ENA nucleic acid pharmaceuticals capable of modifying splicing of mRNA precursors

• US 9,657,050 B2
• Filed: 09/18/2015
• Issued: 05/23/2017
• Est. Priority Date: 11/25/2002
• Status: Active Grant

• Current Assignee: Daiichi Sankyo Company Limited
• Original Assignee: Daiichi Sankyo Company Limited, Matsuo Masafumi, Orphan Disease Treatment Institute Co., Ltd., Takeshima Yasuhiro
• Inventors: Matsuo, Masafumi, Takeshima, Yasuhiro, Koizumi, Makoto
• Primary Examiner(s): SCHNIZER, RICHARD A
• Application Number:
  US14/858,404
• Publication Number:
  US 20160002636A1
• Time in Patent Office: 613 Days
• Field of Search: None
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• 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 21/04 for myasthenia gravis
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• 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 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

Oligonucleotides having a nucleotide sequence complementary to nucleotide numbers such as 2571-2607, 2578-2592, 2571-2592, 2573-2592, 2578-2596, 2578-2601 or 2575-2592 of the dystrophin cDNA (Gene Bank accession No. NM_004006.1) and therapeutic agents for muscular dystrophy comprising such oligonucleotides.

Claims

1. An oligonucleotide having the nucleotide sequence as shown in SEQ ID NO:

75 or 77 in the SEQUENCE LISTING, or a pharmacologically acceptable salt thereof, wherein at least one sugar constituting the oligonucleotide is modified and/or at least one phosphate constituting the oligonucleotide is modified.

2. The oligonucleotide of claim 1, wherein the oligonucleotide has the nucleotide sequence as shown in SEQ ID NO:

75 in the SEQUENCE LISTING, or a pharmacologically acceptable salt thereof, wherein at least one sugar constituting the oligonucleotide is modified and/or at least one phosphate constituting the oligonucleotide is modified.

3. The oligonucleotide of claim 1, wherein the oligonucleotide has the nucleotide sequence as shown in SEQ ID NO:

77 in the SEQUENCE LISTING, or a pharmacologically acceptable salt thereof, wherein at least one sugar constituting the oligonucleotide is modified and/or at least one phosphate constituting the oligonucleotide is modified.

4. The oligonucleotide or a pharmacologically acceptable salt thereof according to claim 1, wherein at least one of sugar constituting the oligonucleotide is modified, the sugar constituting the oligonucleotide is D-ribofuranose, and the modification of the sugar is modification of the hydroxyl group at position 2′

of D-ribofuranose.

5. The oligonucleotide or a pharmacologically acceptable salt thereof according to claim 1, wherein the modification of the sugar is 2′

-O-alkylation and/or 2′

-O,4′

-C-alkylenation of the D-ribofuranose.

6. The oligonucleotide or a pharmacologically acceptable salt thereof according to claim 1, wherein at least one of sugar constituting the oligonucleotide is modified, and said modification is selected from the group consisting of 2′

-O-methylation of D-ribofuranose, 2′

-O-aminoethylation of D-ribofuranose, 2′

-O-propylation of D-ribofuranose, 2′

-O-allylation of D-ribofuranose, 2′

-O-methoxyethylation of D-ribofuranose, 2′

-O-butylation of D-ribofuranose, 2′

-O-pentylation of D-ribofuranose, 2′

-O-propargylation of D-ribofuranose, 2′

-O,4′

-C-ethylenation of D-ribofuranose, 2′

-O,4′

-C-methylenation of D-ribofuranose, 2′

-O,4′

-C-propylenation of D-ribofuranose, 2′

-O,4′

-C-tetramethylation of D-ribofuranose, 2′

-O,4′

-C-pentamethylation of D-ribofuranose, 3′

-deoxy-3′

-amino-2′

-deoxy-D-ribofuranose, and 3′

-deoxy-3′

-amino-2′

-deoxy-2′

-fluoro-D-ribofuranose.

7. The oligonucleotide or a pharmacologically acceptable salt thereof according to claim 1, wherein at least one of phosphate constituting the oligonucleotide is modified and the modification of the phosphate is thioation of the phosphate group.

8. The oligonucleotide or a pharmacologically acceptable salt thereof according to claim 1, wherein at least one sugar constituting the oligonucleotide is modified and at least one phosphate constituting the oligonucleotide is modified.

9. The oligonucleotide or a pharmacologically acceptable salt thereof according to claim 1, wherein at least one sugar constituting the oligonucleotide is modified.

10. The oligonucleotide or a pharmacologically acceptable salt thereof according to claim 1, wherein at least one phosphate constituting the oligonucleotide is modified.

