Methods for delivering compounds into a cell

• US 7,612,033 B2
• Filed: 03/18/2004
• Issued: 11/03/2009
• Est. Priority Date: 11/29/1994
• Status: Expired due to Fees

• Current Assignee: Cerevast Therapeutics, Inc.
• Original Assignee: Imarx Pharmaceutical Corp. (Sycamore Entertainment Group, Inc.)
• Inventors: McCreery, Thomas, Unger, Evan C.
• Primary Examiner(s): Tsang; Cecilia
• Assistant Examiner(s): AUDET, MAURY A
• Application Number:
  US10/802,919
• Publication Number:
  US 20040176473A1
• Time in Patent Office: 2,056 Days
• Field of Search: None
• US Class Current: 514/1.1
• 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 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: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• 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
• 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 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle

Abstract

The present invention is directed, inter alia, to a method for delivering a compound into a cell comprising administering to the cell the compound to be delivered, an organic halide, and/or a carrier. Ultrasound may also be applied, if desired.

Claims

1. A method for delivering a protein into a cell in vivo, comprising:

(a) forming a plurality of protein-halide vesicles comprising the protein to be delivered and an organic halide incorporated in the core of the vesicles; and

(b) administering the protein-halide vesicles to the cell,

wherein the organic halide is selected from the group consisting of 1-bromo-nonafluorobutane, perfluorooctyliodide, perfluoroocytlbromide, 1-chloro-1-fluoro-1-bromomethane, 1,1,1-trichloro-2,2,2-trifluoroethane, 1,2-dichloro-2,2-difluoroethane, 1,1-dichloro-1,2-difluoroethane, 1,2-dichloro-1,1,3-trifluoropropane, 1-bromoperfluorobutane, 2-iodo-1,1,1-trifluoroethane, 5-bromovaleryl chloride, 1,3-dichlorotetrafluoroacetone, 1-bromo-1,1,2,3,3,3-hexafluoropropane, 2-chloro-1,1,1,4,4,4-hexafluoro-2-butene, 2-chloropentafluoro-1,3-butadiene, iodotrifluoroethylene, 1,1,2-trifluoro-2-chloroethane, 1,2-difluorochloroethane, 1,1-difluoro-2-chloroethane, 1,1-dichlorofluoroethane, heptafluoro-2-iodopropane, bromotrifluoroethane, chlorotrifluoromethane, dibromofluoromethane, chloropentafluoroethane, bromochlorodifluoromethane, dichloro-1,1,2,2-tetrafluoroethane, 1,1,1,3,3-pentafluoropentane, perfluorotributylamine, perfluorotripropylamine, 2,2,2-trifluoroethylacrylate, 3-(trifluoromethoxy)-acetophenone, 1,1,2,2,3,3,4,4-octafluorobutane, 1,1,1,3,3-pentafluorobutane, 1-fluorobutane, 1,1,2,2,3,3,4,4-octafluorobutane, 1,1,1,3,3-pentafluorobutane, tetradecaperfluoroheptane, dodecaperfluorocyclohexane, perfluoromethane, perfluoroethane, perfluoropropane, perfluorobutane, perfluoropentane, perfluorohexane, perfluoroheptane, perfluorooctane, perfluorononane, perfluorodecane, perfluorododecane, perfluoro-2-methyl-2-pentene, perfluorocyclohexane, perfluoropropylene, perfluorocyclobutane, perfluoro-2-butyne, perfluoro-2-butene, perfluorobuta-1,3-diene, perfluorobutylethyl ether, bis(perfluoroisopropyl) ether, bis(perfluoropropyl) ether, perfluoromethyl tetrahydrofuran, perfluoro t-butyl methyl ether, perfluoro isobutyl methyl ether;

perfluoro n-butyl methyl ether, perfluoro isopropyl ethyl ether, perfluoro n-propyl ethyl ether, perfluoro cyclobutyl methyl ether, perfluoro cyclopropyl ethyl ether, perfluoro isopropyl methyl ether, perfluoro n-propyl methyl ether, perfluoro diethyl ether, perfluoro cyclopropyl methyl ether, perfluoro methyl ethyl ether, and perfluoro dimethyl ether,

wherein the path of administering of the protein-halide composition to the cell is selected from a group consisting of the administering through a cell membrane, cell wall, and nuclear membrane, or any combination thereof,

to achieve the intracellular delivery of the protein thereby.

2. The method of claim 1, wherein the organic halide is selected from the group consisting of 1-bromo-nonafluorobutane, 1,1,1,3,3-pentafluoropentane, perfluorohexane, perfluorocyclohexane, 1-bromo-1,1,2,3,3,3-hexafluoropropane, heptafluoro-2-iodopropane, 1,1,2,2,3,3,4,4-octafluorobutane, 1-fluorobutane, tetradecaperfluoroheptane, and dodecaperfluorocyclohexane.

3. The method of claim 1, wherein the organic halide is selected from the group consisting of perfluorohexane and perfluorocyclohexane.

4. The method of claim 1, wherein the protein is selected from the group consisting of albumin, collagen, polyarginine, polylysine, polyhistidine, γ

-globulin, and β

-globulin.

5. The method of claim 1, wherein the protein is a cationic protein.

6. The method of claim 5, wherein the cationic protein is selected from the group consisting of polylysine and polyethyleneimine.

7. The method of claim 1, further comprising applying ultrasound to the cell.

8. The method of claim 7, wherein the ultrasound is applied at a frequency between about 40 kHz and 25 MHz, and an energy level between about 500 mW/cm² and 10 W/cm².

9. The method of claim 7, wherein the ultrasound is applied at a frequency between about 200 kHz and 500 kHz, and the energy level is between about 200 mW/cm² and 500 W/cm².

10. The method of claim 7, wherein the ultrasound is applied at a frequency between about 1 MHz and 20 MHz, and the energy level is between about 100 W/cm² and 200 W/cm².

11. The method of claim 10, wherein the ultrasound is applied at a duty cycle between about 1% and 100% of the treatment time.

12. The method of claim 7, wherein the protein and the ultrasound are administered and applied simultaneously.