Design and development of fluorescent probe substrates for carboxylesterase 1 using BODIPY as the basic fluorophore
10.16438/j.0513-4870.2016-1004
- VernacularTitle:基于BODIPY母核的羧酸酯酶1特异性荧光探针底物的设计研发
- Author:
Le-le DING
1
;
Zhen-hao TIAN
1
;
Jie HOU
1
;
Zi-miao WENG
2
;
Jing-nan CUI
1
;
Ling YANG
3
;
Guang-bo GE
3
Author Information
1. State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116023, China
2. Dalian Medical University, Dalian 116011, China
3. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
- Publication Type:SPECIAL REPORTS
- Keywords:
carboxylesterase 1;
boron-dipyrromethene;
fluorescent probe;
specificity;
inhibitors screening
- From:
Acta Pharmaceutica Sinica
2017;52(1):58-65
- CountryChina
- Language:Chinese
-
Abstract:
Carboxylesterase 1 (CE1) is an important serine hydrolase in mammals, which involved in the hydrolysis of a variety of compounds (endogenous substrates like cholesterol and xenobiotic compounds like ester-contain drugs and pesticides). This study aimed to design and develop the fluorescent probe substrates for human carboxylesterase 1 (hCE1), on the basis of the structural features of hCE1 preferred substrates. Four carboxylic esters deriving from BODIPY-8-carboxylic acid were designed and synthesized. After then, reaction phenotyping assays and chemical inhibition assays were used to evaluate the selectivity of these four ester derivatives towards hCE1. Our results clearly demonstrated that the substrate specificity of these ester substrates towards hCE1 would be improved with the decrease of the alcohol group on BODIPY-8-carboxylesters, while BODIPY-8-carboxylesters with small alcohol groups including methyl (BCM) and ethyl (BCE) esters could serve as the ideal probe substrates for hCE1. Given that BCM exhibit rapid hydrolytic rate in hCE1, we further investigate the enzymatic kinetics of this fluorescent probe substrate in both human liver microsomes (HLM) and recombinant hCE1, as well as to explore its potential application in high-throughput screening of hCE1 inhibitors by using HLM as enzyme source. The results showed that the kinetic behaviors and the affinity of BCM in HLM is much closed to those in recombinant hCE1, implying that hCE1 played the key roles in BCM hydrolysis in HLM. Furthermore, the inhibition study demonstrated that BCM could be used for rapid screening and characterization of hCE1 inhibitors, by using HLM to replace recombinant hCE1 as enzyme source.
