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Screening Method for the Discovery of Potential Cancer Chemoprevention Agents Based on Mass Spectrometric Detection of Alkylated Keap1

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    Screening Method for the Discovery of Potential Cancer Chemoprevention Agents Based on Mass Spectrometric Detection of Alkylated Keap1
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    Department of Medicinal Chemistry and Pharmacognosy, University of Illinois College of Pharmacy, 833 South Wood Street, Chicago, Illinois 60612-7231, and Medicinal Chemistry and Molecular Pharmacology, School of Pharmacy, Nursing and Health Sciences, Purdue University, 1330 Heine Pharmacy Building, West Lafayette, Indiana 47907
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    Analytical Chemistry

    Cite this: Anal. Chem. 2005, 77, 19, 6407–6414
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    https://doi.org/10.1021/ac050892r
    Published September 3, 2005
    Copyright © 2005 American Chemical Society
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    Abstract

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    Natural products are important sources of drugs such as cancer chemopreventive agents, but most assays for the discovery of compounds in natural product extracts are low throughput and provide little information about lead compounds in these complex mixtures. The induction of enzymes such as quinone reductase, glucuronyl transferases, glutathione S-transferases, and sulfotransferases can protect cells against the toxic and neoplastic effects of carcinogens. An increase in the concentration of Nrf2 in the nucleus of a cell upregulates the antioxidant response element and induces the expression of these chemopreventive enzymes. Based on the hypothesis that ubiquitination and proteosome-mediated degradation of Nrf2 in the cytoplasm decreases upon the covalent modification of 1 or more of the 27 cysteine sulfhydryl groups on Keap1 (a protein that sequesters Nrf2 in the cytoplasm) and results in higher Nrf2 levels both in the cytoplasm and in the nucleus, a high-throughput mass spectrometry-based screening assay was designed to detect alkylation of sulfhydryl groups of human Keap1. As an initial high-throughput screening step, matrix-assisted laser desorption time-of-flight mass spectrometry was used to determine whether incubation of Keap1 with a botanical sample produced adducts of Keap1. Test extracts found to form adducts with Keap1 were then incubated with the alternative biological nucleophile glutathione and characterized using LC-UV-MS-MS. After validation of the assay using two model alkylating agents, fractions of an extract of hops (Humulus lupulus L.) from the brewing industry were screened, and several compounds were detected as potential chemopreventive agents. Two of these electrophilic hops constituents were identified as xanthohumol and xanthohumol D. In a subsequent cell-based assay, xanthohumol and xanthohumol D were confirmed to be potent inducers of quinone reductase, and reaction with Keap1 was also confirmed. Therefore, this new mass spectrometric screening assay was demonstrated to facilitate the discovery of chemoprevention agents in complex natural product mixtures.

    Copyright © 2005 American Chemical Society

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    †

     University of Illinois.

    ‡

     Purdue University.

    *

     To whom correspondence should be addressed. Telephone: (312)-996-9353. Fax:  (312) 996-7107. E-mail:  [email protected].

