Synthesis and use of anti-reverse phosphorothioate analogs of the messenger RNA cap

US8153773B2 · kind B2 · utility

Assignees

• BOARD OF SUPERVISORS OF LOUISIANA STATE UNIVERSITY AND AGRICULTURAL AND MECHANICAL COLLEGE · US
• University of Warsaw · PL

Inventors

• Jacek Jemielity · Warszawa, PL
• Ewa M. Grudzien-Nogalska · Shreveport, US
• Joanna Kowalska · Radom, PL
• Edward Darzynkiewicz · Białystok, PL
• Robert E. Rhoads · Shreveport, US

Key dates

• Filing date: Jun 19, 2008
• Grant date: Apr 10, 2012
• Expiry date: Jul 13, 2029

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12P21/02
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

New RNA cap analogs are disclosed containing one or more phosphorothioates groups. The analogs also contain modifications at the 2′-O position of 7-methylguanosine that prevent them from being incorporated in the reverse orientation during in vitro synthesis of mRNA and that hence are “anti-reverse cap analogs” (ARCAs). The ARCA modification ensures that the S atom is precisely positioned within the active sites of cap-binding proteins in both the translational and decapping machinery. The new S-ARCA analogs are resistant to in vivo decapping enzymes. Some S-ARCAs have a higher affinity for eIF4E than the corresponding analogs not containing a phosphorothioate group. When mRNAs containing the various S-ARCAs are introduced into cultured cells, some are translated as much as five-fold more efficiently than mRNAs synthesized with the conventional analog m7GpppG.