Details for Patent: 6,521,211

Title:	Methods of imaging and treatment with targeted compositions
Abstract:	Novel ultrasound methods comprising administering to a patient a targeted vesicle composition which comprises vesicles comprising a lipid, protein or polymer, encapsulating a gas, in combination with a targeting ligand, and scanning the patient using ultrasound. The scanning may comprise exposing the patient to a first type of ultrasound energy and then interrogating the patient using a second type of ultrasound energy. The targeting ligand preferably targets tissues, cells or receptors, including myocardial cells, endothelial cells, epithelial cells, tumor cells and the glycoprotein GPIIbIIIa receptor. The methods may be used to detect a thrombus, enhancement of an old or echogenic thrombus, low concentrations of vesicles and vesicles targeted to tissues, cells or receptors.
Inventor(s):	Unger; Evan C. (Tucson, AZ), Wu; Yunqiu (Tucson, AZ)
Assignee:	Bristol-Myers Squibb Medical Imaging, Inc. (Princeton, NJ)
Filing Date:	Feb 03, 1999
Application Number:	09/243,640
Claims:	1. A diagnostic ultrasound method comprising (i) administering to a patient a targeted vesicle composition which comprises vesicles encapsulating a gas, in combination with a targeting ligand; and (ii) scanning the patient using dual frequency ultrasound insonation, wherein said vesicles are selected from the group consisting of liposomes and micelles, and said vesicles comprise a phospholipid selected from the group consisting of dioleoylphosphatidylcholine, dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dioleoylphosphatidylethanolamine, N-succinyldioleoylphosphatidylethanolamine,1 -hexadecyl-2-palmitoylglycerophosphoethanolamine, and phosphatidic acids, and wherein said scanning comprises exposing the patient to a first ultrasound energy having a first insonation frequency to cause the vesicle to oscillate, and then subsequently, while the vesicle is oscillating, exposing the patient to a second ultrasound energy having a second insonation frequency that is different from said first insonation frequency, and detecting the reflected ultrasound signal. 2. A method according to claim 1, wherein the method is used for detection of a thrombus. 3. A method according to claim 2, wherein said thrombus comprises an old or echogenic thrombus. 4. A method according to claim 1, wherein the method is used for detecting vesicles targeted to endothelial tissue. 5. A method according to claim 4, wherein said endothelial tissue includes integrins associated with malignancy or inflammation. 6. A method according to claim 1, comprising the use of insonation frequencies with a bandwidth of about 100 kHz or less. 7. A method according to claim 6, wherein said bandwidth is about 50 kHz or less. 8. A method according to claim 7, wherein said bandwidth is about 10 Hz or less. 9. A method according to claims 8, wherein said bandwidth is less than about 1000 Hz. 10. A method according to claim 1, wherein detecting the reflected ultrasound signal comprises the use of a broadband receiver and a digital filter with multiple center frequencies, wherein said center frequencies are digitally adjusted with respect to an insonating frequency, and filter gates and bandwidth windows are selected and controlled such that said filter rejects signals with center frequencies outside of a pre-selected range. 11. A method according to claim 1, comprising the use of pulsing and phase modulation of said first and second ultrasound energy. 12. A method according to claim 1, wherein the frequency of said first ultrasound energy is lower than the frequency of said second ultrasound energy. 13. A method according to claim 12, wherein said first ultrasound energy has a frequency of about 1 MHz and said second ultrasound energy has a frequency of about 3 MHz. 14. A method according to claim 12, wherein said second ultrasound energy has a frequency at least two times that of said first ultrasound energy. 15. A method according to claim 12, wherein said first ultrasound energy has a frequency of about 100 kHz and said second ultrasound energy has a frequency of about 3 MHz. 16. A method according to claim 12, wherein said first ultrasound energy is administered as a pulse train. 17. A method according to claim 16, wherein said pulse train comprises about 10 or fewer pulses. 18. A method according to claim 12, wherein said first ultrasound energy is administered as a single pulse of low frequency ultrasound, and said second ultrasound energy comprises pulses of higher frequency ultrasound. 19. A method according to claim 12, wherein said first ultrasound energy is a single pulse of about 100 kHz, said second ultrasound energy comprises one or several pulses of higher frequency ultrasound, and said second ultrasound energy is applied within about 40 milliseconds of said first ultrasound energy. 20. A method according to claim 1, comprising forming a summation of several pulses given in rapid succession after an initial stimulation pulse. 