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D-Amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination.

Publication ,  Journal Article
Pruszynski, M; Koumarianou, E; Vaidyanathan, G; Chitneni, S; Zalutsky, MR
Published in: Nucl Med Biol
January 2015

INTRODUCTION: Proteins that undergo receptor-mediated endocytosis are subject to lysosomal degradation, requiring radioiodination methods that minimize loss of radioactivity from tumor cells after this process occurs. To accomplish this, we developed the residualizing radioiodination agent N(ϵ)-(3-[(*)I]iodobenzoyl)-Lys(5)-N(α)-maleimido-Gly(1)-D-GEEEK (Mal-D-GEEEK-[(*)I]IB), which enhanced tumor uptake but also increased kidney activity and necessitates generation of sulfhydryl moieties on the protein. The purpose of the current study was to synthesize and evaluate a new D-amino acid based agent that might avoid these potential problems. METHODS: N(α)-(3-iodobenzoyl)-(5-succinimidyloxycarbonyl)-D-EEEG (NHS-IB-D-EEEG), which contains 3 D-glutamates to provide negative charge and a N-hydroxysuccinimide function to permit conjugation to unmodified proteins, and the corresponding tin precursor were produced by solid phase peptide synthesis and subsequent conjugation with appropriate reagents. Radioiodination of the anti-HER2 antibody trastuzumab using NHS-IB-D-EEEG and Mal-D-GEEEK-IB was compared. Paired-label internalization assays on BT474 breast carcinoma cells and biodistribution studies in athymic mice bearing BT474M1 xenografts were performed to evaluate the two radioiodinated D-peptide trastuzumab conjugates. RESULTS: NHS-[(131)I]IB-D-EEEG was produced in 53.8%±13.4% and conjugated to trastuzumab in 39.5%±7.6% yield. Paired-label internalization assays with trastuzumab-NHS-[(131)I]IB-D-EEEG and trastuzumab-Mal-D-GEEEK-[(125)I]IB demonstrated similar intracellular trapping for both conjugates at 1h ((131)I, 84.4%±6.1%; (125)I, 88.6%±5.2%) through 24h ((131)I, 60.7%±6.8%; (125)I, 64.9%±6.9%). In the biodistribution experiment, tumor uptake peaked at 48 h (trastuzumab-NHS-[(131)I]IB-D-EEEG, 29.8%±3.6%ID/g; trastuzumab-Mal-D-GEEEK-[(125)I]IB, 45.3%±5.3%ID/g) and was significantly higher for (125)I at all time points. In general, normal tissue levels were lower for trastuzumab-NHS-[(131)I]IB-D-EEEG, with the differences being greatest in kidneys ((131)I, 2.2%±0.4%ID/g; (125)I, 16.9%±2.8%ID/g at 144 h). CONCLUSION: NHS-[(131)I]IB-D-EEEG warrants further evaluation as a residualizing radioiodination agent for labeling internalizing antibodies/fragments, particularly for applications where excessive renal accumulation could be problematic.

Duke Scholars

Published In

Nucl Med Biol

DOI

EISSN

1872-9614

Publication Date

January 2015

Volume

42

Issue

1

Start / End Page

19 / 27

Location

United States

Related Subject Headings

  • Trastuzumab
  • Radiochemistry
  • Oligopeptides
  • Nuclear Medicine & Medical Imaging
  • Mice
  • Isotope Labeling
  • Iodine Radioisotopes
  • Humans
  • Female
  • Esters
 

Citation

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Pruszynski, M., Koumarianou, E., Vaidyanathan, G., Chitneni, S., & Zalutsky, M. R. (2015). D-Amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination. Nucl Med Biol, 42(1), 19–27. https://doi.org/10.1016/j.nucmedbio.2014.08.007
Pruszynski, Marek, Eftychia Koumarianou, Ganesan Vaidyanathan, Satish Chitneni, and Michael R. Zalutsky. “D-Amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination.Nucl Med Biol 42, no. 1 (January 2015): 19–27. https://doi.org/10.1016/j.nucmedbio.2014.08.007.
Pruszynski M, Koumarianou E, Vaidyanathan G, Chitneni S, Zalutsky MR. D-Amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination. Nucl Med Biol. 2015 Jan;42(1):19–27.
Pruszynski, Marek, et al. “D-Amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination.Nucl Med Biol, vol. 42, no. 1, Jan. 2015, pp. 19–27. Pubmed, doi:10.1016/j.nucmedbio.2014.08.007.
Pruszynski M, Koumarianou E, Vaidyanathan G, Chitneni S, Zalutsky MR. D-Amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination. Nucl Med Biol. 2015 Jan;42(1):19–27.
Journal cover image

Published In

Nucl Med Biol

DOI

EISSN

1872-9614

Publication Date

January 2015

Volume

42

Issue

1

Start / End Page

19 / 27

Location

United States

Related Subject Headings

  • Trastuzumab
  • Radiochemistry
  • Oligopeptides
  • Nuclear Medicine & Medical Imaging
  • Mice
  • Isotope Labeling
  • Iodine Radioisotopes
  • Humans
  • Female
  • Esters