# US 8,569,474 B2 · Double stranded constructs comprising one or more short strands hybridized to a longer strand

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- Number: US 8,569,474 B2
- Document ID: US8569474B2
- Status: Granted
- Kind: B2
- Application number: 11/072,806
- Filed: 2005-03-04
- Published: 2005-11-03
- Granted: 2013-10-29
- Earliest claimed: 2004-03-08
- Assignees: Isis Pharmaceuticals, Inc.
- Applicants: Brenda F. Baker; Bryan A. Kraynack
- Inventors: Brenda F. Baker; Bryan A. Kraynack
- Web page: https://patentgrep.com/patent/US8569474B2

## CPC

- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/315: Phosphorothioates
- C12N 2310/3183: Diol linkers, e.g. glycols or propanediols
- C12N 2310/321: 2'-O-R Modification
- C12N 2310/322: 2'-R Modification
- C12N 2310/3231: having an additional ring, e.g. LNA, ENA
- C12N 2310/3341: 5-Methylcytosine
- C12N 2310/341: Gapmers, i.e. of the type ===---===
- C12N 2310/346: having a combination of backbone and sugar modifications
- C12N 2310/3519: Fusion with another nucleic acid
- C12N 2310/51: in polymeric form, e.g. multimers, concatemers
- C12N 2330/30: chemically synthesised
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention provides compositions comprising an antisense oligomeric compound hybridized to at least one shorter sense oligomeric compound. The sense oligomeric compounds can be covalently linked to the antisense oligomeric compounds. At least a portion of the antisense oligomeric compound is complementary to and hybridizes with a nucleic acid target. Methods for modulating gene expression are also provided using the compositions of the present invention.

## Claims

### Claim 1

A composition comprising an antisense oligomeric compound and two sense oligomeric compounds wherein:
each of the sense oligomeric compounds is complementary to a separate region of the antisense oligomeric compound and one of the sense oligomeric compounds forms a duplex region with the antisense oligomeric compound starting at the 5′-end of the antisense oligomeric compound and the other of the sense oligomeric compounds forms a duplex region with the antisense oligomeric compound starting at the 3′-end of the antisense oligomeric compound, and the duplex regions are separate and non-overlapping and include each nucleoside of the antisense oligomeric compound; and
the antisense oligomeric compound comprises from 20 to 24 linked nucleosides and each of the sense oligomeric compounds comprises from 10 to 12 linked nucleosides.

### Claim 2 (depends on 1)

The composition of claim 1 wherein the conformational geometry of each of said linked nucleosides is 3′-endo.

### Claim 3 (depends on 2)

The composition of claim 2 wherein each of said nucleosides having 3′-endo conformational geometry comprises a 2′-substituent group.

### Claim 4 (depends on 3)

The composition of claim 3 wherein each 2′-substituent group is, independently, —F, —OH, —O—CH2CH2—O—CH3, —O—CH3, —O—CH2—CH═CH2 or —O—CH2—CH—CH2—NH(Rj) where Rj is H or C1-C10 alkyl.

### Claim 5 (depends on 4)

The composition of claim 4 wherein each of said 2′-substituent groups is, independently, —F, —OH, —O—CH2CH2—O—CH3 or —O—CH3.

### Claim 6 (depends on 2)

The composition of claim 2 wherein at least one of said nucleosides has a 2′-OH group.

### Claim 7 (depends on 2)

The composition of claim 2 wherein each of said nucleosides are modified nucleosides having other than 2′-OH groups.

### Claim 8 (depends on 2)

The composition of claim 2 wherein at least one of said nucleosides is a bicyclic sugar nucleoside having a 2′-O—(CH2)n-4′ bridge wherein n is 1 or 2.

### Claim 9 (depends on 2)

The composition of claim 2 wherein at least one of said nucleosides is a 4′-S sugar modified nucleoside.

### Claim 10 (depends on 2)

The composition of claim 2 wherein each of said linked nucleosides is independently linked by a phosphodiester or a phosphorothioate internucleoside linking group.

### Claim 11 (depends on 1)

The composition of claim 1 wherein at least one of said antisense oligomeric compound and said two sense oligomeric compounds further comprises at least one terminal phosphate group, a terminal stabilizing or capping group, a 3′-overhang or a conjugate group.

### Claim 12 (depends on 11)

The composition of claim 11 wherein the 5′-terminus of said antisense oligomeric compound comprises a 5′-phosphate group.

### Claim 13 (depends on 1)

The composition of claim 1 wherein each of said sense oligomeric compounds is 10 nucleosides in length.

### Claim 14 (depends on 1)

The composition of claim 1 wherein said antisense oligomeric compound is 20 nucleosides in length.

### Claim 15 (depends on 14)

The composition of claim 14 wherein each of said linked nucleosides of said antisense oligomeric compound is linked by a phosphodiester internucleoside linking group.

### Claim 16 (depends on 14)

The composition of claim 14 wherein each of said linked nucleosides of said antisense oligomeric compound independently comprises a 2′-OH or a 2′-F substituent group.

### Claim 17 (depends on 1)

The composition of claim 1 wherein said antisense oligomeric compound is not covalently linked to either of said sense oligomeric compounds.

### Claim 18 (depends on 1)

A method of inhibiting gene expression comprising contacting one or more cells, a tissue or an animal with the composition of claim 1.