Sonochemical induction of ABCA1 expression and compositions therefor

• US 9,101,745 B2
• Filed: 03/14/2013
• Issued: 08/11/2015
• Est. Priority Date: 03/14/2013
• Status: Active Grant

• Current Assignee: SonoThera, Inc.
• Original Assignee: Sonogene, LLC
• Inventors: Coles, Eric, Davidson, Michael
• Primary Examiner(s): Long, Scott
• Application Number:
  US13/826,066
• Publication Number:
  US 20140276363A1
• Time in Patent Office: 880 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 31/7105 Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• 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 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• 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 48/0033 the non-active part being non-polymeric
• CPC Class Codes: A61K 49/223 Microbubbles, hollow microspheres, free gas bubbles, gas microspheres
• CPC Class Codes: A61M 37/0092 using ultrasonic, sonic or infrasonic vibrations, e.g. phonophoresis
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/00 Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00
• CPC Class Codes: C12N 15/85 for animal cells

Abstract

The present invention provides compositions useful for transfecting cells (e.g., liver cells) to express ABCA1. The compositions described herein comprise a pharmaceutically acceptable aqueous carrier containing sonochemically-active microspheres together with a plasmid DNA construct encoding an active form of ABCA1 and at least one promoter for the expression thereof. Preferably, the sonochemically-active microspheres comprise, consist essentially of, or consist of gas bubbles (e.g., a fluorocarbon gas, such as octafluoropropane) encapsulated within protein-containing or lipid-containing shells (e.g., human serum albumin shells). The microspheres are disruptable by exposure to ultrasonic acoustic energy to release the encapsulated gas.

Claims

1. A pharmaceutical composition for increasing HDL-cholesterol levels in the blood of a patient, the composition comprising a mixture of a nucleic acid encoding an expressible open reading frame and sonochemically-active microspheres in a pharmaceutically acceptable aqueous carrier;

wherein the nucleic acid of the mixture consists of a plasmid vector, the expressible open reading frame consists of an expressible open reading frame encoding the active form of ATP-binding cassette transporter A1 (ABCA1), and the plasmid vector also includes at least one sequence adapted to promote expression of the open reading frame in a mammalian cell; and

wherein the sonochemically-active microspheres comprise gas bubbles encapsulated within shells comprising a protein, a lipid, or a combination thereof, the microspheres being disruptable upon exposure to ultrasonic acoustic energy to release the encapsulated gas bubbles; and

wherein the composition transfects liver cells when intravenously administered to the patient while ultrasonically imaging the liver, to elicit production of an increased level of HDL-cholesterol in the blood of the patient compared to the HDL-cholesterol level in the blood of the patient prior to administering the composition.

2. The composition of claim 1 wherein the microspheres have an average particle size in the range of about 0.5 to about 20 micrometers.

3. The composition of claim 1 wherein the gas bubbles comprise a fluorocarbon gas.

4. The composition of claim 1 wherein the shells comprise human serum albumin.

5. The composition of claim 1 wherein the active form of ABCA1 has the amino acid sequence of SEQ ID NO:

2.

6. The composition of claim 1 wherein the open reading frame has the nucleotide sequence of SEQ ID NO:

1.

7. The composition of claim 1 wherein the at least one sequence adapted to promote expression of the open reading frame comprises a cytomegalovirus promoter.

8. The composition of claim 1 wherein the plasmid is present in the composition at a concentration in the range of about 0.5 to about 50 milligrams per milliliter.

9. The composition of claim 1 wherein the microspheres are present in the composition at a concentration in the range of about 10⁸ to about 10⁹ microspheres per milliliter.

10. The composition of claim 1 wherein the aqueous carrier comprises physiological saline, optionally buffered at physiological pH.

11. The composition of claim 1 further comprising a drug for treating a condition relating to lipid metabolism or transport.

12. A pharmaceutical composition for increasing HDL-cholesterol levels in the blood of a patient, the composition comprising a mixture of about 0.5 to about 50 milligrams per milliliter of a nucleic acid encoding an expressible open reading frame and about 108 to about 109 microspheres per milliliter of sonochemically-active microspheres in a pharmaceutically acceptable aqueous carrier;

wherein the nucleic acid of the mixture consists of a plasmid vector, the expressible open reading frame consists of an expressible open reading frame encoding the active form of ATP-binding cassette transporter A1 (ABCA1), and the plasmid vector also includes at least one sequence adapted to promote expression of the open reading frame in a mammalian cell; and

wherein the sonochemically-active microspheres comprise fluorocarbon gas bubbles encapsulated within shells comprising human serum albumin, the microspheres being disruptable upon exposure to ultrasonic acoustic energy to release the encapsulated gas bubbles; and

wherein the composition transfects liver cells when intravenously administered to the patient while ultrasonically imaging the liver, to elicit production of an increased level of HDL-cholesterol in the blood of the patient compared to the HDL-cholesterol level in the blood of the patient prior to administering the composition.

13. The composition of claim 12 wherein the active form of ABCA1 has the amino acid sequence of SEQ ID NO:

2.

14. The composition of claim 12 wherein the open reading frame has the nucleotide sequence of SEQ ID NO:

1.