Methylenetetrahydrofolate reductase inhibitors and use thereof

• US 7,319,145 B2
• Filed: 12/03/2001
• Issued: 01/15/2008
• Est. Priority Date: 12/03/2001
• Status: Expired due to Fees

• Current Assignee: Rima Rozen
• Original Assignee: Rima Rozen
• Inventors: Rozen, Rima, Sekhon, Jaspreet
• Primary Examiner(s): Schnizer; Richard
• Assistant Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US10/433,392
• Publication Number:
  US 20040121972A1
• Time in Patent Office: 2,234 Days
• Field of Search: 435/6, 514/44, 514/12, 424/146.1
• US Class Current: 536/24.5
• CPC Class Codes: A61K 38/44 Oxidoreductases (1)
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C12Y 105/0102 Methylenetetrahydrofolate reductase [NAD(P)H] (1.5.1.20)

Abstract

The present invention provides methylenetetrahydrofolate reductase (MTHFR) inhibitors for use in selective inhibition of cancer cell growth in a mammal. These inhibitors can be a small molecule, an antisense oligonucleotide, a ribozyme, a triple helix forming oligonucleotide, an anti-MTHFR antibody or fragment thereof, an MTHFR mutant or a fragment of MTHFR. The present invention further provides nucleic acids encoding an inhibitor of MTHFR, and vectors comprising these nucleic acids. Also encompassed by the present invention are methods of using the MTHFR inhibitors for selective inhibition of cancer cell growth, and pharmaceutical compositions comprising the MTHFR inhibitors.

Claims

1. An inhibitor of methylenetetrahydrofolate reductase (MTHFR) that selectively inhibits cancer cell growth in a mammal comprising an antisense oligonucleotide of 20 to about 50 nucleotides in length that is at least 90% identical to the complement of a region of exon 5 of a human MTHFR mRNA and comprises the sequence as set forth in SEQ ID NO:

16, wherein said human MTHFR mRNA has the sequence as set forth in SEQ ID NO;

1.

2. A nucleic acid sequence encoding the inhibitor according to claim 1.

3. The nucleic acid sequence according to claim 2, which is operatively associated with a regulatory control element.

4. A vector comprising the nucleic acid sequence according to claim 2 operatively associated with a regulatory control element.

5. A composition comprising the inhibitor according to claim 1 and a pharmaceutically acceptable carrier, diluent or excipient.

6. A method of preparing a medicament comprising admixing the inhibitor according to claim 1 with a pharmaceutically acceptable carrier, wherein the inhibitor reduces or inhibits MTHFR gene expression.

7. The inhibitor according to claim 1, wherein said antisense oligonucleotide comprises a sequence that is about 20 to about 22 nucleotides in length.

8. The inhibitor according to claim 1, wherein said antisense oligonucleotide is a phosphorothioate oligonucleotide.

9. The inhibitor according to claim 1, wherein said antisense oligonucleotide comprises a sequence that is at least 95% identical to the complement of a region of exon 5 of the human MTHFR mRNA.

10. The inhibitor according to claim 1, wherein said antisense oligonucleotide comprises a sequence that is identical to the complement of a region of exon 5 of the human MTHFR mRNA.

11. The inhibitor according to claim 1, wherein said antisense oligonucleotide is a modified oligonucleotide.

12. The inhibitor according to claim 11, wherein said modified oligonucleotide is nuclease resistant, has improved pharmacokinetics relative to an unmodified oligonucleotide, or a combination thereof.

28. A composition comprising the vector according to claim 4 and a pharmaceutically acceptable carrier, diluent or excipient.

30. The inhibitor according to claim 1, wherein said antisense oligonucleotide is a DNA oligonucleotide.

31. The inhibitor according to claim 1, wherein said antisense oligonucleotide is a RNA oligonucleotide.

13. An inhibitor of methylenetetrahydrofolate reductase (MTHFR) that selectively inhibits cancer cell growth in a mammal comprising an antisense oligonucleotide of 20 to about 25 nucleotides in length that is at least 90% identical to the complement of a region of exon 5 of a human MTHFR mRNA and comprises the sequence as set forth in SEQ ID NO:

16, wherein said human MTHFR has the sequence as set forth in SEQ ID NO;

1.

14. The inhibitor according to claim 13, wherein said antisense oligonucleotide is at least 95% identical to the complement of a region of exon 5 of the human MTHFR mRNA.

15. The inhibitor according to claim 13, wherein said antisense oligonucleotide is identical to the complement of a region of exon 5 of a human MTHFR mRNA.

16. The inhibitor according to claim 13, wherein said antisense oligonucleotide comprises a sequence that is about 20 to about 22 nucleotides in length.

17. The inhibitor according to claim 13, wherein said antisense oligonucleotide is a modified oligonucleotide.

18. The inhibitor according to claim 17, wherein said modified oligonucleotide is nuclease resistant, has improved pharmacokinetics relative to an unmodified oligonucleotide, or a combination thereof.

19. The inhibitor according to claim 13, wherein said antisense oligonucleotide is a phosphorothioate oligonucleotide.

20. A composition comprising the inhibitor according to claim 13 and a pharmaceutically acceptable carrier, diluent or excipient.

27. A vector comprising a nucleic acid encoding the inhibitor according to claim 13 operatively associated with a regulatory control element.

29. A composition comprising the vector according to claim 27 and a pharmaceutically acceptable carrier, diluent or excipient.

32. The inhibitor according to claim 13, wherein said antisense oligonucleotide is a DNA oligonucleotide.

33. The inhibitor according to claim 13, wherein said antisense oligonucleotide is a RNA oligonucleotide.

21. An inhibitor of methylenetetrahydrofolate reductase (MTHFR) that selectively inhibits cancer cell growth in a mammal comprising an antisense oligonucleotide of 20 to about 50 nucleotides in length that is complementary to a region of exon 5 of a human MTHFR mRNA and comprises the sequence as set forth in SEQ ID NO:

16, wherein said human MTHFR mRNA has the sequence as set forth in SEQ ID NO;

1, and wherein said antisense oligonucleotide is a modified oligonucleotide.

22. The inhibitor according to claim 21, wherein said modified oligonucleotide is nuclease resistant, has improved pharmacokinetics relative to an unmodified oligonucleotide, or a combination thereof.

23. The inhibitor according to claim 21, wherein said antisense oligonucleotide is about 20 to about 25 nucleotides in length.

24. The inhibitor according to claim 21, wherein said antisense oligonucleotide is about 20 to about 22 nucleotides in length.

25. The inhibitor according to claim 21, wherein said antisense oligonucleotide is a phosphorothioate oligonucleotide.

26. A composition comprising the inhibitor according to claim 21 and a pharmaceutically acceptable carrier, diluent or excipient.

34. The inhibitor according to claim 21, wherein said antisense oligonucleotide is a DNA oligonucleotide.

35. The inhibitor according to claim 21, wherein said antisense oligonucleotide is a RNA oligonucleotide.