21. A method according to claim 1, wherein said first and second ultrasound energy comprise pulses that are phase modulated with respect to each other. 22. A method according to claim, 21, wherein said phase modulation and/or delay is/are selected to maximize vesicle oscillation and collapse. 23. A method according to claim 21, wherein said first ultrasound energy comprises one pulse or a train with a duration from about 10 microseconds to about 10 seconds. 24. A method according to claim 23, wherein said duration id from about 1 millisecond to about 2 seconds. 25. A method according to claim 20, wherein a peak acoustic pressure for a first pulse of about 100 pascals to about 10 megapascals is employed. 26. A method according to claim 25, wherein said peak acoustic pressure is about 1 kilopascal to about 5 megapascals. 27. A method according to claim 26, wherein said peak acoustic pressure is about 10 kilopascals to about 5 megapascals. 28. A method according to claim 1 wherein said phospholipid is dipalmitoylphosphatidylethanolamine. 29. A method according to claim 1 wherein said phosphatidic acid is dipalmitoylphosphatidic acid. 30. A method according to claim 1 wherein said phospholipid further comprises a polymer. 31. A method according to claim 30 wherein said polymer comprises a hydrophilic polymer. 32. A method according to claim 31 wherein said polymer comprises polyethylene glycol. 33. A method according to claim 1, wherein said gas comprises a fluorinated gas. 34. A method according to claim 33, wherein said fluorinated gas comprises a perfluorocarbon gas. 35. A method according to claim 34 wherein said perfluorocarbon gas is selected from the group consisting of perfluoromethane, perfluoroethane, perfluoropropane, perfluorobutane and perfluorocyclobutane. 36. A method according to claim 35 wherein said perfluorocarbon gas is selected from the group consisting of perfluoropropane and perfluorobutane. 37. A method according to claim 36 wherein said perfluorocarbon gas comprises perfluorobutane. 38. A method according to claim 1 wherein said gas is derived, at least in part, from a gaseous precursor. 39. A method according to claim 38 wherein said gaseous precursor has a boiling point of greater than about 37.degree. C. 40. A method according to claim 38 wherein said gaseous precursor comprises a perfluorocarbon. 41. A method according to claim 40 wherein said perfluorocarbon is selected from the group consisting of perfluoropentane and perfluorohexane. 42. A method according to claim 1 wherein said targeting ligand targets cells or receptors selected from the group consisting of myocardial cells, endothelial cells, epithelial cells, tumor cells and the glycoprotein GPIIbIIIa receptor. 43. A method according to claim 42 wherein said targeting ligand is selected from the group consisting of proteins, peptides, saccharides, steroids, steroid analogs, bioactive agents and genetic material. 44. A method according to claim 43 wherein said targeting ligand is selected from the group consisting of proteins, peptides and saccharides. 45. A method according to claim 44 wherein said targeting ligand is selected from the group consisting of proteins and peptides. 46. A method according to claim 45 wherein said targeting ligand comprises a peptide. 47. A method according to claim 46 wherein said peptide comprises a sequence selected from the group consisting of Arg--Gly-Asp and Lys-Gln-Ala-Gly-Asp--Val SEQ ID NO. 1. 48. A method according to claim 1 wherein said targeting ligand is associated with said vesicles covalently. 49. A method according to claim 1 wherein said targeting ligand is unbound. 50. A method according to claim 1 wherein said targeting ligand is associated with said vesicles non-covalently. 51. A method according to claim 48 where said covalent association comprises a covalent bond selected from the group consisting of amide, thioamide, ether, ester, thioester, O--, --S--, --S.sub.n --, wherein n is greater than 1, carbamate, --NH--, --NR--, where R is alkyl of from 1 to about 4 carbons, urethane, and substituted imidate bonds. 52. A method according to claim 51 wherein said covalent association further comprises crosslinking. 53. A method according to claim 48 wherein said targeting ligand is covalently associated with said phospholipid via linking group. 54. A method according to claim 53 wherein said linking group comprises a hydrophilic polymer. 55. A method according to claim 54 wherein said hydrophilic polymer is selected from the group consisting of polyalkyleneoxides, polyvinyl alcohol, polyvinylpyrrolidones, polyacrylamides, polymethacrylamides, polyphosphazenes, poly(hydroxyalkylcarboxylic acids) and polyoxazolidines. 56. A method according to claim 54 wherein said hydrophilic polymer comprises a polyalkyleneoxide. 57. A method according to claim 56 wherein said polyalkyleneoxide is selected from the group consisting of polyethylene glycol and polypropylene glycol. 58. A method according to claim 57 wherein said polyalkyleneoxide comprises polyethylene glycol.