Mammalian transformation complex comprising a lipid carrier and DNA encoding CFTR

• US 6,001,644 A
• Filed: 08/22/1994
• Issued: 12/14/1999
• Est. Priority Date: 12/17/1991
• Status: Expired due to Fees

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Zhu, Ning, Debs, Robert J.
• Primary Examiner(s): Crouch, Deborah
• Assistant Examiner(s): Martin, Jill D.
• Application Number:
  US08/256,004
• Time in Patent Office: 1,940 Days
• Field of Search: 514/44, 935/62, 935/54, 935/55, 435/320.1, 435/172.1, 435/172.3, 435/458, 128/200, 128/14, 128/18, 128/23, 128/24, 424/450
• US Class Current: 435/320.1
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 47/54 the modifying agent being an organic compound
• CPC Class Codes: A61K 47/543 Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
• CPC Class Codes: A61K 47/544 Phospholipids
• CPC Class Codes: A61K 47/547 Chelates, e.g. Gd-DOTA or Zinc-amino acid chelates; Chelate-forming compounds, e.g. DOTA or ethylenediamine being covalently linked or complexed to the pharmacologically- or therapeutically-active agent
• CPC Class Codes: A61K 47/554 the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
• CPC Class Codes: A61K 47/6911 the form being a liposome
• 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 9/0078 for inhalation via a nebulizer such as a jet nebulizer, ultrasonic nebulizer, e.g. in the form of aqueous drug solutions or dispersions
• 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 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 14/4712 Cystic fibrosis
• CPC Class Codes: C07K 14/55 IL-2
• CPC Class Codes: C12N 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle

Abstract

Compositions comprising lipid carrier-CFTR nucleic acid complexes are disclosed.

Claims

1. A mammalian transformation complex, comprising:

a lipid carrier, comprising cationic lipid and a non-cationic lipid, wherein the complex is between about 100 nanometers and 10 microns in diameter; and

a DNA expression cassette comprising a promoter in operable linkage with a polynucleotide encoding a functional cystic fibrosis transmembrane conductance regulator (CFTR), wherein said DNA expression cassette is in a ratio of less than 4;

1 micrograms DNA to nanomoles cationic lipid; and

wherein said complex does not aggregate in vitro.

2. The mammalian transformation complex of claim 1, wherein the non-cationic lipid is a steroid.

3. The mammalian transformation complex of claim 1, wherein the non-cationic lipid is cholesterol.

4. The mammalian transformation complex of claim 1, wherein the cationic lipid and non-cationic lipid are in a molar ratio of about 1:

1.

5. The mammalian transformation complex of claim 1, wherein the lipid carrier is an MLV (multilamellar vesicle).

6. The mammalian transformation complex of claim 1, wherein the lipid carrier is an SUV (small unilamellar vesicle).

7. The mammalian transformation complex of claim 1, wherein the promoter is an inducible promoter.

8. The mammalian transformation complex of claim 1 wherein the promoter is from a cystic fibrosis transmembrane conductance regulator gene.

9. The mammalian transformation complex of claim 1, wherein the complex comprises DOPE and a lipid selected from the group consisting of DOTMA, DDAB, DOTAP and L-PE.

10. The mammalian transformation complex of claim 1, wherein the complex comprises cholesterol and a lipid selected from the group consisting of DOTMA, DDAB, DOTAP, E-PC, CEBA, E-DMPC, L-PE, and DOPE.

11. The mammalian transformation complex of claim 1, wherein the complex comprises a lipid selected from the group consisting of cholesterol, DOTMA, DDAB, DOTAP, E-PC, CEBA, E-DMPC, L-PE, and DOPE.

12. The mammalian transformation complex of claim 1, wherein the complex comprises a lipid other than DOTMA.

13. The mammalian transformation complex of claim 1, wherein the complex does not comprise DOTMA.