal stem cells. The AhR selective antagonists BAY 2416964, GNF351, and CH-223191 were tested individually and in combination with twenty-five small molecule anticancer agents. As single agents, BAY 2416964 and CH-223191 showed minimal activity, whereas GNF351 reduced the viability of some spheroid models at concentrations greater than 1 µM. The activity of most combinations aligned well with the single agent activity of the combined agent, without apparent contributions from the AhR antagonist. All three AhR antagonists sensitized tumor spheroids to TAK-243, an E1 ubiquitin-activating enzyme inhibitor. These combinations were active in spheroids containing bladder, breast, ovary, kidney, pancreas, colon, and lung tumor cell lines. The AhR antagonists also potentiated pevonedistat, a selective inhibitor of the NEDD8-activating enzyme E1 regulatory subunit, in several tumor spheroid models. In contrast, the AhR antagonists did not enhance the cytotoxicity of the proteasome inhibitor bortezomib.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 7","pages":"Article 100186"},"PeriodicalIF":2.7,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142373664","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Design considerations when creating a high throughput screen-compatible in vitro model of osteogenesis","authors":"Stephanie E. Doyle,&nbsp;Courtney N. Cazzola,&nbsp;Cynthia M. Coleman","doi":"10.1016/j.slasd.2024.100184","DOIUrl":"10.1016/j.slasd.2024.100184","url":null,"abstract":"<div><div>Inducing osteogenic differentiation <em>in vitro</em> is useful for the identification and development of bone regeneration therapies as well as modelling bone disorders. To couple <em>in vitro</em> models with high throughput screening techniques retains the assay's relevance in research while increasing its therapeutic impact. Miniaturizing, automating and/or digitalizing <em>in vitro</em> assays will reduce the required quantity of cells, biologic stimulants, culture/output assay reagents, time and cost. This review highlights the design and workflow considerations for creating a high throughput screen-compatible model of osteogenesis, comparing and contrasting osteogenic cell type, assay fabrication and culture methodology, osteogenic induction approach and repurposing existing output techniques.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 7","pages":"Article 100184"},"PeriodicalIF":2.7,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142309323","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an automated 3D high content cell screening platform for organoid phenotyping","authors":"Suleyman B. Bozal ,&nbsp;Greg Sjogren ,&nbsp;Antonio P. Costa ,&nbsp;Joseph S. Brown ,&nbsp;Shannon Roberts ,&nbsp;Dylan Baker ,&nbsp;Paul Gabriel Jr. ,&nbsp;Benjamin T. Ristau ,&nbsp;Michael Samuels ,&nbsp;William F. Flynn ,&nbsp;Paul Robson ,&nbsp;Elise T. Courtois","doi":"10.1016/j.slasd.2024.100182","DOIUrl":"10.1016/j.slasd.2024.100182","url":null,"abstract":"<div><p>The use of organoid models in biomedical research has grown substantially since their inception. As they gain popularity among scientists seeking more complex and biologically relevant systems, there is a direct need to expand and clarify potential uses of such systems in diverse experimental contexts. Herein we outline a high-content screening (HCS) platform that allows researchers to screen drugs or other compounds against three-dimensional (3D) cell culture systems in a multi-well format (384-well). Furthermore, we compare the quality of robotic liquid handling with manual pipetting and characterize and contrast the phenotypic effects detected by confocal imaging and biochemical assays in response to drug treatment. We show that robotic liquid handling is more consistent and amendable to high throughput experimental designs when compared to manual pipetting due to improved precision and automated randomization capabilities. We also show that image-based techniques are more sensitive to detecting phenotypic changes within organoid cultures than traditional biochemical assays that evaluate cell viability, supporting their integration into organoid screening workflows. Finally, we highlight the enhanced capabilities of confocal imaging in this organoid screening platform as they relate to discerning organoid drug responses in single-well co-cultures of organoids derived from primary human biopsies and patient-derived xenograft (PDX) models. Altogether, this platform enables automated, imaging-based HCS of 3D cellular models in a non-destructive manner, opening the path to complementary analysis through integrated downstream methods.