11. An oligonucleotide having the nucleotide sequence as shown in SEQ ID NO:

23, 214, or 215 in the SEQUENCE LISTING, or a pharmacologically acceptable salt thereof, wherein at least one D-ribofuranose constituting the oligonucleotide is 2′

-O,4′

-C-ethylenated, wherein;

the oligonucleotide of SEQ ID NO;

23 has the structure of AO90 or AO91;

AO90;

HO-C^e2p-U^mp-G^mp-C^e2p-U^mp-U^mp-C^e2p-C^e2p-U^mp-C^e2p-C^e2p-A^mp-A^mp-C^e2p-C^e2p-CH₂CH₂OH

AO91;

HO-C^e2s-U^ms-G^ms-C^e2s-U^ms-U^ms-C^e2s-C^e2s-U^ms-C^e2s-C^e2s-A^ms-A^ms-C^e2s-C^e2s-CH₂CH₂OH

the oligonucleotide of SEQ ID NO;

214 has the structure of AO27, AO89, AO92, or AO93;

AO27;

HO-C^e2p-T^e2p-G^e2p-C^e2p-T^e2p-U^mp-C^mp-C^mp-U^mp-C^mp-C^e2p-A^e2p-A^e2p-C^e2p-C^e2p-CH₂CH₂OH

AO89;

HO-C^e2s-T^e2s-G^e2s-C^e2s-T^e2s-U^ms-C^ms-C^ms-U^ms-C^ms-C^e2s-A^e2s-A^e2s-C^e2s-C^e2s-CH₂CH₂OH

AO92;

HO-C^e2p-T^e2p-G^mp-C^e2p-T^e2p-U^mp-C^mp-C^e2p-U^mp-C^mp-C^e2p-A^mp-A^mp-C^e2p-C^e2p-CH₂CH₂OH

AO93;

HO-C^e2s-T^e2s-G^ms-C^e2s-T^e2s-U^ms-C^ms-C^e2s-U^ms-C^ms-C^e2s-A^ms-A^ms-C^e2s-C^e2s-CH₂CH₂OH

the oligonucleotide of SEQ ID NO;

215 has the structure of AO28;

AO28;

HO-G^e2p-T^e2p-T^e2p-A^e2p-T^e2p-C^mp-U^mp-G^mp-C^mp-U^mp-U^mp-C^mp-C^mp-U^mp-C^mp-C^e2p-A^e2p-A^e2p-C^e2p-C^e2p-CH₂CH₂OH

wherein A^e2p, G^e2p, C^e2p, T^e2p, A^mp, G^mp, C^mp, U^mp, A^e2s, G^e2s, C^e2s, T^e2s, A^ms, G^ms, C^ms, and U^ms have the following structures;

12. The oligonucleotide of claim 11, wherein the oligonucleotide has the nucleotide sequence AO90, or a pharmacologically acceptable salt thereof, wherein at least one D-ribofuranose constituting the oligonucleotide is 2′

-O,4′

-C-ethylenated.

13. The oligonucleotide of claim 11, wherein the oligonucleotide has the nucleotide sequence AO91, or a pharmacologically acceptable salt thereof, wherein at least one D-ribofuranose constituting the oligonucleotide is 2′

-O,4′

-C-ethylenated.

14. The oligonucleotide of claim 11, wherein the oligonucleotide has the nucleotide sequence AO27, or a pharmacologically acceptable salt thereof, wherein at least one D-ribofuranose constituting the oligonucleotide is 2′

-O,4′

-C-ethylenated.

15. The oligonucleotide of claim 11, wherein the oligonucleotide has the nucleotide sequence AO89, or a pharmacologically acceptable salt thereof, wherein at least one D-ribofuranose constituting the oligonucleotide is 2′

-O,4′

-C-ethylenated.

16. The oligonucleotide of claim 11, wherein the oligonucleotide has the nucleotide sequence AO92, or a pharmacologically acceptable salt thereof, wherein at least one D-ribofuranose constituting the oligonucleotide is 2′

-O,4′

-C-ethylenated.

17. The oligonucleotide of claim 11, wherein the oligonucleotide has the nucleotide sequence AO93, or a pharmacologically acceptable salt thereof, wherein at least one D-ribofuranose constituting the oligonucleotide is 2′

-O,4′

-C-ethylenated.

18. The oligonucleotide of claim 11, wherein the oligonucleotide has the nucleotide sequence AO28, or a pharmacologically acceptable salt thereof, wherein at least one D-ribofuranose constituting the oligonucleotide is 2′

-O,4′

-C-ethylenated.