Seven new derivatives, 6α-hydroxy-1-methyl-3-oxo-5α-androst-1-en-17-yl acetate (2), 6α,17β-dihydroxy-1-methyl-3-oxo-5α-androst-1-en (3), 7β-hydroxy-1-methyl-3-oxo-5α-androst-1-en-17-yl acetate (4), 15β,20-dihydroxy-1-methyl-3-oxo-5α-androst-1-en-17-yl acetate (5), 15β-hydroxy-1-methyl-3-oxo-5α-androst-1-en-17-yl acetate (6), 12β,17β-dihydroxy-1-methyl-3-oxoandrosta-1,4-dien (11), and 7β,15β,17β-trihydroxy-1-methyl-3-oxo-5α-androst-1-en (14), along with six known metabolites, 17β-hydroxy-1-methyl-3-oxoandrosta-1,4-dien (7), 17β-hydroxy-1-methyl-3-oxo-5α-androst-1-en (8), 17β-hydroxy-1-methyl-3-oxo-5β-androst-1-en (9), 1-methyl-5β-androst-1-en-3,17-dione (10), 1-methyl-3-oxoandrosta-1,4-dien-3,17-dione (12), and 17β-hydroxy-1α-methyl-5α-androstan-3-one (13) of metenolone acetate (1), were synthesized through whole-cell biocatalysis with Rhizopus stolonifer, Aspergillus alliaceous, Fusarium lini, and Cunninghamella elegans. Atamestane (12), an aromatase inhibitor, was synthesized for the first time via F. lini-mediated transformation of 1 as the major product. Hydroxylation, dehydrogenation, and reduction were occurred during biocatalysis. Study indicated that F. lini was able to catalyze dehydrogenation reactions selectively. Structures of compounds 1–14 were determined through NMR, HRFAB-MS, and IR spectroscopic data. Compounds 1–14 were identified as non-cytotoxic against BJ human fibroblast cell line (ATCC CRL-2522). Metabolite 5 (81.0 ± 2.5%) showed a potent activity against TNF-α production, as compared to the substrate 1 (62.5 ± 4.4%). Metabolites 2 (73.4 ± 0.6%), 8 (69.7 ± 1.4%), 10 (73.2 ± 0.3%), 11 (60.1 ± 3.3%), and 12 (71.0 ± 7.2%), also showed a good inhibition of TNF-α production. Compounds 3 (IC₅₀ = 4.4 ± 0.01 µg/mL), and 5 (IC₅₀ = 10.2 ± 0.01 µg/mL) showed a significant activity against T-cell proliferation. Identification of selective inhibitors of TNF-α production, and T-cell proliferation is a step forward towards the development of anti-inflammatory drugs.

七种新衍生物，6 α -hydroxy-1-methyl-3-oxo-5 α -androst-1-en-17-yl acetate ( 2 ), 6 α ,17 β -dihydroxy-1-methyl-3-oxo- 5 α -androst-1-en ( 3 ), 7 β -hydroxy-1-methyl-3-oxo-5 α -androst-1-en-17-yl acetate ( 4 ), 15 β ,20-dihydroxy-1 -methyl-3-oxo-5 α -androst-1-en-17-yl acetate ( 5 ), 15 β -hydroxy-1-methyl-3-oxo-5 α -androst-1-en-17-yl acetate ( 6 ), 12 β ,17 β -dihydroxy-1-methyl-3-oxoandrosta-1,4-dien (11 ) 和 7 β、 15 β、 17 β -trihydroxy-1-methyl-3-oxo-5 α -androst-1-en ( 14 )，以及六种已知代谢物，17 β -hydroxy-1-methyl- 3-oxoandrosta-1,4-dien ( 7 ), 17 β -hydroxy-1-methyl-3-oxo-5 α -androst-1-en ( 8 ), 17 β -hydroxy-1-methyl-3-oxo -5 β -androst-1-en ( 9 ), 1-methyl-5 β -androst-1-en-3,17-dione (10), 1-methyl-3-oxoandrosta-1,4-dien-3 ,17-二酮 ( 12 ) 和 17 β -hydroxy-1 α -methyl-5 α-androstan-3-one ( 13 ) 醋酸美替诺龙 ( 1 ) 是通过与匍匐根霉、葱曲霉、亚麻镰刀菌和Cunninghamella elegans的全细胞生物催化合成的。Atamestane ( 12 ) 是一种芳香酶抑制剂，首次通过 F. lini介导的1作为主要产物的转化而合成。在生物催化过程中发生羟基化、脱氢和还原。研究表明F. lini能够选择性地催化脱氢反应。化合物1 – 14的结构通过 NMR、HRFAB-MS 和 IR 光谱数据确定。化合物1-14被鉴定为对BJ人成纤维细胞系 (ATCC CRL-2522) 无细胞毒性。与底物1 (62.5 ± 4.4%)相比，代谢物5 (81.0 ± 2.5%) 显示出对 TNF -α产生的有效活性。代谢物2 (73.4 ± 0.6%)、8 (69.7 ± 1.4%)、10 (73.2 ± 0.3%)、11 (60.1 ± 3.3%) 和12 (71.0 ± 7.2%) 也表现出良好的 TNF- α 生产。化合物3 (IC ₅₀  = 4.4 ± 0.01 µg/mL) 和5 (IC₅₀  = 10.2 ± 0.01 µg/mL) 显示出显着的抗 T 细胞增殖活性。识别 TNF-α 产生和 T 细胞增殖的选择性抑制剂是朝着开发抗炎药物迈出的一步。