Modulation of transthyretin expression

• US 20050244869A1
• Filed: 04/01/2005
• Published: 11/03/2005
• Est. Priority Date: 04/05/2004
• Status: Abandoned Application

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Jain, Ravi, Brown-Driver, Vickie L.
• Application Number:
  US11/097,928
• Publication Number:
  US 20050244869A1
• Time in Patent Office: Days
• US Class Current: 435/6
• 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 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

Compounds, compositions and methods are provided for modulating the expression of transthyretin. The compositions comprise oligonucleotides, targeted to nucleic acid encoding transthyretin. Methods of using these compounds for modulation of transthyretin expression and for diagnosis and treatment of diseases and conditions associated with expression of transthyretin are provided.

Claims

1. An antisense compound 13 to 80 nucleobases in length targeted to a nucleic acid molecule encoding human transthyretin, wherein said compound inhibits the expression of human transthyretin mRNA by at least 10%.

2. The antisense compound of claim 1 comprising 13 to 50 nucleobases in length.

3. The antisense compound of claim 2 comprising 15 to 30 nucleobases in length.

4. The antisense compound of claim 1 comprising an oligonucleotide.

5. The antisense compound of claim 4 comprising a DNA oligonucleotide.

6. The antisense compound of claim 4 comprising an RNA oligonucleotide.

7. The antisense compound of claim 6 comprising a single-stranded RNA oligonucleotide.

8. The antisense compound of claim 4 comprising a chimeric oligonucleotide.

9. The antisense compound of claim 4 wherein at least a portion of said compound hybridizes with RNA to form an oligonucleotide-RNA duplex.

10. The antisense compound of claim 1 having at least one modified internucleoside linkage, sugar moiety, or nucleobase.

11. The antisense compound of claim 1 having at least one 2′

-O-methoxyethyl sugar moiety.

12. The antisense compound of claim 1 having at least one phosphorothioate internucleoside linkage.

13. The antisense compound of claim 1 wherein at least one cytosine is a 5-methylcytosine.

14. A method of inhibiting the expression of human transthyretin in a cell or tissue comprising contacting said cell or tissue with the antisense compound of claim 1 so that expression of human transthyretin is inhibited.

18. The antisense compound of claim 1, wherein said antisense compound has at least an 13 nucleobase portion of a sequence selected from the group consisting of SEQ ID NOs 15, 20, 24, 29, 30, 34, 35, 37, 39, 42, 44, 47, 49, 50, 53, 54, 59, 60, 61, 62, 63, 64, 65, 66, 67, 69, 70, 71, 72, 73, 74, 75, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87 and 88.

15. A method of screening for a modulator of human transthyretin, the method comprising the steps of:

contacting a preferred target segment of a nucleic acid molecule encoding human transthyretin with one or more candidate modulators of human transthyretin, and

identifying one or more modulators of human transthyretin expression which modulate the expression of human transthyretin.

16. The method of claim 15 wherein the modulator of transthyretin expression comprises an oligonucleotide, an antisense oligonucleotide, a DNA oligonucleotide, an RNA oligonucleotide, an RNA oligonucleotide having at least a portion of said RNA oligonucleotide capable of hybridizing with RNA to form an oligonucleotide-RNA duplex, or a chimeric oligonucleotide.

17. A diagnostic method for identifying a disease state comprising identifying the presence of human transthyretin in a sample using at least one of the primers comprising SEQ ID NOs 5 or 6, or the probe comprising SEQ ID NO:

7.