New Anti-Inflammatory Metabolites by Microbial Transformation of Medrysone

Abstract

Microbial transformation of the anti-inflammatory steroid medrysone (1) was carried out for the first time with the filamentous fungi Cunninghamella blakesleeana (ATCC 8688a), Neurospora crassa (ATCC 18419), and Rhizopus stolonifer (TSY 0471). The objective was to evaluate the anti-inflammatory potential of the substrate (1) and its metabolites. This yielded seven new metabolites, 14α-hydroxy-6α-methylpregn-4-ene-3,11,20-trione (2), 6β-hydroxy-6α-methylpregn-4-ene-3,11,20-trione (3), 15β-hydroxy-6α-methylpregn-4-ene-3,11,20-trione (4), 6β,17α-dihydroxy-6α-methylpregn-4-ene-3,11,20-trione (5), 6β,20S-dihydroxy-6α-methylpregn-4-ene-3,11-dione (6), 11β,16β-dihydroxy-6α-methylpregn-4-ene-3,11-dione (7), and 15β,20R-dihydroxy-6α-methylpregn-4-ene-3,11-dione (8). Single-crystal X-ray diffraction technique unambiguously established the structures of the metabolites 2, 4, 6, and 8. Fungal transformation of 1 yielded oxidation at the C-6β, -11β, -14α, -15β, -16β positions. Various cellular anti-inflammatory assays, including inhibition of phagocyte oxidative burst, T-cell proliferation, and cytokine were performed. Among all the tested compounds, metabolite 6 (IC50 = 30.3 μg/mL) moderately inhibited the reactive oxygen species (ROS) produced from zymosan-induced human whole blood cells. Compounds 1, 4, 5, 7, and 8 strongly inhibited the proliferation of T-cells with IC50 values between <0.2–10.4 μg/mL. Compound 7 was found to be the most potent inhibitor (IC50 < 0.2 μg/mL), whereas compounds 2, 3, and 6 showed moderate levels of inhibition (IC50 = 14.6–20.0 μg/mL). Compounds 1, and 7 also inhibited the production of pro-inflammatory cytokine TNF-α. All these compounds were found to be non-toxic to 3T3 cells (mouse fibroblast), and also showed no activity when tested against HeLa (human epithelial carcinoma), or against PC3 (prostate cancer) cancer cell lines.

Isolation, Biological Evaluation, and Molecular Docking Studies of Compounds from Sophora mollis (Royle) Graham Ex Baker

“…Cytotoxic activity of compounds 1–5 was evaluated in 96-well flat-bottomed microplates by conducting cell viability using the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay according to published procedures. , In Dulbecco’s modified Eagle medium, 3T3 and HeLa cell lines were cultured, treated with streptomycin (100 μg/mL), penicillin (100 IU/mL), and 4% of fetal bovine serum (FBS) and placed in an incubator (37 °C) with (5% CO 2 ). The growing cells were rapidly collected and counted by using a haemocytometer and then diluted with a medium (5% FBS).…”

“…Cell cultures (with a concentration of 1 × 10 4 cells/mL) were prepared, placed in (85 μL/well) into 96-well plates, and incubated overnight. Cells were then treated with various concentrations − (μM) of the tested compounds. In all assays, tested compounds were dissolved in dimethyl sulfoxide immediately before the addition to cell cultures, and equal amounts of the solvent were added to control cells.…”

Cunninghamella blakesleeana-mediated biotransformation of a contraceptive drug, desogestrel, and anti-MDR-Staphylococcus aureus activity of its metabolites

“…In continuation of our work on fungal-mediated synthesis of bioactive steroids [17][18][19][20], including 17a-ethynyl substituted steroidal drugs [21][22][23], an oral contraceptive drug, desogestrel…”

Seven new metabolites of drostanolone heptanoate by using Beauveria bassiana, and Macrophomina phaseolina cell suspension cultures

“…12 Similarly, several metabolites of drostanolone on incubation with cryopreserved human hepatocytes are also reported. 13 In continuation of our structural transformation studies on steroidal drugs, [14][15][16][17][18][19][20] biotransformation of drostanolone heptanoate (1) was carried out. Based on small scale screening results, compound 1 was subjected to biotransformation by using two microbial cell cultures, Beauveria bassiana, and Macrophomina phaseolina, for the rst time, yielding seven new, and two known compounds.…”