Synthesis of some potent immunomodulatory and anti-inflammatory metabolites by fungal transformation of anabolic steroid oxymetholone

• Main Authors: Khan Naik Tameen, Bibi Marium, Yousuf Sammer, Qureshi Izhar Husain, Atta-ur-Rahman, Al-Majid Abdullah Mohammad, Mesaik Muhammad Ahmed, Khalid Ahmed Shukralla, Sattar Samina A, Atia-tul-Wahab, Choudhary M Iqbal
• Format: Article
• Language: English
• Published: BMC 2012-12-01
• Series: Chemistry Central Journal
• Subjects: Oxymetholone; Anabolic steroid; Biotransformation; <it>de novo</it> Hydroxylation; Immunomodulation; T-Cell proliferation inhibition; Anti-inflammation; Inhibition of reactive oxygen species production; 3T3 Fibroblast cells
• Online Access: http://journal.chemistrycentral.com/content/6/1/153

<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>Incubation of oxymetholone (<b>1</b>) with <it>Macrophomina phaseolina</it>, <it>Aspergillus niger</it>, <it>Rhizopus stolonifer</it>, and <it>Fusarium lini</it> produced 17β-hydroxy-2-(hydroxy-methyl)-17α-methyl-5α-androstan-1-en-3-one (<b>2</b>), 2α,17α-di(hydroxyl-methyl)-5α-androstan-3β,17β-diol (<b>3</b>), 17α-methyl-5α-androstan-2α,3β,17β-triol (<b>4</b>), 17β-hydroxy-2-(hydroxymethyl)-17α-methyl-androst-1,4-dien-3-one (<b>5</b>), 17β-hydroxy-2α-(hydroxy-methyl)-17α-methyl-5α-androstan-3-one (<b>6</b>), and 2α-(hydroxymethyl)-17α-methyl-5α-androstan-3β-17β-diol (<b>7</b>). Their structures were deduced by spectral analyses, as well as single-crystal X-ray diffraction studies. Compounds <b>2</b>–<b>5</b> were identified as the new metabolites of <b>1</b>. The immunomodulatory, and anti-inflammatory activities and cytotoxicity of compounds <b>1</b>–<b>7</b> 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.</p> <p>Conclusion</p> <p>Microbial transformation of oxymetholone (<b>1</b>) provides an efficient method for structural transformation of <b>1</b>. The transformed products were obtained as a result of <it>de novo</it> 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.</p>