Gas-filled microvesicles composition for contrast imaging
The present invention relates to a new composition comprising gas-filled microvesicles for contrast imaging which are particularly suitable for providing an effective echo response to at least two selected ultrasound waves having different frequencies. Said composition preferably comprises at least two different preparations of gas-filled microvesicles having respective peaks of non-liner echographic response differing by at least 2 MHz to each other, and preferably have respective size distributions with different mean sizes. In particular, said preparations preferably have size distributions with respective D V50 values differing from each other by at least 0.5 μm, more preferably at least 1.0 μm. Alternatively, said composition has a volume size distribution showing a value of Bowley skewness of 0.16 or higher. According to a preferred embodiment, at least 95% of the total volume of gas contained in said microvesicles, calculated on the population of microvesicles up to a diameter of 10 μm, is contained in microvesicles having a diameter of 8 micron or less.
1. A composition for diagnostic and/or therapeutic imaging which comprises a mixture of at least two different preparations of gas-filled microvesicles, wherein
a) said at least two different preparations are microbubbles stabilized by a film layer of an amphiphilic material and have respective size distributions with different median sizes, said median sizes being defined by a respective first and second median diameter in volume (D V50 ) and said first and second D V50 differing from each other by a value of at least 0.5 μm;
b) said at least two preparations have a size distribution defined by a respective ratio between said mean diameter in volume and a corresponding mean diameter in number (D V /D N ratio), each of said at least two preparations having a D V /D N ratio of from 1.2 to 3;
c) at least 95% of the total volume of gas contained in said microvesicles with a diameter of 10 μm or less is contained in microvesicles having a diameter of 8 μm or less;
d) said at least two different preparations have respective peaks of non-linear echographic response differing by at least 2 MHz to each other wherein first preparation of microvesicles has a peak of non-linear echographic response of from 1.5 to 3 MHz, and a second preparation of microvesicles with a peak of non-linear echographic response of from 5 to 10 MHz; and
e) the volume size distribution of said gas-filled microvesicles, determined on a population of microvesicles with a diameter up to 8 μm, has a Bowley skewness of 0.16 or higher.
2. A composition according to claim 1 wherein said peaks of non-linear echographic response differ by at least 3 MHz to each other.
3. A composition according to claim 1 wherein said first and second D V50 differ from each other by a value of at least 1.0 μm.
4. A composition according to claim 1 , wherein the microbubbles of said at least two sets have a. size distribution defined by a respective ratio between said mean diameter in volume and a corresponding mean diameter in number (D V /D N ), at least one of said sets of gas-filled microbubbles having a D V /D N ratio of from 1.2 to 2.
5. A composition according to claim 1 wherein said amphiphilic material is a phospholipid.
6. A composition according to claim 1 , further comprising a physiologically acceptable aqueous carrier.
7. A composition according to claim 1 , wherein said gas-filled microvesicles are in the form of a dried powder reconstitutable upon contact with a physiologically acceptable aqueous carrier.
8. A composition according to claim 1 , wherein said gas-filled microvesicles comprise a targeting ligand, a diagnostic agent, a bioactive agent or any combination thereof.
9. A composition according to claim 1 , wherein at least one of said preparation of gas-filled microvesicles comprises a targeting ligand, a diagnostic agent, a bioactive agent or any combination thereof.
10. A method of manufacturing a contrast agent according to claim 1 , which comprises admixing at least two different preparations of gas-filled microvesicles or precursors thereof having respective peaks of non-linear echographic response differing by at least 2 MHz to each other.
11. A method according to claim 10 wherein said precursors are in the form of a dried powder forming said microvesicle preparation upon reconstitution in a pharmaceutically acceptable liquid carrier.
12. A method according to claim 11 wherein the at least two different preparations of gas-filled microvesicles or precursors thereof are directly obtained as a combined preparation by applying different process parameter to a same preparation mixture.
13. A method according to claim 10 wherein said precursors are microemulsions obtained by dispersing a phospholipid in an emulsion of water with a water immiscible organic solvent, said emulsion forming microvesicle preparation upon lyophilization in the presence of a lyoprotecting agent and subsequent. reconstitution in a pharmaceutically acceptable liquid carrier.
14. A method of diagnostic and/or therapeutic imaging which comprises administering to a patient an effective amount of a composition according to claim 1 .
15. A diagnostic and/or therapeutic kit comprising a composition according to claim 1 in dried powdered form and a physiologically acceptable aqueous carrier.