Choudhary M Iqbal, Siddiqui Mahwish, Yousuf Sammer, Fatima Narjis, Ahmad Malik S, Choudhry Hani
H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan.
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan.
Front Pharmacol. 2017 Dec 20;8:900. doi: 10.3389/fphar.2017.00900. eCollection 2017.
In search of selective and effective anti-cancer agents, eight metabolites of anti-cancer steroid, drostanolone enanthate (), were synthesized microbial biotransformation. Enzymes such as reductase, oxidase, dehydrogenase, and hydrolase from , and were likely involved in the biotransformation of into new metabolites at pH 7.0 and 26°C, yielding five new metabolites, 2α-methyl-3α,14α,17β-trihydroxy-5α-androstane (), 2α-methyl-7α-hydroxy-5α-androstan-3,17-dione (), 2-methylandrosta-11α-hydroxy-1, 4-diene-3,17-dione (), 2-methylandrosta-14α-hydroxy-1,4-diene-3,17-dione (), and 2-methyl-5α-androsta-7α-hydroxy-1-ene-3,17-dione (), along with three known metabolites, 2α-methyl-3α,17β-dihydroxy-5α-androstane (), 2-methylandrosta-1, 4-diene-3,17-dione (), and 2α-methyl-5α-androsta-17β-hydroxy-3-one (), on the basis of NMR, and HREI-MS data, and single-crystal X-ray diffraction techniques. Interestingly, and were able to catalyze hydroxylation only at alpha positions of . Compounds showed a varying degree of cytotoxicity against HeLa (human cervical carcinoma), PC3 (human prostate carcinoma), H460 (human lung cancer), and HCT116 (human colon cancer) cancer cell lines. Interestingly, metabolites (IC = 49.5 ± 2.2 μM), (IC = 39.8 ± 1.5 μM), (IC = 40.7 ± 0.9 μM), (IC = 43.9 ± 2.4 μM), (IC = 19.6 ± 1.4 μM), and (IC = 25.1 ± 1.6 μM) were found to be more active against HeLa cancer cell line than the substrate (IC = 54.7 ± 1.6 μM). Similarly, metabolites (IC = 84.6 ± 6.4 μM), (IC = 68.1 ± 1.2 μM), (IC = 60.4 ± 0.9 μM), (IC = 84.0 ± 3.1 μM), (IC = 58.4 ± 1.6 μM), (IC = 59.1 ± 2.6 μM), (IC = 51.8 ± 3.4 μM), and (IC = 57.8 ± 3.2 μM) were identified as more active against PC-3 cancer cell line than the substrate (IC = 96.2 ± 3.0 μM). Metabolite (IC = 2.8 ± 0.2 μM) also showed potent anticancer activity against HCT116 cancer cell line than the substrate (IC = 3.1 ± 3.2 μM). In addition, compounds showed no cytotoxicity against 3T3 normal cell line, while compounds (IC = 74.6 ± 3.7 μM), and (IC = 62.1 ± 1.2 μM) were found to be weakly cytotoxic.
为寻找具有选择性且有效的抗癌药物，通过微生物生物转化合成了抗癌甾体药物庚酸羟甲雄酮（）的8种代谢产物。在pH 7.0和26°C条件下，来自、和的还原酶、氧化酶、脱氢酶和水解酶等酶类可能参与了向新代谢产物的生物转化过程，产生了5种新的代谢产物，即2α-甲基-3α,14α,17β-三羟基-5α-雄甾烷（）、2α-甲基-7α-羟基-5α-雄甾烷-3,17-二酮（）、2-甲基雄甾-11α-羟基-1,4-二烯-3,17-二酮（）、2-甲基雄甾-14α-羟基-1,4-二烯-3,17-二酮（）和2-甲基-5α-雄甾-7α-羟基-1-烯-3,17-二酮（），以及3种已知的代谢产物，即2α-甲基-3α,17β-二羟基-5α-雄甾烷（）、2-甲基雄甾-1,4-二烯-3,17-二酮（）和2α-甲基-5α-雄甾-17β-羟基-3-酮（）。基于核磁共振、高分辨电子轰击电离质谱数据和单晶X射线衍射技术确定了这些产物。有趣的是，和仅能催化在的α位发生羟基化反应。化合物对HeLa（人宫颈癌）、PC3（人前列腺癌）、H460（人肺癌）和HCT116（人结肠癌）癌细胞系表现出不同程度的细胞毒性。有趣的是，发现代谢产物（IC = 49.5 ± 2.2 μM）、（IC = 39.8 ± 1.5 μM）、（IC = 40.7 ± 0.9 μM）、（IC = 43.9 ± 2.4 μM）、（IC = 19.6 ± 1.4 μM）和（IC = 25.1 ± 1.6 μM）对HeLa癌细胞系的活性比底物（IC = 54.7 ± 1.6 μM）更高。同样，代谢产物（IC = 84.6 ± 6.4 μM）、（IC = 68.1 ± 1.2 μM）、（IC = 60.4 ± 0.9 μM）、（IC = 84.0 ± 3.1 μM）、（IC = 58.4 ± 1.6 μM）、（IC = 59.1 ± 2.6 μM）、（IC = 51.8 ± 3.4 μM）和（IC = 57.8 ± 3.2 μM）被确定为对PC-3癌细胞系的活性比底物（IC = 96.2 ± 3.0 μM）更高。代谢产物（IC = 2.8 ± 0.2 μM）对HCT116癌细胞系也显示出比对底物（IC = 3.1 ± 3.2 μM）更强的抗癌活性。此外，化合物对3T3正常细胞系无细胞毒性，而化合物（IC = 74.6 ± 3.7 μM）和（IC = 62.1 ± 1.2 μM）具有较弱的细胞毒性。
