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Highly Informative Drug Screening by Overcoming NMR Restrictions

Periodic Reporting for period 1 - HiSCORE (Highly Informative Drug Screening by Overcoming NMR Restrictions)

Okres sprawozdawczy: 2021-05-01 do 2022-10-31

ERC Synergy project 2021-2026

Highly Informative Drug Screening by Overcoming NMR Restrictions (“HiSCORE”)

The need for drug screening with increasingly higher throughput is dictated both by the increasing number of drug targets that are becoming available through genomics and by the increasing number of chemical molecules generated through combinatorial chemistry. Several Boolean high-throughput screening techniques today can scan large compound libraries, but the ever increasing throughput has not translated into a significant increase in late-phase drug candidates. HiSCORE presents a synergistic approach to high-throughput, high-information drug screening that builds on the complementary skills of four laboratories supported by two external experts of drug screening:
(i) Research and build innovative magnetic resonance instrumentation (Kentgens, IMM/RU) that can provide small, hyperpolarized solid samples on a seconds timescale, transfer and dissolve or liquefy these samples with minimum dilution, and acquire multiple high-resolution NMR spectra of the liquefied samples in parallel (Meier, IBG/KIT), using complementary contrast-enhancement methods, in up to 1000 massively parallelized microfluidic detectors (Korvink, IMT/KIT). (ii) Use this instrumentation for binding assays and measure the dissociation constants in the nano to micromolar range, and determine kinetic rates of the association and dissociation for a large number of complexes of putative drug compounds and protein targets (Bodenhausen, ENS) (iii) Use this instrumentation for functional assays, in particular for systems that comprise multiple enzyme steps with intermediate products, and to determine the efficacy of potential inhibitors, while fully exploiting the rich information that can be obtained by fluorine-19 NMR. (iv) Use this instrumentation for metabolomic assays to observe the metabolism of the compounds in cultures in cells in view of identifying potentially toxic side-products. The contrast between compounds that bind to targets and those that fail to bind will be boosted by exploiting long-lived states.

Synergy builds on the know-how of 4 teams and 2 advisors

1. Geoffrey Bodenhausen Paris
2. Benno Meier Karlsruhe
3. Arno Kentgens Nijmegen
4. Jan Korvink Karlsruhe
5. Alvar Gossert Zürich
6 Claudio Dalvit Trento

These 6 groups will contribute complementary expertise:

1. Geoffrey Bodenhausen and his team (Abergel, Bouvignies, Pelupessy, Ferrage, Baudin, Birlirakis) develop NMR methodology, long-lived states, extending life-times of the hyperpolarization, methods for accurate determination of binding parameters, including kinetics, and will develop libraries of compounds, with the advice of Dalvit and Gossert.
2. Benno Meier: Alternatives to dissolution DNP : ballistic transfer of solid hyperpolarized “bullets” with minimal dilution, improved understanding and acceleration of the DNP process, parallelization.
3. Arno Kentgen: Novel miniaturized NMR detectors, Overhauser DNP, rapid-melt DNP, microfluidics, 1 μL samples, supercritical CO2 as a solvent, and SABRE for quantitative identification of nM compounds.
4. Jan Korvink: miniaturization based on microelectromechanical systems (MEMS), complementary metal-oxide-semiconductors (CMOS), massively parallel detection, automatic data analysis, and machine learning.
5. Alvar Gossert: worked for 10 years in the drug development/screening labs of Novartis and moved in 2017 to ETH Zürich.
6. Claudio Dalvit: worked for 9 years in drug development/screening at Novartis in Basel, 9 years at Pharmacia & Upjohn in Milan, at the Italian Institute of Technology (IIT) in Genova, and at the University of Neuchatel.


The project has two objectives:
(A) Development of novel iInstrumentation to produce hyperpolarized solid samples at intervals of 1 minute or less, and to paralleliize 4, 10, 100, … microfluidic micro-coils or strip-line detectors
(B) Chemistry: develop binding assays to measure dissociation constants nM < KD < µM, to determine kinetic rates kon and koff of the association and dissociation ligands and proteins, to develop functional assays to determine IC50 of inhibitors of multiple-step enzyme cascades, and to explore metabolomic assays in cultures of cells

Why should one wish to parallellize 4, 10, 100, 1000 microfluidic NMR detectors? For two reasons:
(A) Different sample compositions by running titrations,e.g. by varying the ratio [L] : [P] = [ligand] : [protein]; and for functional screening by varying the ratio [inhibitor] : [enzyme]
(B) Different experimental parameters: to run complementary experiments (T1, T2, CPMG, CEST, TLLS, TLLC...) to vary pulse intervals and relaxation delays, and to observe 1H, 2H, 13C, 15N, 19F, 31P…
Preliminary work before the start of the contrat on May 1st 2021 has been limited to purchasing instrumentation and appointing staff
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