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Identification of Mediators of T-cell Receptor Signaling via the Screening of Chemical Inhibitor Libraries
Published on: January 22, 2019
Discovery of potential TMPRSS2 inhibitors using integrated computational and experimental screening
Huiru Xie1, Qibo Hu2, Qiuhong Zhang2
1College of Chemistry and Life Science, Beijing University of Technology, Beijing, 100124, China; State Key Laboratory of National Security Specially Needed Medicines, Beijing, 100039, China.
Abstract:
In prostate cancer (PCa), the TMPRSS2-ERG fusion gene drives aberrant activation of androgen receptor (AR) signaling, while TMPRSS2 protease activity contributes to remodeling the tumor microenvironment. Targeting TMPRSS2 can suppress cancer metastasis and represents a potential therapeutic strategy for castration-resistant prostate cancer (CRPC). This study aimed to screen and identify promising TMPRSS2-binding lead candidates via integrated virtual screening and experimental validation. Screening of the ChemDiv, ChemBridge, and TargetMol compound libraries yielded 53 hit compounds. Surface plasmon resonance (SPR) assays were subsequently performed to evaluate binding affinities. Six candidate compounds (15, 26, 29, 31, 41, and 42) exhibited higher affinity for TMPRSS2 compared to the reference inhibitor Nafamostat (KD = 6.09 × 10-5 M). Among them, compound 26 showed the highest affinity (KD = 3.88 × 10-6 M). Subsequent in vitro inhibition assays using LNCaP clone FGC cells demonstrated that compound 26 possessed the strongest anti-proliferative activity (IC50 = 10.95 μM), indicating its therapeutic potential against PCa. Confocal microscopy showed apparent reduced TMPRSS2 staining after 24 h compound 26 treatment, implying potential TMPRSS2 modulation. Further mechanistic insights were gained through molecular dynamics (MD) simulations, alanine scanning mutagenesis, quantum mechanics/molecular mechanics (QM/MM) calculations, and dynamical cross-correlation matrix (DCCM) analysis, which revealed specific ligand-protein interactions in the compound 26-TMPRSS2 complex and reinforced the reliability of our findings. Collectively, these results identify compound 26 as a promising TMPRSS2-binding lead candidate, providing a solid foundation for further biochemical verification and structural optimization.
Insights
Researchers identified compound 26 as a promising therapeutic candidate for prostate cancer (PCa). This TMPRSS2 inhibitor demonstrates strong anti-proliferative effects and potential for treating castration-resistant prostate cancer (CRPC).
Area of Science:
- Biochemistry and Molecular Biology
- Oncology
- Drug Discovery
Background:
- TMPRSS2-ERG fusion gene activates androgen receptor (AR) signaling in prostate cancer (PCa).
- TMPRSS2 protease activity remodels the tumor microenvironment, promoting cancer metastasis.
- Targeting TMPRSS2 offers a therapeutic strategy for castration-resistant prostate cancer (CRPC).
Purpose of the Study:
- To screen and identify novel TMPRSS2-binding lead compounds.
- To evaluate the therapeutic potential of identified compounds against PCa.
Main Methods:
- Integrated virtual screening of compound libraries (ChemDiv, ChemBridge, TargetMol).
- Surface Plasmon Resonance (SPR) assays for binding affinity evaluation.
- In vitro anti-proliferative assays, confocal microscopy, and computational analyses (MD, QM/MM, DCCM).
Main Results:
- 53 hit compounds were identified; six showed higher affinity than Nafamostat.
- Compound 26 exhibited the highest affinity (KD = 3.88 × 10-6 M) and potent anti-proliferative activity (IC50 = 10.95 μM) in LNCaP cells.
- Compound 26 treatment reduced TMPRSS2 staining and mechanistic studies revealed specific ligand-protein interactions.
Conclusions:
- Compound 26 is a promising TMPRSS2-binding lead candidate for prostate cancer therapy.
- The findings provide a foundation for further development and structural optimization of TMPRSS2 inhibitors.
