Claims
- 1. A cell line comprising a recombinant nucleic acid, wherein said recombinant nucleic acid comprises one or more nucleotide sequences encoding one or more peripheral nerve (PN) sodium channels, wherein said PN sodium channel is stably expressed on the surface of said cell line.
- 2. The cell line according to claim 1, wherein said sodium channel comprises the peripheral nerve type sodium channel 1 (PN1).
- 3. The cell line according to claim 1, wherein said sodium channel comprises the peripheral nerve type sodium channel 3 (PN3).
- 4. The cell line according to claim 2, wherein said sodium channel further comprises the peripheral nerve type sodium channel 3 (PN3).
- 5. The cell line according to any one of claims 1 to 4, wherein said recombinant nucleic acid comprises a nucleotide sequence as in Genbank accession number X82835 or a functional derivative or a fragment thereof.
- 6. The cell line according to any one of claims 1 to 4, said recombinant nucleic acid comprises a nucleotide sequence as in Genbank accession number AFI 17907 or a functional derivative or a fragment thereof.
- 7. The cell line according to claim 5, wherein said recombinant nucleic acid further comprises a nucleotide sequence as in Genbank accession number AF117907 or a functional derivative or a fragment thereof.
- 8. The cell line according to any one of claims 1 to 4, wherein said peripheral nerve sodium channel is human.
- 9. The cell line according to any one of claims 1 to 4 which is derived from a CHO cell line.
- 10. The cell line according to claim 5 which is derived from a CHO cell line.
- 11. The cell line according to claim 6 which is derived from a CHO cell line.
- 12. The cell line according to claim 7 which is derived from a CHO cell line.
- 13. A CHO cell line comprising a recombinant nucleic acid, wherein said recombinant nucleic acid comprises a nucleotide sequence encoding human peripheral nerve type 1 (PN1) sodium channel, wherein said human PN1 sodium channel is stably expressed on the surface of said cell line.
- 14. A CHO cell line comprising a recombinant nucleic acid, wherein said recombinant nucleic acid comprises a nucleotide sequence encoding human peripheral nerve type 3 (PN3) sodium channel, wherein said human PN3 sodium channel is stably expressed on the surface of said cell line.
- 15. A nucleic acid comprising one or more nucleotide sequences encoding a recombinant human peripheral nerve (PN) sodium channel or recombinant human PN sodium channels.
- 16. The nucleic acid according to claim 15, wherein said nucleotide sequence comprises the sequence in Genbank accession number X82835 or a functional derivative or a fragment thereof or a variant due to the degenerate code.
- 17. The nucleic acid according to claim 15, wherein said nucleotide sequence comprises the sequence in Genbank accession number AF117907 or a functional derivative or a fragment thereof or a variant due to the degenerate code.
- 18. A method for preparing a cell line according to any one of claims 1 to 4, comprising:
a) constructing an expression vector which comprises a nucleic acid encoding a peripheral nerve (PN) sodium channel; b) transfecting a suitable cell line with said expression vector; and c) screening said cell line for stable transfection employing a suitable selection agent.
- 19. A method for preparing a cell line according to claim 5, comprising:
a) constructing an expression vector which comprises a nucleic acid encoding a peripheral nerve (PN) sodium channel; b) transfecting a suitable cell line with said expression vector; and c) screening said cell line for stable transfection employing a suitable selection agent.
- 20. A method for preparing a cell line according to claim 6, comprising:
a) constructing an expression vector which comprises a nucleic acid encoding a peripheral nerve (PN) sodium channel; b) transfecting a suitable cell line with said expression vector; and c) screening said cell line for stable transfection employing a suitable selection agent.
- 21. A method for preparing a cell line according to claim 7, comprising:
a) constructing an expression vector which comprises a nucleic acid encoding a peripheral nerve (PN) sodium channel; b) transfecting a suitable cell line with said expression vector; and c) screening said cell line for stable transfection employing a suitable selection agent.
- 22. The method for preparing a cell line according to claim 18, wherein said cell line is a CHO cell line.
- 23. The method according to claim 18, wherein said cell line is co-transfected with two or more of said expression vectors each of which comprises a nucleic acid specific for a different peripheral nerve (PN) sodium channel.
