# High-throughput chemoenzymatic synthesis of bioactive compounds

> **NIH NIH F31** · UNIVERSITY OF MICHIGAN AT ANN ARBOR · 2021 · $6,911

## Abstract

Proposal Summary
With drug-resistant infections and cancers becoming more prominent, it is crucial to investigate new classes of
small molecules to combat these growing health crises. Azaphilones, an underexplored class of fungal natural
products, have been shown to exhibit diverse biological properties, making them candidates as a novel class of
therapeutics. Although preliminary studies demonstrate their utility in this regard, further investigation of their
activities has been hindered due to the challenges of constructing their densely functionalized core and
congested stereocenter. Furthermore, selectively accessing either C7 configuration of these scaffolds presents
an addition hurdle to exploring azaphilone structure-activity relationships.
The most concise approach toward this class of molecules relies on the oxidative dearomatization of
prefunctionalized arenes. Current state-of-the-art methods in asymmetric oxidative dearomatization require
superstoichiometric quantities of both an oxidant and expensive chiral ligand. These methods have also only
been demonstrated on a limited substrate scope, can exhibit poor site-selectivity, and require forcing reaction
conditions. Fortunately, Nature has evolved superior catalysts to perform oxidative dearomatization with greater
site- and stereoselectivity than these established chemical methods. Promisingly, we have identified two flavin-
dependent monooxygenase homologs, AzaH and AfoD, which can perform an oxidative dearomatization on
aromatic substrates with the same site-selectivity, but provide the opposite stereochemical configuration in the
subsequent azaphilone product. This pivotal discovery enables their use in the synthesis of various natural
products and synthetic building blocks, providing orthogonal site- and stereoselectivity to more readily access
greater chemical space in an environmentally-benign manner. Furthermore, these catalysts can operate mild
reaction conditions, making them compatible with high-throughput compound generation platforms. This
proposal describes strategies for the use of these biocatalysts in stereodivergent, chemoenzymatic syntheses
for the rapid generation of azaphilone analogs.
In summary, this work aims to investigate azaphilone structure-activity relationships through high-throughput and
strategic diversification of these scaffolds, directed by a fluorescence polarization assay and a cell painting
screen. The methods established herein will provide a means to efficiently develop increasingly potent
azaphilone-based drugs.

## Key facts

- **NIH application ID:** 10218074
- **Project number:** 5F31GM139387-02
- **Recipient organization:** UNIVERSITY OF MICHIGAN AT ANN ARBOR
- **Principal Investigator:** Joshua Pyser
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $6,911
- **Award type:** 5
- **Project period:** 2020-06-19 → 2021-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10218074

## Citation

> US National Institutes of Health, RePORTER application 10218074, High-throughput chemoenzymatic synthesis of bioactive compounds (5F31GM139387-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10218074. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*