title: A phospholipid-PEG2000 conjugate of a vascular endothelial growth factor receptor 2 (VEGFR2)-targeting heterodimer peptide for contrast-enhanced ultrasound imaging of angiogenesis.
abstract: The transition of a targeted ultrasound contrast agent from animal imaging to testing in clinical studies requires considerable chemical development. The nature of the construct changes from an agent that is chemically attached to microbubbles to one where the targeting group is coupled to a phospholipid, for direct incorporation to the bubble surface. We provide an efficient method to attach a heterodimeric peptide to a pegylated phospholipid and show that the resulting construct retains nanomolar affinity for its target, vascular endothelial growth factor receptor 2 (VEGFR2), for both the human (kinase insert domain-containing receptor - KDR) and the mouse (fetal liver kinase 1 - Flk-1) receptors. The purified phospholipid-PEG-peptide isolated from TFA-based eluents is not stable with respect to hydrolysis of the fatty ester moieties. This leads to the time-dependent formation of the lysophospholipid and the phosphoglycerylamide derived from the degradation of the product. Purification of the product using neutral eluent systems provides a stable product. Methods to prepare the lysophospholipid (hydrolysis product) are also included. Biacore binding data demonstrated the retention of binding of the lipopeptide to the KDR receptor. The phospholipid-PEG2000-peptide is smoothly incorporated into gas-filled microbubbles and provides imaging of angiogenesis in a rat tumor model.
authors.affiliation: The Ernst Felder Laboratories, Bracco Research USA, 305 College Road East, Princeton, New Jersey 08540, and Bracco Research SA, Route de la Galaise, 31, CH-1228 Plan-les-Ouates, Geneva, Switzerland.
authors.forename: R
authors.initials: R
authors.lastname: Pillai
authors.forename: E R
authors.initials: ER
authors.lastname: Marinelli
authors.forename: H
authors.initials: H
authors.lastname: Fan
authors.forename: P
authors.initials: P
authors.lastname: Nanjappan
authors.forename: B
authors.initials: B
authors.lastname: Song
authors.forename: M A
authors.initials: MA
authors.lastname: von Wronski
authors.forename: S
authors.initials: S
authors.lastname: Cherkaoui
authors.forename: I
authors.initials: I
authors.lastname: Tardy
authors.forename: S
authors.initials: S
authors.lastname: Pochon
authors.forename: M
authors.initials: M
authors.lastname: Schneider
authors.forename: A D
authors.initials: AD
authors.lastname: Nunn
authors.forename: R E
authors.initials: RE
authors.lastname: Swenson
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D015674:Mammary Neoplasms, Animal / Q000098:blood supply* / Q000378:metabolism; D051379:Mice; D015394:Molecular Structure; D009389:Neovascularization, Pathologic / Q000000981:diagnostic imaging* / Q000473:pathology; D010455:Peptides / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D051381:Rats; D011916:Rats, Inbred F344; D014463:Ultrasonography; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism*
chemical_list: D010455:Peptides; D010743:Phospholipids; D011092:Polyethylene Glycols; D040301:Vascular Endothelial Growth Factor Receptor-2