Potent Inhibition of Human Apurinic/Apyrimidinic Endonuclease 1 by Arylstibonic Acids

Seiple, Lauren; Cardellina, John H.; Akee, Rhone K.; Stivers, James T.

doi:10.1124/mol.107.042622

Cited by 79 publications

(97 citation statements)

References 34 publications

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“…Several small molecules have previously been identified as potential inhibitors of APE1's AP endonuclease activity (42,43,44). Cytotoxicity of these molecules has been evaluated by clonogenic survival, which does not illuminate the mechanism by which the molecules are cytotoxic.…”

Section: Resultsmentioning

confidence: 99%

Single cell trapping and DNA damage analysis using microwell arrays

Wood

Weingeist

Bhatia

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

242

284

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With a direct link to cancer, aging, and heritable diseases as well as a critical role in cancer treatment, the importance of DNA damage is well-established. The intense interest in DNA damage in applications ranging from epidemiology to drug development drives an urgent need for robust, high throughput, and inexpensive tools for objective, quantitative DNA damage analysis. We have developed a simple method for high throughput DNA damage measurements that provides information on multiple lesions and pathways. Our method utilizes single cells captured by gravity into a microwell array with DNA damage revealed morphologically by gel electrophoresis. Spatial encoding enables simultaneous assays of multiple experimental conditions performed in parallel with fully automated analysis. This method also enables novel functionalities, including multiplexed labeling for parallel single cell assays, as well as DNA damage measurement in cell aggregates. We have also developed 24-and 96-well versions, which are applicable to high throughput screening. Using this platform, we have quantified DNA repair capacities of individuals with different genetic backgrounds, and compared the efficacy of potential cancer chemotherapeutics as inhibitors of a critical DNA repair enzyme, human AP endonuclease. This platform enables high throughput assessment of multiple DNA repair pathways and subpathways in parallel, thus enabling new strategies for drug discovery, genotoxicity testing, and environmental health.base excision repair | comet assay | DNA repair D NA damage is a critical risk factor for cancer, aging, and heritable diseases (1-3), and it is the underlying basis for most frontline cancer therapies (4). Increased knowledge about DNA damage and repair would facilitate disease prevention, identification of individuals at increased risk of cancer, discovery of novel therapeutics, and better drug safety testing. Despite their obvious translational impact, most DNA damage assays have not changed significantly in more than two decades, and thus are not compatible with modern high throughput screening technologies. To better assess the biological impact of damage, we need data that reflect the integrated effect of multiple DNA repair pathways and subpathways, in response to a range of DNA lesions, measured in an accurate and high throughput fashion.The single cell gel electrophoresis or "comet" assay is based on the principle that relaxed loops (induced by single strand breaks) and DNA fragments migrate farther in an agarose gel than undamaged DNA (5-8). The alkali comet assay sensitively detects a range of DNA lesions, including strand breaks (single and double), as well as alkali sensitive sites. Although most base lesions are not directly detected, base adducts can be detected when converted to abasic sites or single strand breaks with the addition of purified DNA repair enzymes (7-11). The comet assay has been used in a variety of applications, including genotoxicity testing, human biomonitoring and epidemiology, environmental healt...

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Section: Resultsmentioning

confidence: 99%

Single cell trapping and DNA damage analysis using microwell arrays

Wood

Weingeist

Bhatia

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

242

284

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“…The ethyl linkage seems to be important for their inhibitory activities, given that P6954 and P6966, which have two carboxylic acid groups directly attached to the phenyl ring, are much less active. Similar effects were observed by the Stivers group when arylstibonic acids were screened for apurinic/apyrimidinic endonuclease (Ape1) inhibitors (Seiple et al, 2008), although a different set of compounds was found to be active (see below). In addition, the compounds with carboxylic acid groups on the phenyl ring meta to the stibonic acid group are more active than those with carboxylic acid groups at the para positions (compare NSC13778 and NSC732942 with NSC732943; compare P6981 with P6982; compare NSC13759 with NSC13760).…”

