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In Situ Inhibitor Synthesis and Screening by Fluorescence Polarization: An Efficient Approach for Accelerating Drug

Zhihong Li¹, Yue Wu¹, Shuai Zhen¹

¹State Key Laboratory of Natural Medicines Jiangsu Key Laboratory of Drug Design and Optimization, and Department of Chemistry China Pharmaceutical University Nanjing 211198 China.

Angewandte Chemie (Weinheim an Der Bergstrasse, Germany)

|March 20, 2024

Summary

In situ inhibitor synthesis and screening (ISISS) offers a simpler approach to drug discovery by combining synthesis and screening. This method efficiently identified a novel inhibitor for prolyl hydroxylase 2 (PHD2), a key target in anemia treatment.

Keywords:

Drug Discovery Fluorescence Polarization Hypoxia In Situ Inhibitor Synthesis and Screening PHD2

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Area of Science:

Medicinal Chemistry
Chemical Biology
Drug Discovery

Background:

Target-directed dynamic combinatorial chemistry (DCC) is valuable for hit identification but complex mixture analysis hinders its application.
A need exists for simplified, efficient methods in hit identification and lead optimization.

Purpose of the Study:

To introduce and exemplify the in situ inhibitor synthesis and screening (ISISS) method as an accessible alternative to DCC.
To demonstrate the utility of ISISS for discovering potent and novel inhibitors against therapeutic targets.

Main Methods:

Developed ISISS by integrating high-throughput bioorthogonal synthesis with fluorescence-based target binding screening.
Coupled fluorescence polarization screening with the reaction between acyl-hydrazides and aldehydes.
Applied ISISS to identify inhibitors of human prolyl hydroxylase 2 (PHD2).

Main Results:

Successfully identified a potent and novel acylhydrazone-based inhibitor of PHD2 using the ISISS method.
The discovered inhibitor demonstrated equivalent in vivo potency compared to an existing approved medicine for anemia treatment.
ISISS proved to be an operationally simple and efficient approach for hit discovery.

Conclusions:

ISISS provides a streamlined and effective strategy for hit identification, overcoming limitations of traditional DCC.
The method facilitates the discovery of novel, potent inhibitors for therapeutic targets like PHD2.
ISISS holds promise for accelerating drug discovery pipelines, particularly for targets implicated in diseases such as anemia.