Metabolically activated recombinant viral vectors and methods for their preparation and use

• US 20070087433A1
• Filed: 09/01/2006
• Published: 04/19/2007
• Est. Priority Date: 08/09/1999
• Status: Active Grant

• Current Assignee: Genzyme Corporation (Sanofi )
• Original Assignee: Targeted Genetics Corporation (Innoviva, Inc.)
• Inventors: Carter, Barrie
• Granted Patent: US 7,785,888 B2
• Time in Patent Office: Days
• US Class Current: 435/320.100
• CPC Class Codes: A61K 38/191 Tumor necrosis factors [TNF], e.g. lymphotoxin [LT], i.e. TNF-beta
• 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: C07K 14/70578 NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12P 21/06 produced by the hydrolysis of a peptide bond, e.g. hydrolysate products preparing foodstuffs by protein hydrolysis A23J3/00

Abstract

Recombinant viral vectors, especially parvovirus vectors such as adeno-associated virus (AAV) vectors, capable of enhanced expression of heterologous sequences, and methods for their construction and use, are provided. The vectors have a structure, or are capable of rapidly adopting a structure, which involves intrastrand base pairing of at least one region in a heterologous sequence.

Claims

1. A recombinant viral vector comprising a single-stranded heterologous nucleotide sequence comprising a region which forms intrastrand base pairs such that expression of a coding region of the heterologous sequence is enhanced relative to a recombinant viral vector that lacks sufficient intrastrand base pairing to enhance said expression.

2. The recombinant viral vector of claim 1, wherein the region which forms intrastrand base pairs is a coding region.

3. The recombinant viral vector of claim 1, which is a recombinant parvovirus vector, comprising one or more inverted terminal repeat (ITR) sequences flanking said heterologous sequence.

4. The recombinant viral vector of claim 3, which is a recombinant adeno-associated virus (rAAV) vector.

5. The rAAV vector of claim 4, wherein said rAAV vector is capable of being packaged in an AAV particle.

6. The rAAV vector of claim 4, wherein the sequence complexity of the heterologous sequence is about one-half of the heterologous sequence.

7. The rAAV vector of claim 6, further comprising an ITR flanked on both sides by heterologous sequences.

8. A host cell comprising the recombinant viral vector of claim 1.

9. A host cell comprising the rAAV vector of claim 5.

10. The host cell of claim 9, which is mammalian.

11. A recombinant viral particle comprising the recombinant viral vector of claim 1.

12. A recombinant AAV particle comprising the rAAV vector of claim 5.

13. A recombinant AAV particle comprising the rAAV vector of claim 6.

14. A composition comprising the recombinant viral vector of claim 1.

15. A composition comprising the rAAV vector of claim 5.

16. The composition of claim 15, further comprising a pharmaceutically acceptable excipient.

17. A library comprising a population of recombinant viral vectors according to claim 1.

18. A library comprising a population of rAAV vectors according to claim 5.

19-38. -38. (canceled)