PURPOSE: To design a binding-induced conformation change drug delivery system for integrin-targeted delivery of methotrexate and prove the feasibility of using hairpin peptide structure for binding triggered drug delivery. METHODS: Methotrexate prodrugs were synthesized using solid phase peptide synthesis techniques by conjugating methotrexate to Arg-Gly-Asp (RGD) or a hairpin peptide, RWQYV(D)PGKFTVQRGD (hairpin-RGD). Levels of integrin α(V)β(3) in HUVEC were up-regulated using adenoviral system and knocked down using siRNA. Stability of prodrugs and methotrexate release from prodrugs were evaluated in plasma, in presence or absence of integrin α(V)β(3)-expressing cells. Molecular modeling was performed to support experimental results using MOE. RESULTS: Prodrugs recognized and bound to integrin α(V)β(3)-expressing cells in integrin α(V)β(3) expression level-dependent manner. Prodrug with hairpin peptide could resist Streptomyces griseus-derived glutamic acid-specific endopeptidase (SGPE) and plasma enzyme hydrolysis. Drug release was triggered in presence of HUVEC cells and SGPE. Analysis of conformation energy supported that conformational change in MTX-hairpin-RGD led to exposure of labile link upon binding to integrin α(V)β(3)-expressing cells. CONCLUSIONS: Binding-induced conformation change of hairpin peptide can be used to design integrin-targeted drug delivery system.
PURPOSE: To design a binding-induced conformation change drug delivery system for integrin-targeted delivery of methotrexate and prove the feasibility of using hairpin peptide structure for binding triggered drug delivery. METHODS:Methotrexate prodrugs were synthesized using solid phase peptide synthesis techniques by conjugating methotrexate to Arg-Gly-Asp (RGD) or a hairpin peptide, RWQYV(D)PGKFTVQRGD (hairpin-RGD). Levels of integrin α(V)β(3) in HUVEC were up-regulated using adenoviral system and knocked down using siRNA. Stability of prodrugs and methotrexate release from prodrugs were evaluated in plasma, in presence or absence of integrin α(V)β(3)-expressing cells. Molecular modeling was performed to support experimental results using MOE. RESULTS: Prodrugs recognized and bound to integrin α(V)β(3)-expressing cells in integrin α(V)β(3) expression level-dependent manner. Prodrug with hairpin peptide could resist Streptomyces griseus-derived glutamic acid-specific endopeptidase (SGPE) and plasma enzyme hydrolysis. Drug release was triggered in presence of HUVEC cells and SGPE. Analysis of conformation energy supported that conformational change in MTX-hairpin-RGD led to exposure of labile link upon binding to integrin α(V)β(3)-expressing cells. CONCLUSIONS: Binding-induced conformation change of hairpin peptide can be used to design integrin-targeted drug delivery system.
Authors: Ingrid Dijkgraaf; John A W Kruijtzer; Shuang Liu; Annemieke C Soede; Wim J G Oyen; Frans H M Corstens; Rob M J Liskamp; Otto C Boerman Journal: Eur J Nucl Med Mol Imaging Date: 2006-08-15 Impact factor: 9.236
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Authors: R J Francis; S K Sharma; C Springer; A J Green; L D Hope-Stone; L Sena; J Martin; K L Adamson; A Robbins; L Gumbrell; D O'Malley; E Tsiompanou; H Shahbakhti; S Webley; D Hochhauser; A J Hilson; D Blakey; R H J Begent Journal: Br J Cancer Date: 2002-09-09 Impact factor: 7.640