title: Synthesis of some potent immunomodulatory and anti-inflammatory metabolites by fungal transformation of anabolic steroid oxymetholone.
abstract: 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 17β-hydroxy-2-(hydroxy-methyl)-17α-methyl-5α-androstan-1-en-3-one (2), 2α,17α-di(hydroxyl-methyl)-5α-androstan-3β,17β-diol (3), 17α-methyl-5α-androstan-2α,3β,17β-triol (4), 17β-hydroxy-2-(hydroxymethyl)-17α-methyl-androst-1,4-dien-3-one (5), 17β-hydroxy-2α-(hydroxy-methyl)-17α-methyl-5α-androstan-3-one (6), and 2α-(hydroxymethyl)-17α-methyl-5α-androstan-3β-17β-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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: H, E, J, Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan. iqbal.choudhary@iccs.edu.
authors.forename: Naik Tameen
authors.initials: NT
authors.lastname: Khan
authors.forename: Marium
authors.initials: M
authors.lastname: Bibi
authors.forename: Sammer
authors.initials: S
authors.lastname: Yousuf
authors.forename: Izhar Husain
authors.initials: IH
authors.lastname: Qureshi
authors.forename: Null
authors.initials: N
authors.lastname: Atta-Ur-Rahman
authors.forename: Abdullah Mohammad
authors.initials: AM
authors.lastname: Al-Majid
authors.forename: Muhammad Ahmed
authors.initials: MA
authors.lastname: Mesaik
authors.forename: Ahmed Shukralla
authors.initials: AS
authors.lastname: Khalid
authors.forename: Samina A
authors.initials: SA
authors.lastname: Sattar
authors.forename: Null
authors.initials: N
authors.lastname: Atia-Tul-Wahab
authors.forename: M Iqbal
authors.initials: MI
authors.lastname: Choudhary