Substituted Aryl Benzylamines as Potent and Selective Inhibitors of 17β-Hydroxysteroid Dehydrogenase Type 3.

17β-Hydroxysteroid dehydrogenase type 3 (17β-HSD3) is expressed at high levels in testes and seminal vesicles; it is also present in prostate tissue and involved in gonadal and non-gonadal testosterone biosynthesis. The enzyme is membrane-bound, and a crystal structure is not yet available. Selective aryl benzylamine-based inhibitors were designed and synthesised as potential agents for prostate cancer therapeutics through structure-based design, using a previously built homology model with docking studies. Potent, selective, low nanomolar IC ₅₀ 17β-HSD3 inhibitors were discovered using N -(2-([2-(4-chlorophenoxy)phenylamino]methyl)phenyl)acetamide ( 1 ). The most potent compounds have IC ₅₀ values of approximately 75 nM. Compound 29 , N -[2-(1-Acetylpiperidin-4-ylamino)benzyl]- N -[2-(4-chlorophenoxy)phenyl]acetamide, has an IC ₅₀ of 76 nM, while compound 30 , N -(2-(1-[2-(4-chlorophenoxy)-phenylamino]ethyl)phenyl)acetamide, has an IC ₅₀ of 74 nM. Racemic C -allyl derivative 26 (IC ₅₀ of 520 nM) was easily formed from 1 in good yield and, to determine binding directionality, its enantiomers were separated by chiral chromatography. Absolute configuration was determined using single crystal X-ray crystallography. Only the S -(+)-enantiomer ( 32 ) was active with an IC ₅₀ of 370 nM. Binding directionality was predictable through our in silico docking studies, giving confidence to our model. Importantly, all novel inhibitors are selective over the type 2 isozyme of 17β-HSD2 and show <20% inhibition when tested at 10 µM. Lead compounds from this series are worthy of further optimisation and development as inhibitors of testosterone production by 17β-HSD3 and as inhibitors of prostate cancer cell growth.