Gel-based delivery of recombinant adeno-associated virus vectors

• US 20060078542A1
• Filed: 02/10/2005
• Published: 04/13/2006
• Est. Priority Date: 02/10/2004
• Status: Abandoned Application

• Current Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Original Assignee: University of Florida Research Foundation, Incorporated (State University System of Florida)
• Inventors: Byrne, Barry J., Mah, Cathryn S., Fraites, Thomas J. Jr.
• Application Number:
  US11/055,497
• Publication Number:
  US 20060078542A1
• Time in Patent Office: Days
• US Class Current: 424/93.200
• CPC Class Codes: A61K 38/47 acting on glycosyl compounds (3.2), e.g. cellulases, lactases
• CPC Class Codes: A61K 47/10 Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• CPC Class Codes: A61K 47/14 Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 47/42 Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein oligopeptides having up to five amino acids A61K47/183; polyamino acids A61K47/34
• 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: A61K 48/0008 characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
• CPC Class Codes: A61K 9/0014 Skin, i.e. galenical aspects of topical compositions non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector

Abstract

Disclosed are water-soluble gel-based compositions for the delivery of recombinant adeno-associated virus (rAAV) vectors that express nucleic acid segments encoding therapeutic constructs including peptides, polypeptides, ribozymes, and catalytic RNA molecules, to selected cells and tissues of vertebrate animals. Also disclosed are gel-based rAAV compositions are useful in the treatment of mammalian, and in particular, human diseases, including for example, cardiac disease or dysfunction, and musculoskeletal disorders and congenital myopathies, including, for example, muscular dystrophy, acid maltase deficiency (Pompe'"'"'s disease), and the like. In illustrative embodiments, the invention provides rAAV vectors comprised within a biocompatible gel composition for enhanced viral delivery/transfection to mammalian tissues, and in particular to vertebrate muscle tissues such as a human heart or diaphragm tissue.

Claims

1. A composition comprising:

(a) a recombinant adeno-associated viral vector that comprises a nucleic acid segment that encodes a mammalian therapeutic agent; and

(b) a water-soluble biocompatible gel.

2. The composition of claim 1, wherein said biocompatible gel comprises a sol, a matrix, a biogel, a hydrogel, a polymer, a polysaccharide, an oligosaccharide, or a viscous suspension.

3. The composition of claim 1, wherein said biocompatible gel comprises a polymer, a viscosity agent or a sucrose-based medium.

4. The composition of claim 1, wherein said biocompatible gel comprises iodixanol or sucrose acetate isobutyrate.

5. The composition of claim 1, wherein said biocompatible gel comprises glycerin, gelatin, or alginate.

6. The composition of claim 1, wherein said biocompatible gel comprises SAF-Gel, Duoderm Hydroactive Gel, Nu-Gel;

Carrasyn (V) Acemannan Hydrogel, Elta Hydrogel or K-Y Sterile Gel.

7. The composition of claim 1, wherein said biocompatible gel comprises a cross-linked or a conjugated gel.

8. The composition of claim 1, wherein said recombinant adeno-associated viral vector is present in said composition at a concentration of at least 1×

10¹² AAV particles per milliliter.

9. The composition of claim 1, wherein said biocompatible gel comprises at least about 85% by weight of said composition.

10. The composition of claim 9, wherein said biocompatible gel comprises at least about 95% by weight of said composition.

11. The composition of claim 1, wherein said mammalian therapeutic agent is a mammalian peptide, polypeptide, enzyme, protein, antisense, or ribozyme.

12. The composition of claim 1, wherein said mammalian therapeutic agent is a peptide, polypeptide, enzyme, protein, antisense, or ribozyme that can be expressed in human tissue.

13. The composition of claim 12, wherein said mammalian therapeutic agent is a peptide, polypeptide, enzyme, protein, antisense, or ribozyme that can be expressed in human cardiac or diaphragm muscle tissue.

14. The composition of claim 13, wherein said mammalian therapeutic agent is a biologically-active acid α

-glucosidase (GAA), dystrophin, or α

-1 antitrypsin polypeptide.

15. The composition of claim 1, further comprising a pharmaceutical excipient, buffer, carrier, or diluent.

16. A kit for diagnosing, preventing, treating or ameliorating the symptoms of a diseases or disorder in a mammal comprising:

(i) the composition of claim 1;

and (ii) instructions for using said kit.

17. A method of providing a biologically-effective amount of a therapeutic agent to a tissue site of a mammal in need thereof, said method comprising the step of providing to said mammal, the composition of claim 1, in an amount and for a time effective to provide said biologically-effective amount of said therapeutic agent to said tissue site of said mammal.

18. The method of claim 17, wherein said composition is provided to said mammal by infection, systemic administration, or by direct, indirect, or localized injection to a cell, tissue, or organ of said mammal.

19. The method of claim 17, wherein said mammal is human.

20. The method of claim 19, wherein said mammal is a human that has, is suspected of having, or at risk for developing a musculoskeletal disorder, a glycogen storage disease, or a congenital myopathy.

21. The method of claim 20, wherein said mammal is a human that has, is suspected of having, or at risk for developing muscular dystrophy, cardiac hypertrophy, or acid maltase deficiency (Pompe'"'"'s Disease).

22. A method of treating or preventing a musculoskeletal disease or dysfunction, or a congenital myopathy in a mammal, said method comprising at least the step of providing to said mammal, the composition of claim 1, in an amount and for a time effective to treat or prevent said musculoskeletal disease or dysfunction, or said congenital myopathy in said mammal.

23. The method of claim 22, wherein said mammal is a human that has, is suspected of having, or at risk for developing muscular dystrophy.

24. A method of expressing in cells of a mammalian heart or diaphragm muscle, a nucleic acid segment that encodes an exogenously-provided mammalian therapeutic agent, said method comprising at least the step of directly injecting into said heart or said diaphragm muscle, the composition of claim 1, in an amount and for a time effective to express said exogenously-provided mammalian therapeutic agent.

25. A method for treating or ameliorating the symptoms of a congenital myopathy in a mammal, said method comprising administering to said mammal the composition of claim 1;

in an amount and for a time sufficient to treat or ameliorate the symptoms of said congenital myopathy in said mammal.

26. The method of claim 25, wherein said congenital myopathy is muscular dystrophy.

27. A method for expressing a biologically-effective amount of an exogenously-supplied therapeutic polypeptide in a mammalian diaphragm, heart, or muscle cell, said method comprising:

introducing into a population of said mammalian diaphragm, heart, or muscle cells, an amount of the composition of claim 1, for a time effective to express said biologically-effective amount of said exogenously-supplied therapeutic polypeptide in said mammalian diaphragm, heart or muscle cell.

28. The method of claim 27, wherein said therapeutic polypeptide is an enzyme, a kinase, a protease, a glucosidase, a glycosidase, a nuclease, a growth factor, a tissue factor, a myogenic factor, a neurotrophic factor, a neurotrophin, a dystrophin, an interleukin, or a cytokine.

29. The method of claim 28, wherein said therapeutic polypeptide is acid α

-glucosidase (GAA).

30. The method of claim 27, wherein said composition is introduced into said population of said mammalian diaphragm, heart, or muscle cells by systemic, indirect, or localized infection, or by intramuscular, subcutaneous, intra-abdominal, transpleural, intracardiac, or transperitoneal injection.