Structure‐Guided Synthesis and Evaluation of Small‐Molecule Inhibitors Targeting Protein–Protein Interactions of BRCA1 tBRCT Domain. (9th September 2019)

• Record Type: Journal Article
• Authors: Kurdekar, Vadiraj; Giridharan, Saranya; Subbarao, Jasti; Nijaguna, Mamatha B.; Periasamy, Jayaprakash; Boggaram, Sanjana; Shivange, Amol V.; Sadasivam, Gayathri; Padigaru, Muralidhara; Potluri, Vijay; Venkitaraman, Ashok R.; Bharatham, Kavitha
• Is Part Of: ChemMedChem. Volume 14:Number 18(2019)
• Page Start: 1620
• Page End: 1632
• Publication Date: 2019-09-09
• Subjects: BRCA1 -- BRCT domain -- DNA damage -- GIST -- Homologous recombination -- Protein–protein interactions; Pharmaceutical chemistry -- Periodicals; 615.19005
• DOI: 10.1002/cmdc.201900300
• Languages: English
• ISSNs: 1860-7179

Abstract

Abstract: The tandem BRCT domains (tBRCT) of BRCA1 engage phosphoserine‐containing motifs in target proteins to propagate intracellular signals initiated by DNA damage, thereby controlling cell cycle arrest and DNA repair. Recently, we identified Bractoppin, the first small‐molecule inhibitor of the BRCA1 tBRCT domain, which selectively interrupts BRCA1‐mediated cellular responses evoked by DNA damage. Here, we combine structure‐guided chemical elaboration, protein mutagenesis and cellular assays to define the structural features responsible for Bractoppin's activity. Bractoppin fails to bind mutant forms of BRCA1 tBRCT bearing K1702A, a key residue mediating phosphopeptide recognition, or F1662R or L1701K that adjoin the pSer‐recognition site. However, the M1775R mutation, which engages the Phe residue in the consensus phosphopeptide motif pSer‐X‐X‐Phe, does not affect Bractoppin binding, confirming a binding mode distinct from the substrate phosphopeptide binding. We explored these structural features through structure‐guided chemical elaboration and characterized structure–activity relationships (SARs) in biochemical assays. Two analogues, CCBT2088 and CCBT2103 were effective in abrogating BRCA1 foci formation and inhibiting G2 arrest induced by irradiation of cells. Collectively, our findings reveal structural features underlying the activity of a novel inhibitor of phosphopeptide recognition by the BRCA1 tBRCT domain, providing fresh insights to guide the development of inhibitors that target protein–protein interactions. Abstract : Taming BRCA1 by its tail, tBRCT : Bractoppin blocks the phosphopeptide‐binding site of BRCA1 via structural features distinct from the substrate phosphopeptide. Chemical elaboration conceptualized from mutagenesis and structure‐guided strategies reveal structural features underlying the biochemical potency of Bractoppin analogs against BRCA1 tBRCT domain. This expands the scope of developing inhibitors targeting protein–protein interactions.