RESUMO
BACKGROUND: Biotransformation of organic compounds by using microbial whole cells provides an efficient approach to obtain novel analogues which are often difficult to synthesize chemically. In this manuscript, we report for the first time the microbial transformation of a synthetic anabolic steroidal drug, oxymetholone, by fungal cell cultures. RESULTS: Incubation of oxymetholone (1) with Macrophomina phaseolina, Aspergillus niger, Rhizopus stolonifer, and Fusarium lini produced 17ß-hydroxy-2-(hydroxy-methyl)-17α-methyl-5α-androstan-1-en-3-one (2), 2α,17α-di(hydroxyl-methyl)-5α-androstan-3ß,17ß-diol (3), 17α-methyl-5α-androstan-2α,3ß,17ß-triol (4), 17ß-hydroxy-2-(hydroxymethyl)-17α-methyl-androst-1,4-dien-3-one (5), 17ß-hydroxy-2α-(hydroxy-methyl)-17α-methyl-5α-androstan-3-one (6), and 2α-(hydroxymethyl)-17α-methyl-5α-androstan-3ß-17ß-diol (7). Their structures were deduced by spectral analyses, as well as single-crystal X-ray diffraction studies. Compounds 2-5 were identified as the new metabolites of 1. The immunomodulatory, and anti-inflammatory activities and cytotoxicity of compounds 1-7 were evaluated by observing their effects on T-cell proliferation, reactive oxygen species (ROS) production, and normal cell growth in MTT assays, respectively. These compounds showed immunosuppressant effect in the T-cell proliferation assay with IC50 values between 31.2 to 2.7 µg/mL, while the IC50 values for ROS inhibition, representing anti-inflammatory effect, were in the range of 25.6 to 2.0 µg/mL. All the compounds were found to be non-toxic in a cell-based cytotoxicity assay. CONCLUSION: Microbial transformation of oxymetholone (1) provides an efficient method for structural transformation of 1. The transformed products were obtained as a result of de novo stereoselective reduction of the enone system, isomerization of double bond, insertion of double bond and hydroxylation. The transformed products, which showed significant immunosuppressant and anti-inflammatory activities, can be further studied for their potential as novel drugs.
RESUMO
The biotransformation of dehydroepiandrosterone (1) with Macrophomina phaseolina was investigated. A total of eight metabolites were obtained which were characterized as androstane-3,17-dione (2), androst-4-ene-3,17-dione (3), androst-4-ene-17ß-ol-3-one (4), androst-4,6-diene-17ß-ol-3-one (5), androst-5-ene-3ß,17ß-diol (6), androst-4-ene-3ß-ol-6,17-dione (7), androst-4-ene-3ß,7ß,17ß-triol (8), and androst-5-ene-3ß,7α,17ß-triol (9). All the transformed products were screened for enzyme inhibition, among which four were found to inhibit the ß-glucuronidase enzyme, while none inhibited the α-chymotrypsin enzyme.
Assuntos
Ascomicetos/metabolismo , Desidroepiandrosterona/metabolismo , Desidroepiandrosterona/farmacologia , Inibidores Enzimáticos/metabolismo , Inibidores Enzimáticos/farmacologia , Glucuronidase/antagonistas & inibidores , Biotransformação , Desidroepiandrosterona/química , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/química , Glucuronidase/metabolismo , Conformação Molecular , Relação Estrutura-AtividadeRESUMO
Microbial transformation of (20S)-20-hydroxymethylpregna-1,4-dien-3-one (1) by four filamentous fungi, Cunninghamella elegans, Macrophomina phaseolina, Rhizopus stolonifer, and Gibberella fujikuroi, afforded nine new, and two known metabolites 2-12. The structures of these metabolites were characterized through detailed spectroscopic analysis. These metabolites were obtained as a result of biohydroxylation of 1 at C-6ß, -7ß, -11α, -14α, -15ß, -16ß, and -17α positions, except metabolite 2 which contain an O-acetyl group at C-22. These fungal strains demonstrated to be efficient biocatalysts for 11α-hydroxylation. Compound 1, and its metabolites were evaluated for the first time for their cytotoxicity against the HeLa cancer cell lines, and some interesting results were obtained.
Assuntos
Fungos/metabolismo , Pregnadienos/metabolismo , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Ascomicetos/metabolismo , Biotransformação , Cunninghamella/metabolismo , Gibberella/metabolismo , Células HeLa , Humanos , Hidroxilação , Estrutura Molecular , Pregnadienos/química , Rhizopus/metabolismo , Análise Espectral , EstereoisomerismoRESUMO
The microbial transformation of an oral contraceptive, mestranol (1) by Cunninghamella elegans yielded two hydroxylated metabolites, 6beta-hydroxymestranol (2) and 6beta,12beta-dihydroxymestranol (3). Metabolite 3 was found to be a new compound. These metabolites were structurally characterized on the basis of spectroscopic techniques.
