Targeted site specific antisense oligodeoxynucleotide delivery method

• US 6,245,747 B1
• Filed: 07/13/1998
• Issued: 06/12/2001
• Est. Priority Date: 03/12/1996
• Status: Expired due to Term

• Current Assignee: Board of Regents of the University of Nebraska
• Original Assignee: Board of Regents of the University of Nebraska
• Inventors: Iversen, Patrick L., Meyer, Gary D., Porter, Thomas R.
• Primary Examiner(s): Martin, Jill D.
• Application Number:
  US09/114,399
• Time in Patent Office: 1,065 Days
• Field of Search: 514/44, 435/455, 435/320.1, 435/173.1, 424/9.5, 424/9.52, 536/24.31
• US Class Current: 514/44.00A
• CPC Class Codes: A61K 41/0028 Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
• CPC Class Codes: A61K 47/6925 the form being a microcapsule, nanocapsule, microbubble or nanobubble
• CPC Class Codes: A61K 9/0009 involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis microelectromechanical systems A61K9/0097
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/5015 Organic compounds, e.g. fats, sugars
• CPC Class Codes: A61K 9/5052 Proteins, e.g. albumin
• CPC Class Codes: C12N 15/1135 against oncogenes or tumor suppressor genes
• 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 2310/315 Phosphorothioates

Abstract

The invention relates to a new and improved pharmaceutical composition and method for delivery of therapeutic agents. The methods and composition of the invention can be used with several therapeutic agents and can achieve site specific delivery of a therapeutic substance. This can allow for lower doses and for improved efficacy with drugs which traditionally reach targeted sites and can result in utility for agents such as oligonucleotides which are plagued with problems in reaching targeted sites in necessary therapeutic levels.

The delivery system includes gas-filled microbubbles formed in a nitrogen-free environment. Microbubbles formed through sonication in a nitrogen-free environment are smaller and more stable than microbubbles sonicated in the presence of room air.

Claims

1. A method of inhibiting carotid or coronary artery stenosis formation, comprising:

intravenously administering to an animal a suspension of human serum albumin encapsulated perfluorocarbon gas-filled microbubbles, said microbubbles conjugated to an antisense olioodeoxynucleotide to c-myc, wherein the gas filling the microbubbles has a decreased partial pressure of nitrogen compared to that of microbubbles formed by room air sonication, and

exposing the carotid or coronary artery of said animal to ultrasound so that said antisense oligodeoxvnucieotide to c-myc is released at said carotid or coronary artery, such that said antisense oligodeoxynucleotide to c-myc inhibits the expression of endogenous c-myc, thereby inhibiting stenosis formation in said carotid or coronary artery of said animal.

2. The method of claim 1, wherein said microbubbles are formed by sonication in a nitrogen free environment.

3. The method of claim 2, wherein said environment is 100% oxygen.

4. The method of claim 1 wherein said microbubbles are formed by the steps of:

mixing an aqueous solution comprising about 2% to about 10% by weight of human serum albumin diluted about two-fold to about eight-fold with 5% to 50% by weight of dextrose; and

exposing said solution to a sonication horn in a nitrogen-free environment to create cavitation at particulate sites in said solution thereby generating stable microbubbles from about 0.1 to 10 microns in diameter.

5. The method of claim 4, wherein the solution is exposed to a nitrogen-free environment comprising 100% oxygen.

6. The method of claim 5, wherein the 100% oxygen is blown into interface between the sonicating horn and the solution.