# US 2013/0158097 A1 · Compositions and methods directed to treating liver fibrosis

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- Number: US 2013/0158097 A1
- Document ID: US20130158097A1
- Status: Published application
- Kind: A1
- Application number: 13/701,551
- Filed: 2011-06-02
- Published: 2013-06-20
- Earliest claimed: 2010-06-02
- Assignees: Alnylam Pharmaceuticals, Inc.
- Inventors: Greg Hinkle; Victor Kotelianski; Brian Bettencourt; Alfica Sehgal; Tatiana Novobrantseva
- Web page: https://patentgrep.com/patent/US20130158097A1

## CPC

- C07H 21/02: with ribosyl as saccharide radical
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 19/04: for non-specific disorders of the connective tissue
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 14/4705: {stimulating, promoting or activating activity}
- C07K 14/495: Transforming growth factor [TGF]
- C07K 14/78: Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
- 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)}
- C12N 2310/14: interfering nucleic acids [NA]
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The invention relates to double-stranded ribonucleic acid (dsRNA) compositions targeting the COL1A1, TGF-β, and SMAD2/3 genes, and methods of using such dsRNA compositions to inhibit expression of COL1A1, TGF-β, and SMAD2/3.

## Claims

### Claim 1

A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of COL1A1, wherein said dsRNA comprises a sense strand and an antisense strand, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: 53 and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the corresponding antisense nucleotide sequence of SEQ ID NO: 54.

### Claim 2

A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of COL1A1, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a COL1A1 RNA transcript, which antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from one of the antisense sequences of SEQ ID NOs: 1-62 or SEQ ID NOs: 185-2792.

### Claim 3

(canceled)

### Claim 4

(canceled)

### Claim 5

A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of TGFβ, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a TGFβ RNA transcript, which antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from one of the antisense sequences of SEQ ID NOs: 63-130 or SEQ ID NOs: 2793-2860.

### Claim 6

(canceled)

### Claim 7

A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of SMAD2/3, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a SMAD2/3 RNA transcript, which antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from one of the antisense sequences of SEQ ID NOs: 132-167.

### Claim 8 (depends on 2)

The dsRNA of claim 2, wherein said dsRNA comprises at least one modified nucleotide.

### Claim 9 (depends on 8)

The dsRNA of claim 8, wherein at least one of said modified nucleotides is chosen from the group consisting of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group, a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.

### Claim 10

(canceled)

### Claim 11 (depends on 2)

The dsRNA of claim 2, wherein the region of complementarity is 19 to 21 nucleotides in length.

### Claim 12 (depends on 2)

The dsRNA of claim 2, wherein each strand is no more than 30 nucleotides in length.

### Claim 13 (depends on 2)

The dsRNA of claim 2, wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.

### Claim 14 (depends on 2)

The dsRNA of claim 2, further comprising a ligand.

### Claim 15

(canceled)

### Claim 16 (depends on 2)

The dsRNA of claim 2, wherein the region of complementarity consists of one of the antisense sequences of SEQ ID NOs: 1-62 or SEQ ID NOs: 185-2792.

### Claim 17 (depends on 2)

The dsRNA of claim 2, wherein the dsRNA comprises a sense strand consisting of a sense strand sequence selected from SEQ ID NOs: 1-62 or SEQ ID NOs: 185-2792 and an antisense strand consisting of an antisense sequence selected from SEQ ID NOs: 1-62 or SEQ ID NOs: 185-2792.

### Claim 18

(canceled)

### Claim 19

(canceled)

### Claim 20

(canceled)

### Claim 21

(canceled)

### Claim 22 (depends on 2)

A cell containing the dsRNA of claim 2.

### Claim 23

(canceled)

### Claim 24

(canceled)

### Claim 25

(canceled)

### Claim 26 (depends on 2)

The dsRNA of claim 2, further comprising a lipid formulation.

### Claim 27

(canceled)

### Claim 28

A method of inhibiting COL1A1 expression in a cell, the method comprising:
a. introducing into the cell a double-stranded ribonucleic acid (dsRNA) for inhibiting expression of COL1A1, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a COL1A1 RNA transcript, which comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from one of the antisense sequences of SEQ ID NOs: 1-62 or SEQ ID NOs: 185-2792; and
b. maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a COL1A1 gene, thereby inhibiting expression of the COL1A1 gene in the cell.

### Claim 29 (depends on 28)

The method of claim 28, wherein the cell is a hepatic stellate cell.

### Claim 30

(canceled)

### Claim 31

A method of treating liver fibrosis comprising administering to a human in need of such treatment a therapeutically effective amount of a double-stranded ribonucleic acid (dsRNA) for inhibiting expression of COL1A1, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a COL1A1 RNA transcript, which comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from one of the antisense sequences of SEQ ID NOs: 1-62 or SEQ ID NOs: 185-2792 or a pharmaceutical composition thereof.

### Claim 32

(canceled)

### Claim 33

(canceled)

### Claim 34

A vector encoding at least one strand of a dsRNA, wherein said dsRNA comprises a region of complementarity to at least a part of an mRNA encoding COL1A1, wherein said dsRNA is base pairs or less in length, and wherein said dsRNA targets a said mRNA for cleavage.

### Claim 35

(canceled)

### Claim 36

(canceled)

### Claim 37

(canceled)

### Claim 38

(canceled)

### Claim 39

A method of treating a chronic liver disease characterized by liver fibrosis comprising administering to a human in need of such treatment a therapeutically effective amount of a double-stranded ribonucleic acid (dsRNA) for inhibiting expression of COL1A1, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a COL1A1 RNA transcript, which comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from one of the antisense sequences of SEQ ID NOs: 1-62 or SEQ ID NOs: 185-2792 or a pharmaceutical composition thereof.

### Claim 40

(canceled)