# US 9,518,260 B2 · Pain treatment

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- Number: US 9,518,260 B2
- Document ID: US9518260B2
- Status: Granted
- Kind: B2
- Application number: 14/382,206
- Filed: 2013-02-28
- Published: 2015-02-12
- Granted: 2016-12-13
- Earliest claimed: 2012-02-29
- Assignees: Benitec Biopharma Limited
- Applicants: Benitec Biopharma Limited
- Inventors: Peter William French; Michael Wayne Graham
- Web page: https://patentgrep.com/patent/US9518260B2

## CPC

- 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)}
- A61P 25/04: Centrally acting analgesics, e.g. opioids
- C12N 15/1137: {against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138)}
- C12Y 104/03003: D-Amino-acid oxidase (1.4.3.3)
- C12Y 207/11013: Protein kinase C (2.7.11.13)
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/141: MicroRNAs, miRNAs
- C12N 2310/51: in polymeric form, e.g. multimers, concatemers
- C12N 2310/531: Stem-loop; Hairpin
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12Y: ENZYMES

## Abstract

The disclosure relates to an RNA interference (RNAi) agent and the use of that RNAi agent to treat chronic pain in individuals, as well as pharmaceutical compositions containing the RNAi agents of the invention. The DNA-directed RNA interference (ddRNAi) agent for inhibiting expression of one or more target sequences in a pain associated gene comprises, one or more effector sequence sequences and effector complement sequences of at least 17 nucleotides in length, wherein the effector sequence is substantially complementary to the predicted transcript of a target sequence within a pain associated gene.

## Claims

### Claim 1

A DNA-directed RNA interference (ddRNAi) agent for inhibiting expression of one or more target sequences in a pain associated gene, the ddRNAi agent comprising, in a 5′ to 3′ direction:
an effector sequence set forth in SEQ ID NO: 23; and
an effector complement sequence, wherein the effector complement sequence is substantially complementary to the effector sequence.

### Claim 2 (depends on 1)

A ddRNAi agent according to claim 1, wherein the pain associated gene is protein kinase C-γ (PKCγ).

### Claim 3 (depends on 1)

A ddRNAi agent according to claim 1, comprising, in a 5′ to 3′ direction:
a first effector sequence of at least 17 nucleotides in length;
a second effector sequence of at least 17 nucleotides in length;
a second effector complement sequence, wherein the second effector complement sequence is substantially complementary to the second effector sequence; and
a first effector complement sequence, wherein the first effector complement sequence is substantially complementary to the first effector sequence;
wherein one of the first and second effector sequences is the sequence set forth in SEQ ID NO: 23 and the other effector sequence is substantially complementary to a predicted transcript of a region of the pain-associated.

### Claim 4 (depends on 1)

A ddRNAi agent according to claim 1, comprising, in a 5′ to 3′ direction:
a first effector sequence of at least 17 nucleotides in length;
a first effector complement sequence, wherein the first effector complement sequence is substantially complementary to the first effector sequence;
a second effector sequence of at least 17 nucleotides in length; and
a second effector complement sequence, wherein the second effector complement sequence is substantially complementary to the second effector sequence;
wherein one of the first and second effector sequences is the sequence set forth in SEQ ID NO: 23 and the other effector sequence is substantially complementary to a predicted transcript of a region of the pain-associated gene.

### Claim 5 (depends on 1)

A ddRNAi agent according to claim 1, wherein the target sequence in the pain-associated gene is set forth in SEQ ID NO: 5.

### Claim 6 (depends on 1)

A ddRNAi expression cassette for expressing a ddRNAi agent according to claim 1, the expression cassette comprising:
one or more promoter sequences;
one or more DNA sequences that encode for one or more effector sequences; and
one or more DNA sequences that encode for one or more effector complement sequences;
and optionally:
one or more DNA sequences that encode for a loop sequence;
one or more terminator sequences; and/or
one or more enhancer sequences.

### Claim 7 (depends on 6)

A ddRNAi expression cassette according to claim 6 further comprising miRNA expressing sequences, and wherein the ddRNAi agent is expressed within, and as part of, a miRNA structure.

### Claim 8 (depends on 6)

A ddRNAi expression construct comprising a ddRNAi expression cassette according to claim 6.

### Claim 9 (depends on 8)

A method of treating or preventing pain in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of claim 8.

### Claim 10 (depends on 8)

A method of reducing severity of pain in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct of claim 8.

### Claim 11 (depends on 9)

A method according to claim 9, wherein the pain is chronic pain.

### Claim 12 (depends on 9)

A method according to claim 9, wherein the pain is of neuropathic origin.

### Claim 13 (depends on 8)

A pharmaceutical composition comprising a ddRNAi expression construct of claim 8 and a pharmaceutically acceptable carrier or diluent.

### Claim 14 (depends on 6)

A ddRNAi expression construct comprising a ddRNAi expression cassette according to claim 6.

### Claim 15 (depends on 14)

A method of treating or preventing pain, or reducing severity of pain, in a subject comprising administering a therapeutically effective amount of a ddRNAi expression construct according to claim 14.

### Claim 16 (depends on 10)

A method according to claim 10, wherein the pain is chronic pain.

### Claim 17 (depends on 15)

A method according to claim 15, wherein the pain is chronic pain.

### Claim 18 (depends on 10)

A method according to claim 10, wherein the pain is of neuropathic origin.

### Claim 19 (depends on 15)

A method according to claim 15, wherein the pain is of neuropathic origin.

### Claim 20 (depends on 14)

A pharmaceutical composition comprising a ddRNAi expression construct of claim 14 and a pharmaceutically acceptable carrier.

### Claim 21

A DNA-directed RNA interference (ddRNAi) agent for inhibiting expression of one or more target sequences in a pain associated gene, the ddRNAi agent comprising, in a 5′ to 3′ direction:
an effector sequence of at least 17 nucleotides in length; and
an effector complement sequence, wherein the effector complement sequence is substantially complementary to the effector sequence; and
wherein the effector sequence is substantially complementary to a predicted transcript of the target sequence set forth in SEQ ID NO: 5.

### Claim 22 (depends on 21)

A ddRNAi agent according to claim 21, wherein the pain-associated gene is protein kinase C-γ (PKCγ).

### Claim 23 (depends on 21)

A ddRNAi expression cassette for expressing a ddRNAi agent according to claim 21, the expression cassette comprising:
one or more promoter sequences;
one or more DNA sequences that encode for one or more effector sequences; and
one or more DNA sequences that encode for one or more effector complement sequences;
and optionally:
one or more DNA sequences that encode for a loop sequences;
one or more terminator sequences;
one or more enhancer sequences; and/or
one or more miRNA expressing sequences, such that ddRNAi agent is expressed within, and as part of, a miRNA structure.

### Claim 24 (depends on 23)

A ddRNAi expression construct comprising a ddRNAi expression cassette according to claim 23.

### Claim 25 (depends on 24)

A pharmaceutical composition comprising a ddRNAi expression construct of claim 24 and a pharmaceutically acceptable carrier or diluent.