Unlock instant, AI-driven research and patent intelligence for your innovation.

Cell line and method for detecting biological activity of dopamine D4 target compound

A technology of biological activity and detection method, which is applied in the field of detection of biological activity of dopamine D4 target compounds and cell lines for detection of biological activity of dopamine D4 target compounds, can solve the problems of lack of methods for detection of biological activity, and achieve High accuracy, high signal window and good repeatability

Pending Publication Date: 2022-05-13
SHUJING BIOPHARMA CO LTD +1
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the defect in the prior art that there is no method for detecting the biological activity of drugs targeting dopamine D4, the present invention provides a cell line and a detection method using it to detect targeting dopamine D4

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Cell line and method for detecting biological activity of dopamine D4 target compound
  • Cell line and method for detecting biological activity of dopamine D4 target compound
  • Cell line and method for detecting biological activity of dopamine D4 target compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Example 1 is designed as a functional validation of HEK293 / D4 cells.

[0098] 1.1 Purpose of the experiment

[0099] HEK293 / D4 cells were constructed and their activated cAMP responses were verified.

[0100] 1.2 Experimental steps

[0101] 1. Construction of HEK293 / D4 cells

[0102] (1) Digest HEK293 cells with trypsin and plate them into 6-well plates, 1x10 per well 5 cells at 37°C, 5% CO 2 grow overnight in a cell culture incubator;

[0103] (2) Take out a tube of D4 receptor lentivirus from liquid nitrogen, put it on ice to thaw, and spin it at 1000rpm after thawing to make the solution adhering to the tube wall sink to the bottom of the tube; the suitable MOI for infection of HEK293 cells is 5 , calculate the required lentivirus volume according to the number of plated cells and the titer of lentivirus;

[0104] (3) Take out the HEK293 cells from the cell culture incubator, replace them with fresh growth medium, and then add a certain volume of D4 receptor le...

Embodiment 5

[0193] Example 5 is designed as a repeatability test for detecting the biological activity of dopamine D4 target compounds on CHO-K1 / D4 / Nlucp / CRE cells.

[0194] 5.1 Purpose of the experiment Use cryopreserved CHO-K1 / D4 / Nlucp / CRE cells to detect the biological activity of dopamine D4 target compounds, and evaluate the repeatability of the method.

[0195] 5.2 Cell information

[0196]

[0197] 5.3 Experimental steps

[0198] Dopamine agonist and Brilaroxazine (RP5603) adopt the same experimental procedure as in Example 4 to repeat the operation at different time periods and calculate the EC of dopamine agonist and Brilaroxazine 50 ; Wherein Brilaroxazine structural formula is as follows:

[0199]

[0200] 5.4 Results and Analysis

[0201] Table 5: The repeatability evaluation results of the detection method for the biological activity of drugs targeting dopamine D4

[0202] Experiment time DA-D4R EC 50 value

Brilaroxazine EC 50 value

2021091...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a cell line and a method for detecting biological activity of a dopamine D4 target compound. The cell line comprises polynucleotide for coding dopamine D4 receptor protein and polynucleotide for coding luciferase of which expression is controlled by a CRE transcriptional regulatory element. The invention also discloses a method for detecting the biological activity of a dopamine D4 target compound by using the cell line. The cell line disclosed by the invention can be used for rapidly, accurately and efficiently detecting the biological activity of the dopamine D4 target therapeutic drug. In addition, the detection method provided by the invention can completely meet the biological activity detection of the small molecule compound aiming at the dopamine D4 receptor without complicated instruments or operation steps.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular, the invention relates to a polynucleotide encoding a dopamine D4 receptor protein and a polynucleotide encoding a luciferase expressed under the control of a CRE transcriptional regulatory element for detecting dopamine A cell line for the biological activity of a D4 target compound; the present invention also relates to a method for detecting the biological activity of a dopamine D4 target compound using the cell line. Background technique [0002] The dopaminergic system plays an important role in neuromodulation, such as motor control, motivation, reward, cognitive function, maternal and reproductive behavior. Dopamine is a neurotransmitter that is produced in neuronal terminals through the continuous hydroxylation and decarboxylation of tyrosine, and loaded into synaptic vesicles by vesicular monoamine transporter 2 for exocytotic release. Released dopamine can be reuptake...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/10C12N15/12C12N15/53C12Q1/66
CPCC07K14/70571C12N9/0004C12N5/0682C12Q1/66C12N2510/00
Inventor 叶飘钱芸吴鸣葛玉强张艺婷胡志京万泽红
Owner SHUJING BIOPHARMA CO LTD