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Cited by 158Protein Kinase Regulation and GTPase SignalingProtein Tyrosine PhosphatasesMelanoma and MAPK Pathways

KRAS(G12D) can be targeted by potent inhibitors via formation of salt bridge

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Mao, Zhongwei, Xiao, Hongying, Shen, Panpan, Yang, Yu, Xue, Jing, Yang, Yunyun, Shang, Yanguo, Zhang, Lilan +6 more·2022·Cell Discovery
DOI 10.1038/s41421-021-00368-wPMC PMC8786924
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Abstract

KRAS mutation occurs in nearly 30% of human cancers, yet the most prevalent and oncogenic KRAS(G12D) variant still lacks inhibitors. Herein, we designed a series of potent inhibitors that can form a salt bridge with KRAS’s Asp12 residue. Our ITC results show that these inhibitors have similar binding affinity with both GDP-bound and GTP-bound KRAS(G12D), and our crystallographic studies reveal the structural basis of inhibitor binding-induced switch-II pocket in KRAS(G12D), experimentally confirming the formation of a salt bridge between the piperazine moiety of the inhibitors and the Asp12 residue of the mutant protein. Among KRAS family proteins and mutants, both ITC and enzymatic assays demonstrate the selectivity of the inhibitors for KRAS(G12D); and the inhibitors disrupt the KRAS–CRAF interaction. We also observed the inhibition of cancer cell proliferation as well as MAPK signaling by a representative inhibitor (TH-Z835). However, since the inhibition was not fully dependent on KRAS mutation status, it is possible that our inhibitors may have off-target effects via targeting non-KRAS small GTPases. Experiments with mouse xenograft models of pancreatic cancer showed that TH-Z835 significantly reduced tumor volume and synergized with an anti-PD-1 antibody. Collectively, our study demonstrates proof-of-concept for a strategy based on salt-bridge and induced-fit pocket formation for KRAS(G12D) targeting, which warrants future medicinal chemistry efforts for optimal efficacy and minimized off-target effects.

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  • A variant compound significantly reduced tumor volume in KRAS G12D mouse xenograft models.

    “A variant compound significantly reduced tumor volume in KRAS G12D mouse xenograft models.”

    From KRAS: Biology, Inhibition, and Mechanisms of Inhibitor Resistance (2024) · cites this paper as [55]

  • This compound also displayed activity in off-target cell lines, suggesting other small GTPases such as Rho, Ran, Arf, and/or Rab may have also been targeted.

    “However, this compound also displayed activity in off-target cell lines, suggesting other small GTPases such as Rho, Ran, Arf, and/or Rab may have also been targeted [55].”

    From KRAS: Biology, Inhibition, and Mechanisms of Inhibitor Resistance (2024) · cites this paper as [55]

  • The TH-Z8 series works by forming a salt bridge interaction between the KRAS Asp12 residue and the compound's alkyl amine moiety, inducing the formation of an allosteric drug-binding pocket.

    “The methyl-substituted piperazine inhibitor, the TH-Z8 series, works by forming a salt bridge interaction between the KRAS Asp12 residue and the compound’s alkyl amine moiety, inducing the formation of an allosteric drug-binding pocket.”

    From KRAS: Biology, Inhibition, and Mechanisms of Inhibitor Resistance (2024) · cites this paper as [55]

  • Mao et al. developed a series of G12D-selective inhibitor compounds.

    “Starting with the scaffold of Mirati’s G12C inhibitors, Mao et al. developed a series of G12D-selective inhibitor compounds [55].”

    From KRAS: Biology, Inhibition, and Mechanisms of Inhibitor Resistance (2024) · cites this paper as [55]

  • The TH-Z8 molecule did not discriminate between GDP- and GTP-bound KRAS.

    “Notably, this molecule did not discriminate between GDP- and GTP-bound KRAS.”

    From KRAS: Biology, Inhibition, and Mechanisms of Inhibitor Resistance (2024) · cites this paper as [55]

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Research works printed but not linked: 25

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Cited by

Sorted by citations

Authors
Mao, Z., Xiao, H., Shen, P., Yang, Y., Xue, J., Yang, Y., Shang, Y., Zhang, L., Li, X., Zhang, Y., Du, Y., Chen, C.-C., Guo, R.-T., & Zhang, Y.
Affiliations
  • Center for Life Sciences (CN)
  • National Institute of Biological Sciences, Beijing (CN)
  • State Key Laboratory of Biocatalysis and Enzyme Engineering (CN)
Corresponding author
  • Zhang, Yuying
  • Guo, Rey-Ting
  • Zhang, Yonghui
ORCID
Venue
Cell Discovery
Publisher
Springer Science and Business Media LLC
Published
2022-01-25
Volume
8
Issue
1
Pages
5-5
ISSN
2056-5968 (ISSN-L)
DOI
10.1038/s41421-021-00368-w
PMC
PMC8786924
OpenAlex
W4207041243
Cited by
158
References
18
Citations by year
2026: 33 · 2025: 39 · 2024: 42 · 2023: 33 · 2022: 11
FWCI
10.29
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell Discovery (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Natural Science Foundation of Beijing Municipality (Z190015)
  • National Natural Science Foundation of China (81991492)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Melanoma and MAPK Pathways
Keywords
KRAS G12D, KRAS inhibitors, salt bridges, switch-II pocket, induced fit, piperazine, MAPK signaling, pancreatic cancer, small GTPases, off-target effects, tumor cell proliferation

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