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Improved pharmacokinetic and biodistribution properties of the selective urokinase inhibitor PAI-2 (SerpinB2) by site-specific PEGylation: implications for drug delivery

Journal Article


Abstract


  • Purpose

    Overexpression of the serine protease urokinase (uPA) is recognised as an important biomarker of metastatic disease and a druggable anticancer target. Plasminogen activator inhibitor type-2 (PAI-2/SerpinB2) is a specific uPA inhibitor with proven potential for use in targeted therapy. However, PAI-2 is rapidly cleared via the renal system which impairs tumor uptake and efficacy. Here we aimed to improve the pharmacological properties of PAI-2 by site-specific PEGylation.

    Methods

    Several cysteine to serine substitution mutants were generated for PEGylation with PEG-maleimide (size range 12–30 kDa) and the physico-chemical and biochemical properties of the PEG-PAI-2 conjugates characterised. Radiolabeled proteins were used for evaluation of blood clearance and tissue uptake profiles in an orthotopic breast tumor xenograft mouse model.

    Results

    PEGylation of the PAI-2C161S mutant gave a predominant mono-PEGylated-PAI-2 product (~90%) with full uPA inhibitory activity, despite a significant increase in hydrodynamic radius. Compared to un-PEGylated protein the plasma half-life and AUC for PEG20-PAI-2C161S were significantly increased. This translated to a 10-fold increase in tumor retention after 24 h compared to PAI-2C161S, an effect not seen in non-target organs.

    Conclusions

    Our data underscores the potential for PEG20-PAI-2C161S drug conjugates to be further developed as anti-uPA targeted therapeutics with enhanced tumor retention.

Authors


  •   Vine, Kara L.
  •   Lobov, Sergei A. (external author)
  •   Indira Chandran, Vineesh (external author)
  •   Harris, Nathanial (external author)
  •   Ranson, Marie

Publication Date


  • 2015

Citation


  • Vine, K. Lea., Lobov, S., Indira Chandran, V., Harris, N. Lachlan Ewart. & Ranson, M. (2015). Improved pharmacokinetic and biodistribution properties of the selective urokinase inhibitor PAI-2 (SerpinB2) by site-specific PEGylation: implications for drug delivery. Pharmaceutical Research, 32 (3), 1045-1054.

Scopus Eid


  • 2-s2.0-84922998623

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/507

Number Of Pages


  • 9

Start Page


  • 1045

End Page


  • 1054

Volume


  • 32

Issue


  • 3

Abstract


  • Purpose

    Overexpression of the serine protease urokinase (uPA) is recognised as an important biomarker of metastatic disease and a druggable anticancer target. Plasminogen activator inhibitor type-2 (PAI-2/SerpinB2) is a specific uPA inhibitor with proven potential for use in targeted therapy. However, PAI-2 is rapidly cleared via the renal system which impairs tumor uptake and efficacy. Here we aimed to improve the pharmacological properties of PAI-2 by site-specific PEGylation.

    Methods

    Several cysteine to serine substitution mutants were generated for PEGylation with PEG-maleimide (size range 12–30 kDa) and the physico-chemical and biochemical properties of the PEG-PAI-2 conjugates characterised. Radiolabeled proteins were used for evaluation of blood clearance and tissue uptake profiles in an orthotopic breast tumor xenograft mouse model.

    Results

    PEGylation of the PAI-2C161S mutant gave a predominant mono-PEGylated-PAI-2 product (~90%) with full uPA inhibitory activity, despite a significant increase in hydrodynamic radius. Compared to un-PEGylated protein the plasma half-life and AUC for PEG20-PAI-2C161S were significantly increased. This translated to a 10-fold increase in tumor retention after 24 h compared to PAI-2C161S, an effect not seen in non-target organs.

    Conclusions

    Our data underscores the potential for PEG20-PAI-2C161S drug conjugates to be further developed as anti-uPA targeted therapeutics with enhanced tumor retention.

Authors


  •   Vine, Kara L.
  •   Lobov, Sergei A. (external author)
  •   Indira Chandran, Vineesh (external author)
  •   Harris, Nathanial (external author)
  •   Ranson, Marie

Publication Date


  • 2015

Citation


  • Vine, K. Lea., Lobov, S., Indira Chandran, V., Harris, N. Lachlan Ewart. & Ranson, M. (2015). Improved pharmacokinetic and biodistribution properties of the selective urokinase inhibitor PAI-2 (SerpinB2) by site-specific PEGylation: implications for drug delivery. Pharmaceutical Research, 32 (3), 1045-1054.

Scopus Eid


  • 2-s2.0-84922998623

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/507

Number Of Pages


  • 9

Start Page


  • 1045

End Page


  • 1054

Volume


  • 32

Issue


  • 3