Expression of exogenous polynucleotide sequences cardiac muscle of a mammal

• US 5,693,622 A
• Filed: 06/07/1995
• Issued: 12/02/1997
• Est. Priority Date: 03/21/1989
• Status: Expired due to Term

• Current Assignee: Vical, Inc. (Fresh Tracks Therapeutics, Inc.), Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Vical, Inc. (Fresh Tracks Therapeutics, Inc.), Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Wolff, Jon A., Felgner, Philip L., Duke, David J.
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US08/480,039
• Time in Patent Office: 909 Days
• Field of Search: 514/44, 935/53, 935/55, 935/56, 935/60
• US Class Current: 514/44.00R
• 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: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

The present invention provides a method for delivering a pharmaceutical polypeptide to the interior of a cardiac cell of a vertebrate in vivo, comprising the step of introducing a preparation comprising a pharmaceutically acceptable injectable carrier and naked polynucleotide operatively coding for the polypeptide into the interstitial space of the heart, whereby the naked polynucleotide is taken up into the interior of the cell and has a pharmacological effect on the vertebrate. In a preferred embodiment wherein the polynucleotide encodes polypeptide immunologically foreign to the vertebrate, the delivery method preferably comprises delivering an immunosuppressive agent to the vertebrate to limit immune responses directed to the polypeptide.

Claims

1. A method of delivering a desired physiologically active protein, polypeptide or peptide to a mammal, comprising:

injecting into the cardiac muscle of the mammal a non-infectious, non-integrating DNA sequence encoding said protein, polypeptide or peptide operably linked to a promoter,

wherein said DNA sequence is free from association with transfection-facilitating proteins, viral particles, liposomal formulations,, charged lipids and calcium phosphate precipitating agents, whereby said DNA sequence is expressed.

2. The method of claim 1, wherein said DNA sequence is further free from any transfection facilitating material.

3. The method of claim 1 wherein said protein, polypeptide or peptide is immunologically native to said host.

4. The method of claim 1 wherein said protein, polypeptide or peptide is immunologically foreign to said host.

5. The method of claim 4 wherein said method additionally comprises administering an immunosuppressive agent to said host to limit immune responses directed to said protein, polypeptide or peptide.

6. The method of claim 5 wherein said host is immunosuppressed prior to said injection of said DNA sequence.

7. The method of claim 5 wherein said host is immunosuppressed simultaneous with said injection of said DNA sequence.

8. The method of claim 5 wherein said immunosuppressive agent is administered intravenously.

9. The method of claim 5 wherein said immunosuppressive agent is administered to the heart.

10. The method of claim 1 wherein said promoter is a muscle specific promoter.

11. The method of claim 1 wherein said promoter is a cardiac specific promoter.

12. The method of claim 1 wherein said DNA sequence contains a sequence operatively coding for the secretion of said protein, polypeptide or peptide.

13. The method of claim 1 wherein the production of said protein, polypeptide or peptide is transitory.

14. The method of claim 1 wherein said protein, polypeptide or peptide is an enzyme.

15. The method of claim 1 wherein said protein, polypeptide or peptide is a hormone.

16. The method of claim 1 wherein said protein, polypeptide or peptide is a growth factor.

17. The method of claim 1 wherein said protein, polypeptide or peptide is a regulatory sequence.

18. The method of claim 1 wherein said protein, polypeptide or peptide is an immunoregulator.

19. The method of claim 1 wherein said DNA sequence is injected myocardially.

20. The method of claim 1 wherein said DNA sequence is injected into the ventricular wall of the heart.

21. The method of claim 1 wherein said DNA sequence is injected in combination with a chelating agent.

22. The method of claim 21 wherein said chelating agent is EDTA.

23. A method of delivering firefly luceriferase to the interior of a cardiac muscle cell of a rat, comprising injecting into heart of said rat a DNA plasmid constructed of sequence encoding said firefly luceriferase operably linked to a RSV promoter in an amount sufficient for said firefly luceriferase to be detected inside said cardiac muscle cell, wherein said plasmid is free from association with transfection-facilitating viral capsid proteins, liposomes, cationic lipids, and calcium phosphate precipitating agents.