Material for Screening For Compound Acting on ION Channel and Use Thereof

• US 20190331664A1
• Filed: 11/01/2017
• Published: 10/31/2019
• Est. Priority Date: 11/01/2016
• Status: Active Grant

• Current Assignee: Channelosearch Technology, Public University Corporation Nagoya City University
• Original Assignee: Public University Corporation Nagoya City University
• Inventors: IMAIZUMI, Yuji, YAMAMURA, Hisao, SUZUKI, Yoshiaki, KAWASAKI, Keisuke, NARITA, Hiroshi
• Granted Patent: US 11,828,750 B2
• Time in Patent Office: Days
• CPC Class Codes: C12M 1/34 Measuring or testing with condition measuring or sensing means, e.g. colony counters
• CPC Class Codes: C12N 5/10 Cells modified by introduction of foreign genetic material
• CPC Class Codes: C12Q 1/025 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics antimicrobial activity C12Q1/18
• CPC Class Codes: G01N 2500/02 Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
• CPC Class Codes: G01N 2500/10 involving cells
• CPC Class Codes: G01N 33/5014 for testing toxicity
• CPC Class Codes: G01N 33/5032 on intercellular interactions
• CPC Class Codes: G01N 33/5058 Neurological cells
• CPC Class Codes: G01N 33/6872 Intracellular protein regulatory factors and their receptors, e.g. including ion channels

Abstract

A screening system provided with a potential-dependent Na ion channel that extends the duration of the action potential associated with depolarization, and a K ion channel that deepens the resting membrane potential in the negative direction, said screening system furthermore including cells provided with ion channels that contribute to deepening the resting membrane potential in the negative direction and/or shortening the duration of the action potential as target ion channels. By such cells, the action of a test compound on the target ion channel can be easily evaluated by providing an inhibitor for the K ion channel to control the probability of cell death.

Claims

1. A screening material for agonists or inhibitors for a target ion channel, wherein the screening material contains cells comprising:

a voltage-dependent Na ion channel that prolongs the duration of an action potential associated with depolarization,

a K ion channel that deepens a resting membrane potential in the negative direction, and

a target ion channel that is an ion channel that contributes to deepening the resting membrane potential in the negative direction and/or that shortens the duration of the action potential associated with depolarization,

wherein the Na ion channel, the K ion channel, and the target ion channel constitute a cell death induction system with which cell death is induced by the induction of depolarization in the presence of an inhibitor for the K ion channel.

2. The screening material according to claim 1, wherein the target ion channel has an inhibitory action on the cell death induced by the cell death induction system.

3. The screening material according to claim 1, that uses a selective inhibitor of the K ion channel.

4. The screening material according to claim 3, wherein the target ion channel has a lower sensitivity to the K ion channel inhibitor than the K ion channel.

5. The screening material according to claim 1, wherein the target ion channel is an ion channel that contributes to deepening the resting membrane potential in the negative direction and is another K ion channel that is different from the aforementioned K ion channel.

6. The screening material according to claim 5, wherein the another K ion channel is a K2P channel.

7. The screening material according to claim 1, wherein the target ion channel is an ion channel that contributes to a shortening of the duration of the action potential associated with depolarization.

8. A method for screening for an agonist or inhibitor of a target ion channel, wherein the method

uses cells provided with a voltage-dependent Na ion channel that prolongs the duration of an action potential associated with depolarization, a K ion channel that deepens the resting membrane potential in the negative direction, and a target ion channel that is an ion channel that contributes to deepening the resting membrane potential in the negative direction and/or that shortens the duration of the action potential associated with depolarization, wherein the Na ion channel, the K ion channel, and the target ion channel constitute a cell death induction system with which cell death is induced by the induction of depolarization in the presence of an inhibitor for the K ion channel,

and is provided with

a step of supplying the inhibitor to the cells;

a step of supplying the cells with a test compound having the potential to inhibit or activate the target ion channel; and

a step of evaluating the effect on cell death of the cells due to the supply of the test compound.

9. A kit for screening for an agonist or inhibitor of a target ion channel, the kit being provided with

cells provided with a voltage-dependent Na ion channel that prolongs the duration of an action potential associated with depolarization, a K ion channel that deepens the resting membrane potential in the negative direction, and a target ion channel that is an ion channel that contributes to deepening the resting membrane potential in the negative direction and/or that shortens the duration of the action potential associated with depolarization, wherein the Na ion channel, the K ion channel, and the target ion channel constitute a cell death induction system with which cell death is induced by the induction of depolarization in the presence of an inhibitor for the K ion channel, and

the inhibitor.