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Early drug screening method based on fluorescence

A drug and fluorescence technology, used in fluorescence/phosphorescence, material analysis by optical means, measurement devices, etc., can solve problems such as difficulties, achieve stable data, avoid adverse reactions and off-target effects, and achieve good repeatability.

Pending Publication Date: 2019-12-03
ICE BIOSCI INC +1
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Problems solved by technology

Therefore, it is also difficult to detect the effect of compounds on each subtype at the same time.

Method used

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  • Early drug screening method based on fluorescence
  • Early drug screening method based on fluorescence
  • Early drug screening method based on fluorescence

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Experimental program
Comparison scheme
Effect test

Embodiment

[0046] 1. Materials and methods

[0047] 1.1 Materials

[0048] 1.1.1 Cells and reagents GABAα1-6-HEK293 stably transfected cell line was constructed according to the steps in the following experimental method; ScreenQuestMembranePotentialAssayKit*RedFluorescence*membrane potential detection kit was purchased from AAT Company in the United States; FLIPRMembranePotentialRed, Explorer was purchased from Molecular Devices (MD ) company; HBSS, FBS, 0.25% Trypsin-EDTA were purchased from gibco company; DMEM was purchased from Corning company; GABA, DMSO, Poly-L-lysinehydrobromide were purchased from sigma company; Bicuculline was purchased from Santa Cruz company in the United States; transfection reagent: X- tremeGENEHPDNATransf.Reag., purchased from Roche Company; Antibiotics G418, HygromycinB, Zeocin Table 1: Stable cell lines and deposit numbers

[0049]

[0050] 1.1.2 The main instruments Synergy4 multi-functional microplate reader was purchased from BioTek in the United S...

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Abstract

The invention relates to an early drug screening method based on fluorescence. The method comprises the following steps of: 1, providing target cells which stably express ion channel targets; 2, carrying out incubation on a voltage-sensitive dye and drugs with different concentrations together with the target cells; 3, measuring a fluorescence baseline value and a time-varying value of the incubated cells by utilizing a fluorescence measurement device; and 4, carrying out data fitting on the obtained values and determining whether the drugs have an exciting effect or an inhibiting effect on the ion channel targets. By utilizing the method, dozens of ion channel targets can be subjected to drug screening with an intermediate flux through utilizing a microplate reader with low cost; and theearly drug screening method is particularly applicable to early drug screening.

Description

technical field [0001] The invention relates to a drug screening method, in particular to a fluorescence-based early drug screening method. Background technique [0002] Ion channels are the basis of central nervous system activities and are closely related to various activities of the central nervous system. They are also important drug screening targets. Drugs act non-specifically on ion channels in the central nervous system, which is the main cause of adverse drug reactions in the central nervous system. Therefore, early screening of non-specific effects of drugs on the central nervous system is an important part of secondary pharmacology screening and safety pharmacology of the central nervous system, and an important tool for early detection of drug central nervous system toxicity in the process of new drug development. [0003] The traditional central nervous system ion channel target screening technology has certain defects in the screening of toxic targets, such as...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6486
Inventor 郑双佳魏成喜张旭赵明李英骥闫励
Owner ICE BIOSCI INC
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