三七(Panax notoginseng (Burk.) F. H. Chen) 属于五加科（Araliaceae）植物，被中国东北地区的传统民间使用了数百年，具有降糖、抗氧化、抗肿瘤等作用。植物内生或根际微生物在植物防御机制中起着关键作用，对于药物和有价值的新次生代谢产物的发现也是至关重要的。特别是传统药用植物内生或根际微生物。为了从三七根系土壤链霉菌Streptomyces sp. SYP-A7185中发现具有生物活性和靶标的有价值的新次生代谢物。代谢产物经过色谱层析法得到并经过多重光谱分析鉴定。通过使用3-[4，5-二甲基噻唑-2-基]-2，5-二苯基四唑溴化物（MTT），2,2-二苯基-1-亚硝基肼（DPPH），96孔浊度法和α-葡萄糖苷酶抑制试验来测试分离代谢物的抗肿瘤、抗氧化、抗菌和抗糖化效应。通过网络药理学方法预测可能的抗肿瘤靶标。代谢物与靶标之间的相互作用通过分子对接和生物层干涉（BLI）实验来验证。细胞验证实验证明了基于网络药理学的假设。从Streptomyces sp. SYP-A7185中鉴定出五种不同的蒽醌类衍生物（1-10），包括六个新化合物（3，6-10）。化合物1-6和9在五种人类癌细胞株(A549、HepG2、MCF-7、MDA-MD-231和MGC-803)中显示出中等到强的细胞毒性。此外，全面的网络药理学分析预测基质金属蛋白酶（MMP2）是代谢物1-6和9的潜在抗肿瘤靶标。而BLI实验显示代谢物和抗肿瘤代谢物之间存在强烈的分子间相互作用，分子对接结果显示代谢物1-6和9与MMP2的相互作用取决于关键氨基酸残基LEU-83、ALA-84、LEU-117、HIS-131、PRO-135、GLY-136、ALA-140、PRO-141、TYR-143和THR-144。这些结果暗示代谢物（1-6和9）除了抑制癌细胞的增殖外，还能抑制癌细胞的迁移。随后，细胞创面愈合实验表明，在A549和MCF-7中，化合物1和3的治疗后，细胞迁移过程也被抑制了。此外，反转录定量聚合酶链反应（RT-qPCR）和免疫印迹（WB）结果证明，在A549和MCF-7中，化合物1和3的处理后，MMP2的基因表达水平降低了。此外，化合物2表现出中等抗氧化活性（EC50，27.43 μM），化合物3和6表现出中等抗菌活性，而化合物3的α-葡萄糖苷酶抑制活性的IC50值为13.10μM。三七根系土壤链霉菌Streptomyces sp.代谢产物具有抗肿瘤、抗氧化、抗菌和抗糖化活性。此外，化合物1和3通过下调MMP2抑制了癌细胞的迁移。Copyright © 2023. Published by Elsevier B.V.

Panax notoginseng (Burk.) F. H. Chen belongs to the Araliaceae family. It has been used by traditional Chinese people in Northeast Asia for centuries as an antidiabetic, antioxidant, antitumor agent, etc. Endophytic or rhizospheric microorganisms play key roles in plant defense mechanisms, and they are essential in the discovery of pharmaceuticals and valuable new secondary metabolites. In particular, endophytic or rhizospheric microorganisms of traditional medicinal plants.To discover valuable new secondary metabolites from rhizosphere soil Streptomyces sp. SYP-A7185 of P. notoginseng, and to explore potential bioactivities and targets of metabolites protrusive function.The metabolites were obtained via column chromatography and identified by multiple spectroscopic analyses. The antitumor, antioxidant, antibacterial, and antiglycosidases effects of isolated metabolites were tested using 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyltetazolium bromide (MTT), 2,2-diphenyl-1-picrylhydrazyl (DPPH), 96-well turbidimetric, and α-glucosidase inhibitory assays. The potential antitumor targets were predicted through network pharmacological approaches. The interactions between metabolites and target were verified by molecular docking and biolayer interferometry (BLI) assay. The effects of cancer cells migration were detected through wound healing assays in A549 and MCF-7. Other cellular validation experiments including reverse transcription-quantitative PCR (RT‒qPCR) and western blotting (WB) were used to confirm the hypothesis of network pharmacology.Five different chemotypes of anthraquinone derivatives (1-10), including six new compounds (3, 6-10), were identified from Streptomyces sp. SYP-A7185. Compounds 1-6 and 9 displayed moderate to strong cytotoxicity on five human cancer cell lines (A549, HepG2, MCF-7, MDA-MD-231, and MGC-803). Moreover, matrix metalloproteinase-2 (MMP2) were predicted as a potential antitumor target of metabolites 1-6 and 9 by comprehensive network pharmacology analysis. Later, BLI assays revealed strong intermolecular interactions between MMP2 and antitumor metabolites, and molecular docking results showed the interaction of metabolites 1-6 and 9 with MMP2 was dependent on the crucial amino acid residues of LEU-83, ALA-84, LEU-117, HIS-131, PRO-135, GLY-136, ALA-140, PRO-141, TYR-143, and THR-144. These results implied that metabolites (1-6 and 9) might inhibit cancer cell migration besides cancer cell proliferation. After that, the cell wound healing assay showed that the cell migration processes were also inhibited after the treatments of compounds 1 and 3 in A549 and MCF-7 cells. In addition, the RT‒qPCR and WB results demonstrated that the gene expression levels of MMP2 were decreased after the treatment with compounds 1 and 3 in A549 and MCF-7 cells. Besides, compound 2 displayed moderate antioxidant activity (EC50, 27.43 μM), compounds 3 and 6 exhibited moderate antibacterial activity, and compound 3 inhibited α-glucosidase with an IC50 value of 13.10 μM.Anthraquinone metabolites, from rhizosphere soil Streptomyces sp. of P. notoginseng, possess antitumor, antioxidant, antibacterial, and antiglycosidase activities. Moreover, metabolites 1 and 3 inhibit cancer cells migration through downregulating MMP2.Copyright © 2023. Published by Elsevier B.V.