Inhibitors of Fumarylacetoacetate Hydrolase Domain Containing Protein 1 (FAHD1).

Alexander K. H. Weiss,Johannes R. Loeffler,Richard Wurzer,Hubert Gstach,Stefania Monteleone,Manuel Philip Eder,Pidder Jansen-Dürr,Susanne von Grafenstein,Klaus R. Liedl,Patrycia Klapec

doi:10.3390/molecules26165009

Alexander K. H. Weiss, Johannes R. Loeffler + Show 8 more

Open Access

https://doi.org/10.3390/molecules26165009

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Abstract

FAH domain containing protein 1 (FAHD1) acts as oxaloacetate decarboxylase in mitochondria, contributing to the regulation of the tricarboxylic acid cycle. Guided by a high-resolution X-ray structure of FAHD1 liganded by oxalate, the enzymatic mechanism of substrate processing is analyzed in detail. Taking the chemical features of the FAHD1 substrate oxaloacetate into account, the potential inhibitor structures are deduced. The synthesis of drug-like scaffolds afforded first-generation FAHD1-inhibitors with activities in the low micromolar IC50 range. The investigations disclosed structures competing with the substrate for binding to the metal cofactor, as well as scaffolds, which may have a novel binding mode to FAHD1.

Highlights

FAH domain containing protein 1 (FAHD1) acts as bifunctional oxaloacetate (OAA) decarboxylase (ODx) [1] and acylpyruvate hydrolase (ApH) [2] in mitochondria
While its ApH activity is only of minor biological relevance, FAHD1 is involved as ODx in the regulation of the tricarboxylic acid (TCA) cycle flux [5,6]
Our working model refers to a mitochondrial dysfunction associated senescence (MiDAS)-like phenotype [6], where the mitochondrial OAA levels are tightly regulated by FAHD1 activity

Summary

Introduction

FAH domain containing protein 1 (FAHD1) acts as bifunctional oxaloacetate (OAA) decarboxylase (ODx) [1] and acylpyruvate hydrolase (ApH) [2] in mitochondria. A further residue in the catalytic cavity of FAHD1 must exist for control of the conformation of the bound OAA keto form. The lid of FAHD1 would not open for product release (equivalent to inhibition by the product) At this point of the discussion, it must be added that pyruvate enol is a high-energy molecule storing ~ 7 kcal/moL of free energy relative to the keto form [14]. )); quench oEf 3t3h-He 3p0rainmdacrayptuernedolwaatteeromf opleycruulev; daotettebdyblKac1k2li3netso: hpyydrrougveantbeo–nedninogl; fialnledd bCoOnd2sraereleinasfreon(rt eandd, p(Coi)n)t;tobwluared:scatalytic dy the viewer)

Design Steps of First Generation of FAHD1 Inhibitors

Materials and Methods

Inhibitor Assay on the 96-Well Plate