Lipid-mediated polynucleotide administration to deliver a biologically active peptide and to induce a cellular immune response

• US 7,250,404 B2
• Filed: 07/26/2002
• Issued: 07/31/2007
• 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, Wolff, Jon A, Malone, Robert Wallace, Rhodes, Gary H
• Primary Examiner(s): PRIEBE, SCOTT DAVID
• Application Number:
  US10/202,858
• Publication Number:
  US 20030032615A1
• Time in Patent Office: 1,831 Days
• Field of Search: 514/44, 424/450, 435/69.3
• 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 of producing an immune response in a vertebrate, comprising:

administering in vivo into a tissue of said vertebrate a composition comprising a polynucleotide encoding an immunogen, said polynucleotide being complexed with a cationic lipid;

wherein said polynucleotide is taken up into the cells of said vertebrate;

wherein said immunogen is expressed in an amount sufficient to produce said immune response;

wherein said polynucleotide is messenger RNA or is nonintegrating, non-replicating DNA comprising a promoter operably linked to a sequence encoding said immunogen;

wherein said immunogen is a peptide associated with a tumor cell; and

wherein said immune response is selected from the group consisting of a detectable humoral response, a detectable cellular response, or a combination thereof.

2. The method of claim 1, wherein said polynucleotide is DNA comprising a promoter operably linked to a sequence encoding said immunogen.

3. The method of claim 2, wherein said promoter is selected from the group consisting of a Rous sarcoma virus long terminal repeat (RSV LTR), a mycloproliferative sarcoma virus long terminal repeat (MPSV LTR), a simian virus 40 immediate early promoter (SV40IEP), a metallothionein promoter, and a human cytomegalovirus immediate early promoter (CMV IEP).

4. The method of claim 2, wherein said polynucleotide is a plasmid.

5. The method of claim 1, wherein said composition is administered to the interstitial space of one or more tissues selected from the group consisting of muscle, skin, liver, spleen, and blood.

6. The method of claim 1, wherein said composition is administered by injection.

7. The method of claim 6, wherein said composition is injected into muscle.

8. The method of claim 1, wherein said polynucleotide encodes an immunogenic polypeptide, wherein said immunogenic polypeptide is presented in the context of a class I major histocompatibility antigen, a class II major histocompatibility antigen, or a combination of class I and class II major histocompatibility antigens.

9. The method of claim 1, wherein said vertebrate is a mammal, a fish, or a bird.

10. The method of claim 1, wherein said vertebrate is a mammal.

11. The method of claim 10, wherein said mammal is a human.

12. The method of claim 1, wherein said cationic lipid comprises one or more saturated or unsaturated alkyl groups having from 6 to 30 carbon atoms.

13. The method of claim 1, wherein said cationic lipid comprises a quaternary ammonium group.

14. The method of claim 13, wherein said cationic lipid has a formula selected from the group consisting of:

15. The method of claim 14, wherein said cationic lipid is selected from the group consisting of N-(2,3-di-(9-(Z)-octadecenyloxy))-prop-1-yl-N,N,N-trimethylammonium chloride (DOTMA) and 1,2-bis(oleoyloxy)-3-(trimethylammonio)propane (DOTAP).