Antisense agents combining strongly bound base-modified oligonucleotide and artificial nuclease

• US 7,786,292 B2
• Filed: 04/30/2007
• Issued: 08/31/2010
• Est. Priority Date: 05/03/2006
• Status: Expired due to Fees

• Current Assignee: Baltic Technology Development, Ltd.
• Original Assignee: Baltic Technology Development, Ltd.
• Inventors: Karelson, Mati, Pilv, Mehis, Saarma, Mart
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US11/742,384
• Publication Number:
  US 20070259830A1
• Time in Patent Office: 1,219 Days
• Field of Search: None
• US Class Current: 536/24.5
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/333 Modified A
• CPC Class Codes: C12N 2310/334 Modified C
• CPC Class Codes: C12N 2310/335 Modified T or U
• CPC Class Codes: C12N 2310/336 Modified G
• CPC Class Codes: C12N 2310/3511 intercalating or cleaving agent
• CPC Class Codes: C12N 2320/51 modulating the chemical stability, e.g. nuclease-resistance

Abstract

The present invention provides compounds having a chelating moiety and an oligonucleotide sequence wherein the oligonucleotide includes one or more modified nucleobases, such as hydroxynucleobases. The disclosed compounds are suitable for antisense therapy. The chelating moiety can be complexed to an ion of a lanthanide metal. These compounds are efficient translation inhibitors of nucleic acids and have increased binding affinity for target nucleic acids. The invention also includes compositions and methods of using these compositions as antisense therapy.

Claims

1. A compound comprising an oligonucleotide having from 5 to 150 nucleobases and a chelating moiety attached to the oligonucleotide, wherein at least 10% of said nucleobases are a modified nucleobase selected from the group consisting of:

5-mercaptocytosine, 5-mercaptouracil, 8-mercaptoguanine, 8-mercaptoadenine, 5-hydroxycytosine, 5-hydroxyuracil, 8-hydroxyadenine and 8-hydroxyguanine.

2. The compound of claim 1, wherein the modified nucleobase is a hydroxynucleobase selected from the group consisting of 5-hydroxycytosine, 5-hydroxyuracil, 8-hydroxyadenine and 8-hydroxyguanine.

3. The compound of claim 1 comprising from 10 to 100 nucleobases.

4. The compound of claim 1 comprising from 10 to 50 nucleobases.

5. The compound of claim 1 comprising from 20 to 30 nucleobases.

6. The compound according to claim 1, wherein at least 2 of the nucleobases are hydroxynucleobases selected from the group consisting of 5-hydroxycytosine, 5-hydroxyuracil, 8-hydroxyadenine and 8-hydroxyguanine.

7. The compound according to claim 6, wherein from 10% to 20% of the nucleobases are the hydroxynucleobases.

8. The compound according to claim 1, wherein the chelating moiety has a formula:

9. The compound according to claim 1, wherein the chelating moiety has a formula:

10. The compound according to claim 1, wherein the chelating moiety has a formula:

11. The compound according to claim 1, wherein the chelating moiety has a formula:

12. The compound of claim 11, wherein R¹ are independently selected from hydrogen, —

C(O)CF₃ and —

CH₂Phenyl, and wherein Phenyl is substituted with H, OH, C(O)Oheterocycloalkyl, C(O)Oalkyl, or alkyl.

13. The compound according to claim 1, wherein the chelating moiety has a formula:

14. The compound according to claim 1, wherein the chelating moiety has a formula:

15. The compound according to claim 1, wherein the chelating moiety has a formula:

16. The compound according to claim 1, wherein the chelating moiety has a formula:

17. The compound of claim 1, further comprising an ion of a metal, wherein the metal is selected from the group consisting of lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium.

18. The compound of claim 17, wherein the metal is europium or lanthanum.

19. A composition comprising the compound according to claim 1 and a pharmaceutically acceptable carrier.

20. The composition of claim 19 further comprising a delivery vehicle.

21. The composition of claim 20, wherein the delivery vehicle comprises a liposome, wherein the compound is contained within the liposome.

22. A method of inhibiting translation of a target nucleic acid comprising contacting the target nucleic acid with a compound according to claim 1, or a composition according to claim 19, under conditions that permit hybridizing of the compound to the target nucleic acid, wherein the hybridized compound inhibits translation of the target nucleic acid.

23. The method of claim 22 wherein the target nucleic acid is in an organism.

24. The method of claim 23 wherein the contacting comprises administering to the organism a composition that comprises the compound and a pharmaceutically acceptable carrier.

25. The method of claim 23, wherein the organism is a human or animal subject.

26. The method of claim 23, wherein the contacting comprises mixing the compound with a biological sample that comprises the target nucleic acid.

27. The method according to claim 22, wherein the hybridizing induces a cleavage of the target nucleic acid.

28. The method of claim 22, wherein the target nucleic acid is mRNA.

29. The method of claim 22, wherein the compound cleaves a bond of the target nucleic acid.

30. The method of claim 25, wherein the human or animal subject suffers from a viral infection, bacterial infection, microbial infection, fungal infection, or cancer.

31. A method of inhibiting translation of a nucleic acid in an organism, comprising:

predicting or determining a nucleotide sequence of a target nucleic acid in an organism; and

administering to the organism a composition according to claim 19, wherein the compound comprises a nucleotide sequence, wherein, under physiological conditions of the organism, said compound is sufficiently complementary to the nucleotide sequence of the target nucleic acid to hybridize thereto in the organism, thereby inhibiting translation of the nucleic acid.

32. The method according to claim 26, wherein the nucleotide sequence of the compound is fully complementary to all or a portion of the nucleotide sequence of the target nucleic acid.

37. The compound according to claim 1, wherein the chelating moiety has a formula selected from the group consisting of:

33. A method of making a compound to inhibit translation of a nucleic acid of an organism under physiological conditions of the organism, comprising:

a) determining a nucleotide sequence of a target nucleic acid;

b) synthesizing a compound that comprises a chelating moiety attached to an oligonucleotide that comprises a nucleotide sequence that is sufficiently complementary to at least part of the nucleotide sequence of the target nucleic acid to permit the hybridization, wherein the oligonucleotide comprises from 5 to 150 nucleobases and wherein at least 10% of the nucleobases of the oligonucleotide are a modified nucleobase selected from the group consisting of;

5-mercaptocytosine, 5-mercaptouracil, 8-mercaptoguanine, 8-mercaptoadenine, 5-hydroxycytosine, 5-hydroxyuracil, 8-hydroxyadenine and 8-hydroxyguanine; and

c) mixing said compound with an ion of a metal selected from the group consisting of lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium.

34. The method of claim 33, wherein the modified nucleobase is a hydroxynucleobase and is selected from the group consisting of 5-hydroxycytosine, 5-hydroxyuracil, 8-hydroxyadenine and 8-hydroxyguanine.

35. The method of claim 33 wherein the chelating moiety has a formula selected from the group consisting of:

36. The method according to claim 33, wherein the conditions comprise human physiological conditions.