کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2153407 1090181 2015 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
D-amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی تحقیقات سرطان
پیش نمایش صفحه اول مقاله
D-amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination
چکیده انگلیسی

IntroductionProteins 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 Nϵ-(3-[⁎I]iodobenzoyl)-Lys5-Nα-maleimido-Gly1-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.MethodsNα-(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.ResultsNHS-[131I]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-[131I]IB-d-EEEG and trastuzumab-Mal-d-GEEEK-[125I]IB demonstrated similar intracellular trapping for both conjugates at 1 h (131I, 84.4% ± 6.1%; 125I, 88.6% ± 5.2%) through 24 h (131I, 60.7% ± 6.8%; 125I, 64.9% ± 6.9%). In the biodistribution experiment, tumor uptake peaked at 48 h (trastuzumab-NHS-[131I]IB-d-EEEG, 29.8% ± 3.6%ID/g; trastuzumab-Mal-d-GEEEK-[125I]IB, 45.3% ± 5.3%ID/g) and was significantly higher for 125I at all time points. In general, normal tissue levels were lower for trastuzumab-NHS-[131I]IB-d-EEEG, with the differences being greatest in kidneys (131I, 2.2% ± 0.4%ID/g; 125I, 16.9% ± 2.8%ID/g at 144 h).ConclusionNHS-[131I]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.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Nuclear Medicine and Biology - Volume 42, Issue 1, January 2015, Pages 19–27
نویسندگان
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