Measuring the pharmacodynamic effects of a novel Hsp90 inhibitor on HER2/neu expression in mice using Zr-DFO-trastuzumab.

Abstract

BACKGROUND:

The positron-emitting radionuclide (89)Zr (t(1/2) = 3.17 days) was used to prepare (89)Zr-radiolabeled trastuzumab for use as a radiotracer for characterizing HER2/neu-positive breast tumors. In addition, pharmacodynamic studies on HER2/neu expression levels in response to therapeutic doses of PU-H71 (a specific inhibitor of heat-shock protein 90 [Hsp90]) were conducted.

METHODOLOGY/PRINCIPAL FINDINGS:

Trastuzumab was functionalized with desferrioxamine B (DFO) and radiolabeled with [(89)Zr]Zr-oxalate at room temperature using modified literature methods. ImmunoPET and biodistribution experiments in female, athymic nu/nu mice bearing sub-cutaneous BT-474 (HER2/neu positive) and/or MDA-MB-468 (HER2/neu negative) tumor xenografts were conducted. The change in (89)Zr-DFO-trastuzumab tissue uptake in response to high- and low-specific-activity formulations and co-administration of PU-H71 was evaluated by biodistribution studies, Western blot analysis and immunoPET. (89)Zr-DFO-trastuzumab radiolabeling proceeded in high radiochemical yield and specific-activity 104.3+/-2.1 MBq/mg (2.82+/-0.05 mCi/mg of mAb). In vitro assays demonstrated >99% radiochemical purity with an immunoreactive fraction of 0.87+/-0.07. In vivo biodistribution experiments revealed high specific BT-474 uptake after 24, 48 and 72 h (64.68+/-13.06%ID/g; 71.71+/-10.35%ID/g and 85.18+/-11.10%ID/g, respectively) with retention of activity for over 120 h. Pre-treatment with PU-H71 was followed by biodistribution studies and immunoPET of (89)Zr-DFO-trastuzumab. Expression levels of HER2/neu were modulated during the first 24 and 48 h post-administration (29.75+/-4.43%ID/g and 41.42+/-3.64%ID/g, respectively). By 72 h radiotracer uptake (73.64+/-12.17%ID/g) and Western blot analysis demonstrated that HER2/neu expression recovered to baseline levels.

CONCLUSIONS/SIGNIFICANCE:

The results indicate that (89)Zr-DFO-trastuzumab provides quantitative and highly-specific delineation of HER2/neu positive tumors, and has potential to be used to measure the efficacy of long-term treatment with Hsp90 inhibitors, like PU-H71, which display extended pharmacodynamic profiles.

• Authors: Jason P Holland, Eloisi Caldas-Lopes, Vadim Divilov, Valerie A Longo, Tony Taldone, Danuta Zatorska, Gabriela Chiosis, Jason S Lewis
• Journal: PloS one (PLoS One) Vol. 5 Issue 1 Pg. e8859 (Jan 25 2010) ISSN: 1932-6203 [Electronic] United States
• PMID: 20111600 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Antibodies, Monoclonal
• Antibodies, Monoclonal, Humanized
• Benzodioxoles
• HSP90 Heat-Shock Proteins
• Purines
• 9H-purine-9-propanamine, 6-amino-8-((6-iodo-1,3-benzodioxol-5-yl)thio)-N-(1-methylethyl)-
• Zirconium
• Deferoxamine
• Trastuzumab

Topics

• Animals
• Antibodies, Monoclonal (chemistry, pharmacology)
• Antibodies, Monoclonal, Humanized
• Benzodioxoles (pharmacokinetics, pharmacology)
• Cell Line, Tumor
• Deferoxamine (chemistry)
• Female
• Genes, erbB-2
• HSP90 Heat-Shock Proteins (antagonists & inhibitors)
• Humans
• Mice
• Mice, Nude
• Purines (pharmacokinetics, pharmacology)
• Tissue Distribution
• Trastuzumab
• Zirconium (chemistry)