Methods of delivering a physiologically active polypeptide to a mammal

• US 6,413,942 B1
• Filed: 12/02/1999
• Issued: 07/02/2002
• Est. Priority Date: 03/21/1989
• Status: Expired due to Fees

• 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: Carson, Dennis A., Felgner, Philip L., Malone, Robert Wallace, Wolff, Jon Asher, Rhodes, Gary H.
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US09/449,923
• Time in Patent Office: 943 Days
• Field of Search: 514/44
• 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: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 14/61 Growth hormone [GH], i.e. somatotropin
• 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 2740/16122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 9/0069 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: C12N 9/1033 Chloramphenicol O-acetyltransferase (2.3.1.28)
• CPC Class Codes: C12N 9/1247 DNA-directed RNA polymerase (2.7.7.6)
• CPC Class Codes: C12N 9/78 acting on carbon to nitrogen bonds other than peptide bonds (3.5)
• CPC Class Codes: C12Y 113/12007 Photinus-luciferin 4-monooxygenase (ATP-hydrolysing) (1.13.12.7), i.e. firefly-luciferase

Abstract

A method for delivering an isolated polynucleotide to the interior of a cell in a vertebrate, comprising the interstitial introduction of an isolated polynucleotide into a tissue of the vertebrate where the polynucleotide is taken up by the cells of the tissue and exerts a therapeutic effect on the vertebrate. The method can be used to deliver a therapeutic polypeptide to the cells of the vertebrate, to provide an immune response upon in vivo translation of the polynucleotide, to deliver antisense polynucleotides, to deliver receptors to the cells of the vertebrate, or to provide transitory gene therapy.

Claims

1. A method for delivering a physiologically active polypeptide to a mammal, comprising:

administering in vivo into a tissue of a mammal a composition comprising a polynucleotide which directs synthesis of said polypeptide in mammalian cells, and a pharmaceutically acceptable carrier;

wherein said tissue is selected from the group consisting of muscle and skin;

wherein said polynucleotide is a DNA plasmid operably encoding said physiologically active polypeptide through association with a promoter;

wherein said polynucleotide is free from association with liposomal formulations and charged lipids;

wherein a sufficient amount of said composition is administered to allow incorporation of said polynucleotide into the cells of said mammal; and

wherein said polypeptide is expressed.

2. The method of claim 1, wherein said promoter is selected from the group consisting of a Rous sarcoma virus long terminal repeat (RSV LTR), al myeloproliferative sarcoma virus long terminal repeat (MPSVLTR), a simian virus 40 immediate early promoter (SV40 IEP), a metallothionein promoter, and a human cytomegalovirus immediate early promoter (CMV IEP).

3. The method of claim 1, wherein said composition is injected into muscle.

4. The method of claim 1, wherein said polypeptide is selected from the group consisting of an enzyme, a hormone, a lymphokine, a receptor, a growth factor, a regulatory protein, and an immunoregulator.

5. The method of claim 1, wherein said polypeptide is selected from the group consisting of growth hormone, insulin, interleukin, interferon, granulocyte macrophage colony stimulating factor (GMCSF), and erythropoietin.

6. The method of claim 1, wherein said physiologically active polypeptide is a therapeutic polypeptide.

7. The method of claim 1, wherein activity of said physiologically active polypeptide is attained.

8. A method for delivering a physiologically active polypeptide to a mammal, comprising:

administering in vivo into a tissue of a mammal in need of said polypeptide a composition consisting essentially of a DNA plasmid operably encoding said polypeptide through association with a promoter, and a pharmaceutically acceptable carrier;

wherein a sufficient amount of said composition is administered to allow uptake of said plasmid into the cells of said mammal; and

wherein said polypeptide is expressed.

9. The method of claim 8, wherein said promoter is selected from the group consisting of a Rous sarcoma virus long terminal repeat (RSV LTR), a myeloproliferative sarcoma virus long terminal repeat (MPSV LTR), a simian virus 40 immediate early promoter (SV40 IEP), a metallothionein promoter, and a human cytomegalovirus immediate early promoter (CMV IEP).

10. The method of claim 8, wherein said tissue is selected from the group consisting of muscle, skin, and blood.

11. The method of claim 10, wherein said composition is injected into muscle.

12. The method of claim 8, wherein said polypeptide is selected from the group consisting of an enzyme, a hormone, a lymphokine, a receptor, a growth factor, a regulatory protein, and an immunoregulator.

13. The method of claim 8, wherein said polypeptide is selected from the group consisting of growth hormone, insulin, interleukin, interferon, GMCSF, and erythropoietin.

