A bioluminescent sensor for highly selective and sensitive detection of human carboxylesterase 1 in complex biological samples

Wang, Dandan; Jin, Qiang; Zou, Li-Wei; Hou, Jie; Lv, Xia; Lei, Wei; Cheng, Hai‐Ling Margaret; Ge, Guang-Bo; Yang, Ling

doi:10.1039/c5cc09874b

Cited by 83 publications

(57 citation statements)

References 35 publications

(5 reference statements)

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“…Human liver microsomes (HLMs) were obtained from Celsis (Shanghai, China). The specific probes DME (hCE1) and DDAB (hCE2) were synthesized in our lab as described previously (Jin et al, 2016; Wang et al, 2016). Millipore water (Millipore, Bedford, USA), HPLC grade acetonitrile, methanol, and formic acid (Tedia company, USA) were employed throughout the study.…”

Section: Methodsmentioning

confidence: 99%

“…Thus, it is urgently necessary to find more potent and selective hCE1 inhibitors from phytochemicals. More recently, we have developed a highly specific bioluminescent probe substrate (termed DME) for hCE1 and a highly selective near-infrared fluorescent probe (termed DDAB) for hCE2, which have been successfully used for rapid screening and characterization of inhibitors against hCEs using cell or tissue preparations as enzyme sources (Jin et al, 2016; Wang et al, 2016). In the present study, DME and DDAB were used as the highly selective optical substrates for human CEs to rapidly screen hCE1 inhibitors from natural triterpenoid compounds.…”

Section: Introductionmentioning

confidence: 99%

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Structure-Activity Relationships of Pentacyclic Triterpenoids as Potent and Selective Inhibitors against Human Carboxylesterase 1

et al. 2017

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Human carboxylesterase 1 (hCE1), one of the most important serine hydrolases distributed in liver and adipocytes, plays key roles in endobiotic homeostasis and xenobiotic metabolism. This study aimed to find potent and selective inhibitors against hCE1 from phytochemicals and their derivatives. To this end, a series of natural triterpenoids were collected and their inhibitory effects against human carboxylesterases (hCEs) were assayed using D-Luciferin methyl ester (DME) and 6,8-dichloro-9,9-dimethyl-7-oxo-7,9-dihydroacridin-2-yl benzoate (DDAB) as specific optical substrate for hCE1, and hCE2, respectively. Following screening of a series of natural triterpenoids, oleanolic acid (OA), and ursolic acid (UA) were found with strong inhibitory effects on hCE1 and relative high selectivity over hCE2. In order to get the highly selective and potent inhibitors of hCE1, a series of OA and UA derivatives were synthesized from OA and UA by chemical modifications including oxidation, reduction, esterification, and amidation. The inhibitory effects of these derivatives on hCEs were assayed and the structure-activity relationships of tested triterpenoids as hCE1 inhibitors were carefully investigated. The results demonstrated that the carbonyl group at the C-28 site is essential for hCE1 inhibition, the modifications of OA or UA at this site including esters, amides and alcohols are unbeneficial for hCE1 inhibition. In contrast, the structural modifications on OA and UA at other sites, such as converting the C-3 hydroxy group to 3-O-β-carboxypropionyl (compounds 20 and 22), led to a dramatically increase of the inhibitory effects against hCE1 and very high selectivity over hCE2. 3D-QSAR analysis of all tested triterpenoids including OA and UA derivatives provide new insights into the fine relationships linking between the inhibitory effects on hCE1 and the steric-electrostatic properties of triterpenoids. Furthermore, both inhibition kinetic analyses and docking simulations demonstrated that compound 22 was a potent competitive inhibitor against hCE1-mediated DME hydrolysis. All these findings are very helpful for medicinal chemists to design and develop highly selective and more potent hCE1 inhibitors for biomedical applications.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Structure-Activity Relationships of Pentacyclic Triterpenoids as Potent and Selective Inhibitors against Human Carboxylesterase 1

et al. 2017

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“…By contrast, fluorescence‐based biochemical assays have attracted great attention in biological and medical related fields, due to their inherent advantages including non‐destructive, highly sensitive, and applicable to HTS assays . Over the past decade, a wide range of fluorescence probes for serine hydrolases and other phase I metabolizing enzymes have been developed and widely used for evaluating enzyme activities and screening of enzyme modulators . However, the design and development of fluorescence probes for UGTs is a very difficult task, due to the following three reasons.…”