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 7","pages":"Article 100182"},"PeriodicalIF":2.7,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000443/pdfft?md5=95bb4a25e3d1035eb2e9758c7aafa3a7&pid=1-s2.0-S2472555224000443-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142156865","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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 for SARS-CoV-2 helicase inhibitors","authors":"Yuka Otsuka ,&nbsp;Eunjung Kim ,&nbsp;Austin Krueger ,&nbsp;Justin Shumate ,&nbsp;Chao Wang ,&nbsp;Bilel Bdiri ,&nbsp;Sultan Ullah ,&nbsp;HaJeung Park ,&nbsp;Louis Scampavia ,&nbsp;Thomas D. Bannister ,&nbsp;Donghoon Chung ,&nbsp;Timothy P. Spicer","doi":"10.1016/j.slasd.2024.100180","DOIUrl":"10.1016/j.slasd.2024.100180","url":null,"abstract":"<div><p>Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for nearly 7 million deaths worldwide since its outbreak in late 2019. Even with the rapid development and production of vaccines and intensive research, there is still a huge need for specific anti-viral drugs that address the rapidly arising new variants. To address this concern, the National Institute of Allergy and Infectious Diseases (NIAID) established nine Antiviral Drug Discovery (AViDD) Centers, tasked with exploring approaches to target pathogens with pandemic potential, including SARS-CoV-2. In this study, we sought inhibitors of SARS-CoV2 non-structural protein 13 (nsP13) as potential antivirals, first developing a HTS-compatible assay to measure SARS-CoV2 nsP13 helicase activity. Here we present our effort in implementing the assay in a 1,536 well-plate format and in identifying nsP13 inhibitor hit compounds from a ∼650,000 compound library. The primary screen was robust (average Z’ = 0.86 ± 0.05) and resulted in 7,009 primary hits. 1,763 of these compounds upon repeated retests were further confirmed, showing consistent inhibition. Following in-silico analysis, an additional orthogonal assay and titration assays, we identified 674 compounds with IC₅₀ &lt;10 μM. We confirmed activity of independent compound batches from de novo powders while also incorporating multiple counterscreen assays. Our study highlights the potential of this assay for use on HTS platforms to discover novel compounds inhibiting SARS-CoV2 nsP13, which merit further development as an effective SARS-CoV2 antiviral.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 6","pages":"Article 100180"},"PeriodicalIF":2.7,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S247255522400042X/pdfft?md5=1a67cf248af10a04f27581fcaef8915e&pid=1-s2.0-S247255522400042X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142037948","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SARS-CoV-2 Mpro inhibitor identification using a cellular gain-of-signal assay for high-throughput screening","authors":"Renee Delgado ,&nbsp;Jyoti Vishwakarma ,&nbsp;Seyed Arad Moghadasi ,&nbsp;Yuka Otsuka ,&nbsp;Justin Shumate ,&nbsp;Ashley Cuell ,&nbsp;Megan Tansiongco ,&nbsp;Christina B. Cooley ,&nbsp;Yanjun Chen ,&nbsp;Agnieszka Dabrowska ,&nbsp;Rahul Basu ,&nbsp;Paulina Duhita Anindita ,&nbsp;Dahai Luo ,&nbsp;Peter I. Dosa ,&nbsp;Daniel A. Harki ,&nbsp;Thomas Bannister ,&nbsp;Louis Scampavia ,&nbsp;Timothy P. Spicer ,&nbsp;Reuben S. Harris","doi":"10.1016/j.slasd.2024.100181","DOIUrl":"10.1016/j.slasd.2024.100181","url":null,"abstract":"<div><p>Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2, SARS2) is responsible for the COVID-19 pandemic and infections that continue to affect the lives of millions of people worldwide, especially those who are older and/or immunocompromised. The SARS2 main protease enzyme, M^pro (also called 3C-like protease, 3CL^pro), is a <em>bona fide</em> drug target as evidenced by potent inhibition with nirmatrelvir and ensitrelvir, the active components of the drugs Paxlovid and Xocova, respectively. However, the existence of nirmatrelvir and ensitrelvir-resistant isolates underscores the need to develop next-generation drugs with different resistance profiles and/or distinct mechanisms of action. Here, we report the results of a high-throughput screen of 649,568 compounds using a cellular gain-of-signal assay. In this assay, M^pro inhibits expression of a luciferase reporter, and 8,777 small molecules were considered hits by causing a gain in luciferase activity 3x SD above the sample field activity (6.8% gain-of-signal relative to 100 µM GC376). Single concentration and dose-response gain-of-signal experiments confirmed 3,522/8,762 compounds as candidate inhibitors. In parallel, all initial high-throughput screening hits were tested in a peptide cleavage assay with purified M^pro and only 39/8,762 showed inhibition. Importantly, 19/39 compounds (49%) re-tested positive in both SARS2 assays, including two previously reported M^pro inhibitors, demonstrating the efficacy of the overall screening strategy. This approach led to the rediscovery of known M^pro inhibitors such as calpain inhibitor II, as well as to the discovery of novel compounds that provide chemical information for future drug development efforts.