Section: Discussionmentioning

confidence: 50%

“…NSC13778 was reported to have anti-human immunodeficiency virus activity, mediated by binding to CD4 on cell surfaces and thus interfering with viral gp120 binding and virus entry (Yang et al, 2005). Stivers and colleagues recently showed that arylstibonic acids inhibit type I DNA topoisomerases (human and poxvirus Topo) and Ape1 (Kim et al, 2008;Seiple et al, 2008). Different arylstibonic acids showed different activities in these screens.…”

Section: Discussionmentioning

confidence: 99%

P6981, An Arylstibonic Acid, Is a Novel Low Nanomolar Inhibitor of cAMP Response Element-Binding Protein Binding to DNA

Zhao

Stagno²,

Varticovski³

et al. 2012

Mol Pharmacol

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Several basic leucine zipper (B-ZIP) transcription factors have been implicated in cancer, substance abuse, and other pathological conditions. We previously identified arylstibonic acids that bind to B-ZIP proteins and inhibit their interaction with DNA. In this study, we used electrophoretic mobility shift assay to analyze 46 arylstibonic acids for their activity to disrupt the DNA binding of three B-ZIP [CCAAT/enhancer-binding protein ␣, cyclic AMP-response element-binding protein (CREB), and vitellogenin gene-binding protein (VBP)] and two basic helixloop-helix leucine zipper (B-HLH-ZIP) [USF (upstream stimulating factor) and Mitf] proteins. Twenty-five arylstibonic acids showed activity at micromolar concentrations. The most active compound, P6981 [2-(3-stibonophenyl)malonic acid], had halfmaximal inhibition at ϳ5 nM for CREB. Circular dichroism thermal denaturation studies indicated that P6981 binds both the B-ZIP domain and the leucine zipper. The crystal structure of an arylstibonic acid, NSC13778, bound to the VBP leucine zipper identified electrostatic interactions between both the stibonic and carboxylic acid groups of NSC13778 [(E)-3-(3-stibonophenyl)acrylic acid] and arginine side chains of VBP, which is also involved in interhelical salt bridges in the leucine zipper. P6981 induced GFP-B-ZIP chimeric proteins to partially localize to the cytoplasm, demonstrating that it is active in cells. P6981 inhibited the growth of a patient-derived clear cell sarcoma cell line whose oncogenic potential is driven by a chimeric protein EWS-ATF1 (Ewing's sarcoma protein-activating transcription factor 1), which contains the DNA binding domain of ATF1, a B-ZIP protein. NSC13778 inhibited the growth of xenografted clear cell sarcoma, and no toxicity was observed. These experiments suggest that antimony containing arylstibonic acids are promising leads for suppression of DNA binding activities of B-ZIP and B-HLH-ZIP transcription factors.

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“…Zawahir et al performed an in silico pharmacophore model-based screen to identify 21 potent and specific inhibitors. The most potent of the inhibitors identified to date share common features of two carboxylate groups arranged around a hydrophobic core, bearing structural similarity to the 3'-and 5'-deoxyribosephosphate groups on abasic DNA (Seiple et al 2008;Zawahir et al 2009). The topoisomerase II inhibitor lucanthone has also been identified as a potential inhibitor of APE1 DNA repair activity, without impact on redox function (Bases and Mendez 1997;Luo and Kelley 2004).…”

Section: Ape1 Dna Repair Domain Inhibitorsmentioning

confidence: 99%

“…A number of other groups have been working on the development of APE1 DNA repair inhibitors. Seiple et al have identified the potential of arylstibonic acids, which possess inherent inhibitory activity on an initial fluorescence-based high-throughput screen (Seiple et al 2008). Simeonov et al utilised a similar fluorescence screening assay to identify three compounds (6-hydroxy-DL-DOPA, Reactive Blue 2 and myricetin) which potentiate MMS cytotoxicity in HeLa cells associated with a quantifiable increase in AP site accumulation.…”

Section: Ape1 Dna Repair Domain Inhibitorsmentioning

confidence: 99%