siRNA SILENCING OF APOLIPOPROTEIN B

• US 20110189300A1
• Filed: 05/20/2010
• Published: 08/04/2011
• Est. Priority Date: 11/17/2004
• Status: Abandoned Application

• Current Assignee: Protiva Biotherapeutics Inc (Arbutus Biopharma Corporation)
• Original Assignee: Protiva Biotherapeutics Inc (Arbutus Biopharma Corporation)
• Inventors: Sood, Vandana, Lee, Amy C.H., Jeffs, Lloyd B., Judge, Adam, MacLachlan, Ian, Palmer, Lorne R.
• Application Number:
  US12/784,402
• Publication Number:
  US 20110189300A1
• Time in Patent Office: Days
• US Class Current: 424/498
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: 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
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/14 of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• 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 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific

Abstract

The present invention provides nucleic acid-lipid particles comprising siRNA molecules that silence ApoB expression and methods of using such nucleic acid-lipid particles to silence ApoB expression.

Claims

1. A method for the in vivo delivery of an siRNA molecule that silences Apolipoprotein B (ApoB) expression, said method comprising:

administering to a mammal a nucleic acid-lipid particle, wherein said particle comprises an siRNA molecule that silences ApoB expression, a cationic lipid, a non-cationic lipid, and a conjugated lipid that inhibits aggregation of said particle.

2. The method of claim 1, wherein said siRNA molecule is a double-stranded siRNA molecule formed by two complementary strands.

3. The method of claim 2, wherein each strand of said siRNA molecule is 19 to 25 nucleotides in length.

4. The method of claim 2, wherein each strand of said siRNA molecule comprises a 3′

overhang.

5. The method of claim 1, wherein said siRNA molecule in said particle is resistant in aqueous solution to degradation with a nuclease.

6. The method of claim 1, wherein said siRNA molecule silences ApoB expression by at least 50% in a test mammal relative to the level of ApoB expression in a control mammal not administered said siRNA molecule.

7. The method of claim 1, wherein said siRNA molecule silences ApoB expression by at least 60% in a test mammal relative to the level of ApoB expression in a control mammal not administered said siRNA molecule.

8. The method of claim 1, wherein said siRNA molecule silences ApoB expression by at least 70% in a test mammal relative to the level of ApoB expression in a control mammal not administered said siRNA molecule.

9. The method of claim 1, wherein said siRNA molecule silences ApoB expression by at least 80% in a test mammal relative to the level of ApoB expression in a control mammal not administered said siRNA molecule.

10. The method of any of claims 6-9, wherein said test mammal and said control mammal are both mice.

11. The method of claim 1, wherein said siRNA molecule silences ApoB mRNA levels, ApoB protein levels, or a combination thereof.

12. The method of claim 1, wherein said siRNA molecule targets SEQ ID NO:

48.

13. The method of claim 1, wherein said siRNA molecule comprises a sense strand sequence consisting of nucleotides 3-23 of SEQ ID NO:

48.

14. The method of claim 1, wherein said siRNA molecule is fully encapsulated in said particle.

15. The method of claim 1, wherein said siRNA molecule comprises at least one modified nucleotide.

16. The method of claim 15, wherein said modified nucleotide is a 2′

-O-methyl (2′

OMe) nucleotide.

17. The method of claim 1, wherein said non-cationic lipid comprises a mixture of a phospholipid and cholesterol.

18. The method of claim 1, wherein said conjugated lipid that inhibits aggregation of said particle comprises a polyethyleneglycol (PEG)-lipid conjugate.

19. The method of claim 18, wherein said PEG-lipid conjugate is selected from the group consisting of a PEG-dialkyloxypropyl (PEG-DAA) conjugate, a PEG-diacylglycerol (PEG-DAG) conjugate, a PEG-phospholipid conjugate, a PEG-ceramide (PEG-Cer) conjugate, and a mixture thereof.

20. The method of claim 1, wherein said cationic lipid comprises from about 2 mol % to about 60 mol % of the total lipid present in said particle.

21. The method of claim 1, wherein said non-cationic lipid comprises from about 5 mol % to about 90 mol % of the total lipid present in said particle.

22. The method of claim 1, wherein said conjugated lipid that inhibits aggregation of said particle comprises from about 0.5 mol % to about 20 mol % of the total lipid present in said particle.

23. The method of claim 1, wherein said particle is administered by a route selected from the group consisting of intravenous, subcutaneous, and intraperitoneal.

24. The method of claim 1, wherein said mammal is a human.

25. The method of claim 24, wherein said human has a disease or disorder associated with ApoB expression or overexpression.

26. The method of claim 24, wherein said human has a disease or disorder selected from the group consisting of atherosclerosis, angina pectoris, high blood pressure, diabetes, and hypothyroidism.

27. The method of claim 24, wherein said human has a disease or disorder which involves hypercholesterolemia and wherein serum cholesterol levels are lowered when ApoB expression is silenced by said siRNA molecule.

28. The method of claim 27, wherein said disease or disorder which involves hypercholesterolemia is selected from the group consisting of atherosclerosis, angina pectoris, and high blood pressure.