OLIGONUCLEOTIDE COMPRISING AN INOSINE FOR TREATING DMD

• US 20170204414A1
• Filed: 03/24/2017
• Published: 07/20/2017
• Est. Priority Date: 04/24/2009
• Status: Active Grant

• Current Assignee: BioMarin Technologies B.V.
• Original Assignee: BioMarin Technologies B.V.
• Inventors: De KIMPE, Josephus Johannes, PLATENBURG, Gerard Johannes, Van DEUTEKOM, Judith Christina Theodora
• Granted Patent: US 10,533,171 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• 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 2310/11 Antisense
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/3181 Peptide nucleic acid, PNA
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/331 Universal or degenerate base
• CPC Class Codes: C12N 2310/333 Modified A
• CPC Class Codes: C12N 2310/336 Modified G
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

The invention provides an oligonucleotide comprising an inosine, and/or a nucleotide containing a base able to form a wobble base pair or a functional equivalent thereof, wherein the oligonucleotide, or a functional equivalent thereof, comprises a sequence which is complementary to at least part of a dystrophin pre-m RNA exon or at least part of a non-exon region of a dystrophin pre-m RNA said part being a contiguous stretch comprising at least 8 nucleotides. The invention further provides the use of said oligonucleotide for preventing or treating DMD or BMD.

Claims

1. An isolated antisense oligonucleotide fully complementary to a contiguous sequence of an of exon of dystrophin pre-mRNA, wherein said contiguous sequence is fully complementary to at least 21 nucleotides of a base sequence, said oligonucleotide comprises at least one position wherein a guanosine base is substituted with an inosine base, wherein said oligonucleotide comprises a modification and is capable of inducing skipping of said exon of human dystrophin pre-mRNA.

2. The oligonucleotide of claim 1, said oligonucleotide being 25-50 nucleotides in length.

3. The oligonucleotide of claim 1, said oligonucleotide being 25-30 nucleotides in length.

4. The oligonucleotide of claim 1, said oligonucleotide comprising from one to four inosine bases.

5. The oligonucleotide of claim 1, further comprising a base and/or sugar modification.

6. The oligonucleotide of claim 1, wherein said oligonucleotide is a 2′

-O-methyl phosphorothioate oligonucleotide.

7. The oligonucleotide of claim 1, wherein said exon is selected from the group consisting of 51, 45, 53, 44, 46, 52, 50, 43 and 55.

8. The oligonucleotide of claim 1, wherein said base sequence is selected from the group consisting of:

SEQ ID NO;

2-473, 539-576.

9. The oligonucleotide of claim 8, further comprising a base and/or sugar modification.

10. The oligonucleotide of claim 1, wherein said oligonucleotide is a 2′

-O-methyl phosphorothioate oligonucleotide.

11. The oligonucleotide of claim 1, which is a locked nucleic acid oligonucleotide (LNA).

12. The oligonucleotide of claim 1, which is a peptide nucleic acid oligonucleotide (PNA).

13. The oligonucleotide of claim 1, which is a morpholino phosphorodiamidate oligonucleotide (PMO).

14. The oligonucleotide of claim 1, further comprising modified internucleoside linkage.

15. A pharmaceutical composition comprising the oligonucleotide of claim 1 and a pharmaceutically acceptable carrier.

16. A method for inducing skipping of an exon of human dystrophin pre-mRNA in a muscle cell, the method comprising contacting said cell with an oligonucleotide of claim 1 for a time and under conditions which permit exon skipping.

17. A method for inducing skipping of an exon of human dystrophin pre-mRNA in a human subject, the method comprising administering an oligonucleotide of claim 1 to said subject in an amount and for a time which is effective to induce exon skipping.

18. A method for alleviating one or more symptom(s) of Duchenne Muscular Dystrophy or Becker Muscular Dystrophy in an individual, the method comprising administering to said individual an oligonucleotide of claim 1.

19. A method for inducing and/or promoting skipping of exon 43, 44, 45, 46, 50, 51, 52 or 53 of the dystrophin pre-mRNA in a patient, the method comprising administering an oligonucleotide of claim 3 to said patient.