title: Synthesis, purification, and characterization of oligoribonucleotides that act as agonists of TLR7 and/or TLR8.
abstract: Viral single-stranded (ss) RNA is the natural ligand for TLR7 and TLR8. Synthetic ssRNA has been shown to act as a ligand for TLR7 and TLR8. We have previously reported a novel RNA structure, referred to as stabilized immune modulatory RNA (SIMRA), in which two short phosphorothioate oligoribonucleotides were linked through their 3'-ends via a linker. SIMRA compounds had greater stability in serum than unmodified ssRNA and induced immune responses via TLR7 and/or TLR8. SIMRA compounds were synthesized using phosphoramidite chemistry on controlled-pore glass solid support derivatized with a linker. After cleavage from the solid support and removal of protecting groups, SIMRA compounds were purified on an anion-exchange HPLC followed by desalting/dialysis, and lyophilization. SIMRA compounds were characterized for their purity and sequence integrity by anion-exchange HPLC, capillary gel electrophoresis, polyacrylamide gel electrophoresis, and MALDI-TOF mass spectrophotometric analysis. As SIMRA compounds induce TLR7- and/or TLR8-mediated Th1-type immune responses, they have potential utility as therapeutic agents for a broad range of diseases, including cancer, infectious diseases, asthma, and allergies, and as adjuvants with vaccines.
authors: Idera Pharmaceuticals, Inc., Cambridge, MA, USA. tlan@iderapharma.com
authors: Tao
authors: T
authors: Lan
authors: Ekambar R
authors: ER
authors: Kandimalla
mesh_terms: D000276:Adjuvants, Immunologic / Q000138:chemical synthesis* / Q000302:isolation & purification / Q000494:pharmacology; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D003141:Communicable Diseases / Q000188:drug therapy / Q000276:immunology / Q000473:pathology; D004355:Drug Stability; D019075:Electrophoresis, Capillary; D006801:Humans; D006967:Hypersensitivity / Q000188:drug therapy / Q000276:immunology / Q000473:pathology; D007155:Immunologic Factors / Q000138:chemical synthesis* / Q000302:isolation & purification / Q000494:pharmacology; D009369:Neoplasms / Q000188:drug therapy / Q000276:immunology / Q000473:pathology; D009843:Oligoribonucleotides / Q000138:chemical synthesis* / Q000302:isolation & purification / Q000494:pharmacology; D054735:Phosphorothioate Oligonucleotides / Q000138:chemical synthesis* / Q000302:isolation & purification / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D051199:Toll-Like Receptor 7 / Q000819:agonists* / Q000276:immunology / Q000378:metabolism; D051200:Toll-Like Receptor 8 / Q000819:agonists* / Q000276:immunology / Q000378:metabolism
chemical_list: D000276:Adjuvants, Immunologic; D007155:Immunologic Factors; D009843:Oligoribonucleotides; D054735:Phosphorothioate Oligonucleotides; D051199:Toll-Like Receptor 7; D051200:Toll-Like Receptor 8