Targeted gas-filled microvesicles formulation
Suspension of gas-filled microvesicles comprising a targeting ligand for binding to KDR or VEGF/KDR complex. The suspension is obtained by reconstituting a freeze-dried residue with a carbohydrate-containing solution in the presence of a physiologically acceptable gas and is stabilized by the presence of histidine.
1. A physiologically acceptable aqueous suspension of gas-filled microvesicles, said gas-filled microvesicles comprising a at least one lipid and a lipopeptide of formula II:
wherein said lipopeptide is present in a molar amount of 0.5% to by molar ratio with respect to the at least one lipid, and wherein the at least one lipid comprises
distearoyl-phosphatidyl-choline (DSPC) and palmitic acid in a molar ratio of 95:5 and DPPE-PEG5000 in a molar amount of 0.5% by molar ratio with respect to the at least one lipid and the lipopeptide, said aqueous suspension further comprising
glucose at a concentration of 5% (w/w);
and a histidine at a concentration of from 1.5 mM to 20 mM.
2. The aqueous suspension according to claim 1 , wherein said gas-filled microvesicles comprise a fluorinated physiologically acceptable gas.
3. The aqueous suspension according to claim 1 , wherein the concentration of the histidine in the aqueous suspension is from 2.5 mM to 10 mM.
4. A pharmaceutical kit for preparing a reconstituted aqueous suspension of gas-filled microvesicles comprising:
(a) A first container comprising a physiologically acceptable gas in contact with a freeze-dried formulation comprising (i) at least one lipid, wherein the at least one lipid comprises distearoyl-phosphatidyl-choline (DSPC) and palmitic acid in a molar ratio of 95:5, (ii) a lipopeptide of formula II:
wherein said lipopeptide is present in a molar amount of 0.5% by molar ratio with respect to the at least one lipid, and wherein the at least one lipid further comprises DPPE-PEG5000, wherein the DPPE-PEG5000 is present in a molar amount of 0.5% by molar ratio with respect to the at least one lipid and the lipopeptide; and
(iii) a histidine, said histidine being in an amount capable of reconstitution to a concentration of from 1.5 mM to 20 mM in the reconstituted aqueous suspension of gas-filled microvesicles; and
(b) a second container comprising a glucose-containing physiologically acceptable solution, at a glucose concentration of 5% (w/w), for reconstituting the freeze-dried formulation into the reconstituted aqueous suspension of gas-filled microvesicles.
5. The pharmaceutical kit according to claim 4 , wherein the amount of the histidine is such that the concentration of the histidine in the reconstituted aqueous suspension of gas-filled microvesicles is from 2.5 mM to 10 mM.
6. A method for imaging a region of interest in a subject comprising administering an effective amount of the aqueous suspension of gas-filled microvesicles according to claim 1 to the subject,
submitting said region of interest to ultrasound irradiation, and
collecting respective echo signals from said region of interest.
7. A pharmaceutical kit for preparing a reconstituted aqueous suspension of gas-filled microvesicles comprising:
(a) A first container comprising a physiologically acceptable gas in contact with a freeze-dried formulation comprising (i) at least one lipid, wherein the at least one lipid comprises distearoyl-phosphatidyl-choline (DSPC) and palmitic acid in a molar ratio of 95:5, (ii) a lipopeptide of formula II:
wherein said lipopeptide is present in a molar amount of 0.5% by molar ratio with respect to the at least one lipid, and wherein the at least one lipid further comprises DPPE-PEG5000, wherein the DPPE-PEG5000 is present in a molar amount of 0.5% by molar ratio with respect to the at least one lipid and the lipopeptide;
(iii) a histidine, said histidine being in an amount capable of reconstitution to a concentration of from 2.5 mM to 10 mM in the reconstituted aqueous suspension of gas-filled microvesicles; and
(iv) PEG4000; and
(b) a second container comprising a glucose-containing physiologically acceptable solution, at a glucose concentration of 5% (w/w), for reconstituting the freeze-dried formulation into the reconstituted aqueous suspension of gas-filled microvesicles.