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1. WO2020118115 - COMPOSITIONS ET MÉTHODES POUR TRAITER UNE DÉFICIENCE EN ORNITHINE TRANSCARBAMYLASE

Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

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CLAIMS

What is claimed is:

1. An ornithine transcarbamylase (OTC) protein comprising the amino acid sequence of SEQ ID NO:4 and having OTC enzymatic activity.

2. A polynucleotide encoding the protein of claim 1.

3. The polynucleotide of claim 2 comprising an optimized coding region encoding SEQ ID NO: 4.

4. The polynucleotide of claim 3, wherein the polynucleotide is DNA.

5. The polynucleotide of claim 3, wherein the polynucleotide is an mRNA.

6. The polynucleotide of claim 5 comprising a 3’poly A tail.

7. The polynucleotide of claim 6, wherein the 3’ poly A tail comprises from about 60 sequential adenine nucleotides to about 125 sequential adenine nucleotides.

8. The polynucleotide of claim 7, wherein the 3’ poly A tail comprises from about 100 sequential adenine nucleotides to about 125 sequential adenine nucleotides.

9. The polynucleotide of claim 5 comprising a non-human 5’ untranslated region (5’UTR).

10. The polynucleotide of claim 9, wherein the 5’UTR is derived from a gene expressed by Arabidopsis thaliana.

11. The polynucleotide of claim 10, wherein the 5’ UTR comprises SEQ ID NO: 6, SEQ ID NOS: 125-127 or SEQ ID NOS: 227-247.

12. The polynucleotide of claim 11, wherein the 5’UTR comprises SEQ ID NO: 6.

13. The polynucleotide of claim 5 comprising a 3’ UTR.

14. The polynucleotide of claim 13, wherein the 3’ UTR is selected from the group consisting of: SEQ ID NOS: 16-22.

15. The polynucleotide of claim 5 comprising a Kozak sequence of SEQ ID NO: 23 or a partial Kozak sequence of SEQ ID NO: 24.

16. The polynucleotide of claim 5 comprising a 5’ cap.

17. The polynucleotide of claim 16, wherein the 5’ cap is m7GpppGm having the following structure:


18. The polynucleotide of claim 4, wherein the optimized coding region comprises SEQ ID NO: 221.

19. The polynucleotide of claim 5 comprising SEQ ID NO: 119.

20. The polynucleotide of claim 5, wherein the percentage of uracil nucleobases in the coding region of the polynucleotide is reduced with respect to the percentage of uracil nucleobases in the wild-type OTC nucleic acid sequence.

21. The polynucleotide of claim 5, wherein 1-100% of the uridines are modified uridine analogs.

22. The polynucleotide of claim 21, wherein 100% of the uridine nucleotides are 5-methoxy undine.

23. The polynucleotide of claim 22, wherein 100% of the uridine nucleotides are N1-methylpseudouridine.

24. The polynucleotide of claim 5, comprising SEQ ID NO: 251 or SEQ ID NO: 252.

25. The polynucleotide of claim 19 or 24, wherein 1-100% of the uridines are modified uridine analogs.

26. The polynucleotide of claim 19 or 24, wherein 100% of the uridine nucleotides are 5-metlioxyuridme.

27. The polynucleotide of claim 19 or 24, wherein 100% of the uridine nucleotides are N1-methylpseudouridine.

28. A pharmaceutical composition comprising the polynucleotide of claim 19 or 24.

29. A pharmaceutical composition comprising the polynucleotide of claim 25.

30. A pharmaceutical composition comprising the polynucleotide of claim 26.

31. A pharmaceutical composition comprising the polynucleotide of claim 27.

32. The pharmaceutical composition of claim 28 comprising a pharmaceutically acceptable carrier wherein the pharmaceutically acceptable carrier is a lipid formulation.

33. The pharmaceutical composition of claim 29 comprising a pharmaceutically acceptable carrier wherein the pharmaceutically acceptable carrier is a lipid formulation.

34. The pharmaceutical composition of claim 30 comprising a pharmaceutically acceptable carrier wherein the pharmaceutically acceptable carrier is a lipid formulation.

35. The pharmaceutical composition of claim 31 comprising a pharmaceutically acceptable carrier wherein the pharmaceutically acceptable carrier is a lipid formulation.

36. A method of treating OTC deficiency in a patient identified as suffering from OTC deficiency, comprising administering to the patient, a pharmaceutical composition of any one of claims 28-35, wherein upon administration of the pharmaceutical composition to the patient, the protein of SEQ ID NO: 4 is expressed in the patient.

37. A method of treating OTC deficiency in a patient identified as suffering from OTC deficiency, comprising administering to the patient a polynucleotide of claim 5, wherein the polynucleotide expresses the protein of SEQ ID NO: 4 in the patient.

38. A vector comprising a polynucleotide of claim 4.

39. A host cell transformed with a vector of claim 38.

40. An mRNA encoding an OTC protein having the amino acid sequence of SEQ ID NO: 4, wherein the mRNA has the polynucleotide sequence of SEQ ID NO: 119, SEQ ID NO:

251 or SEQ ID NO: 252 and wherein the encoded protein has OTC enzymatic activity.