14. The method of claim 8, wherein said physiologically active polypeptide is a therapeutic polypeptide.

15. The method of claim 8, wherein activity of said physiologically active polypeptide is attained.

16. A method for delivering a physiologically active polypeptide to a mammal, comprising:

administering in vivo into muscle of a mammal a composition consisting essentially of a messenger RNA operably encoding said polypeptide and a pharmaceutically acceptable carrier;

wherein a sufficient amount of said composition is administered to allow uptake of said mRNA into the cells of said mammal; and

wherein said polypeptide is expressed.

17. The method of claim 16, wherein said polypeptide is selected from the group consisting of an enzyme, a hormone, a lymphokine, a receptor, a growth factor, a regulatory protein, and an immunoregulator.

18. The method of claim 16, wherein said polypeptide is selected from the group consisting of growth hormone, insulin, interleukin, interferon, GMCSF, and erythropoietin.

19. The method of claim 16, wherein activity of said physiologically active polypeptide is attained.

20. A method for delivering a physiologically active polypeptide to a mammal, comprising:

administering in vivo into a tissue of a mammal in need of said polypeptide a composition consisting of a DNA plasmid operably encoding said polypeptide through association with a promoter, and a pharmaceutically acceptable carrier;

wherein a sufficient amount of said composition is administered to allow uptake of said plasmid into the cells of said mammal; and

wherein said polypeptide is expressed.

21. The method of claim 20, wherein said promoter is selected from the group consisting of a Rous sarcoma virus long terminal repeat (RSV LTR), a myeloproliferative sarcoma virus long terminal repeat (MPSV LTR), a simian virus 40 immediate early promoter (SV40 IEP), a metallothionein promoter, and a human cytomegalovirus immediate early promoter (CMV IEP).

22. The method of claim 20, wherein said tissue is selected from the group consisting of muscle, skin, and blood.

23. The method of claim 22, wherein said composition is injected into muscle.

24. The method of claim 20, wherein said polypeptide is selected from the group consisting of an enzyme, a hormone, a lymphokine, a receptor, a growth factor, a regulatory protein, and an immunoregulator.

25. The method of claim 20, wherein said polypeptide is selected from the group consisting of growth hormone, insulin, interleukin, interferon, GMCSF, and erythropoietin.

26. The method of claim 20, wherein said physiologically active polypeptide is a therapeutic polypeptide.

27. The method of claim 20, wherein activity of said physiologically active polypeptide is attained.

28. A method for delivering a physiologically active polypeptide to a mammal, comprising:

administering in vivo into muscle of a mammal a composition consisting of a messenger RNA operably encoding said polypeptide, and a pharmaceutically acceptable carrier;

wherein a sufficient amount of said composition is administered to allow uptake of said mRNA into the cells of said mammal; and

wherein said polypeptide is expressed.

29. The method of claim 28, wherein said polypeptide is selected from the group consisting of an enzyme, a hormone, a lymphokine, a receptor, a growth factor, a regulatory protein, and an immunoregulator.

30. The method of claim 28, wherein said polypeptide is selected from the group consisting of growth hormone, insulin, interleukin, interferon, GMCSF, and erythropoietin.

31. The method of claim 28, wherein activity of said physiologically active polypeptide is attained.

32. A method for delivering a therapeutic polypeptide to the interior of a mammalian cell in vivo, comprising the steps of:

obtaining an injectable preparation consisting of a pharmaceutically acceptable injectable carrier containing a DNA plasmid coding for said therapeutic polypeptide;

introducing said injectable preparation into muscle tissue of a mammal and permitting said polynucleotide to be incorporated into a cell, wherein said therapeutic polypeptide is formed as the translation product of said polynucleotide.

33. The method of claim 32, wherein said therapeutic polypeptide is selected from the group consisting of an enzyme, a hormone, a lymphokine, a receptor, a growth factor, a regulatory protein, and an immunoregulator.

34. The method of claim 32, wherein said therapeutic polypeptide is selected from the group consisting of growth hormone, insulin, interleukin, interferon, GMCSF, and erythropoietin.

35. A method of delivering human growth hormone to a mouse, comprising injecting into muscle of said mouse a composition consisting essentially of a DNA plasmid encoding said human growth hormone operably associated with a metallothionein promoter, and a pharmaceutically acceptable carrier;

wherein a sufficient amount of said composition is administered to allow incorporation of said DNA plasmid into the cells of said mouse; and

wherein said human growth hormone is detectable in the serum of said mouse having been fed zinc sulfate.