Methods and compositions for liver specific delivery of therapeutic molecules using recombinant AAV vectors

• US 20030166284A1
• Filed: 03/28/2002
• Published: 09/04/2003
• Est. Priority Date: 09/06/1996
• Status: Abandoned Application

• Current Assignee: Chiron Corporation (Bayer AG)
• Original Assignee: Chiron Corporation (Bayer AG)
• Inventors: Ponnazhagan, Selvarangan, Zhou, Shang-Zhen, Schloemer, Robert H., Escobedo, Jaime, Srivastava, Arun, Yoder, Mervin C., Dwarki, Varavani, Wang, Xu-Shan
• Application Number:
  US10/109,799
• Publication Number:
  US 20030166284A1
• Time in Patent Office: Days
• US Class Current: 435/456
• 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: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector

Abstract

Provided are methods for selectively expressing therapeutic molecules, such as secretory proteins, antisense molecules and ribozymes, in the liver. The methods find use in treating hepatic diseases or conditions. The methods also find use in treating any disease or condition in which systemic administration of the therapeutic substance, for example, a secretory protein, is desired. The methods involve administering to a mammalian patient having a need for liver expression of a therapeutic molecule an AAV vector containing a therapeutically effective amount of the therapeutic molecule. Also provided are novel vectors employable in these methods.

Claims

1. A method for realizing liver specific delivery of a therapeutic molecule in a mammalian patient comprising administering to said mammalian patient a therapeutically effective amount of an AAV vector containing said therapeutic molecule.

2. A method according to claim 1 wherein said therapeutic molecule is a nucleic acid sequence encoding a secretory protein, an antisense molecule or a ribozyme.

3. A method according to claim 1 wherein the AAV vector is selected from the group consisting of pD-5, pD-10, pD-15, pD-20.

4. A method according to claim 1 wherein said AAV vector is administered by intravenous or intraportal injection.

5. A method according to claim 1 wherein said AAV vector is administered ex vivo.

6. A method according to claim 1 wherein said AAV vector is administered by direct injection.

7. A method according to claim 1 wherein said therapeutic molecule is a nucleic acid encoding the LDL receptor.

8. A method according to claim 1 wherein said AAV vector additionally contains a liver specific promoter.

9. A method according to claim 8 wherein said AAV vector additionally contains a liver specific enhancer.

10. A method according claim 8 wherein said promoter is selected from the group consisting of the hepatitis B virus X gene promoter, the hepatitis B virus core protein promoter, the AFP gene promoter, the albumin gene promoter, the α

-1 antitrypsin gene promoter, the fibrinogen gene promoter, the APO-A1 gene promoter and the promoter genes for liver transference enzymes.

11. A method according to claim 10 wherein said AAV vector additionally contains a liver specific enhancer.

12. A hybrid helper AAV vector comprising a DNA sequence encoding hepatitis B virus surface antigen or a functional fragment thereof linked to a DNA sequence encoding the AAV VP-1 protein to form a chimeric DNA sequence.

13. A recombinant AAV vector comprising two AAV ITRs (inverted terminal repeats) in which the native D-sequences of each of said ITRs are modified by the substitution of nucleotides such that at least 5 native nucleotides and up to 18 native nucleotides are retained and the remaining nucleotides of the D-sequence are deleted or replaced with non-native nucleotides.

14. A recombinant AAV vector according to claim 13 wherein said at least 5 native nucleotides are 5′

CTCCA 3′

.

15. A recombinant AAV vector comprising two AAV ITRs (inverted terminal repeats) in which the native D-sequences of each of said ITRs are modified by the substitution of nucleotides such that at least 10 native nucleotides up to 18 native nucleotides are retained and the remaining nucleotides of the D-sequence are deleted or replaced with non-native nucleotides.

16. A recombinant AAV vector comprising two AAV ITRs (inverted terminal repeats) in which the native D-sequences of each of said ITRs are modified by the substitution of nucleotides such that 10 native nucleotides are retained and the remaining nucleotides of the D-sequence are deleted or replaced with non-native nucleotides.

17. A recombinant AAV vector according to claim 16 wherein said 10 native nucleotides comprise nucleotides 5′

CTCCA 3′ and

five other native nucleotides of said D-sequence.

18. A recombinant AAV vector selected from the group consisting of pD-5, pD-10, pD-15 and pD-20.