The chemical synthesis, stability, and activity of MAIT cell prodrug agonists that access MR1 in recycling endosomes

Journal article

Mucosal-associated invariant T (MAIT) cells are antibacterial effector T cells that react to pyrimidines derived from bacterial riboflavin synthesis presented by the monomorphic molecule MR1. A major challenge in MAIT cell research is that the commonly used MAIT agonist precursor, 5-amino-6-d-ribitylaminouracil (5-A-RU), is labile to autoxidation, resulting in a loss of biological activity. Here, we characterize two independent autoxidation processes by LCMS. To overcome the marked instability, we report the synthesis of a 5-A-RU prodrug generated by modification of the 5-amino group with a cleavable valine-citrulline-p-aminobenzyl carbamate. The compound is stable in prodrug form, with the parent amine (i.e., 5-A-RU) released only after enzymatic cleavage. Analysis of the prodrug in vitro and in vivo showed an enhanced MAIT cell activation profile compared to 5-A-RU, which was associated with preferential loading within recycling endosomes, a route used by some natural agonists. This prodrug design therefore overcomes the difficulties associated with 5-A-RU in biological studies and provides an opportunity to explore different presentation pathways.

Authors: Lange, J; Anderson, R; Marshall, A; Chan, S; Bilbrough, T

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: American Chemical Society
• Journal: ACS Chemical Biology
• Volume: 15
• Issue: 2
• Pages: 437-445
• Publication date: 2020-01-07
• Acceptance date: 2020-01-01
• DOI: 10.1021/acschembio.9b00902
• EISSN: 1554-8937
• ISSN: 1554-8929
• Language: English
• Keywords: FFR