Targeting oncogenic KRasG13C with nucleotide-based covalent inhibitors

Abstract

Mutations within Ras proteins represent major drivers in human cancer. In this study, we report the structure-based design, synthesis, as well as biochemical and cellular evaluation of nucleotide-based covalent inhibitors for KRasG13C, an important oncogenic mutant of Ras that has not been successfully addressed in the past. Mass spectrometry experiments and kinetic studies reveal promising molecular properties of these covalent inhibitors, and X-ray crystallographic analysis has yielded the first reported crystal structures of KRasG13C covalently locked with these GDP analogues. Importantly, KRasG13C covalently modified with these inhibitors can no longer undergo SOS-catalysed nucleotide exchange. As a final proof-of-concept, we show that in contrast to KRasG13C, the covalently locked protein is unable to induce oncogenic signalling in cells, further highlighting the possibility of using nucleotide-based inhibitors with covalent warheads in KRasG13C-driven cancer.

Date of acceptance

2023

Autoren

Lisa Goebel
Tonia Kirschner
Sandra Koska
Amrita Rai
Petra Janning
Stefano Maffini
Helge Vatheuer
Paul Czodrowski
Roger S Goody
Matthias P Müller
Daniel Rauh

DOI

10.7554/elife.82184

eISSN

2050-084X

Zeitschrift

eLife

Sprache

en

Online publication date

2023

Status

Published online

Herausgeber

eLife Sciences Publications, Ltd

Herausgeber URL

http://dx.doi.org/10.7554/elife.82184

Datum der Datenerfassung

2023

Titel

Targeting oncogenic KRasG13C with nucleotide-based covalent inhibitors

Ausgabe der Zeitschrift

12

Datenquelle: Crossref

Abstract

Mutations within Ras proteins represent major drivers in human cancer. In this study, we report the structure-based design, synthesis, as well as biochemical and cellular evaluation of nucleotide-based covalent inhibitors for KRasG13C, an important oncogenic mutant of Ras that has not been successfully addressed in the past. Mass spectrometry experiments and kinetic studies reveal promising molecular properties of these covalent inhibitors, and X-ray crystallographic analysis has yielded the first reported crystal structures of KRasG13C covalently locked with these GDP analogues. Importantly, KRasG13C covalently modified with these inhibitors can no longer undergo SOS-catalysed nucleotide exchange. As a final proof-of-concept, we show that in contrast to KRasG13C, the covalently locked protein is unable to induce oncogenic signalling in cells, further highlighting the possibility of using nucleotide-based inhibitors with covalent warheads in KRasG13C-driven cancer.

Addresses

Department of Chemistry and Chemical Biology, TU Dortmund University, Dortmund, Germany.

Autoren

Lisa Goebel
Tonia Kirschner
Sandra Koska
Amrita Rai
Petra Janning
Stefano Maffini
Helge Vatheuer
Paul Czodrowski
Roger S Goody
Matthias P Müller
Daniel Rauh

DOI

10.7554/elife.82184

eISSN

2050-084X

Externe Identifier

PubMed Identifier: 36972177
PubMed Central ID: PMC10042540

Funding acknowledgements

Deutsche Forschungsgemeinschaft: RA 1055/5-1
Deutsche Forschungsgemeinschaft: GO 284/10-1
TU Dortmund University:

Open access

true

ISSN

2050-084X

Zeitschrift

eLife

Schlüsselwörter

Humans
Neoplasms
ras Proteins
Nucleotides
Signal Transduction
Kinetics

Sprache

eng

Medium

Electronic

Online publication date

2023

Open access status

Open Access

Paginierung

e82184

Datum der Veröffentlichung

2023

Status

Published

Publisher licence

CC BY

Datum der Datenerfassung

2023

Titel

Targeting oncogenic KRasG13C with nucleotide-based covalent inhibitors.

Sub types

Research Support, Non-U.S. Gov't
research-article
Journal Article

Ausgabe der Zeitschrift

12

Files

https://europepmc.org/articles/PMC10042540?pdf=render

Datenquelle: Europe PubMed Central

Abstract

Mutations within Ras proteins represent major drivers in human cancer. In this study, we report the structure-based design, synthesis, as well as biochemical and cellular evaluation of nucleotide-based covalent inhibitors for KRasG13C, an important oncogenic mutant of Ras that has not been successfully addressed in the past. Mass spectrometry experiments and kinetic studies reveal promising molecular properties of these covalent inhibitors, and X-ray crystallographic analysis has yielded the first reported crystal structures of KRasG13C covalently locked with these GDP analogues. Importantly, KRasG13C covalently modified with these inhibitors can no longer undergo SOS-catalysed nucleotide exchange. As a final proof-of-concept, we show that in contrast to KRasG13C, the covalently locked protein is unable to induce oncogenic signalling in cells, further highlighting the possibility of using nucleotide-based inhibitors with covalent warheads in KRasG13C-driven cancer.

Date of acceptance

2023

Autoren

Lisa Goebel
Tonia Kirschner
Sandra Koska
Amrita Rai
Petra Janning
Stefano Maffini
Helge Vatheuer
Paul Czodrowski
Roger S Goody
Matthias P Müller
Daniel Rauh

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/36972177

DOI

10.7554/eLife.82184

eISSN

2050-084X

Externe Identifier

PubMed Central ID: PMC10042540

Zeitschrift

Elife

Schlüsselwörter

E. coli
G13C
Ras
biochemistry
cancer
chemical biology
covalent inhibitors
nucleotide analogues
Humans
Nucleotides
Kinetics
ras Proteins
Signal Transduction
Neoplasms

Sprache

eng

Country

England

PII

82184

Datum der Veröffentlichung

2023

Status

Published online

Datum, an dem der Datensatz öffentlich gemacht wurde

2023

Titel

Targeting oncogenic KRasG13C with nucleotide-based covalent inhibitors.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

12

Datenquelle: PubMed

Targeting oncogenic KRasG13C with nucleotide-based covalent inhibitors

Files

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