Targeting vector-phospholipid conjugates
View Patent ↗Peptide vectors having high KDR binding affinity and processes for making such vectors are provided. The peptide vectors may be conjugated to phospholipids and included in ultrasound contrast agent compositions. Such ultrasound contrast agents are particularly useful in therapeutic and diagnostic methods, such as in imaging KDR-containing tissue and in the evaluation and treatment of angiogenic processes associated with neoplastic conditions. The present invention also provides processes for the large scale production of highly pure dimeric and monomeric peptide phospholipid conjugates as well as precursor materials used to form the conjugates. The present invention further provides processes for the large scale production of highly pure peptide phospholipid conjugates which contain very low levels of TFA.
1. A peptide-phospholipid conjugate selected from the group consisting of:
2. An ultrasound contrast agent composition comprising a conjugate of claim 1 , wherein the contrast agent comprises a gas-filled microvesicle.
3. The composition of claim 2 , wherein the gas filled microvesicle comprises a phospholipid.
4. The composition of claim 2 , wherein the gas-filled microvesicle further comprises two or more components selected form the group consisting of: DSPC, DPPG, DPPA, DSPA, DPPE, DSPL-PEG1000, DSPL-PEG2000, palmitic acid and stearic acid.
5. The composition of claim 4 , wherein the conjugate comprises Ac-AGPTWCEDDWYYCWLFGTGGGK{Ac-VCWEDSWGGEVCFRYDP-GGGK[-Adoa-Adoa-Glut-K(DSPE-PEG2000-NH-Glut)]-NH2 cyclic (2-12) disulfide}-NH2 cyclic (6-13) disulfide and the contrast agent further comprises DSPC and DPPG.
6. The composition of claim 4 , wherein the conjugate comprises Ac-AGPTWCEDDWYYCWLFGTGGGK{Ac-VCWEDSWGGEVCFRYDP-GGGK[-Adoa-Adoa-Glut-K(DSPE-PEG2000-NH-Glut)]-NH2 cyclic (2-12) disulfide}-NH2 cyclic (6-13) disulfide and the contrast agent further comprises DSPE-PEG1000, DPPE and DPPG.
7. The composition of claim 4 , wherein the conjugate comprises Ac-AGPTWCEDDWYYCWLFGTGGGK{Ac-VCWEDSWGGEVCFRYDP-GGGK[-Adoa-Adoa-Glut-K(DSPE-PEG2000-NH-Glut)]-NH2 cyclic (2-12) disulfide}-NH2 cyclic (6-13) disulfide and the contrast agent further comprises DSPE-PEG1000, DSPC and DSPA.
8. The composition of claim 4 , wherein the conjugate comprises Ac-AGPTWCEDDWYYCWLFGTGGGK{Ac-VCWEDSWGGEVCFRYDP-GGGK[-Adoa-Adoa-Glut-K(DSPE-PEG2000-NH-Glut)]-NH2 cyclic (2-12) disulfide}-NH2 cyclic (6-13) disulfide and the contrast agent further comprises DSPE-PEG2000, DSPC and stearate.
9. The composition of claim 4 , wherein the conjugate comprises Ac-AGPTWCEDDWYYCWLFGTGGGK{Ac-VCWEDSWGGEVCFRYDP-GGGK[-Adoa-Adoa-Glut-K(DSPE-PEG2000-NH-Glut)]-NH2 cyclic (2-12) disulfide}-NH2 cyclic (6-13) disulfide and the contrast agent further comprises DSPC and stearate.
10. The composition of claim 4 , wherein the conjugate comprises Ac-AGPTWCEDDWYYCWLFGTGGGK{Ac-VCWEDSWGGEVCFRYDP-GGGK[-Adoa-Adoa-Glut-K(DSPE-PEG2000-NH-Glut)]-NH2 cyclic (2-12) disulfide}-NH2 cyclic (6-13) disulfide and the contrast agent further comprises DSPE-PEG1000, DSPC and stearate.
11. The composition of claim 4 further comprising a component selected from the group consisting of: sugars, polysaccharides and polyols.
12. The composition of claim 11 , wherein said component is selected from mannitol, dextran and polyethyleneglycol.
13. The composition of claim 11 , wherein said gas comprises a fluorinated gas.
14. The composition any one of claims 2 , 6 , 7 , 8 , 9 , 10 , or 11 wherein the gas comprises C3F8, C4F10 or SF6, optionally in admixture with air, nitrogen, oxygen or carbon dioxide.
15. A method for imaging KDR-containing tissue in a mammal comprising administering an effective amount of a composition of claim 2 to the mammal and imaging the mammal.
16. A method for detecting or imaging angiogenic processes in a mammal comprising administering an effective amount of a composition of claim 2 to the mammal and imaging the mammal.
17. A method for detecting or imaging tumor tissue containing KDR in a mammal comprising administering an effective amount of a composition of claim 2 to the mammal and imaging the mammal.
18. An ultrasound contrast agent composition comprising one or more monomers selected from the group consisting of Ac-Arg-Ala-Gln-Asp-Trp-Tyr-Tyr-Asp-Glu-Ile-Leu-Ser-Met-Ala-Asp-Gln-Leu-Arg-His-Ala-Phe-Leu-Ser-Gly-Gly-Gly-Gly-Gly-Lys-NH2 (SEQ ID NO 5); Ac-Ala-Gln-Asp-Trp-Tyr-Tyr-Asp-Glu-Ile-Leu-Ser-Met-Ala-Asp-Gln-Leu-Arg-His-Ala-Phe-Leu-Ser-Gly-Gly-Gly-Gly-Gly-Lys-NH2 (SEQ ID NO 6); Ac-Ala-Gly-Pro-Thr-Trp-Cys-Glu-Asp-Asp-Trp-Tyr-Tyr-Cys-Trp-Leu-Phe-Gly-Thr-Gly-Gly-Gly-Lys(ivDde)-NH2 cyclic (6-13) disulfide (SEQ ID NO 7), and Ac-Val-Cys-Trp-Glu-Asp-Ser-Trp-Gly-Gly-Glu-Val-Cys-Phe-Arg-Tyr-Asp-Pro-Gly-Gly-Gly-Lys(Adoa-Adoa)-NH2 cyclic (2-12) disulfide (SEQ ID NO 8).
19. The composition of claim 18 , wherein the contrast agent comprises a gas filled microvesicle.
20. The composition of claim 19 , wherein said gas comprises a fluorinated gas.
21. The composition of claim 20 , further comprising a phospholipid.
22. The composition of claim 21 , further comprising two or more components selected from the group consisting of: DSPC, DPPG, DPPA, DSPA, DPPE, DSPE-PEG1000, DSPE-PEG2000, palmitic acid and stearic acid.
23. The composition of claim 22 , wherein the gas the gas comprises C3F8, C4F10 or SF6, optionally in admixture with air, nitrogen, oxygen or carbon dioxide.
24. A method for imaging KDR-containing tissue in a mammal comprising administering an effective amount of a composition of claim 18 to the mammal and imaging the mammal.
25. A method for detecting or imaging angiogenic processes in a mammal comprising administering an effective amount of a composition of claim 18 to the mammal and imaging the mammal.
26. A method for detecting or imaging tumor tissue containing KDR in a mammal comprising administering an effective amount of a composition of claim 18 to the mammal and imaging the mammal.