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    17. Jan F. Stevens. Xanthohumol and Structurally Related Prenylflavonoids for Cancer Chemoprevention and Control. 2020, 319-350. https://doi.org/10.1007/978-3-030-39855-2_10
    18. Manuel Matzinger, Katrin Fischhuber, Elke H. Heiss. Activation of Nrf2 signaling by natural products-can it alleviate diabetes?. Biotechnology Advances 2018, 36 (6) , 1738-1767. https://doi.org/10.1016/j.biotechadv.2017.12.015
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    37. Zhongjun Ma, Min Zhang, Zhihang Song. Characterization of tanshinones with quinone reductase induction activity from Radix Salvia miltiorrhiza by liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry 2009, 23 (18) , 2857-2866. https://doi.org/10.1002/rcm.4195
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    39. Zhongjun Ma, Xiaoyu Zhang, Lin Cheng, Peng Zhang. Three lignans and one coumarinolignoid with quinone reductase activity from Eurycorymbus cavaleriei. Fitoterapia 2009, 80 (6) , 320-326. https://doi.org/10.1016/j.fitote.2009.04.003
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    41. Lin Cheng, Xiaoyu Zhang, Min Zhang, Peng Zhang, Zhihang Song, Zhongjun Ma, Yiyu Cheng, Haibin Qu. Characterization of chemopreventive agents from the dichloromethane extract of Eurycorymbus cavaleriei by liquid chromatography–ion trap mass spectrometry. Journal of Chromatography A 2009, 1216 (24) , 4859-4867. https://doi.org/10.1016/j.chroma.2009.04.031
    42. Lichuan Yang, Noel Y. Calingasan, Bobby Thomas, Rajnish K. Chaturvedi, Mahmoud Kiaei, Elizabeth J. Wille, Karen T. Liby, Charlotte Williams, Darlene Royce, Renee Risingsong, Eric S. Musiek, Jason D. Morrow, Michael Sporn, M. Flint Beal, . Neuroprotective Effects of the Triterpenoid, CDDO Methyl Amide, a Potent Inducer of Nrf2-Mediated Transcription. PLoS ONE 2009, 4 (6) , e5757. https://doi.org/10.1371/journal.pone.0005757
    43. Soona Shin, Thomas W. Kensler. Dietary Factors Regulate Metabolism of Carcinogens through Transcriptional Signaling Pathways. 2009, 109-120. https://doi.org/10.1002/9783527626588.ch7
    44. Hajime Nozawa, Feng Zhao, Keiji Kondo. Hops Derived Inhibitors of Nitric Oxide. 2009, 759-764. https://doi.org/10.1016/B978-0-12-373891-2.00077-8
    45. Jie Xing, Zijuan Yang, Beibei Lv, Lan Xiang. Rapid screening for cyclo‐dopa and diketopiperazine alkaloids in crude extracts of Portulaca oleracea L. using liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry 2008, 22 (9) , 1415-1422. https://doi.org/10.1002/rcm.3526
    46. Ian M. Copple, Christopher E. Goldring, Neil R. Kitteringham, B. Kevin Park. The Nrf2–Keap1 defence pathway: Role in protection against drug-induced toxicity. Toxicology 2008, 246 (1) , 24-33. https://doi.org/10.1016/j.tox.2007.10.029
    47. Jana Nagel, Lana K. Culley, Yuping Lu, Enwu Liu, Paul D. Matthews, Jan F. Stevens, Jonathan E. Page. EST Analysis of Hop Glandular Trichomes Identifies an O -Methyltransferase That Catalyzes the Biosynthesis of Xanthohumol. The Plant Cell 2008, 20 (1) , 186-200. https://doi.org/10.1105/tpc.107.055178
    48. Norman R Farnsworth, Elizabeth C Krause, Judy L Bolton, Guido F Pauli, Richard B van Breemen, James G Graham. The University of Illinois at Chicago/National Institutes of Health Center for Botanical Dietary Supplements Research for Women's Health: from plant to clinical use. The American Journal of Clinical Nutrition 2008, 87 (2) , 504S-508S. https://doi.org/10.1093/ajcn/87.2.504S
    49. Sung Ho Lee, Hyun Jung Kim, Jung Sun Lee, Ik-Soo Lee, Bok Yun Kang. Inhibition of topoisomerase I activity and efflux drug transporters’ expression by xanthohumol from hops. Archives of Pharmacal Research 2007, 30 (11) , 1435-1439. https://doi.org/10.1007/BF02977368
    50. Jie Xing, Chunfeng Xie, Jianbo Qu, Huaifang Guo, Beibei Lv, Hongxiang Lou. Rapid screening for bisbibenzyls in bryophyte crude extracts using liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry 2007, 21 (15) , 2467-2476. https://doi.org/10.1002/rcm.3115
    51. Emily C. Colgate, Cristobal L. Miranda, Jan F. Stevens, Tammy M. Bray, Emily Ho. Xanthohumol, a prenylflavonoid derived from hops induces apoptosis and inhibits NF-kappaB activation in prostate epithelial cells. Cancer Letters 2007, 246 (1-2) , 201-209. https://doi.org/10.1016/j.canlet.2006.02.015
    52. Dwight D. Baker, Min Chu, Uma Oza, Vineet Rajgarhia. The value of natural products to future pharmaceutical discovery. Natural Product Reports 2007, 24 (6) , 1225. https://doi.org/10.1039/b602241n
    53. Chris Goldring, Neil Kitteringham, Rosalind Jenkins, Ian Copple, Jean-Francois Jeannin, B. Kevin Park. Plasticity in cell defence: access to and reactivity of critical protein residues and DNA response elements. Journal of Experimental Biology 2006, 209 (12) , 2337-2343. https://doi.org/10.1242/jeb.02209
    54. . Current literature in mass spectrometry. Journal of Mass Spectrometry 2006, 693-704. https://doi.org/10.1002/jms.952
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    Cite this: Anal. Chem. 2005, 77, 19, 6407–6414
    Click to copy citationCitation copied!
    https://doi.org/10.1021/ac050892r
    Published September 3, 2005
    Copyright © 2005 American Chemical Society
    Request reuse permissions

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