OLIGOMERS

• US 20160024500A1
• Filed: 08/24/2015
• Published: 01/28/2016
• Est. Priority Date: 09/11/2008
• Status: Active Grant

• Current Assignee: Royal Holloway University of London
• Original Assignee: Royal Holloway University of London
• Inventors: POPPLEWELL, Linda, GRAHAM, Ian, DICKSON, John George
• Granted Patent: US 9,650,632 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/115 Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12N 2320/31 Combination therapy
• CPC Class Codes: C12N 2320/35 based on a specific dosage / administration regimen

Abstract

Molecules are provided for inducing or facilitating exon skipping in forming spliced mRNA products from pre-mRNA molecules in cells. The molecules may be provided directly as oligonucleotides or expression products of vectors that are administered to a subject. High rates of skipping can be achieved. High rates of skipping reduce the severity of a disease like Duchene Muscular Dystrophy so that the disease is more like Becker Muscular Dystrophy. This is a severe reduction in symptom severity and mortality.

Claims

1-12. -12. (canceled)

13. A method of ameliorating Duchenne muscular dystrophy, the method comprising administering at least one oligomer to a patient, wherein the at least one oligomer comprises a sequence selected from the group consisting of:

14. The method according to claim 13, wherein at least two oligomers are administered to the patient and each of the at least two oligomers causes skipping of a different exon.

15. The method according to claim 13, wherein at least two oligomers are administered to the patient and each of the at least two oligomers causes skipping of the same exon.

16. The method according to claim 13, wherein the at least one oligomer includes at least one modified nucleotide.

17. The method according to claim 16, wherein the at least one modified nucleotide is modified at the 2′

position of the ribose.

18. The method according to claim 17, wherein the at least one modified nucleotide is a 2′

-O-methyl oligonucleotide.

19. The method according to claim 16, wherein the at least one modified nucleotide is a phosphorodiamidate morpholino oligonucleotide.

20. The method according to claim 13, wherein the at least one oligomer is conjugated to or complexed with a targeting protein.

21. The method according to claim 13, wherein the at least one oligomer is conjugated to or complexed with a drug for treating Duchenne muscular dystrophy.

22. The method according to claim 13, wherein the at least one oligomer is conjugated to or complexed with an arginine-rich cell penetrating peptide.

23. The method according to claim 13, comprising administering the at least one oligomer to a patient by injection.

24. The method according to claim 23, wherein the injection is an intramuscular, intravenous or subcutaneous injection.

25. The method according to claim 13, comprising administering the at least one oligomer to a patient by oral administration.

26. The method according to claim 25, wherein the oral administration includes administration using a capsule, a tablet, an aqueous suspension and/or an aqueous solution.

27. The method according to claim 13, wherein administration of the at least one oligomer results in increased expression of a truncated dystrophin protein in a muscle of the patient.

28. A vector for ameliorating Duchenne muscular dystrophy, the vector encoding an oligomer comprising a sequence selected from the group consisting of:

29. The vector according to claim 28, wherein the oligomer can bind to a target site to cause exon skipping of an exon of the dystrophin gene at a rate of at least 50%.

30. The vector according to claim 28, wherein the oligomer is between 25 and 35 bases in length.

31. The vector according to claim 28, wherein the oligomer is 30 bases in length.

32. A pharmaceutical composition for ameliorating Duchenne muscular dystrophy, the composition comprising the vector according to claim 28 or the oligonucleotide encoded by the vector according to claim 28, and a pharmaceutically acceptable carrier, adjuvant or vehicle.

33. A pharmaceutical composition for ameliorating Duchenne muscular dystrophy, the composition comprising a plurality of vectors encoding oligomers including the vector according to claim 28 and/or a plurality of oligomers including the oligomer encoded by the vector according to claim 28, or a combination of the oligomers and vectors, wherein the each oligomer encoded by a vector in the plurality of vectors causes skipping of an exon, and each oligomer in the plurality of oligomers in the pharmaceutical composition cause causes skipping of an exon.

34. The pharmaceutical composition according to claim 33, wherein each oligomer encoded by a vector in the plurality of vectors causes skipping of a different exon, and each oligomer in the plurality of oligomers in the pharmaceutical composition cause causes skipping of a different exon.

35. A pharmaceutical composition comprising at least one oligomer, wherein the at least one oligomer comprises a sequence selected from the group consisting of:

36. The pharmaceutical composition according to claim 35, wherein the pharmaceutical composition includes at least two oligomers, and the at least two oligomers each cause skipping of a different exon.

37. The pharmaceutical composition according to claim 35, wherein the pharmaceutical composition includes at least two oligomers, and the at least two oligomers cause skipping of the same exon.

38. The pharmaceutical composition according to claim 35, wherein the at least one modified nucleotide is modified at the 2′

position of the ribose.

39. The pharmaceutical composition according to claim 35, wherein the at least one modified nucleotide is 2′

-O-methyl oligonucleotide.

40. The pharmaceutical composition according to claim 35, wherein the at least one modified nucleotide is a phosphorodiamidate morpholino oligonucleotide.

41. The pharmaceutical composition according to claim 35, wherein the at least one oligomer is conjugated to or complexed with a targeting protein.

42. The pharmaceutical composition according to claim 35, wherein the at least one oligomer is conjugated to or complexed with a drug for treating Duchenne muscular dystrophy.

43. The pharmaceutical composition according to claim 35, wherein the at least one oligomer is conjugated to or complexed with an arginine-rich cell penetrating peptide.

44. The pharmaceutical composition according to claim 35, wherein the pharmaceutical composition is in the form of a sterile injectable preparation.

45. An oligomer comprising a sequence selected from the group consisting of:

46. The oligomer according to claim 45, wherein the oligomer can bind to a target site to cause exon skipping of an exon of the dystrophin gene at a rate of at least 50%.

47. The oligomer according to claim 45, wherein the oligomer can bind to a target site to cause exon skipping of an exon of the dystrophin gene at a rate of at least 70%.

48. The oligomer according to claim 45, wherein the oligomer can bind to a target site to cause exon skipping of an exon of the dystrophin gene at a rate of at least 90%.

49. The oligomer according to claim 45, wherein the at least one modified nucleotide is modified at the 2′

position of the ribose.

50. The oligomer according to claim 49, wherein the at least one modified nucleotide is 2′

-O-methyl oligonucleotide.

51. The oligomer according to claim 45, wherein the at least one modified nucleotide is a phosphorodiamidate morpholino oligonucleotide.

52. The oligomer according to claim 45, wherein the oligomer is at least 28 and at most 32 bases in length.

53. The oligomer according to claim 52, wherein the oligomer is at least 29 and at most 31 bases in length.

54. The oligomer according to claim 45, wherein the oligomer is conjugated to or complexed with a targeting protein.

55. The oligomer according to claim 45, wherein the oligomer is conjugated to or complexed with a drug for treating Duchenne muscular dystrophy.

56. The oligomer according to claim 45, wherein the oligomer is conjugated to or complexed with an arginine-rich cell penetrating peptide.