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 6","pages":"Article 100181"},"PeriodicalIF":2.7,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000431/pdfft?md5=9fdb1c3b7ba672d8f03e70671b9bc52f&pid=1-s2.0-S2472555224000431-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142037949","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a time-resolved fluorescence resonance energy transfer ultra-high throughput screening assay targeting SYK and FCER1G interaction","authors":"Yuhong Du ,&nbsp;Dongxue Wang ,&nbsp;Vittorio L. Katis ,&nbsp;Elizabeth L. Zoeller ,&nbsp;Min Qui ,&nbsp;Allan I. Levey ,&nbsp;Opher Gileadi ,&nbsp;Haian Fu ,&nbsp;the Emory-SAGE-SGC TREAT-AD Center","doi":"10.1016/j.slasd.2024.100177","DOIUrl":"10.1016/j.slasd.2024.100177","url":null,"abstract":"<div><p>The spleen tyrosine kinase (SYK) and high affinity immunoglobulin epsilon receptor subunit gamma (FCER1G) interaction has a major role in the normal innate and adaptive immune responses, but dysregulation of this interaction is implicated in several human diseases, including autoimmune disorders, hematological malignancies, and Alzheimer's Disease. Development of small molecule chemical probes could aid in studying this pathway both in normal and aberrant contexts. Herein, we describe the miniaturization of a time-resolved fluorescence resonance energy transfer (TR-FRET) assay to measure the interaction between SYK and FCER1G in a 1536-well ultrahigh throughput screening (uHTS) format. The assay utilizes the His-SH2 domains of SYK, which are indirectly labeled with anti-His-terbium to serve as a TR-FRET donor and a FITC-conjugated phosphorylated ITAM domain peptide of FCER1G to serve as an acceptor. We have optimized the assay into a 384-well HTS format and further miniaturized the assay into a 1536-well uHTS format. Robust assay performance has been achieved with a Z’ factor &gt; 0.8 and signal-to-background (S/B) ratio &gt; 15. The utilization of this uHTS TR-FRET assay for compound screening has been validated by a pilot screening of 2,036 FDA-approved and bioactive compounds library. Several primary hits have been identified from the pilot uHTS. One compound, hematoxylin, was confirmed to disrupt the SYK/FECR1G interaction in an orthogonal protein–protein interaction assay. Thus, our optimized and miniaturized uHTS assay could be applied to future scaling up of a screening campaign to identify small molecule inhibitors targeting the SYK and FCER1G interaction.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 6","pages":"Article 100177"},"PeriodicalIF":2.7,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S247255522400039X/pdfft?md5=7e901da804daafea39d23b46e08f7bdf&pid=1-s2.0-S247255522400039X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142001528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simultaneous screening for selective SARS-CoV-2, Lassa, and Machupo virus entry inhibitors","authors":"Yuka Otsuka ,&nbsp;Lizhou Zhang ,&nbsp;Huihui Mou ,&nbsp;Justin Shumate ,&nbsp;Claire E. Kitzmiller ,&nbsp;Louis Scampavia ,&nbsp;Thomas D. Bannister ,&nbsp;Michael Farzan ,&nbsp;Hyeryun Choe ,&nbsp;Timothy P. Spicer","doi":"10.1016/j.slasd.2024.100178","DOIUrl":"10.1016/j.slasd.2024.100178","url":null,"abstract":"<div><p>Emerging highly pathogenic viruses can pose profound impacts on global health, the economy, and society. To meet that challenge, the National Institute of Allergy and Infectious Diseases (NIAID) established nine Antiviral Drug Discovery (AViDD) centers for early-stage identification and validation of novel antiviral drug candidates against viruses with pandemic potential. As part of this initiative, we established paired entry assays that simultaneously screen for inhibitors specifically targeting SARS-CoV-2 (SARS2), Lassa virus (LASV) and Machupo virus (MACV) entry. To do so we employed a dual pseudotyped virus (PV) infection system allowing us to screen ∼650,000 compounds efficiently and cost-effectively. Adaptation of these paired assays into 1536 well-plate format for ultra-high throughput screening (uHTS) resulted in the largest screening ever conducted in our facility, with over 2.4 million wells completed. The paired infection system allowed us to detect two PV infections simultaneously: LASV + MACV, MACV + SARS2, and SARS2 + LASV. Each PV contains a different luciferase reporter gene which enabled us to measure the infection of each PV exclusively, albeit in the same well. Each PV was screened at least twice utilizing different reporters, which allowed us to select the inhibitors specific to a particular PV and to exclude those that hit off targets, including cellular components or the reporter proteins. All assays were robust with an average Z’ value ranging from 0.5 to 0.8. The primary screening of ∼650,000 compounds resulted in 1812, 1506, and 2586 unique hits for LASV, MACV, and SARS2, respectively. The confirmation screening narrowed this list further to 60, 40, and 90 compounds that are unique to LASV, MACV, and SARS2, respectively. Of these compounds, 8, 35, and 50 compounds showed IC₅₀ value &lt; 10 μM, some of which have much greater potency and excellent antiviral activity profiles specific to LASV, MACV, and SARS2, and none are cytotoxic. These selected compounds are currently being studied for their mechanism of action and to improve their specificity and potency through chemical modification.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 6","pages":"Article 100178"},"PeriodicalIF":2.7,"publicationDate":"2024-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000406/pdfft?md5=fc5c2087563791142a95d69edf0d38c3&pid=1-s2.0-S2472555224000406-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142006028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a sensitive high-throughput enzymatic assay capable of measuring sub-nanomolar inhibitors of SARS-CoV2 Mpro","authors":"Peter Kovar,&nbsp;Paul L Richardson,&nbsp;Alla Korepanova,&nbsp;Gustavo A Afanador,&nbsp;Vladimir Stojkovic,&nbsp;Tao Li,&nbsp;Michael R Schrimpf,&nbsp;Teresa I Ng,&nbsp;David A Degoey,&nbsp;Sujatha M Gopalakrishnan,&nbsp;Jun Chen","doi":"10.1016/j.slasd.2024.100179","DOIUrl":"10.1016/j.slasd.2024.100179","url":null,"abstract":"<div><p>The SARS-CoV-2 main protease (Mpro) is essential for viral replication because it is responsible for the processing of most of the non-structural proteins encoded by the virus. Inhibition of Mpro prevents viral replication and therefore constitutes an attractive antiviral strategy. We set out to develop a high-throughput Mpro enzymatic activity assay using fluorescently labeled peptide substrates. A library of fluorogenic substrates of various lengths, sequences and dye/quencher positions was prepared and tested against full length SARS-CoV-2 Mpro enzyme for optimal activity. The addition of buffers containing strongly hydrated kosmotropic anion salts, such as citrate, from the Hofmeister series significantly boosted the enzyme activity and enhanced the assay detection limit, enabling the ranking of sub-nanomolar inhibitors without relying on the low-throughput Morrison equation method. By comparing cooperativity in citrate or non-citrate buffer while titrating the Mpro enzyme concentration, we found full positive cooperativity of Mpro with citrate buffer at less than one nanomolar (nM), but at a much higher enzyme concentration (∼320 nM) with non-citrate buffer. In addition, using a tight binding Mpro inhibitor, we confirmed there was only one active catalytical site in each Mpro monomer. Since cooperativity requires at least two binding sites, we hypothesized that citrate facilitates dimerization of Mpro at sub-nanomolar concentration as one of the mechanisms enhances Mpro catalytic efficiency. This assay has been used in high-throughput screening and structure activity relationship (SAR) studies to support medicinal chemistry efforts. IC₅₀ values determined in this assay correlates well with EC₅₀ values generated by a SARS-CoV-2 antiviral assay after adjusted for cell penetration.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 6","pages":"Article 100179"},"PeriodicalIF":2.7,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000418/pdfft?md5=b7b058077a624412d931d004dd0e213e&pid=1-s2.0-S2472555224000418-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141997052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"From labs to legislation: The interplay between academic research and political realities","authors":"Saumya Gopalkrishnan Ph.D","doi":"10.1016/j.slasd.2024.100175","DOIUrl":"10.1016/j.slasd.2024.100175","url":null,"abstract":"","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 6","pages":"Article 100175"},"PeriodicalIF":2.7,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000376/pdfft?md5=74ee6b11020cedb38ee53e6fd094975a&pid=1-s2.0-S2472555224000376-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141908597","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}