Nanodroplet Array Platform for Integrated Synthesis and Screening of MEK Inhibitors: a Miniaturized Approach to Early Drug Discovery
Abstract
Abstract Early‐stage drug discovery relies on high‐throughput screenings, which are costly and time‐intensive, limiting access for academic laboratories and small companies. A key bottleneck is the lack of miniaturization and the separation of compound synthesis from screening. We present a nanoliter droplet array platform integrating synthesis, characterization, and cell‐based screening of 325 MEK (mitogen‐activated protein kinase kinase) inhibitors, targeting the MAPK/ERK (mitogen‐activated protein kinase/extracellular signal‐regulated kinase) pathway, implicated in colorectal and pancreatic cancer. The platform enables on‐chip synthesis, MALDI‐MSI (matrix‐assisted laser desorption/ionization‐mass spectrometry imaging) characterization, and cell‐based screening within 200 nL droplets containing 20 nmol starting material (∼4 ng final compound), and only 300 cells per droplet. Screening identified 46 compounds with higher cytotoxicity than mirdametinib, a clinically approved MEK inhibitor. Molecular docking revealed a shared allosteric binding mechanism, indicating non‐competitive ATP inhibition. Synthesis and screening of all 325 compounds were completed within 7 days, requiring <10 mg of reactants, <250 µL solvent, and ∼100 µL of cell suspension (∼100,000 cells in total). Our results demonstrate that integrating miniaturized combinatorial synthesis and biological screening in a single platform can accelerate early‐stage drug discovery while reducing cost and resource use.
Article Details
Authors (11)
Maximilian Seifermann
Institute of Biological and Chemical Systems‐Functional Molecular Systems (IBCS‐FMS), Karlsruhe Institute of Technology (KIT) Hermann‐von Helmholtz‐Platz 1 Eggenstein‐Leopoldshafen 76344 Germany
Julius Höpfner
Institute of Biological and Chemical Systems‐Functional Molecular Systems (IBCS‐FMS), Karlsruhe Institute of Technology (KIT) Hermann‐von Helmholtz‐Platz 1 Eggenstein‐Leopoldshafen 76344 Germany
Liana Bauer
Institute of Biological and Chemical Systems‐Functional Molecular Systems (IBCS‐FMS), Karlsruhe Institute of Technology (KIT) Hermann‐von Helmholtz‐Platz 1 Eggenstein‐Leopoldshafen 76344 Germany
Divya Varadharajan
Scivalon, Bruckenäcker 9 Stuttgart 70565 Germany
Stefan Schmidt
Björn Fröhlich
Center for Mass Spectrometry and Optical Spectroscopy (CeMOS) Technische Hochschule Mannheim Paul‐Wittsack‐Straße 10 Mannheim 68163 Germany
Benjamin Wellenhofer
Institute of Biological and Chemical Systems‐Functional Molecular Systems (IBCS‐FMS), Karlsruhe Institute of Technology (KIT) Hermann‐von Helmholtz‐Platz 1 Eggenstein‐Leopoldshafen 76344 Germany
Charlotte Luchena
Institute of Biological and Chemical Systems‐Functional Molecular Systems (IBCS‐FMS), Karlsruhe Institute of Technology (KIT) Hermann‐von Helmholtz‐Platz 1 Eggenstein‐Leopoldshafen 76344 Germany
Carsten Hopf
Anna A. Popova
Institute of Biological and Chemical Systems‐Functional Molecular Systems (IBCS‐FMS), Karlsruhe Institute of Technology (KIT) Hermann‐von Helmholtz‐Platz 1 Eggenstein‐Leopoldshafen 76344 Germany
Pavel A. Levkin
Institute of Biological and Chemical Systems‐Functional Molecular Systems (IBCS‐FMS), Karlsruhe Institute of Technology (KIT) Hermann‐von Helmholtz‐Platz 1 Eggenstein‐Leopoldshafen 76344 Germany