Synthesis and properties of mRNA cap analogs containing imidodiphosphate moiety—fairly mimicking natural cap structure, yet resistant to enzymatic hydrolysis

• Published in: Bioorganic & medicinal chemistry Vol. 20; no. 5; pp. 1699 - 1710
• Main Authors: Rydzik, Anna M., Kulis, Marta, Lukaszewicz, Maciej, Kowalska, Joanna, Zuberek, Joanna, Darzynkiewicz, Zbigniew M., Darzynkiewicz, Edward, Jemielity, Jacek
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Ltd 01.03.2012; Elsevier
• Subjects: binding proteins; Biological and medical sciences; Biomimetic Materials - chemical synthesis; Biomimetic Materials - chemistry; Biomimetic Materials - metabolism; Bioorganic chemistry; cell free system; chemistry; Chloride; Diphosphonates - chemistry; enzymatic hydrolysis; Enzymatic resistance; Enzymes; Humans; Hydrolysis; Imidodiphosphate; Medical sciences; messenger RNA; Metals; Mimicry; mRNA cap; nucleosides; Nucleotides; Pharmacology. Drug treatments; Protein Binding; Protein Biosynthesis; pyrophosphatase; RNA Cap Analogs - chemical synthesis; RNA Cap Analogs - chemistry; RNA Cap Analogs - metabolism; translation (genetics); Translation inhibition; Translation initiation; DQGOKZCCDMDPNS-WKHPZNKTSA-O; Enzymatic resistance; UTWHLHAJSNQWET-WKHPZNKTSA-O; HUUHNLWHWKBSBJ-BILJAGFLSA-O; Nucleotides; Bioorganic chemistry; Imidodiphosphate; MVLPCPQCYLKPPQ-WKHPZNKTSA-O; Translation inhibition; OTDSBYOQHAECNA-BILJAGFLSA-O; mRNA cap; TYNSCSFUWYGBRA-WKHPZNKTSA-O; VGMWHHNGQBDEBP-BILJAGFLSA-O; GSOLLBGXRSSEFN-BILJAGFLSA-O; Treatment resistance; Chemical structure; Translation inhibitor; Messenger RNA; Cap structure; Nucleotide; Chemical synthesis
• ISSN: 0968-0896; 1464-3391
• DOI: 10.1016/j.bmc.2012.01.013

We describe synthesis and properties of eight dinucleotide mRNA 5′ cap analogs containing imidodiphosphate moiety within 5′,5′-tri- or tetraphosphate bridge (NH-analogs). The compounds were obtained by coupling an appropriate nucleoside 5′-imidodiphosphate with nucleotide P-imidazolide mediated by divalent metal chloride in anhydrous DMF. To evaluate the novel compounds as tools for studying cap-dependent processes, we determined their binding affinities for eukaryotic translation initiation factor 4E, susceptibilities to decapping pyrophosphatase DcpS and, for non-hydrolysable analogs, binding affinities to this enzyme. The results indicate that the O to NH substitution in selected positions of oligophosphate bridge ensures resistance to enzymatic decapping and suggest that interactions of NH-analogs with cap binding proteins fairly mimic interactions of unmodified parent compounds. Finally, we identified NH-analogs as potent inhibitors of cap-dependent translation in cell free system, and evaluated their utility as reagents for obtaining 5′ capped mRNAs in vitro to be rather moderate.