Section: Fluorescent Probes For Ugtsmentioning

confidence: 99%

Chemical Probes for Human UDP‐Glucuronosyltransferases: A Comprehensive Review

Zhang

Hou

et al. 2018

Biotechnology Journal

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UGTs play crucial roles in the metabolism and detoxification of both endogenous and xenobiotic compounds. The key roles of UGTs in human health have garnered great interest in the design and development of specific probes for human UGTs. However, in contrast to other human enzymes, the probe substrates for human UGTs are rarely reported, owing to the highly overlapping substrate specificities of UGTs and the lack of the integrated crystal structures of UGTs. Over the past decades, many efforts are made to develop specific probe substrates for UGTs and use them in both basic research and drug discovery. This review focuses on recent progress in the development of probe substrates for UGTs and their biomedical applications. A long list of chemical probes for UGTs, including non-fluorescent and fluorescent probes along with their structural information and kinetic parameters, are prepared and analyzed. Additionally, challenges and future directions in this field are highlighted in the final section. All information and knowledge presented in this review provide practical tools/methods for measuring UGT activities in complex biological samples, which will be very helpful for rapid screening and characterization of UGT modulators, and for exploring the relevance of UGT enzymes to human diseases.

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“…It is well-known that the enzyme kinetic behaviour is very crucial for the quantitative applications of the activity-based fluorescent probes (Ge et al, 2013;Liu et al, 2014a, Liu et al, 2014bFeng et al, 2015;Lv et al, 2015;D.D. Wang et al, 2016;Y.…”

Section: Enzymatic Kinetics Of Dpp-iv Mediated Gp-ban Hydrolysismentioning

confidence: 99%

“…Although several fluorescent probes for DPP-IV have been developed and used in vitro bioassays, these probes do not have enough isoform selectivity to precisely evaluate the real activities of DPP-IV in complex biological systems, such as living cells and body fluids (Grant et al, 2002;Ho et al, 2006;Kawaguchi et al, 2010;Y. Wang et al, 2016;D.D. Wang et al, 2016;Gong et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

A highly specific ratiometric two-photon fluorescent probe to detect dipeptidyl peptidase IV in plasma and living systems

Zou

Wang

Qian

et al. 2017

Biosensors and Bioelectronics

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In this study, a highly specific ratiometric two-photon fluorescent probe GP-BAN was developed and well-characterized to monitor dipeptidyl peptidase IV in plasma and living systems. GP-BAN was designed on the basis of the catalytic properties and substrate preference of DPP-IV, and it could be readily hydrolyzed upon addition of DPP-IV under physiological conditions. Both reaction phenotyping and inhibition assays demonstrated that GP-BAN displayed good reactivity and high selectivity towards DPP-IV over other human serine hydrolases including FAP, DPP-VIII, and DPP-IX. The probe was successfully used to monitor the real activities of DPP-IV in complex biological systems including diluted plasma, while it could be used for high throughput screening of DPP-IV inhibitors by using human plasma or tissue preparations as enzyme sources. As a two-photon fluorescent probe, GP-BAN was also successfully used for two-photon imaging of endogenous DPP-IV in living cells and tissues, and showed high ratiometric imaging resolution and deep-tissue penetration ability. Taken together, a ratiometric two-photon fluorescent probe GP-BAN was developed and well-characterized for highly selective and sensitive detection of DPP-IV in complex biological systems, which could serve as a promising imaging tool to explore the biological functions and physiological roles of this key enzyme in living systems.

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A bioluminescent sensor for highly selective and sensitive detection of human carboxylesterase 1 in complex biological samples

Abstract: A highly selective and sensitive bioluminescent sensor (DME) for human carboxylesterase 1 (hCE1) has been developed and well characterized. DME could be used for real-time monitoring of hCE1 activities in complex biological samples and for bio-imaging of endogenous hCE1 in living cells.

Cited by 83 publications

References 35 publications

Structure-Activity Relationships of Pentacyclic Triterpenoids as Potent and Selective Inhibitors against Human Carboxylesterase 1

Structure-Activity Relationships of Pentacyclic Triterpenoids as Potent and Selective Inhibitors against Human Carboxylesterase 1

Chemical Probes for Human UDP‐Glucuronosyltransferases: A Comprehensive Review

A highly specific ratiometric two-photon fluorescent probe to detect dipeptidyl peptidase IV in plasma and living systems

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