Targeting Steroid-Metabolizing Enzymes with 15β-Substituted Estrone Analogues: Dual Discovery of AKR1C2/17β-HSD1 Inhibitors and a Fluorescent 17β-HSD1 Ligand.

BACKGROUND/

OBJECTIVES

Aldo-keto reductase isoforms AKR1C1-3 and 17β-hydroxysteroid dehydrogenase 1 and 2 (17β-HSD1 and 17β-HSD2) are key enzymes in steroid metabolism and validated targets in hormone-dependent cancers.

METHODS

In this study, Δ15- and 15β-substituted estrone derivatives were evaluated as inhibitors of AKR1C1-3 and 17β-HSD1 using enzymatic assays, cell viability assaysand computational modeling. Cellular uptake of the fluorescent estrone-based inhibitor was investigated using confocal microscopy.

RESULTS

The Δ15-estrone derivative showed potent and selective inhibition of 17β-HSD1 in the low nanomolar range, while 15β-O-propargyl and 15β-azide derivatives exhibited dual inhibitory activity against 17β-HSD1 and AKR1C2. The Δ15- and 15β-azide derivatives reduced cell viability in hormone-dependent breast, endometrial, and ovarian cancer cell lines in the sub- to low-micromolar range. A BODIPY-labeled 15β-O-propargyl analogue retained submicromolar inhibitory potency toward 17β-HSD1, representing the first fluorescent estrane-based inhibitor with preserved biological activity. Confocal microscopy confirmed efficient cellular uptake and predominant cytosolic localization in MCF-7 cells.

CONCLUSIONS

These findings identify Δ15- and 15β-modified estrone derivatives as promising single- and dual-target inhibitors and introduce a fluorescent probe suitable for investigating intracellular steroid metabolism in hormone-dependent malignancies.