REGULATION OF NEUROTRANSMITTER RELEASE THROUGH ANION CHANNELS

• US 20110201668A1
• Filed: 01/30/2008
• Published: 08/18/2011
• Est. Priority Date: 01/30/2008
• Status: Abandoned Application

• Current Assignee: Korea Advanced Institute of Science and Technology
• Original Assignee: Korea Advanced Institute of Science and Technology
• Inventors: Woo, Dong-Ho, Park, Hyung-Ju, Park, Hye-Kyung, Lee, Justin Changjoon
• Application Number:
  US12/865,126
• Publication Number:
  US 20110201668A1
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed

Abstract

A novel use of anion channels, preferably Ca2+-activated anion channels (CAACs), in regulating release of neurotransmitters from neurons and/or astrocytes is provided. More specifically, CAAC activity regulators, agents for regulating neurotransmitter release comprising such CAAC activity regulators, and methods of screening agents for regulating neurotransmitter release using CAAC as a target.

Claims

1. A composition for regulating release of an excitatory neurotransmitter through a Ca2+-activated anion channel (CAAC) from neurons and astrocytes, containing a CAAC activity regulator as an active ingredient.

2. The composition for regulating release of an excitatory neurotransmitter according to claim 1, wherein said CAAC is encoded by Bestrophin 1 gene.

3. The composition for regulating release of an excitatory neurotransmitter according to claim 1, wherein said excitatory neurotransmitter is one or more selected from the group consisting of acetyl choline, aspartic acid, D-serine, glutamate, enkephalin, and histamine.

4. The composition for regulating release of an excitatory neurotransmitter according to claim 1, wherein said CAAC activity regulator is a CAAC inhibitor comprising one or more selected from the group consisting of anion channel blocking agents and antisense RNA or shRNA against a CAAC-coding nucleotide, and has an inhibiting activity against neurotransmitter release through a CAAC from neurons or astrocytes.

5. The composition for regulating release of an excitatory neurotransmitter according to claim 4, wherein said anion channel blocking agent is one or more selected from the group consisting of niflumic acid, flumenamic acid, 5-nitro-2(3-phenylpropylamino)-benzoic acid (NPPB), and 4,4′

-diisothiocyanatostilbene-2,2′

-disulfonic acid (DIDS).

6. The composition for regulating release of an excitatory neurotransmitter according to claim 4, wherein said antisense RNA is against Bestrophin 1 gene having the sequence of SEQ ID NO:

1 or 2.

7. The composition for regulating release of an excitatory neurotransmitter according to claim 4, wherein said shRNA is one or more selected from the group consisting of sequences of SEQ ID NOs:

3 to 5.

8. The composition for regulating release of an excitatory neurotransmitter according to claim 1, wherein said CAAC activity regulator is a CAAC activator and has activity of promoting release of neurotransmitter through a CAAC from neurons or astrocytes.

19. The composition for regulating release of an excitatory neurotransmitter according to claim 2, wherein said CAAC activity regulator is a CAAC inhibitor comprising one or more selected from the group consisting of anion channel blocking agents and antisense RNA or shRNA against a CAAC-coding nucleotide, and has an inhibiting activity against neurotransmitter release through a CAAC from neurons or astrocytes.

20. The composition for regulating release of an excitatory neurotransmitter according to claim 3, wherein said CAAC activity regulator is a CAAC inhibitor comprising one or more selected from the group consisting of anion channel blocking agents and antisense RNA or shRNA against a CAAC-coding nucleotide, and has an inhibiting activity against neurotransmitter release through a CAAC from neurons or astrocytes.

9. A composition for preventing or treating diseases caused by over-release of an excitatory neurotransmitter containing one or more selected from the group consisting of a channel blocking agent against a Ca2+-activated anion channel (CAAC) and an antisense RNA or a shRNA against a CAAC-coding nucleotide, as an active ingredient, wherein the diseases caused by over-release of an excitatory neurotransmitter is one or more selected from the group consisting of epileptic seizures, neurotransmitter-induced excitotoxicity, ischemia, brain stroke, brain hemorrhage, epilepsy, traumatic brain injury, and hypoxia.

10. The composition according to claim 9, wherein said channel blocking agent against CAAC is one or more selected from the group consisting of niflumic acid, flumenamic acid, 5-nitro-2(3-phenylpropylamino)-benzoic acid (NPPB), and 4,4′

-diisothiocyanatostilbene-2,2′

-disulfonic acid (DIDS).

11. The composition according to claim 9, wherein said antisense RNA is against Bestrophin 1 gene having the sequence of SEQ ID NO:

1 or 2.

12. The composition according to claim 9, wherein said shRNA is selected from the group consisting of sequences of SEQ ID NOs:

3 to 5.

13. A composition for improving recognition, cognition, movement, memory, or learning capabilities, said composition comprising a Ca2+-activated anion channel (CAAC) activator as an active ingredient.

14. A preparing method of a channel for an excitatory neurotransmitter release on neurons or astrocytes, comprising the steps of transfecting a mammalian neuron or astrocyte with Bestrophin 1 gene having the sequence of SEQ ID NO:

1 or 2, and expressing the transfected gene.

15. The preparing method according to claim 14, wherein said excitatory neurotransmitter is one or more selected from the group consisting of acetyl choline, aspartic acid, D-serine, glutamate, enkephalin, and histamine.

16. A screening method for a neuroregulatory agent comprising the steps of:

preparing a sample of neuron and/or astrocyte;

contacting said sample with a candidate substance;

testing whether or not a Ca2+-activated anion channel (CAAC) on neuron and/or astrocyte is activated; and

determining said candidate substance as a neurotransmission promoting agent when the CAAC is activated, or determining said candidate substance as a neuroprotective agent when the CAAC is not activated.

17. The screening method according to claim 16, wherein said CAAC is encoded by Bestrophin 1 gene.

18. The screening method according to claim 16, wherein said step of whether or not a CAAC on neuron and/or astrocyte is activated is conducted by measuring inward current change through a sniffer patch method.