Target engagement and drug residence time can be observed in living cells with BRET

被引:188
作者
Robers, Matthew B. [1 ]
Dart, Melanie L. [1 ]
Woodroofe, Carolyn C. [2 ]
Zimprich, Chad A. [1 ]
Kirkland, Thomas A. [2 ]
Machleidt, Thomas [1 ]
Kupcho, Kevin R. [1 ]
Levin, Sergiy [2 ]
Hartnett, James R. [1 ]
Zimmerman, Kristopher [1 ]
Niles, Andrew L. [1 ]
Ohana, Rachel Friedman [1 ]
Daniels, Danette L. [1 ]
Slater, Michael [1 ]
Wood, Monika G. [1 ]
Cong, Mei [1 ]
Cheng, Yi-Qiang [3 ]
Wood, Keith V. [1 ]
机构
[1] Promega Corp, Fitchburg, WI 53711 USA
[2] Promega Biosci Inc, San Luis Obispo, CA 93401 USA
[3] Univ N Texas, Hlth Sci Ctr, Dept Pharmaceut Sci, UNT Syst Coll Pharm, Ft Worth, TX USA
来源
NATURE COMMUNICATIONS | 2015年 / 6卷
关键词
Chemical sciences; Medicinal chemistry; Chemical biology; HISTONE DEACETYLASE INHIBITOR; ENERGY-TRANSFER METHODS; 2 CATALYTIC DOMAINS; IN-VITRO; HDAC INHIBITORS; LIGAND-BINDING; ASSAY; THAILANDEPSINS; DISPLACEMENT; DEPSIPEPTIDE;
D O I
10.1038/ncomms10091
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The therapeutic action of drugs is predicated on their physical engagement with cellular targets. Here we describe a broadly applicable method using bioluminescence resonance energy transfer (BRET) to reveal the binding characteristics of a drug with selected targets within intact cells. Cell-permeable fluorescent tracers are used in a competitive binding format to quantify drug engagement with the target proteins fused to Nanoluc luciferase. The approach enabled us to profile isozyme-specific engagement and binding kinetics for a panel of histone deacetylase (HDAC) inhibitors. Our analysis was directed particularly to the clinically approved prodrug FK228 (Istodax/Romidepsin) because of its unique and largely unexplained mechanism of sustained intracellular action. Analysis of the binding kinetics by BRET revealed remarkably long intracellular residence times for FK228 at HDAC1, explaining the protracted intracellular behaviour of this prodrug. Our results demonstrate a novel application of BRET for assessing target engagement within the complex milieu of the intracellular environment.
引用
收藏
页数:24
相关论文
共 55 条
[1]   Chemoproteomics profiling of HDAC inhibitors reveals selective targeting of HDAC complexes [J].
Bantscheff, Marcus ;
Hopf, Carsten ;
Savitski, Mikhail M. ;
Dittmann, Antje ;
Grandi, Paola ;
Michon, Anne-Marie ;
Schlegl, Judith ;
Abraham, Yann ;
Becher, Isabelle ;
Bergamini, Giovanna ;
Boesche, Markus ;
Delling, Manja ;
Duempelfeld, Birgit ;
Eberhard, Dirk ;
Huthmacher, Carola ;
Mathieson, Toby ;
Poeckel, Daniel ;
Reader, Valerie ;
Strunk, Katja ;
Sweetman, Gavain ;
Kruse, Ulrich ;
Neubauer, Gitte ;
Ramsden, Nigel G. ;
Drewes, Gerard .
NATURE BIOTECHNOLOGY, 2011, 29 (03) :255-U124
[2]   Chemoproteomics Reveals Time-Dependent Binding of Histone Deacetylase Inhibitors to Endogenous Repressor Complexes [J].
Becher, Isabelle ;
Dittmann, Antje ;
Savitski, Mikhail M. ;
Hopf, Carsten ;
Drewes, Gerard ;
Bantscheff, Marcus .
ACS CHEMICAL BIOLOGY, 2014, 9 (08) :1736-1746
[3]   Evaluation of the pharmacodynamic effects of MGCD0103 from preclinical models to human using a novel HDAC enzyme assay [J].
Bonfils, Claire ;
Kalita, Ann ;
Dubay, Marja ;
Siu, Lillian L. ;
Carducci, Michael A. ;
Reid, Gregory ;
Martell, Robert E. ;
Besterman, Jeffrey M. ;
Li, Zuomei .
CLINICAL CANCER RESEARCH, 2008, 14 (11) :3441-3449
[4]   Chemical phylogenetics of histone deacetylases [J].
Bradner, James E. ;
West, Nathan ;
Grachan, Melissa L. ;
Greenberg, Edward F. ;
Haggarty, Stephen J. ;
Warnow, Tandy ;
Mazitschek, Ralph .
NATURE CHEMICAL BIOLOGY, 2010, 6 (03) :238-243
[5]  
CHENG Y, 1973, BIOCHEM PHARMACOL, V22, P3099
[6]   Opinion - Drug-target residence time and its implications for lead optimization [J].
Copeland, Robert A. ;
Pompliano, David L. ;
Meek, Thomas D. .
NATURE REVIEWS DRUG DISCOVERY, 2006, 5 (09) :730-739
[7]  
Couturier Cyril, 2012, Front Endocrinol (Lausanne), V3, P100, DOI 10.3389/fendo.2012.00100
[8]   Characterisation of the in vitro activity of the depsipeptide histone deacetylase inhibitor spiruchostatin A [J].
Crabb, Simon J. ;
Howell, Melanie ;
Rogers, Helen ;
Ishfaq, Muhammad ;
Yurek-George, Alexander ;
Carey, Krystle ;
Pickering, Becky M. ;
East, Phil ;
Mitter, Richard ;
Maeda, Satoko ;
Johnson, Peter W. M. ;
Townsend, Paul ;
Shin-ya, Kazuo ;
Yoshida, Minoru ;
Ganesan, A. ;
Packham, Graham .
BIOCHEMICAL PHARMACOLOGY, 2008, 76 (04) :463-475
[9]   Direct comparison of fluorescence- and bioluminescence-based resonance energy transfer methods for real-time monitoring of thrombin-catalysed proteolytic cleavage [J].
Dacres, H. ;
Dumancic, M. M. ;
Horne, I. ;
Trowell, S. C. .
BIOSENSORS & BIOELECTRONICS, 2009, 24 (05) :1164-1170
[10]   Direct comparison of bioluminescence-based resonance energy transfer methods for monitoring of proteolytic cleavage [J].
Dacres, Helen ;
Dumancic, Mira M. ;
Horne, Irene ;
Trowell, Stephen C. .
ANALYTICAL BIOCHEMISTRY, 2009, 385 (02) :194-202