Radiometal-Labeled Peptide−PNA Conjugates for Targeting bcl-2 Expression: Preparation, Characterization, and in Vitro mRNA Binding
| Autor: | Fabio Gallazzi, Fang Jia, Susan Z. Lever, Nalini Shenoy, Jiuli Zhang, Mark Hannink, Michael R. Lewis, Yi Wang |
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| Rok vydání: | 2002 |
| Předmět: |
Peptide Nucleic Acids
Biomedical Engineering Pharmaceutical Science Antineoplastic Agents Bioengineering Peptide chemistry.chemical_compound Drug Delivery Systems DOTA Yttrium Radioisotopes RNA Messenger Northern blot Pharmacology chemistry.chemical_classification Messenger RNA Base Sequence Indium Radioisotopes Organic Chemistry Molecular biology In vitro Genes bcl-2 Gene Expression Regulation Neoplastic Biochemistry chemistry Drug Design Nucleic acid Peptides DNA Biotechnology Conjugate |
| Zdroj: | Bioconjugate Chemistry. 13:1176-1180 |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/bc025591s |
| Popis: | A new antisense peptide-peptide nucleic acid (peptide-PNA) conjugate, designed for targeting bcl-2 expression, has been radiolabeled, characterized, and evaluated for bcl-2 mRNA binding in a cell-free system. A PNA complementary to the first six codons of the bcl-2 gene was synthesized by standard solid-phase Fmoc chemistry and conjugated to a new derivative of 1,4,7,10-tetraazacyclododecaneN,N',N'',N'''-tetraacetic acid (DOTA) that allows macrocyclic radiometal chelates to be incorporated into any sequence position of a peptide-PNA conjugate. The DOTA-PNA conjugate was then coupled to a membrane-permeating transduction peptide, PTD-4, designed for intracellular delivery of the radiolabeled PNA. The conjugate was characterized by HPLC and ESI-MS and labeled with 1 1 1 In and 9 0 Y to high specific activities (> 1000 Ci/mmol) with high radiochemical purity. Northern blot analysis showed that 9 0 Y-PTD-4-K(DOTA)-anti-bcl-2-PNA bound specifically to as little as 50 fmol of bcl-2 mRNA, a result equivalent to that obtained with the analogous 3 2 P-labeled DNA antisense oligonucleotide. Thus, the mRNA targeting properties of 1 1 1 In- and 9 0 Y-PTD-4-K(DOTA)-anti-bcl-2-PNA demonstrate potential for diagnostic imaging and targeted radiotherapy applications in bcl-2-positive cancers. |
| Databáze: | OpenAIRE |
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