CN109690316A - 测定细胞内配体靶蛋白相互作用持久力的方法 - Google Patents

测定细胞内配体靶蛋白相互作用持久力的方法 Download PDF

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CN109690316A
CN109690316A CN201680003155.8A CN201680003155A CN109690316A CN 109690316 A CN109690316 A CN 109690316A CN 201680003155 A CN201680003155 A CN 201680003155A CN 109690316 A CN109690316 A CN 109690316A
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target protein
ligand
cell
intracellular
endurance
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王帅
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
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  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
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  • Biotechnology (AREA)
  • Analytical Chemistry (AREA)
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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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Abstract

本发明公开了一种测定细胞内配体靶蛋白相互作用持久力的方法，包括：(1)用所述配体处理所述细胞；(2)去除所述细胞外的所述配体；(3)收集去除所述配体后不同时间点的所述细胞；(4)加热所述细胞；(5)裂解所述细胞；(6)分析细胞裂解液中可溶的或不可溶的所述靶蛋白的水平随时间变化的函数。应用本发明的方法可确定细胞内配体与靶蛋白相互作用的解离速率常数，滞留时间和解离半衰期以及配体的治疗持续时间范围，定量的描述细胞内配体靶蛋白相互作用的持久力以及预测细胞内药理作用的持续时间。本发明的方法可用于新药研发，发现具有期望的结合持久力的药物。本发明的方法可用于设计给药方案，尤其是设计给药的时间间隔。

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PCT国内申请，说明书已公开。

Claims (13)

  1. PCT国内申请，权利要求书已公开。
CN201680003155.8A 2016-05-31 2016-05-31 测定细胞内配体靶蛋白相互作用持久力的方法 Withdrawn CN109690316A (zh)

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PCT/CN2016/083998 WO2017206041A1 (zh) 2016-05-31 2016-05-31 测定细胞内配体靶蛋白相互作用持久力的方法

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112924693A (zh) * 2019-12-06 2021-06-08 中国科学院大连化学物理研究所 一种上清和沉淀协同定量的蛋白质热稳定性分析方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103733067A (zh) * 2011-04-18 2014-04-16 艾维特拉普罗特玛公司 使用热转移试验测定配体与靶蛋白结合的方法
CN104897826A (zh) * 2015-06-16 2015-09-09 广西师范大学 一种用于细胞原位检测小分子化合物与靶蛋白相互作用的方法

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EP1734367A1 (en) * 2005-06-14 2006-12-20 Cellzome Ag Process for the identification of novel enzyme interacting compounds
KR102292431B1 (ko) * 2012-12-12 2021-08-20 프로메가 코포레이션 세포내 생물발광 공명 에너지 전이를 이용하여 생물활성제에 의한 세포 표적 결합의 인식
CN103645324B (zh) * 2013-12-03 2017-02-15 胡朝阳 基于细胞热转移技术的高通量药物靶点筛选方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103733067A (zh) * 2011-04-18 2014-04-16 艾维特拉普罗特玛公司 使用热转移试验测定配体与靶蛋白结合的方法
CN104897826A (zh) * 2015-06-16 2015-09-09 广西师范大学 一种用于细胞原位检测小分子化合物与靶蛋白相互作用的方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ROBERT A. COPELAND 等: "Drug–target residence time and its implications for lead optimization", 《NATURE REVIEWS DRUG DISCOVERY》 *
ROZBEH JAFARI 等: "The cellular thermal shift assay for evaluating drug target interactions in cells", 《NATURE PROTOCOLS》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112924693A (zh) * 2019-12-06 2021-06-08 中国科学院大连化学物理研究所 一种上清和沉淀协同定量的蛋白质热稳定性分析方法
CN112924693B (zh) * 2019-12-06 2022-09-27 中国科学院大连化学物理研究所 一种上清和沉淀协同定量的蛋白质热稳定性分析方法

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