- 24. The method according to claim 22, wherein said cell line is co-transfected with two or more of said expression vectors each of which comprises a nucleic acid specific for a different peripheral nerve (PN) sodium channel.
- 25. A method for identifying an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel comprising:
a) incubating a cell line according to any one of claims 1 to 4 with a test substance; b) measuring a membrane current of said cell line after incubation with test substance; c) incubating said cell line with a known control or in the absence of said test substance; d) measuring a membrane current of said cell line after incubation with a known control or in the absence of said test substance; and e) comparing said membrane current in step (b) with said membrane current obtained in step (d).
- 26. A method for identifying an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel comprising:
a) incubating a cell line according to claim 5 with a test substance; b) measuring a membrane current of said cell line after incubation with test substance; c) incubating said cell line with a known control or in the absence of said test substance; d) measuring a membrane current of said cell line after incubation with a known control or in the absence of said test substance; and e) comparing said membrane current in step (b) with said membrane current obtained in step (d).
- 27. A method for identifying an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel comprising:
a) incubating a cell line according to claim 6 with a test substance; b) measuring a membrane current of said cell line after incubation with test substance; c) incubating said cell line with a known control or in the absence of said test substance; d) measuring a membrane current of said cell line after incubation with a known control or in the absence of said test substance; and e) comparing said membrane current in step (b) with said membrane current obtained in step (d).
- 28. A method for identifying an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel comprising:
a) incubating a cell line according to claim 7 with a test substance; b) measuring a membrane current of said cell line after incubation with test substance; c) incubating said cell line with a known control or in the absence of said test substance; d) measuring a membrane current of said cell line after incubation with a known control or in the absence of said test substance; and e) comparing said membrane current in step (b) with said membrane current obtained in step (d).
- 29. The method according to claim 25 wherein said cell line is derived from a CHO cell line.
- 30. A method for identifying an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel comprising:
a) incubating a cell line according to any one of claims 1 to 4 with a known agonist which is coupled to a reporter; b) incubating said cell line with a test substance; and c) measuring the displacement of the agonist coupled to the reporter by the test substance.
- 31. A method for identifying an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel comprising:
a) incubating a cell line according to claim 5 with a known agonist which is coupled to a reporter; b) incubating said cell line with a test substance; and c) measuring the displacement of the agonist coupled to the reporter by the test substance.
- 32. A method for identifying an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel comprising:
a) incubating a cell line according to claim 6 with a known agonist which is coupled to a reporter; b) incubating said cell line with a test substance; and c) measuring the displacement of the agonist coupled to the reporter by the test substance.
- 33. A method for identifying an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel comprising:
a) incubating a cell line according to claim 7 with a known agonist which is coupled to a reporter; b) incubating said cell line with a test substance; and c) measuring the displacement of the agonist coupled to the reporter by the test substance.
- 34. The method according to claim 25, wherein said method is a high throughput screening assay (HTS).
- 35. The method according to claim 30, wherein said method is a high throughput screening assay (HTS).
- 36. The method according to claim 30, wherein said reporter is a fluorescent molecule or a radioisotope.
- 37. The method according to any one of claims 31 to 33, wherein said reporter is a fluorescent molecule or a radioisotope.
- 38. An agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel identifiable according to the method of claim 25.
- 39. An agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel identifiable according to the method of claim 30.
- 40. The agonist, antagonist or modulator according to claim 38, wherein said agonist, antagonist or modulator is not selected from the group of lidocaine, mexiletine and TTX.
- 41. The agonist, antagonist or modulator according to claim 39, wherein said agonist, antagonist or modulator is not selected from the group of lidocaine, mexiletine and TTX.
- 42. A pharmaceutical composition comprising an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel according to claim 38; and a pharmaceutically acceptable carrier.
- 43. A pharmaceutical composition comprising an agonist, antagonist or modulator of a peripheral nerve (PN) sodium channel according to claim 39; and a pharmaceutically acceptable carrier.
Priority Claims (1)
Number |
Date |
Country |
Kind |
DE 100 54 878.4 |
Nov 2000 |
DE |
|
RELATED APPLICATION
[0001] The benefit of prior U.S. provisional application Ser. No. 60/249,932, filed Nov. 17, 2000 is hereby claimed.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60249932 |
Nov 2000 |
US |