Biology subjects

Fursmidt, R.

Publications and source records attributed to Fursmidt, R..

3 recordsLinked to original sources

bioRxiv2025

Phenotypic Screening Coupled with AI-Driven Target Deconvolution Identifies α-Terthienyl as a Dual DPP-IV/HSD17β13 Modulator with Efficacy in a Mouse Model of MASLD

BackgroundMetabolic dysfunction-associated steatotic liver disease (MASLD) is a highly prevalent condition characterized by fat build-up in the liver and ranges from benign steatosis to progression to metabolic dysfunction-associated steatohepatitis (MASH), fibrosis, cirrhosis and end-stage liver disease including hepatocellular carcinoma, representing a significant cause of chronic liver disease globally.1,2 Current treatment options are limited, primarily relying on lifestyle modifications, highlighting an urgent need for novel therapeutic strategies. MethodsA Cell Painting-style high-content screening phenotypic assay was employed using the PH5CH8 human hepatocyte cell line to identify small molecules capable of modulating induced hepatic steatosis. The Plex Research artificial intelligence (AI) platform was utilized for target deconvolution of the lead hit compound, -terthienyl. In vivo efficacy was assessed in a diet-induced obesity (DIO) C57BL/6J mouse model of MASLD. Biochemical assays and molecular docking simulations were performed to validate predicted target interactions. ResultsPhenotypic screening identified 15 chemical probes/drugs that elicit dose-responsive inhibition of steatosis, and 16 that exacerbate steatosis, which could contribute to worsening of the disease clinically. -terthienyl, a plant-derived natural product, was identified as a potent and non-toxic inhibitor of steatosis in PH5CH8 cells with an EC50 of 106 nM. In vivo, -terthienyl administration to diet-induced obesity (60% fat diet) mice significantly reduced hepatic steatosis histologically, improved glucose tolerance, and favorably modulated serum biomarkers including ALT and AST. AI-driven analysis predicted dipeptidyl peptidase 4 (DPP-IV) and 17-beta hydroxysteroid dehydrogenase 13 (HSD17{beta}13) as potential molecular targets of -terthienyl. Biochemical inhibition of DPP-IV was observed and an oxidized -terthienyl analog inhibited HSD17{beta}13. Molecular docking supported these predictions, indicating binding to DPP-IV and HSD17{beta}13. ConclusionThis study demonstrates the successful application of phenotypic screening integrated with AI-driven target deconvolution to identify compounds and drugs that ameliorate or exacerbate hepatic steatosis. -terthienyl was identified as a novel modulator of hepatic steatosis with in vivo efficacy in a MASLD model. Our findings suggest a dual-target mechanism involving DPP-IV and HSD17{beta}13, potentially engaged by the parent compound and its metabolite, respectively, offering a promising polypharmacological approach for MASLD treatment.

pharmacology and toxicology↗

bioRxiv2022

In Vitro Evaluation and Mitigation of Niclosamide Liabilities as a COVID-19 Treatment.

Niclosamide, an FDA-approved oral anthelmintic drug, has broad biological activity including anticancer, antibacterial, and antiviral properties. Niclosamide has also been identified as a potent inhibitor of SARS-CoV-2 infection in vitro, generating interest in its use for the treatment or prevention of COVID-19. Unfortunately, there are several potential issues with using niclosamide for COVID-19, including low bioavailability, significant polypharmacology, high cellular toxicity, and unknown efficacy against emerging SARS-CoV-2 variants of concern. In this study, we used high-content imaging-based immunofluorescence assays in two different cell models to assess these limitations and evaluate the potential for using niclosamide as a COVID-19 antiviral. We show that despite promising preliminary reports, the antiviral efficacy of niclosamide overlaps with its cytotoxicity giving it a poor in vitro selectivity index for anti-SARS-CoV-2 inhibition. We also show that niclosamide has significantly variable potency against the different SARS-CoV-2 variants of concern and is most potent against variants with enhanced cell-to-cell spread including B.1.1.7. Finally, we report the activity of 33 niclosamide analogs, several of which have reduced cytotoxicity and increased potency relative to niclosamide. A preliminary structure-activity relationship analysis reveals dependence on a protonophore for antiviral efficacy, which implicates nonspecific endolysosomal neutralization as a dominant mechanism of action. Further single-cell morphological profiling suggests niclosamide also inhibits viral entry and cell-to-cell spread by syncytia. Altogether, our results suggest that niclosamide is not an ideal candidate for the treatment of COVID-19, but that there is potential for developing improved analogs with higher clinical translational potential in the future. ImportanceThere is still an urgent need for effective anti-SARS-CoV-2 therapeutics due to waning vaccine efficacy, the emergence of variants of concern, and limited efficacy of existing antivirals. One potential therapeutic option is niclosamide, an FDA approved anthelmintic compound that has shown promising anti-SARS-CoV-2 activity in cell-based assays. Unfortunately, there are significant barriers for the clinical utility of niclosamide as a COVID-19 therapeutic. Our work emphasizes these limitations by showing that niclosamide has high cytotoxicity at antiviral concentrations, variable potency against variants of concern, and significant polypharmacology as a result of its activity as a nonspecific protonophore. Some of these clinical limitations can be mitigated, however, through structural modifications to the niclosamide scaffold, which we demonstrate through a preliminary structure activity relationship analysis. Overall, we show that niclosamide is not a suitable candidate for the treatment of COVID-19, but that structural analogs with improved drug properties may have higher clinical-translational potential.

microbiology↗

bioRxiv2020

Morphological Cell Profiling of SARS-CoV-2 Infection Identifies Drug Repurposing Candidates for COVID-19

The global spread of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and the associated disease COVID-19, requires therapeutic interventions that can be rapidly identified and translated to clinical care. Traditional drug discovery methods have a >90% failure rate and can take 10-15 years from target identification to clinical use. In contrast, drug repurposing can significantly accelerate translation. We developed a quantitative high-throughput screen to identify efficacious agents against SARS-CoV-2. From a library of 1,425 FDA-approved compounds and clinical candidates, we identified 17 dose-responsive compounds with in vitro antiviral efficacy in human liver Huh7 cells and confirmed antiviral efficacy in human colon carcinoma Caco-2, human prostate adenocarcinoma LNCaP, and in a physiologic relevant model of alveolar epithelial type 2 cells (iAEC2s). Additionally, we found that inhibitors of the Ras/Raf/MEK/ERK signaling pathway exacerbate SARS-CoV-2 infection in vitro. Notably, we discovered that lactoferrin, a glycoprotein classically found in secretory fluids, including mammalian milk, inhibits SARS-CoV-2 infection in the nanomolar range in all cell models with multiple modes of action, including blockage of virus attachment to cellular heparan sulfate and enhancement of interferon responses. Given its safety profile, lactoferrin is a readily translatable therapeutic option for the management of COVID-19. IMPORTANCESince its emergence in China in December 2019, SARS-CoV-2 has caused a global pandemic. Repurposing of FDA-approved drugs is a promising strategy for identifying rapidly deployable treatments for COVID-19. Herein, we developed a pipeline for quantitative high-throughput image-based screening of SARS-CoV-2 infection in human cells that led to the identification of several FDA-approved drugs and clinical candidates with in vitro antiviral activity.

cell biology↗