Chimeric modified ion channels and their use for the treatment of trigeminal nerve disorders

Abstract

This disclosure provides an AAV vector encoding a modified chimeric ligand-opening ion channel, and a method for treating trigeminal neuralgia in subjects requiring such a vector. In certain aspects of this disclosure, the disclosure relates to nucleic acid sequences and vectors comprising an LGIC, wherein the LGIC comprises a nucleotide sequence having at least 80% sequence identity with any one of SEQ ID NOs: 1-13 or any sequence listed in Table 1.

Claims

1. Nucleic acids containing SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO:

7.

2. A nucleic acid having at least 90% sequence identity with SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO:

7.

3. A nucleic acid having at least 95% sequence identity with SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO:

7.

4. A nucleic acid having at least 98% sequence identity with SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO:

7.

5. A nucleic acid having at least 99% sequence identity with SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO:

7.

6. The nucleic acid of sequence number 4, sequence number 5, sequence number 6, or sequence number 7.

7. (i) 5'ITR; (ii) Human synapsin promoter; (iii) Transgene sequences encoding a ligand-opening ion channel (LGIC) containing a human α7-nicotinic acetylcholine receptor ligand-binding domain (α7-nAChR LBD) and a glycine receptor ion pore domain (GlyR IPD); (iv) PolyA sequence; and (v) 3'ITR Polynucleotides containing these nucleotides.

8. The polynucleotide according to claim 7, further comprising an intron sequence located between the human synapsin promoter and the transgene sequence encoding the LGIC.

9. (i) The 5'ITR comprises a sequence having at least 90%, 95%, 98%, 99%, or 100% identity with sequence number 10; (ii) The human synapsin promoter comprises a sequence having at least 90%, 95%, 98%, 99%, or 100% identity with SEQ ID NO: 11; (iii) The introduced gene sequence codes for the polypeptide of Sequence ID No. 1; (iv) The polyA sequence includes a sequence that has at least 90%, 95%, 98%, 99%, or 100% identity with sequence number 3; (v) The 3'ITR comprises a sequence having at least 90%, 95%, 98%, 99%, or 100% identity with sequence number 13; (vi) If necessary, the intron includes a sequence having at least 90%, 95%, 98%, 99%, or 100% identity with sequence number 12. The polynucleotide according to claim 7 or 8.

10. A polynucleotide containing the nucleic acid of SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 7, which encodes the sequence of SEQ ID NO:

1.

11. A polynucleotide having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity with a polynucleotide containing the nucleic acid of SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 7, which encodes the sequence of SEQ ID NO:

1.

12. An expression vector comprising a nucleic acid according to any one of claims 1 to 6, or a polynucleotide according to any one of claims 7 to 11.

13. The expression vector according to claim 12, which is a viral expression vector.

14. The expression vector according to claim 13, which is an adeno-associated virus (AAV) expression vector.

15. Cells comprising the expression vector according to claims 12 to 14.

16. The cells according to claim 15, which are isolated cells under culture.

17. An expression vector according to any one of claims 12 to 14, and AAV particles comprising an AAV capsid protein.

18. The AAV particle according to claim 17, wherein the AAV capsid protein is AAV5 capsid protein.

19. The AAV particle according to claim 17 or 18, wherein the AAV capsid protein comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO:

9.

20. The AAV particle according to any one of claims 17 to 19, wherein the AAV capsid protein comprises an amino acid sequence having at least 95% sequence identity with SEQ ID NO:

9.

21. The AAV particle according to any one of claims 17 to 20, wherein the AAV capsid protein comprises an amino acid sequence having at least 97% sequence identity with SEQ ID NO:

9.

22. The AAV particle according to any one of claims 17 to 21, wherein the AAV capsid protein comprises an amino acid sequence having at least 98% sequence identity with SEQ ID NO:

9.

23. The AAV particle according to any one of claims 17 to 22, wherein the AAV capsid protein comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO:

9.

24. The AAV particle according to any one of claims 17 to 23, wherein the AAV capsid protein has the amino acid sequence of SEQ ID NO:

9.

25. A composition comprising a nucleic acid according to any one of claims 1 to 6, a polynucleotide according to any one of claims 7 to 11, an expression vector according to any one of claims 12 to 14, or AAV particles according to any one of claims 17 to 24.

