Abstract
The DNA repair enzyme AAG has been shown in mice to promote tissue necrosis in response to ischaemic reperfusion or treatment with alkylating agents. A chemical probe inhibitor is required for investigations of the biological mechanism causing this phenomenon and as a lead for drugs that are potentially protective against tissue damage from organ failure and transplantation, and alkylative chemotherapy. Herein, we describe the rationale behind the choice of arylmethylpyrrolidines as appropriate aza-nucleoside mimics for an inhibitor followed by their synthesis and the first use of a microplate-based assay for quantification of their inhibition of AAG. We finally report the discovery of an imidazol-4-ylmethylpyrrolidine as a fragment-sized, weak inhibitor of AAG.
Highlights
Alkyladenine glycosylase (AAG; known as methylpurine DNA glycosylase, MPG, and alkyl-N–purine glycosylase, ANPG, EC: 3.2.2.21) is one of several DNA glycosylases that can initiate the base excision repair (BER) pathway by hydrolysis of a range of alkylated, oxidised or deaminated purine bases from the DNA backbone.[1]
Both alkylation- and ischaemia reperfusion-induced AAGdependent tissue necrosis is hypothesised to result from AAG activity on substantial numbers of alkylated and oxidised DNA bases leading to the accumulation of toxic repair intermediates in the DNA, which overwhelm the repair capacity of downstream BER enzymes.[3]
We have shown that the small molecule aza-nucleoside mimic 6b possesses weak inhibitory activity against the DNA base excision repair enzyme AAG
Summary
Alkyladenine glycosylase (AAG; known as methylpurine DNA glycosylase, MPG, and alkyl-N–purine glycosylase, ANPG, EC: 3.2.2.21) is one of several DNA glycosylases that can initiate the base excision repair (BER) pathway by hydrolysis of a range of alkylated, oxidised or deaminated purine bases from the DNA backbone.[1]. AAG has been shown to drive alkylation-induced cytotoxicity in several mouse tissues, namely the cerebellum, spleen, thymus, bone marrow, pancreas and retina, with tissue damage reduced or absent in Aagknockout mice and dramatically increased in Aag-overexpressing transgenic mice.[2,3] in ischemia reperfusion mouse models, Aag-knockout mice display reduced tissue necrosis in liver, kidney and brain when compared to wild type.[4] Ischemia reperfusion models mimic ischaemic stroke, liver and kidney failure, and organ transplantation In these events the tissues are temporarily starved of their blood supply before sudden reperfusion results in a burst of reactive oxygen and nitrogen species (RONS). Consortium for their Target 2035 program to make chemical or antibody probes available for the entire human proteome.[10,11] these are a biochemical assay Kd or IC50 < 100 nM, a cellular assay IC50 or EC50 < 1 μM and > 30-fold selectivity against other mammalian BER enzymes and enzymes known to be inhibited by structurally related molecules, such as purine nucleoside phosphorylase (PNP)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
