Abstract
The quality of the data generated in a high throughput screening (HTS) run is fundamental for selecting bona fide inhibitors and for ensuring the capture of the full richness of inhibitors present in a chemical library. For this purpose a quality control filter based on three one dimensional (1D) proton NMR experiments is proposed. The approach called SPAM (Solubility, Purity and Aggregation of the Molecule) Filter requires the acquisition of a 1D reference spectrum, a WaterLOGSY spectrum and/or a selective longitudinal relaxation filter spectrum for the identified hits dissolved in aqueous solution and in the presence of a water soluble reference molecule. This palette of experiments permits the rapid characterization of the identity, purity, solubility and aggregation state of the active compound. This knowledge is crucial for deriving accurate IC50 and KI values of the inhibitors, for identifying false negatives and for detecting promiscuous inhibitors. Only compounds that pass through the SPAM Filter can be considered as starting points for medicinal chemistry efforts directed toward lead optimization. Examples of this approach in the identification of false positives in a screening run against the enzyme thymidine phosphorylase (TP) and the rescue of a false negative in a screening run against the Ser/Thr kinase AKT1 are presented.
Keywords: Promiscuous compounds, false positives, false negatives, NMR, HTS, enzyme inhibitors
Current Drug Discovery Technologies
Title: NMR-Based Quality Control Approach for the Identification of False Positives and False Negatives in High Throughput Screening
Volume: 3 Issue: 2
Author(s): Claudio Dalvit, Dannica Caronni, Nicola Mongelli, Marina Veronesi and Anna Vulpetti
Affiliation:
Keywords: Promiscuous compounds, false positives, false negatives, NMR, HTS, enzyme inhibitors
Abstract: The quality of the data generated in a high throughput screening (HTS) run is fundamental for selecting bona fide inhibitors and for ensuring the capture of the full richness of inhibitors present in a chemical library. For this purpose a quality control filter based on three one dimensional (1D) proton NMR experiments is proposed. The approach called SPAM (Solubility, Purity and Aggregation of the Molecule) Filter requires the acquisition of a 1D reference spectrum, a WaterLOGSY spectrum and/or a selective longitudinal relaxation filter spectrum for the identified hits dissolved in aqueous solution and in the presence of a water soluble reference molecule. This palette of experiments permits the rapid characterization of the identity, purity, solubility and aggregation state of the active compound. This knowledge is crucial for deriving accurate IC50 and KI values of the inhibitors, for identifying false negatives and for detecting promiscuous inhibitors. Only compounds that pass through the SPAM Filter can be considered as starting points for medicinal chemistry efforts directed toward lead optimization. Examples of this approach in the identification of false positives in a screening run against the enzyme thymidine phosphorylase (TP) and the rescue of a false negative in a screening run against the Ser/Thr kinase AKT1 are presented.
Export Options
About this article
Cite this article as:
Dalvit Claudio, Caronni Dannica, Mongelli Nicola, Veronesi Marina and Vulpetti Anna, NMR-Based Quality Control Approach for the Identification of False Positives and False Negatives in High Throughput Screening, Current Drug Discovery Technologies 2006; 3 (2) . https://dx.doi.org/10.2174/157016306778108875
DOI https://dx.doi.org/10.2174/157016306778108875 |
Print ISSN 1570-1638 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-6220 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
- Announcements
Related Articles
-
Differentiation Between Osteoporotic and Neoplastic Vertebral Fractures: State of The Art and Future Perspectives
Current Medical Imaging Analysis of Chinese Herbal Formulae Recommended for COVID-19 in Different Schemes in China: A Data Mining Approach
Combinatorial Chemistry & High Throughput Screening Revisiting the Antiviral Efficacy of Terpenoids: Plausible Adjunct Therapeutics for Novel SARS-CoV-2?
Endocrine, Metabolic & Immune Disorders - Drug Targets Tacrine Derivatives and Alzheimers Disease
Current Medicinal Chemistry Medicinal Research Progress of Natural Coumarin and its Derivatives
The Natural Products Journal Bifunctional Anti-HIV/TB Inhibitors: Perspective from In-Silico Design and Molecular Dynamics Simulations
Letters in Drug Design & Discovery 8-(Heteroaryl)phenalkyl-1-Phenyl-1,3,8-triazaspiro[4.5]decan-4-ones as Opioid Receptor Modulators
Medicinal Chemistry Biologically Active Peptides Interacting with the G Protein-Coupled Formylpeptide Receptors
Protein & Peptide Letters Computer Aided Drug Design for Combating Diseases (Part-III)
Current Topics in Medicinal Chemistry Recent Advances of Natural and Synthetic β-Carbolines as Anticancer Agents
Anti-Cancer Agents in Medicinal Chemistry Laparoscopic Approach to a Patient with Omental Infarction: A Case Report
New Emirates Medical Journal Utilities of Some Carbon Nucleophiles in Heterocyclic Synthesis
Current Organic Chemistry Mechanisms Involved in the Genesis of Diabetic Nephropathy
Current Diabetes Reviews Chalcone-Coumarin Derivatives as Potential Anti-Cancer Drugs: An in vitro and in vivo Investigation
Anti-Cancer Agents in Medicinal Chemistry Quassinoids from Brucea Javanica Seeds Inhibiting the Replication of Tobacco Mosaic Virus
Current Bioactive Compounds Epigenetic Remodeling of Chromatin Architecture: Exploring Tumor Differentiation Therapies in Mesenchymal Stem Cells and Sarcomas
Current Stem Cell Research & Therapy A Comprehensive Review on <i>In Silico</i>-predicted Potential Phytochemicals against SARS-CoV-2: Food for Thought for Researchers
Letters in Organic Chemistry Placebos Used in Clinical Trials for Chinese Herbal Medicine
Recent Patents on Inflammation & Allergy Drug Discovery Post-Translational Modification of Orphan Nuclear Receptors TR2 and TR4 – Ligand Independent Activities
Current Proteomics Oxidative Status in Multiple Sclerosis and Off-Targets of Antioxidants: The Case of Edaravone
Current Medicinal Chemistry