dsRNA molecules comprising oligonucleotide analogs having modified intersubunit linkages and/or terminal groups

• US 10,017,763 B2
• Filed: 09/02/2011
• Issued: 07/10/2018
• Est. Priority Date: 09/03/2010
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: Iversen, Patrick L.
• Primary Examiner(s): Lau, Jonathan S
• Application Number:
  US13/819,634
• Publication Number:
  US 20130288369A1
• Time in Patent Office: 2,503 Days
• 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/344 Position-specific modifications, e.g. on every purine, at the 3'-end
• CPC Class Codes: C12N 2310/3513 Protein; Peptide

Abstract

Morpholino oligomers containing modified intersubunit linkages and/or terminal groups are provided for use within dsRNA molecules. The oligomers are oligonucleotide analogs containing predetermined sequences of base-pairing moieties. Also provided are such oligomers conjugated to peptide transporter moieties, where the transporters are preferably composed of arginine subunits, or arginine dimers, alternating with neutral amino acid subunits.

Claims

1. A double-stranded ribonucleic acid (dsRNA) molecule for inhibiting the expression of a target gene in a cell, wherein said dsRNA has a length of about 10 to 30 base pairs and comprises at least two sequences that are complementary to each other and wherein a sense strand comprises a first sequence and an antisense strand comprises a second sequence comprising a region of complementarity which is substantially complementary to at least a part of a target gene mRNA, wherein said region of complementarity is less than about 30 nucleotides in length, wherein one or both of the sense or antisense strands comprise an oligomer backbone comprising at least one morpholino subunit joined by intersubunit linkages, wherein each morpholino subunit supports a base-pairing moiety, and wherein the intersubunit linkages comprise a structure selected from the group consisting of (A) and (B) below:

(A) an intersubunit linkage having the following structure (A-I);

2. The dsRNA molecule of claim 1, wherein said morpholino subunits have the following structure:

3. The dsRNA molecule of claim 1, wherein each of R¹ and R², in linkages of type (a1), is methyl.

4. The dsRNA molecule of claim 1, wherein said oligomer includes at least two consecutive linkages of type (a1).

5. The dsRNA molecule of claim 1, having a length of about 15 to 25 base pairs.

6. The dsRNA molecule of claim 1, wherein said cationic linkages are of type (b1), where each R is H, R⁴ is H, CH₃, or an electron pair, and R³ is selected from H, CH₃, C(═

NH)NH₂, and C(O)-L-NHC(═

NH)NH₂.

7. The dsRNA molecule of claim 1, wherein said cationic linkages are of type (b1), where each R is H, and each of R³ and R⁴ is independently H or CH₃.

8. The dsRNA molecule of claim 7, wherein each of R³ and R⁴ is H.

9. The dsRNA molecule of claim 1, wherein said cationic linkages are of type (b2), where L is a linker up to 12 atoms in length having bonds selected from alkyl and alkylamino.

10. The dsRNA molecule of claim 1, wherein W and Y are each O.

11. The dsRNA molecule of claim 1, wherein linkage (b4) has one of the following structures (B-III), (B-IV), (B-V) or (B-VI):

12. The dsRNA molecule of claim 1, wherein one of the sense and antisense strands has the following structure (C-I):

13. A method of inhibiting production of a protein, comprising exposing a nucleic acid encoding the protein to a dsRNA molecule of claim 1, wherein the nucleic acid encoding the protein comprises the at least a part of the target gene mRNA.