Phorbol
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| Compound | Phorbol |
| Animal species | human intestinal microflora |
| Metabolism parameters | |
| Metabolites |
Phorbol
Isophorbol Deoxyphorbol (3aS,6aS,7R,10R,10aR,10bS)-3a,10a-dihydroxy-5-(hydroxymethyl)-2,10-dimethyl-7-(prop-1-en-2-yl)-3a,4,6a,7,10,10a-hexahydrobenzo[e]azulene-3,8(9H,10bH)-dione (3aR,6aR,10R,10aR,10bS)-3a,10a-dihydroxy-5-(hydroxymethyl)-2,10-dimethyl-3a,4,6a,7,10,10a-hexahydrobenzo[e]azulene-3,8(9H,10bH)-dione (3aR,6bS,8aS,9R,9aR,9bS)-3a,9a-dihydroxy-5-(hydroxymethyl)-2,7,7,9-tetramethyl-3a,4,6b,7,8a,9,9a,9b-octahydro-3H-cyclobuta[a]cyclopenta[e]azulene-3,8(6aH)-dione |
| Crude drug | Croton Seed |
| References | 1) Abdel-Hafez A. A., Nakamura N., and Hattori M.: Biotransformation of phorbol by human intestinal bacteria. Cmem. Pharm. Bull., 50, 160-164 (2002). |
| Remarks | ※Preparation of a human intestinal bacterial (HIB) mixture A fresh feces (5 g) obtained from a healthy subject was mixed with 50 mM K-phosphate buffer (50 ml, pH 7.3), and the sediments were removed by decantation. The suspension was made up to 100 ml with the same buffer, and was used as an HIB mixture in this experiment. [Abdel-Hafez et al., Cmem. Pharm. Bull., 50, 160-164 (2002)] ※Transformation of 1 by an HIB mixture A precultured bacterial suspension (300 ml) was added to GAM broth (3000 ml) and the mixture was cultured for 24 h at 37 °C under anaerobic conditions. The culture was centrifuged at 8000 × g for 10 min. The pellets were washed with saline and suspended in GAM broth (1000 ml). Phorbol (1, 1 g in 2 ml MeOH) was added to the bacterial suspension, and the mixture was anaerobically incubated at 37 °C for 3 d. The reaction mixture was extracted with ethyl acetate (500 ml × 4). The residue (2.4 g) after evaporation of ethyl acetate in vacuo was chromatographed over silica gel eluted with CHCl3-MeOH (9:1) to give fractions A (150 mg) and B (315 mg). Fraction A was separated by MPLC using a LiChroprep RP-18 column with MeOH-H2O (4 :6) as a mobile phase to yield compounds 2 (9 mg) and 3 (5 mg). Fraction B was also separated by MPLC with RP-18 (MeOH-H2O, 3 : 7) to afford compounds 4 (28 mg), 5 (16 mg) and 6 (4 mg). [Abdel-Hafez et al., Cmem. Pharm. Bull., 50, 160-164 (2002)] ※Time Course of the metabolism of phorbol (1) by a human intestinal bacterial mixture from human faces To a suspension of the precultured bacterial mixture (3 ml) in GAM broth (10 ml), phorbol (10 mg) was added, and the mixture was anaerobically incubated at 37°C. A portion (1 ml) of it was taken each day for 4 d and extracted with ethyl acetate (1 ml). An aliquot (10 ml) of the ethyl acetate extract was applied to a TLC plate (Merk Kieselgel F245, 0.25 mm thickness) and the plate was developed with CHCl3-MeOH (9:1) and analyzed with a TLC scanner at 260 nm. The amounts of phorbol and its metabolites were determined by using calibration lines prepared with authentic samples. [Abdel-Hafez et al., Cmem. Pharm. Bull., 50, 160-164 (2002)] ※Screening of Bacterial Strains for Their Abilities to Metabolize Phorbol (1) Each precultured bacterial suspension (0.2 ml) was added to GAM broth (10 ml) and cultured for 24 h at 37°C under anaerobic conditions. A portion (3 ml) of each culture was diluted to 10 ml with the same medium and incubated for further 24 h. Phorbol (1, 10 mg) was added to each culture and the mixture was incubated for 3 d at 37°C under anaerobic conditions. The incubation mixture was extracted with EtOAc (10 ml x 3). The extract was evaporated in vacuo to afford a residue. The residue was dissolved in MeOH (1 ml) and analyzed quantitatively by TLC-densitometry. [Abdel-Hafez et al., Cmem. Pharm. Bull., 50, 160-164 (2002)] |
