Last Updated: 24/05/2023
Biocatalytic enantioselective synthesis of non-biaryl and hetero-biaryl atropisomers and testing of their antimalarial properties
Objectives
To develop a convergent biocatalytic approach using engineered P450 variants to streamline the synthesis of non-biaryl and hetero-biaryl atropisomeric compounds which are typically challenging to make through known synthetic routes.
Atropisomerism, or stereoisomerism arising as a consequence of hindered bond rotation, has traditionally been avoided as a design strategy in drug design. As a result of this, catalytic methods for atroposelective synthesis are underdeveloped, despite the growing evidence that atropisomerism can be strategically applied to improve the potency and selectivity of a potential drug by increasing its binding specificity to a biological target. In particular, the synthesis of non-biaryl and hetero-biaryl atropisomers containing chiral C–O and C–N axes often relies on the functionalization of a substrate with the C–X bond already installed, thus limiting the utility of these methods in designing convergent syntheses. Here, I propose the development of a biocatalytic approach for the atroposelective synthesis of diarylethers, C,N-coupled naphthylisoquinoline alkaloids, and N-aryl indoles. These classes of compounds were chosen based on the current limitations for their syntheses and their potential therapeutic properties, with the proposed research aiming to address both points. To achieve this goal, we will take a three stage approach consisting of screening wild-type enzymes, biocatalyst engineering, and combinatorial synthesis and biological testing of compound libraries. In the first stage, monomers will be screened against a library of wild-type P450 enzymes. This library was constructed using a bioinformatics approach to identify P450 enzymes with sequence similarity to those with known reactivity. In this stage, reactions will be analyzed for evidence of reactivity and detection of the target product. Upon identification of a suitable starting point for engineering, directed evolution of the enzyme will be conducted to improve formation of the target product. Finally, once a suitable P450 variant is identified, a library of compounds will be constructed through combinatorial synthesis in 96 well plates. These compounds will then undergo biological testing to evaluate their antimalarial potential by analyzing their impact on transmission and viability. The research proposed above will be facilitated by high-thoroughput experimentation and reaction analysis, in conjunction with high-thoroughput platform for biological testing. We anticipate that that this research will streamline the synthesis of molecule classes previously challenging to access. This will accelerate preparation and evaluation of potential therapeutic compounds, expediting the identification of antimalarial drug targets.
Aug 2022 — Aug 2025
$66,790