26. The composition according to claim 25, further comprising a nonionic copolymer.

27. The composition according to claim 26, wherein the nonionic copolymer is a poloxamer.

28. The composition according to claim 27, wherein the nonionic copolymer is poloxamer 188.

29. The composition according to claim 28, comprising approximately 0.0005% to approximately 0.005% of poloxamer 188.

30. The composition according to claim 27 or 28, comprising approximately 0.001% poloxamer 188.

31. The composition according to any one of claims 25 to 30, further comprising a sodium phosphate buffer.

32. The composition according to claim 31, comprising approximately 1 mM to approximately 10 mM sodium phosphate buffer.

33. The composition according to claim 32, comprising approximately 10 mM sodium phosphate buffer.

34. The composition according to any one of claims 25 to 33, further comprising sodium chloride.

35. The composition according to claim 34, comprising approximately 120 mM to approximately 240 mM sodium chloride.

36. The composition according to claim 35, comprising approximately 180 mM sodium chloride.

37. A composition according to any one of claims 25 to 36, having a pH of approximately 6.8 to approximately 7.

8.

38. The composition according to claim 37, having a pH of 7.3 ± 0.

2.

39. Approximately 0.8×10 １３ vg / ml ~ approx. 5.0 x 10 １３ The composition according to any one of claims 25 to 38, comprising the AAV particles in a concentration of vg / ml.

40. Approximately 2×10 ９ vg / ml ~ approx. 2×10 １０ The composition according to any one of claims 25 to 39, comprising the AAV particles in a concentration of vg / ml.

41. A method for treating a disease or condition caused by excessive excitability of the trigeminal nerve, comprising administering a composition according to any one of claims 25 to 40 to a subject in need thereof.

42. The method according to claim 41, wherein the disease or condition caused by excessive excitability of the trigeminal nerve is trigeminal neuralgia, trigeminal-autonomic headache, episodic cluster headache, chronic cluster headache; afferent pathway deprivation pain of the trigeminal nerve; oral burning syndrome; or post-traumatic trigeminal neuropathic pain.

43. The method according to claim 41, wherein the disease or condition is SUNCT or SUNA.

44. The method according to claim 42, wherein the disease or condition is trigeminal neuralgia.

45. The method according to any one of claims 41 to 44, further comprising administering a low molecular weight agonist to the subject.

46. The method according to claim 45, wherein the low molecular weight agonist is varenicline.

47. The method according to any one of claims 41 to 46, wherein the composition is administered parenterally to the subject.

48. The method according to any one of claims 41 to 46, wherein the low molecular weight agonist is administered orally to the subject.

49. The method according to any one of claims 41 to 46, wherein the composition is administered by transdermal injection.

50. The method according to claim 49, wherein the composition is administered by transdermal injection into the trigeminal ganglion.

51. By percutaneous injection into the trigeminal ganglion, about 6×10 ７ 、2×10 ８ 、6×10 ８ 、2×10 ９ 、6×10 ９ 、2×10 １０ 、6×10 １０ 、2×10 １１ 、6×10 １１ 、2×10 １２ The method according to claim 49 or 50, comprising administering 20 AAV particles.

52. Transcutaneous injection into the trigeminal ganglion delivers approximately 2 x 10⁻¹⁶ ９ ~Approx. 2×10 １１ The method according to any one of claims 49 to 51, comprising administering a certain number of AAV particles.

53. The method according to any one of claims 41 to 52, further comprising orally administering 0.5 mg or 1 mg of varenicline to the subject.

54. The method according to any one of claims 41 to 53, wherein the composition is administered before varenicline.

55. The method according to any one of claims 41 to 53, wherein the composition is administered together with varenicline.

56. The method according to any one of claims 41 to 53, wherein the composition is administered after varenicline.

57. The method according to any one of claims 41 to 56, wherein the subject has acute trigeminal neuralgia.

58. The method according to any one of claims 41 to 56, wherein the subject has chronic trigeminal neuralgia.

59. The method according to any one of claims 41 to 53 or 56 to 58, wherein varenicline is administered about 7 to about 31 days after administration of the composition.

60. The method according to any one of claims 41 to 53, 56 to 58, or 59, wherein varenicline is administered about two weeks after administration of the composition.

61. The method according to any one of claims 41 to 60, wherein the composition is administered once, and varenicline is administered two or more times.

62. The method according to any one of claims 41 to 61, wherein the composition is administered once, and varenicline is administered once daily for about 2 days to about 120 days.

63. The method according to any one of claims 41 to 62, wherein the composition is administered once, and varenicline is administered once daily until the subject no longer experiences trigeminal neuralgia.

64. Approximately 1.85×10 ９ AAV vector genome ~ approximately 1.85 × 10⁻⁶ １１ The method according to any one of claims 41 to 63, comprising administering an AAV vector genome to the subject.

65. The method according to any one of claims 41 to 64, wherein the subject is a human.

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