Antisense oligonucleotide directed removal of proteolytic cleavage sites, the HCHWA-D mutation, and trinucleotide repeat expansions

• US 10,590,421 B2
• Filed: 04/21/2017
• Issued: 03/17/2020
• Est. Priority Date: 08/05/2010
• Status: Active Grant

• Current Assignee: Academisch Ziekenhuis Leiden H.O.D.N. Lumc
• Original Assignee: Academisch Ziekenhuis Leiden H.O.D.N. Lumc
• Inventors: van Roon-Mom, Wilhelmina M. C., Evers, Melvin Maurice, Pepers, Barry Antonius, Aartsma-Rus, Annemieke, Van Ommen, Garrit-Jan Boudewijn
• Primary Examiner(s): Hill, Kevin K
• Application Number:
  US15/494,052
• Publication Number:
  US 20170226521A1
• Time in Patent Office: 1,061 Days
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/33 Alteration of splicing
• CPC Class Codes: C12Y 304/19012 Ubiquitinyl hydrolase 1 (3.4.19.12)

Abstract

Described are methods for removing a proteolytic cleavage site, the HCHWA-D mutation or the amino acids encoded by a trinucleotide repeat expansion from a protein comprising providing a cell that expresses pre-mRNA encoding the protein with an anti-sense oligonucleotide that induces skipping of the exonic sequence that comprises the proteolytic cleavage site, HCHWA-D mutation or trinucleotide repeat expansion, respectively, the method further comprising allowing translation of mRNA produced from the pre-mRNA.

Claims

1. At least one isolated oligonucleotide of between 14-40 nucleotides that induces skipping of an exonic sequence that comprises the HCHWA-D mutation from mutant human APP protein, wherein the oligonucleotide induces skipping of an exonic sequence corresponding to exon 16 of APP751, wherein the oligonucleotide is selected from the group consisting of SEQ ID NO:

20, SEQ ID NO;

22, SEQ ID NO;

24, SEQ ID NO;

26, SEQ ID NO;

28, and SEQ ID NO;

30.

2. The at least one isolated oligonucleotide of claim 1, wherein the oligonucleotide is SEQ ID NO:

20.

3. The at least one isolated oligonucleotide of claim 1, wherein the oligonucleotide is SEQ ID NO:

22.

4. The at least one isolated oligonucleotide of claim 1, wherein the oligonucleotide is SEQ ID NO:

24.

5. The at least one isolated oligonucleotide of claim 1, wherein the oligonucleotide is SEQ ID NO:

26.

6. The at least one isolated oligonucleotide of claim 1, wherein the oligonucleotide is SEQ ID NO:

28.

7. The at least one isolated oligonucleotide of claim 1, wherein the oligonucleotide is SEQ ID NO:

30.