Modulation of apolipoprotein (a) expression

• US 7,259,150 B2
• Filed: 10/15/2003
• Issued: 08/21/2007
• Est. Priority Date: 08/07/2001
• Status: Expired due to Term

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Graham, Mark J., Crooke, Rosanne M.
• Primary Examiner(s): Angell; Jon E.
• Assistant Examiner(s): BOWMAN, AMY HUDSON
• Application Number:
  US10/684,440
• Publication Number:
  US 20040138164A1
• Time in Patent Office: 1,406 Days
• Field of Search: 536/23.1, 536/24.5, 514/44, 435/6, 435/91.1, 435/91.3, 435/325, 435/375
• US Class Current: 514/44.00A
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• 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 15/113 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 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: C12N 2320/30 Special therapeutic applications

Abstract

Compounds, compositions and methods are provided for modulating the expression of apolipoprotein(a). The compositions comprise oligonucleotides, targeted to nucleic acid encoding apolipoprotein(a). Methods of using these compounds for modulation of apolipoprotein(a) expression and for diagnosis and treatment of disease associated with expression of apolipoprotein(a) are provided.

Claims

1. A method of inhibiting the expression of apolipoprotein(a) in a cell or tissue in vivo, said method comprising contacting said cell or tissue with an antisense oligonucleotide 12 to 30 nucleobases in length targeted to SEQ ID NO:

4, wherein said antisense oligonucleotide comprises at least 8 consecutive nucleobases of SEQ ID NO;

23 and is 100% complementary to SEQ ID NO;

4, thereby inhibiting the expression of apolipoprotein(a).

2. The method of claim 1, wherein said antisense oligonucleotide is a chimeric antisense oligonucleotide.

3. The method of claim 1, wherein said antisense oligonucleotide comprises at least one modified internucleoside linkage, modified sugar moiety, or modified nucleobase.

4. The method of claim 3, wherein said modified sugar moiety is a 2′

-O-methoxyethyl sugar moiety.

5. The method of claim 3, wherein said modified internucleoside linkage is a phosphorothioate internucleoside linkage.

6. The method of claim 3, wherein said modified nucleobase is a 5-methylcytosine.

18. The method of claim 1, wherein said antisense oligonucleotide has a nucleobase sequence comprising SEQ ID NO:

23.

19. The method of claim 1, wherein said antisense oligonucleotide consists of SEQ ID NO:

23.

25. The method of claim 1, wherein said antisense oligonucleotide consists of SEQ ID NO:

23, wherein nucleotides 1–

5 and 16–

20 are 2′

-O-methoxyethyl nucleotides, wherein nucleotides 6–

15 are 2′

-deoxynucleotides, and wherein each cytosine is a 5-methylcytosine.

7. A method of treating a human having a disease or condition treatable by the reduction of apolipoprotein(a), said method comprising

selecting a human having elevated plasma lipoprotein(a) levels;

and

administering to said human a therapeutically effective amount of an antisense oligonucleotide 12 to 30 nucleobases in length targeted to SEQ ID NO;

4, wherein said antisense oligonucleotide comprises at least 8 consecutive nucleobases of SEQ ID NO;

23 and is 100% complementary to SEQ ID NO;

4.

8. The method of claim 7, wherein the disease or condition is atherosclerosis or hypercholesterolemia.

9. The method of claim 7, wherein said antisense oligonucleotide is a chimeric antisense oligonucleotide.

10. The method of claim 7, wherein said antisense oligonucleotide comprises at least one modified internucleoside linkage, modified sugar moiety, or modified nucleobase.

11. The method of claim 10, wherein said modified sugar moiety is a 2′

-O-methoxyethyl sugar moiety.

12. The method of claim 10, wherein said modified internucleoside linkage is a phosphorothioate internucleoside linkage.

13. The method of claim 10, wherein said modified nucleobase is a 5-methylcytosine.

14. The method of claim 7, further comprising measuring lipoprotein(a) levels in the serum of said human following said administering.

15. The method of claim 8, wherein said chimeric antisense oligonucleotide comprises a gap segment of ten 2′

-deoxynucleotides positioned between 5′ and

3′

wing segments of five 2′

-O-methoxyethyl modified nucleosides.

16. The method of claim 15, wherein each intemucleoside linkage of said chimeric antisense oligonucleotide is a phosphorothioate internucleoside linkage.

17. The method of claim 16, wherein each cytosine of said chimeric antisense oligonucleotide is a 5-methylcytosine.

20. The method of claim 9, wherein said chimeric antisense oligonucleotide comprises a gap segment of ten 2′

-deoxynucleotides positioned between 5′ and

3′

wing segments of five 2′

-O-methoxyethyl modified nucleosides.

21. The method of claim 20, wherein each internucleoside linkage of said chimeric antisense oligonucleotide is a phosphorothioate internucleoside linkage.

22. The method of claim 21, wherein each cytosine of said chimeric antisense oligonucleotide is a 5-methylcytosine.

23. The method of claim 7, wherein said antisense oligonucleotide has a nucleobase sequence comprising SEQ ID NO:

23.

24. The method of claim 7, wherein said antisense oligonucleotide consists of SEQ ID NO:

23.

26. The method of claim 7, wherein said antisense oligonucleotide consists of SEQ ID NO:

23, wherein nucleotides 1–

5 and 16–

20 are 2′

-O-methoxyethyl nucleotides, wherein nucleotides 6–

15 are 2′

-deoxynucleotides, and wherein each cytosine is a 5-methylcytosine.