# US 2014/0194493 A1 · Compositions and methods for inhibiting expression of transthyretin

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- Number: US 2014/0194493 A1
- Document ID: US20140194493A1
- Status: Published application
- Kind: A1
- Application number: 14/220,829
- Filed: 2014-03-20
- Published: 2014-07-10
- Earliest claimed: 2008-10-20
- Assignees: Alnylam Pharmaceuitcals, Inc.
- Applicants: Alnylam Pharmaceuitcals, Inc.
- Inventors: Dinah Wen-Yee Sah; Gregory Hinkle; Rene Alvarez; Stuart Milstein; Qingmin Chen
- Web page: https://patentgrep.com/patent/US20140194493A1

## 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)}
- A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 31/713: Double-stranded nucleic acids or oligonucleotides
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 5/14: of the thyroid hormones, e.g. T3, T4
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 13/12: of the kidneys
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
- A61P 27/02: Ophthalmic agents
- A61P 35/00: Antineoplastic agents
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07F 9/6533: Six-membered rings
- C07H 21/04: with deoxyribosyl as saccharide radical
- 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 2310/321: 2'-O-R Modification
- C12N 2310/335: Modified T or U
- C12N 2310/3515: Lipophilic moiety, e.g. cholesterol
- C12N 2320/30: Special therapeutic applications
- C12N 2320/32: Special delivery means, e.g. tissue-specific
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07F: ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM (metal-containing porphyrins C07D487/22)
- 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)

## Abstract

The invention relates to a double-stranded ribonucleic acid (dsRNA) targeting a transthyretin (TTR) gene, and methods of using the dsRNA to inhibit expression of TTR.

## Claims

### Claim 1

A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of transthyretin (TTR), wherein the dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region complementary to a part of a mRNA encoding transthyretin (TTR), wherein the region of complementarity is less than 30 nucleotides in length and the antisense strand comprises 15 or more contiguous nucleotides of an antisense strand selected from Tables 3A, 3B, 4, 6A, 6B, 7, and 16 or SEQ ID NO:170, SEQ ID NO:450, SEQ ID NO:730, or SEQ ID NO:1010.

### Claim 2 (depends on 1)

The dsRNA of claim 1, wherein the region of complementarity is 19 nucleotides in length.

### Claim 3 (depends on 1)

The dsRNA of claim 1, wherein each strand of the dsRNA is 19, 20, 21, 22, 23, or 24 nucleotides in length.

### Claim 4 (depends on 1)

The dsRNA of claim 1, wherein the dsRNA comprises at least one modified nucleotide.

### Claim 5 (depends on 1)

The dsRNA of claim 1, wherein the dsRNA comprises at least one modified nucleotide selected 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, a 2′-amino-modified nucleotide, a 2′-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.

### Claim 6 (depends on 1)

The dsRNA of claim 1, wherein the dsRNA is conjugated to a ligand.

### Claim 7 (depends on 1)

The dsRNA of claim 1, wherein the dsRNA is formulated in a lipid formulation.

### Claim 8 (depends on 1)

The dsRNA of claim 1, wherein administration of the dsRNA to a cell results in about 95% inhibition of TTR mRNA expression as measured by a real time PCR assay, wherein the cell is a HepG2 cell or a Hep3B cell, and wherein the concentration of the dsRNA is 10 nM.

### Claim 9 (depends on 1)

The dsRNA of claim 1, wherein the dsRNA has an IC50 of less than 10 pM in a HepG2 cell, and wherein the concentration of the dsRNA is 10 nM.

### Claim 10 (depends on 1)

The dsRNA of claim 1, wherein the dsRNA has an ED50 of about 1 mg/kg.

### Claim 11 (depends on 1)

The dsRNA of claim 1, wherein administration of the dsRNA does not result in immunostimulatory activity in human peripheral blood mononuclear cells (PBMCs) as measured by IFN-alpha and TNF-alpha ELISA assays.

### Claim 12 (depends on 1)

A cell containing the dsRNA of claim 1.

### Claim 13 (depends on 1)

A pharmaceutical composition for inhibiting expression of a TTR gene comprising the dsRNA of claim 1 and a pharmaceutically acceptable carrier.

### Claim 14 (depends on 1)

A method of inhibiting TTR expression in a cell, the method comprising:
(a) contacting the cell with the dsRNA of claim 1; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of a TTR gene, thereby inhibiting expression of the TTR gene in the cell.

### Claim 15 (depends on 1)

A method of treating a disorder mediated by TTR expression comprising administering to a human in need of such treatment a therapeutically effective amount of the dsRNA of claim 1.

### Claim 16 (depends on 15)

The method of claim 15, wherein the human has transthyretin amyloidosis, a liver disorder, or is further provided a liver transplant.