Pharmaceutical Research

, Volume 28, Issue 12, pp 3208–3219 | Cite as

Novel Integrin-Targeted Binding-Triggered Drug Delivery System for Methotrexate

  • Phanidhara Kotamraj
  • Wade A. Russu
  • Bhaskara Jasti
  • Jay Wu
  • Xiaoling Li
Research Paper



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.


Methotrexate prodrugs were synthesized using solid phase peptide synthesis techniques by conjugating methotrexate to Arg-Gly-Asp (RGD) or a hairpin peptide, RWQYVDPGKFTVQRGD (hairpin-RGD). Levels of integrin αVβ3 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 αVβ3-expressing cells. Molecular modeling was performed to support experimental results using MOE.


Prodrugs recognized and bound to integrin αVβ3-expressing cells in integrin αVβ3 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 αVβ3-expressing cells.


Binding-induced conformation change of hairpin peptide can be used to design integrin-targeted drug delivery system.


hairpin peptide integrin αVβ3 methotrexate targeted delivery triggered release 



The authors would like to thank Dr. William Chan and Dr. Shiladitya Bhattacharya for their contributions and supports for the development of AdEasy viral vector.

Supplementary material

11095_2011_495_MOESM1_ESM.doc (234 kb)
Supplementary 1 (DOC 234 kb)


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Phanidhara Kotamraj
    • 1
    • 3
  • Wade A. Russu
    • 1
  • Bhaskara Jasti
    • 1
  • Jay Wu
    • 2
  • Xiaoling Li
    • 1
  1. 1.Department of Pharmaceutics & Medicinal Chemistry, T. J. Long School of Pharmacy & Health SciencesUniversity of the PacificStocktonUSA
  2. 2.VM DiscoveryFremontUSA
  3. 3.Abon PharmaceuticalsNorthvaleUSA

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