1. Volume 73, Issue 1, Pages 4-8 (January 2018)
Moving Towards Precision Urologic Oncology: Targeting Enzalutamide-resistant Prostate Cancer and Mutated Forms of the Androgen Receptor Using the Novel Inhibitor Darolutamide (ODM-201) 
Hendrik Borgmann, Nada Lallous, Deniz Ozistanbullu, Eliana Beraldi, Naman Paul, Kush Dalal, Ladan Fazli, Axel Haferkamp, Pascale Lejeune, Artem Cherkasov, Martin E. Gleave 
European Urology 
Volume 73, Issue 1, Pages 4-8 (January 2018)
DOI: /j.eururo
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2. Fig. 1
Darolutamide (ODM-201) targeting ENZ (enzalutamide)-resistant prostate cancer in vitro and in vivo. (A) Proliferation of MR49F cells assessed with MTS assay after 72h of treatment with darolutamide or ENZ in ENZ-resistant MR49F cells at indicated doses. Pooled means of triplicate experiments are plotted plus or minus the standard error of the mean. (B) Androgen receptor (AR) transactivation as assessed using luciferase assay after 24h of treatment with darolutamide or ENZ in ENZ-resistant MR49F cells at indicated doses. Pooled means of triplicate experiments are plotted plus or minus the standard error of the mean. (C) Down-regulation of prostate-specific antigen (PSA) gene expression by darolutamide and up-regulation by ENZ. MR49F cells were treated with darolutamide or ENZ for 24h at indicated doses. Total messenger RNA (mRNA) was extracted, reverse transcribed into complementary DNA and analyzed using real-time polymerase chain reaction. (D) PSA protein levels in MR49F cells following treatment with darolutamide or ENZ for 48h at indicated doses. (E) Change in mean tumor volume (left) and serum PSA level (right) in mice bearing ENZ-resistant MR49F tumors treated with darolutamide 50mg/kg twice daily versus ENZ 20mg/kg once daily and versus 50mg/kg vehicle once daily. Treatment was started when tumors reached 200mm3. (F) Kaplan-Meier survival curves for mice treated with darolutamide, ENZ, and vehicle. (G) Waterfall plots showing individual responses for each mouse in tumor volume (left) and serum PSA change from baseline (right) between groups after 3 wk of treatment.
DMSO=dimethyl sulfoxide; ns=not significant.
European Urology  , 4-8DOI: ( /j.eururo )
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3. Fig. 2
Effect of enzalutamide (ENZ) and darolutamide (ODM-201) on castration-resistant prostate cancer-associated androgen receptor (AR) mutations and cheminformatics in silico modeling of drug's mode of action. AR transactivation as assessed using luciferase assay 24h after treatment with darolutamide or ENZ in AR-mutants F877L, F877L/T878A, and H875Y/T878A at indicated doses (A). PC3 cells were transfected with the mutated AR construct and a reporter plasmid pARR3-tk-luciferase. Pooled means of triplicate experiments are plotted plus or minus the standard error of the mean. (B) Binding pose of ENZ (left) versus darolutamide (right) within the F877L mutant's androgen-binding-site, reveals how altered conformations within the pocket lead to distinct protein–ligand interactions. (C) Root mean squared deviation (RMSD) analysis comparing the binding conformations of ENZ and darolutamide within the androgen-binding-site pocket of F877L mutant. (D) Binding pose of darolutamide in the androgen binding site of the previously uncharacterized AR mutant T878G. (E) Darolutamide inhibited the transcriptional activity of T878G mutant previously described to induce agonism to ENZ, bicalutamide, and hydroxyflutamide.
WT=wild type.
European Urology  , 4-8DOI: ( /j.eururo )
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