Runlan Wan,Xi Kong,Youzhe Yang,Siwen Tao,Youyou Chen,Alexander Tobias Teichmann,Frank Heinrich Wieland
The clinical use of the steroidal aromatase inhibitor Formestane (4-hydroxandrostenedione, 4-OHA) in the treatment of advanced ER+ breast cancer has been discontinued, and therefore, interest in this remarkable drug has vanished. As a C-19 sterol, 4-OHA can undergo extensive intracellular metabolism depending on the expression of specific enzymes in the corresponding cells. We used the metabolites 4β-hydroxyandrosterone, 4β-hydroxyepiandrosterone and its 17β-reduced derivative as standards for the proof of catalytic activity present in the cell culture medium and expressed by the isolated enzymes. All of the aldo-keto reductases AKR1C1, AKR1C2, AKR1C3 and AKR1C4 catalysed the reduction of the 3-keto-group and the Δ 4,5 double bond of 4-OHA at the same time. Molecular docking experiments using microscale thermophoresis and the examination of the kinetic behaviour of the isolated enzymes with the substrate 4-OHA proved that AKR1C3 had the highest affinity for the substrate, whereas AKR1C1 was the most efficient enzyme. Both enzymes (AKR1C1and AKR1C3) are highly expressed in adipose tissue and lungs, exhibiting 3β-HSD activity. The possibility that 4-OHA generates biologically active derivatives such as the androgen 4-hydroxytestosterone or some 17β-hydroxy derivatives of the 5α-reduced metabolites may reawaken interest in Formestane, provided that a suitable method of administration can be developed, avoiding oral or intramuscular depot-injection administration.
In this study, an untargeted metabolomics method based on LC-MS was used to investigate the differential metabolites of coix seed after fermentation by Monascus purpureus . The compounds with highly correlated variable importance in projection (VIP) values of orthogonal partial least squares discriminant analysis (OPLS-DA) were selected as the important metabolites of coix seed samples. Coix seed was fermented by M. purpureus for 9 d. The color value units (CVU) of pigment in fermented coix seed reached 157.47 CVU/g, and the contents of total phenols and total flavonoids were 0.32 mg/g and 1.2 mg/g, respectively. A total of 270 differential metabolites were detected, including amino acids, peptides, fatty acids, phenols, steroids, and their derivatives, of which the relative contents of 187 metabolites showed a significant upward trend (p < 0.05). Amino acids, peptides and analogues accounted for the largest proportion at 26%, among which most metabolites were enriched. For example, N-alpha-acetyllysine, l -tryptophan and l -arginine exhibited a fold change (FC) of 6200.87, 330.29, and 35.59, respectively. In particular, the detected steroid metabolites accounted for 3% of the total metabolites, and they all participated in the same pathway, in which androsterone, 2-methoxyestrone and etiocholanolone were upregulated. The synthesis of these steroids was related to the precursors and enzymes in the pathway, such as dehydroepiandrosterone, androstenedione, and 3beta-hydroxy-delta5-steroid dehydrogenase (HSD3B). The results of this study reveal M. purpureus fermentation can successfully transform coix seed, improving the functional activities of metabolites. Thus, this process has high development value with important applications in the food industry.
危险品化学品经营许可证（带存储）