Enhancement of transfection of dna into the liver

• US 20040248832A1
• Filed: 08/04/2004
• Published: 12/09/2004
• Est. Priority Date: 07/10/2001
• Status: Active Grant

• Current Assignee: Sonogene, LLC
• Original Assignee: Sonogene, LLC
• Inventors: Davidson, Michael
• Granted Patent: US 7,211,248 B2
• Time in Patent Office: Days
• US Class Current: 514/44
• 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 48/0075 characterised by an aspect of the delivery route, e.g. oral, subcutaneous
• CPC Class Codes: A61K 48/0083 characterised by an aspect of the administration regime
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Abstract

A method for transfection of a DNA or RNA, such as the apo A1 gene into the liver to regulate lipid metabolism comprises injection of a solution containing the DNA or RNA gene into the liver using gas microbubbles or microspheres and using ultrasound to induce the expression of the gene in the liver

Claims

1. A method for transfection of DNA or RNA into the liver of a mammal comprising the steps of

a) delivering a solution containing DNA or RNA to the vasculature of the liver;

and

b) applying ultrasound to the liver during exposure to the solution to induce the expression of the DNA in the liver.

2. A method according to claim 1, wherein the solution is delivered vascularly with increased injection pressure.

3. A method according to claim 1, wherein the solution additionally comprises microbubbles of a gas.

4. A method according to claim 3, wherein the gas is air, nitrogen, or oxygen.

5. A method according to claim 1, wherein the DNA is any DNA that will be actively translated when incorporated into a cell.

6. A method according to claim 3, wherein the gas bubbles are encapsulated gas microspheres or gas bubbles stabilized by one or more surfactants, lipids, proteins, lipoproteins, or polymers.

7. The method according to claim 3, wherein the microspheres are injected into the blood peripherally.

8. The method according to claim 1, wherein the transfected DNA alters cholesterol metabolism of the liver.

9. The method according to claim 1, wherein the DNA encodes apo A1, operably linked to a promoter.

10. The method according to claim 1, wherein the DNA comprises naked plasmid DNA encoding apo A1.

11. The method of claim 3, wherein the DNA containing solution is delivered under high injection pressure.

12. The method of claim 1, wherein the ultrasound is applied to localize the area of liver treatment.

13. A method for increasing apo A1 secretion from the liver comprising the steps of:

delivering a solution comprising DNA encoding for apo A1 to the vasculature of the liver under high intravascular pressure.

14. A method for the transfection of the apo A1 gene in the liver to regulate cholesterol comprising the steps of:

a) forming microspheres containing the apo A1 gene;

b) delivering the microspheres into the liver, and

c) applying ultrasound to the microspheres to induce the expression of the apo A1 gene in the liver.

15. The method according to claim 14, wherein the microspheres are injected into the blood peripherally.

16. The method according to claim 14, wherein the apo A1 gene is naked apo A1 plasmid DNA.

17. A composition for transfection of the liver comprising DNA and stabilized gas bubbles in a pharmacologically acceptable solution.

18. The composition of claim 17, wherein the DNA comprises plasmid DNA.

19. The composition of claim 17, wherein the DNA comprises plasmid DNA encoding apo A1, operably linked to a promoter.

20. The composition of claim 17, wherein the stabilized gas bubbles comprise gas filled microspheres.