Novel integrin-targeted binding-triggered drug delivery system for methotrexate.

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.