Structure-activity relationship analysis of perfluoroalkyl carbonic acids on human and rat placental 3β-hydroxysteroid dehydrogenase activity

Toxicology. 2022 Oct:480:153334. doi: 10.1016/j.tox.2022.153334. Epub 2022 Sep 16.

Abstract

Placenta contains 3β-hydroxysteroid dehydrogenase/steroid Δ5,4-isomerase (HSD3B), which catalyzes pregnenolone to progesterone for maintaining pregnancy. Perfluoroalkyl carbonic acids (PFC) are subclass of perfluoroalkyl substances containing 4-14 carbons (C4-C14) in the carbon backbone and are potential endocrine disruptors. Whether PFC inhibit HSD3B and structure-activity relationship (SAR) remains unclear. Herein, we screened 11 PFC for inhibiting human type I HSD3B (HSD3B1) and rat type IV HSD3B (HSD3B4) activities and determined SAR and mode of inhibition. HSD3B was measured by converting pregnenolone to progesterone assisted by NAD+ in placental microsomes. Of the 11 PFC, C9-C14 significantly inhibited human HSD3B1 activity at 100 μM. Half-maximal inhibitory concentration (IC50) values of C9-C14 compounds were 363.56 ± 12.14, 12.78 ± 0.69, 6.54 ± 0.65, 20.88 ± 0.41, 118.35 ± 0.16, and 149.26 ± 21.67 μM, respectively. We determined Ki values and mode of inhibition of three most potent PFC (C10-C12), and found that they were mixed inhibitors against pregnenolone, with Ki values of 5.57 ± 4.37, 2.04 ± 2.26, and 9.93 ± 7.71, respectively. Docking analysis showed that they bound steroid-binding site. Effects of PFC on rat placental HSD3B4 were performed. Of the 11 PFC, C10-C12 significantly inhibited rat HSD3B4 activity at 100 μM. IC50 values of C10-C12 compounds were 45.85 ± 1.49, 36.08 ± 1.50, and 88.74 ± 1.99 µM, respectively. Ki values and inhibition modes of the three most potent PFC (C10-C12) were studied. It was found that they were mixed inhibitors against pregnenolone, with Ki values of 48.16 ± 20.44, 36.28 ± 53.07, and 91.79 ± 21.75 μM, respectively. Docking analysis showed that they bound steroid-binding site of rat HSD3B4. In conclusion, PFC showed significant SAR differences. The potency of inhibiting HSD3B activity increased from C9 to C11, and then declined. Human HSD3B1 was more sensitive to the inhibition of rat HSD3B4.

Keywords: 3β-hydroxysteroid dehydrogenase 1/steroid Δ(5,4)-isomerase; Mixed inhibitors; Perfluoroalkyl carbonic acids; Pregnenolone; Progesterone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3-Hydroxysteroid Dehydrogenases / metabolism
  • Animals
  • Carbon / metabolism
  • Carbonic Acid
  • Endocrine Disruptors* / metabolism
  • Female
  • Fluorocarbons* / metabolism
  • Fluorocarbons* / toxicity
  • Humans
  • Isomerases / metabolism
  • Isomerases / pharmacology
  • Multienzyme Complexes
  • NAD / metabolism
  • Placenta / metabolism
  • Pregnancy
  • Pregnenolone / metabolism
  • Pregnenolone / pharmacology
  • Progesterone
  • Rats
  • Structure-Activity Relationship

Substances

  • Endocrine Disruptors
  • Fluorocarbons
  • Multienzyme Complexes
  • NAD
  • Carbonic Acid
  • Progesterone
  • Pregnenolone
  • Carbon
  • 3-Hydroxysteroid Dehydrogenases
  • Isomerases