41. The mRNA of claim 40, wherein 1-100% of the uridines are modified uridine analogs.

42. The mRNA of claim 41, wherein 100% of the uridine nucleotides are 5-methoxyundine.

43. The mRNA of claim 41, wherein 100% of the uridine nucleotides are N1-methylpseudouridine.

44. A pharmaceutical composition comprising the mRNA of any one claims 40, 41, 42, or 43.

45. The pharmaceutical composition of claim 44 comprising a pharmaceutically acceptable carrier wherein the pharmaceutically acceptable carrier is a lipid formulation.

46. A method of treating OTC deficiency in a patient identified as suffering from OTC deficiency comprising, administering to the patient an mRNA of claim 40, wherein the mRNA expresses the protein of SEQ ID NO: 4 in the patient.

47. An mRNA comprising a codon-optimized coding region encoding a human ornithine transcarbamylase (OTC) protein of SEQ ID NO: 3 and having OTC protein enzymatic activity wherein the mRNA comprises a 5’UTR derived from a gene expressed by

Arabidopsis thaliana and wherein the mRNA is selected from the group consisting of SEQ ID NOS: 62, 67, 68, 69, 73, 113-118, and 121-124.

48. The mRNA of claim 47 comprising a 3’Poly A tail wherein the 3’ Poly A tail comprises from about 100 nucleotides to about 125 nucleotides.

49. The mRNA of claim 47, wherein 1-100% of the uridine nucleotides are modified uridine analogs.

50. The mRNA of claim 49, wherein the modified uridine analogs are independently selected from 5-methoxyuridine and N'-methyl pseudouridine.

51. The mRNA of claim 50, wherein 100% of the uridine nucleotides are 5-methoxyuridine.

52. The mRNA of claim 50, wherein 100% of the uridine nucleotides are N1-methylpseudouridine.

53. A pharmaceutical composition comprising an mRNA of any one of claims 47-52.

54. The pharmaceutical composition of claim 53 comprising a pharmaceutically acceptable carrier wherein the pharmaceutically acceptable carrier is a lipid formulation.

55. A method of treating OTC deficiency in a patient identified as suffering from OTC deficiency, comprising administering to the patient, a pharmaceutical composition of claim 54, wherein upon administration of the pharmaceutical composition to the patient, the human OTC protein of SEQ ID NO: 3 is expressed in the patient.

56. A method of treating OTC deficiency in a patient identified as suffering from OTC deficiency, comprising administering to the patient an mRNA of any one of claims 47-52, wherein the mRNA expresses the human OTC protein of SEQ ID NO: 3 in the patient.

57. The mRNA of claim 47, wherein the mRNA comprises SEQ ID NO: 73.

58. The mRNA of claim 57 comprising a 3’ Poly A tail wherein the 3’ Poly A tail comprises from about 60 nucleotides to about 125 nucleotides.

59. The mRNA of claim 58, wherein the 3’ Poly A tail comprises from about 100 nucleotides to about 125 nucleotides.

60. The mRNA of claim 57, wherein 1-100% of the uridine nucleotides are modified uridine analogs.

61. The mRNA of claim 60, wherein the modified uridine analogs are independently selected from the group consisting of: 5 -methoxy uridine and N '-methyl pseudouridine.

62. The mRNA of claim 61, wherein 100% of the uridine nucleotides are 5-methoxyuridine

63. The mRNA of claim 61, wherein 100% of the uridine nucleotides are N1-methylpseudouridine.

64. A pharmaceutical composition comprising an mRNA of any one of claims 57-63.

65. The pharmaceutical composition of claim 64 comprising a pharmaceutically acceptable carrier wherein the pharmaceutically acceptable carrier is a lipid formulation.

66. A method of treating OTC deficiency in a patient identified as suffering from OTC deficiency, comprising administering to the patient, a pharmaceutical composition of claim 65, wherein upon administration of the pharmaceutical composition to the patient, the protein of SEQ ID NO: 3 is expressed in the patient.

67. A method of treating OTC deficiency in a patient identified as suffering from OTC deficiency, comprising administering to the patient an mRNA of any one of claims 57-63, wherein the mRNA expresses the protein of SEQ ID NO: 3 in the patient.

68. The mRNA of claim 53, wherein the mRNA comprises SEQ ID NO: 253.

69. The mRNA of claim 68, wherein 1-100% of the uridine nucleotides are modified uridine analogs.

70. The mRNA of claim 69, wherein the modified uridine analogs are independently selected from the group consisting of: 5 -methoxy uridine and N '-methyl pseudouridine.

71. The mRNA of claim 70, wherein 100% of the uridine nucleotides are 5-methoxyundine.

72. The mRNA of claim 70, wherein 100% of the uridine nucleotides are N1-methylpseudouridine.

73. A pharmaceutical composition comprising an mRNA of any one of claims 68-72.

74. The pharmaceutical composition of claim 73 comprising a pharmaceutically acceptable carrier wherein the pharmaceutically acceptable carrier is a lipid formulation.

75. A method of treating OTC deficiency in a patient identified as suffering from OTC deficiency, comprising administering to the patient, a pharmaceutical composition of claim 74, wherein upon administration of the pharmaceutical composition to the patient, the protein of SEQ ID NO: 3 is expressed in the patient.

76. A method of treating OTC deficiency in a patient identified as suffering from OTC deficiency, comprising administering to the patient an mRNA of any one of claims 68-72, wherein the mRNA expresses the protein of SEQ ID NO: 3 in the patient.