Development of a High-Throughput Fluorescence Polarization DNA Cleavage Assay for the Identification of FEN1 Inhibitors

McWhirter, Claire; Tonge, Michael; Plant, Helen; Hardern, Ian; Nissink, J. Willem M.; Durant, Stephen T.

doi:10.1177/1087057113476551

Cited by 20 publications

(19 citation statements)

References 26 publications

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“…Several attempts to develop FEN1 inhibitors have been described (Tumey et al, 2004, Tumey et al, 2005, Mcwhirter et al, 2013, Wadhwa and Jadhav, 2015, Dorjsuren et al, 2011). Tumey et al reported that N-hydroxy urea derivatives are FEN1 inhibitors, and compound #20 has an impressive IC50 value of 3 nM (Tumey et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Targeting DNA Flap Endonuclease 1 to Impede Breast Cancer Progression

et al. 2016

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DNA flap endonuclease 1 (FEN1) plays critical roles in maintaining genome stability and integrity by participating in both DNA replication and repair. Suppression of FEN1 in cells leads to the retardation of DNA replication and accumulation of unrepaired DNA intermediates, resulting in DNA double strand breaks (DSBs) and apoptosis. Therefore, targeting FEN1 could serve as a potent strategy for cancer therapy. In this study, we demonstrated that FEN1 is overexpressed in breast cancers and is essential for rapid proliferation of cancer cells. We showed that manipulating FEN1 levels in cells alters the response of cancer cells to chemotherapeutic drugs. Furthermore, we identified a small molecular compound, SC13 that specifically inhibits FEN1 activity, thereby interfering with DNA replication and repair in vitro and in cells. SC13 suppresses cancer cell proliferation and induces chromosome instability and cytotoxicity in cells. Importantly, SC13 sensitizes cancer cells to DNA damage-inducing therapeutic modalities and impedes cancer progression in a mouse model. These findings could establish a paradigm for the treatment of breast cancer and other cancers as well.

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Section: Discussionmentioning

confidence: 99%

Targeting DNA Flap Endonuclease 1 to Impede Breast Cancer Progression

et al. 2016

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“…Furthermore, a number of highthroughput, robust screening techniques have successfully identified potential novel inhibitors of DNA repair proteins to be taken forward. 99,100 The studies discussed here thus far suggest that targeting DNA repair endonucleases in combination with DNA-damaging agents may be a promising anticancer strategy.…”

Section: Dna Repair Endonucleases As Targets For Cancer Therapymentioning

confidence: 99%

“…In 2013, AstraZeneca (London, UK) conducted a high-throughput screen of 850,000 compounds using a fluorescent DNA cleavage assay to determine FEN1 inhibition activity. 99 This screen resulted in the identification of 6261 compounds with FEN1 inhibition activity at an IC 50 of less than 100 µM. 99 A study in colorectal cancer cell lines revealed that FEN1 inhibition by a number of small molecules had a synthetically lethal effect in cells deficient in the ubiquitin ligase CDC4.…”

Section: Fen1 Inhibitorsmentioning

confidence: 99%

“…99 This screen resulted in the identification of 6261 compounds with FEN1 inhibition activity at an IC 50 of less than 100 µM. 99 A study in colorectal cancer cell lines revealed that FEN1 inhibition by a number of small molecules had a synthetically lethal effect in cells deficient in the ubiquitin ligase CDC4. 115 Interestingly, in a recent study, curcumin, the active ingredient in turmeric, has been shown to restrict cell proliferation in breast cancer cell lines through inhibition of FEN1 activity.…”

Section: Fen1 Inhibitorsmentioning

confidence: 99%

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DNA Repair Endonucleases: Physiological Roles and Potential as Drug Targets

Doherty

Madhusudan

2015

SLAS Discovery

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Genomic DNA is constantly exposed to endogenous and exogenous damaging agents. To overcome these damaging effects and maintain genomic stability, cells have robust coping mechanisms in place, including repair of the damaged DNA. There are a number of DNA repair pathways available to cells dependent on the type of damage induced. The removal of damaged DNA is essential to allow successful repair. Removal of DNA strands is achieved by nucleases. Exonucleases are those that progressively cut from DNA ends, and endonucleases make single incisions within strands of DNA. This review focuses on the group of endonucleases involved in DNA repair pathways, their mechanistic functions, roles in cancer development, and how targeting these enzymes is proving to be an exciting new strategy for personalized therapy in cancer.

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“…Standardized annotation of in-house HTS assays using BAO is demonstrated by previously published in-house screening assay development experiments ( Table 1). [10][11][12] The assay design and detection methods of the in-house HTS assays have been analyzed together with 233 primary PubChem HTS assays. 7 Annotated PubChem assays have been provided by the BAO team and are mainly biochemical assays, G protein-coupled receptor (GPCR) assays, and various assays detected by luminescence.…”

Section: Assay Annotationmentioning

confidence: 99%

Using the BioAssay Ontology for Analyzing High-Throughput Screening Data

et al. 2015

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High-throughput screening (HTS) is the main starting point for hit identification in drug discovery programs. This has led to a rapid increase of available screening data both within pharmaceutical companies and the public domain. We have used the BioAssay Ontology (BAO) 2.0 for assay annotation within AstraZeneca to enable comparison with external HTS methods. The annotated assays have been analyzed to identify technology gaps, evaluate new methods, verify active hits, and compare compound activity between in-house and PubChem assays. As an example, the binding of a fluorescent ligand to formyl peptide receptor 1 (FPR1, involved in inflammation, for example) in an in-house HTS was measured by fluorescence intensity. In total, 155 active compounds were also tested in an external ligand binding flow cytometry assay, a method not used for in-house HTS detection. Twelve percent of the 155 compounds were found active in both assays. By the annotation of assay protocols using BAO terms, internal and external assays can easily be identified and method comparison facilitated. They can be used to evaluate the effectiveness of different assay methods, design appropriate confirmatory and counterassays, and analyze the activity of compounds for identification of technology artifacts.

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Development of a High-Throughput Fluorescence Polarization DNA Cleavage Assay for the Identification of FEN1 Inhibitors

Cited by 20 publications

References 26 publications

Targeting DNA Flap Endonuclease 1 to Impede Breast Cancer Progression

Targeting DNA Flap Endonuclease 1 to Impede Breast Cancer Progression

DNA Repair Endonucleases: Physiological Roles and Potential as Drug Targets

Using the BioAssay Ontology for Analyzing High-Throughput Screening Data

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