Abstract
The Polo-like kinase (Plk) family comprises four cell cycle serine/threonine kinases, Plk1-4. Among these, Plk1 has been most thoroughly characterized; it contains a conserved kinase domain and a C-terminal docking site for S/T-phosphorylated proteins (polo-box domain, PBD). Polo-like kinases are deregulated in oncogenesis and therefore constitute a therapeutic target for cancer. A high throughput screening campaign was carried out by the Pittsburgh Molecular Library Screening Center (PMLSC), using a fluorescence polarization assay with recombinant Plk1-PBD to monitor the inhibition of binding of an optimal phosphopeptide substrate motif with recombinant Plk1-PBD. Screening of 97,090 small molecule library samples provided by the NIH Small Molecule Repository distributed by DPI Galapagos led to 11 confirmed hits. The Pittsburgh MLSCN Chemistry Core selected one of the structurally most tractable hits, SID 861574, for chemical hit-to-probe development. A broad chemistry program was initiated that developed new strategies for 6-amino- and 6-hydroxy uracil synthesis as well as acylanilides, and generated a total of 70 analogs. Out of 46 analogues tested, none, nor the resynthesized hit, showed affinity to Plk1-PBD in the follow up assays. In contrast, reassays of the original screening materials displayed activities similar to the original HTS assay. We ultimately concluded that an impurity in the commercial material led to the positive screening artifact. This case study highlights our development of a synthesis of 6-position functionalized uracil analogs, but also illustrates the importance of careful quality and compound stability monitoring of screening collections.
Keywords: Polo-like kinases, polo-box domain, high-throughput screening, probe development, Molecular Library Screening Center Network, medicinal chemistry, heterocycle synthesis, screening artifact, anticancer drug target
Current Topics in Medicinal Chemistry
Title: A Case Study from the Chemistry Core of the Pittsburgh Molecular Library Screening Center: The Polo-like Kinase Polo-Box Domain (Plk1- PBD)
Volume: 9 Issue: 13
Author(s): Peter Wipf, David Arnold, Karen Carter, Shuzhi Dong, Paul A. Johnston, Elizabeth Sharlow, John S. Lazo and Donna H. Huryn
Affiliation:
Keywords: Polo-like kinases, polo-box domain, high-throughput screening, probe development, Molecular Library Screening Center Network, medicinal chemistry, heterocycle synthesis, screening artifact, anticancer drug target
Abstract: The Polo-like kinase (Plk) family comprises four cell cycle serine/threonine kinases, Plk1-4. Among these, Plk1 has been most thoroughly characterized; it contains a conserved kinase domain and a C-terminal docking site for S/T-phosphorylated proteins (polo-box domain, PBD). Polo-like kinases are deregulated in oncogenesis and therefore constitute a therapeutic target for cancer. A high throughput screening campaign was carried out by the Pittsburgh Molecular Library Screening Center (PMLSC), using a fluorescence polarization assay with recombinant Plk1-PBD to monitor the inhibition of binding of an optimal phosphopeptide substrate motif with recombinant Plk1-PBD. Screening of 97,090 small molecule library samples provided by the NIH Small Molecule Repository distributed by DPI Galapagos led to 11 confirmed hits. The Pittsburgh MLSCN Chemistry Core selected one of the structurally most tractable hits, SID 861574, for chemical hit-to-probe development. A broad chemistry program was initiated that developed new strategies for 6-amino- and 6-hydroxy uracil synthesis as well as acylanilides, and generated a total of 70 analogs. Out of 46 analogues tested, none, nor the resynthesized hit, showed affinity to Plk1-PBD in the follow up assays. In contrast, reassays of the original screening materials displayed activities similar to the original HTS assay. We ultimately concluded that an impurity in the commercial material led to the positive screening artifact. This case study highlights our development of a synthesis of 6-position functionalized uracil analogs, but also illustrates the importance of careful quality and compound stability monitoring of screening collections.
