Oligomers

• US 10,457,944 B2
• Filed: 04/10/2018
• Issued: 10/29/2019
• Est. Priority Date: 09/11/2008
• Status: Active Grant

• Current Assignee: Royal Holloway University of London (University of London)
• Original Assignee: Royal Holloway University of London (University of London)
• Inventors: Popplewell, Linda, Graham, Ian, Dickson, John George
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US15/949,405
• Publication Number:
  US 20190112601A1
• Time in Patent Office: 567 Days
• Field of Search: None
• 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. A method of ameliorating Duchenne muscular dystrophy, the method comprising administering an oligomer to a patient, wherein the oligomer comprises at least 25 nucleotides and has a nucleotide sequence that is:

a) identical to at least 25 contiguous nucleotides of CXG XXG CCX CCG GXX CXG AAG GXG XXC XXG (SEQ ID NO;

     10);

or

b) identical to at least 25 contiguous nucleotides of XXG CCX CCG GXX CXG AAG GXG XXC XXG XAC (SEQ ID NO;

     12),

wherein X=U or T.

2. The method according to claim 1, wherein the oligomer comprises at least 25 nucleotides and has a nucleotide sequence that is identical to at least 25 contiguous nucleotides of CXG XXG CCX CCG GXX CXG AAG GXG XXC XXG (SEQ ID NO:

10).

3. The method according to claim 1, wherein the oligomer comprises at least 25 nucleotides and has a nucleotide sequence that is identical to at least 25 contiguous nucleotides of XXG CCX CCG GXX CXG AAG GXG XXC XXG XAC (SEQ ID NO:

12).

4. The method according to claim 1, wherein the oligomer is between 25 and 30 nucleotides in length.

5. The method according to claim 1, wherein the oligomer is 25 nucleotides in length.

6. The method according to claim 1, wherein the oligomer is 30 nucleotides in length.

7. The method according to claim 1, wherein the method includes administering at least two oligomers and each of the at least two oligomers causes skipping of a different exon.

8. The method according to claim 1, wherein the method includes administering at least two oligomers and each of the at least two oligomers causes skipping of the same exon.

9. The method according to claim 1, wherein the oligomer includes at least one modified nucleotide.

10. The method according to claim 9, wherein the at least one modified nucleotide is modified at the 2′

position of the ribose.

11. The method according to claim 10, wherein the at least one modified nucleotide is a 2′

-O-methyl oligonucleotide.

12. The method according to claim 9, wherein the at least one modified nucleotide is a phosphorodiamidate morpholino oligonucleotide.

13. The method according to claim 1, wherein the oligomer is conjugated to or complexed with a targeting protein, wherein the targeting protein targets the at least one oligomer to muscle tissue.

14. The method according to claim 1, wherein the oligomer is conjugated to or complexed with an arginine-rich cell penetrating peptide.

15. The method according to claim 1, comprising administering the oligomer to a patient by injection.

16. The method according to claim 15, wherein the injection is an intramuscular, intravenous or subcutaneous injection.

17. The method according to claim 1, comprising administering the oligomer to a patient by oral administration.

18. The method according to claim 17, wherein the oral administration includes administration using a capsule, a tablet, an aqueous suspension and/or an aqueous solution.

19. The method according to claim 1, wherein administration of the oligomer results in increased expression of a truncated dystrophin protein in a muscle of the patient as compared to expression of the truncated dystrophin protein in the muscle of the patient prior to administration of the oligomer.