Export Options
About this article
Cite this article as:
Wipf Peter, Arnold David, Carter Karen, Dong Shuzhi, Johnston A. Paul, Sharlow Elizabeth, Lazo S. John and Huryn H. Donna, A Case Study from the Chemistry Core of the Pittsburgh Molecular Library Screening Center: The Polo-like Kinase Polo-Box Domain (Plk1- PBD), Current Topics in Medicinal Chemistry 2009; 9 (13) . https://dx.doi.org/10.2174/156802609789753590
DOI https://dx.doi.org/10.2174/156802609789753590 |
Print ISSN 1568-0266 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4294 |
Call for Papers in Thematic Issues
Chemistry Based on Natural Products for Therapeutic Purposes
The development of new pharmaceuticals for a wide range of medical conditions has long relied on the identification of promising natural products (NPs). There are over sixty percent of cancer, infectious illness, and CNS disease medications that include an NP pharmacophore, according to the Food and Drug Administration. Since NP ...read more
Current Trends in Drug Discovery Based on Artificial Intelligence and Computer-Aided Drug Design
Drug development discovery has faced several challenges over the years. In fact, the evolution of classical approaches to modern methods using computational methods, or Computer-Aided Drug Design (CADD), has shown promising and essential results in any drug discovery campaign. Among these methods, molecular docking is one of the most notable ...read more
Drug Discovery in the Age of Artificial Intelligence
In the age of artificial intelligence (AI), we have witnessed a significant boom in AI techniques for drug discovery. AI techniques are increasingly integrated and accelerating the drug discovery process. These developments have not only attracted the attention of academia and industry but also raised important questions regarding the selection ...read more
From Biodiversity to Chemical Diversity: Focus of Flavonoids
Flavonoids are the largest group of polyphenols, plant secondary metabolites arising from the essential aromatic amino acid phenylalanine (or more rarely from tyrosine) via the phenylpropanoid pathway. The flavan nucleus is the basic 15-carbon skeleton of flavonoids (C6-C3-C6), which consists of two phenyl rings (A and B) and a heterocyclic ...read more
- 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
-
Trafficking of Neuronal Two Pore Domain Potassium Channels
Current Neuropharmacology Interleukin-6 In Rheumatoid Arthritis - From The Laboratory To The Bedside
Current Pharmaceutical Design New Light on An Old Friend: Targeting PUMA in Radioprotection and Therapy of Cardiovascular and Neurodegenerative Diseases
Current Drug Targets Tipping the Balance Between Life and Death: Targeting Histone Acetylation for Cancer Therapy
Drug Delivery Letters Current and Emerging Therapeutic Approaches in HCV-Related Mixed Cryoglobulinemia
Current Medicinal Chemistry Nanotoxicity: The Toxicity Research Progress of Metal and Metal- Containing Nanoparticles
Mini-Reviews in Medicinal Chemistry Thiosemicarbazones as Potent Anticancer Agents and their Modes of Action
Mini-Reviews in Medicinal Chemistry Application of Metabolomics in Drug Discovery, Development and Theranostics
Current Metabolomics Personalized Medicine, Bioethics and Social Responsibilities: Re-thinking the Pharmaceutical Industry to Remedy Inequities in Patient Care and International Health
Current Pharmacogenomics and Personalized Medicine Promising Activity of Mammalian Target of Rapamycin Inhibitors in Hematologic Malignancies Therapy
Current Signal Transduction Therapy Current Perspectives of Healthy Mitochondrial Function for Healthy Neurons
Current Drug Targets Gene and Cancer Therapy - Pseudorabies Virus: A Novel Research and Therapeutic Tool?
Current Gene Therapy Clinical, Prognostic and Therapeutic Significance of Heat Shock Proteins in Cancer
Current Drug Targets Allosteric Targeting of Aurora A Kinase Using Small Molecules: A Step Forward Towards Next Generation Medicines?
Current Medicinal Chemistry Recent Advances in the Therapeutic Perspectives of Nutlin-3
Current Pharmaceutical Design UPA and PAI-1 Analysis from Fixed Tissues – New Perspectives for a Known Set of Predictive Markers
Current Medicinal Chemistry Damage and Recovery of the Bone Marrow Microenvironment Induced by Cancer Chemotherapy – Potential Regulatory Role of Chemokine CXCL12/Receptor CXCR4 Signalling
Current Molecular Medicine Membrane Tyrosine Kinase Receptors Kit and FLT3 are an Important Targets for the Therapy of Acute Myeloid Leukemia
Current Cancer Therapy Reviews Anti-tumor Drug Targets Analysis: Current Insight and Future Prospect
Current Drug Targets Lipids as Biomarkers of Cancer and Bacterial Infections
Current Medicinal Chemistry