Background

Novel compounds and more efficient treatment options are urgently needed for the treatment of non-small cell lung cancer (NSCLC). The decoction of Sophora flavescens has been used to treat NSCLC in the clinic, and matrine-type alkaloids are generally considered to be the key pharmacodynamic material basis. But the previous study showed that common matrine-type alkaloids exhibit significant cytotoxicity only when at concentrations close to the millimolar (mM) level. The key antitumor alkaloids in S. flavescens seem to have not yet been revealed.

Purpose

The aim of this study was to screen water-soluble matrine alkaloid with novel skeleton and enhanced activity from S. flavescens, and to reveal the pharmacological mechanism of its therapeutic effect on NSCLC.

Methods

Alkaloid was obtained from S. flavescens by chromatographic separation methods. The structure of alkaloid was determined by spectroscopic methods, and single-crystal X-ray diffraction. The mechanism of anti-NSCLC in vitro with cellular models was evaluated by MTT assay, western blotting, cell migration and invasion assay, plate colony-formation assay, tube formation assay, immunohistochemistry assay, hematoxylin and eosin staining. The antitumor efficacy in vivo was test in NSCLC xenograft models.

Results

A novel water-soluble matrine-derived alkaloid incorporating 6/8/6/6 tetracyclic ring system, named sophflarine A (SFA), was isolated from the roots of S. flavescens. SFA had significantly enhanced cytotoxicity compared with the common matrine-type alkaloids, having an IC₅₀ value of 11.3 μM in A549 and 11.5 μM in H820 cells at 48h. Mechanistically, SFA promoted NSCLC cell death by inducing pyroptosis via activating the NLRP3/caspase-1/GSDMD signaling pathway, and inhibited cancer cell proliferation by increasing the ROS production to activate autophagy via blocking the PI3K/AKT/mTOR signaling pathway. Additionally, SFA also inhibited NSCLC cell migration and invasion by suppressing EMT pathway, and inhibited cancer cell colony formation and human umbilical vein endothelial cell angiogenesis. In concordance with the above results, SFA treatment blocked tumor growth in an A549 cell-bearing orthotopic mouse model.

Conclusion

This study revealed a potential therapeutic mechanism of a novel matrine-derived alkaloid, which not only described a rational explanation for the clinical utilization of S. flavescens, but also provided a potential candidate compound for NSCLC treatment.

中文翻译：

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Sophflarine A，一种来自苦参的新型苦参碱衍生生物碱，具有通过 ROS 介导的细胞焦亡和自噬治疗非小细胞肺癌的潜力

背景

治疗非小细胞肺癌 (NSCLC) 迫切需要新的化合物和更有效的治疗方案。苦参煎剂在临床上已被用于治疗NSCLC，苦参碱类生物碱被普遍认为是关键的药效物质基础。但之前的研究表明，常见的苦参碱类生物碱只有在浓度接近毫摩尔 (mM) 水平时才会表现出显着的细胞毒性。苦参中的关键抗肿瘤生物碱似乎尚未被发现。

目的

本研究旨在从苦参中筛选出骨架结构新颖、活性增强的水溶性苦参碱，揭示其治疗NSCLC的药理机制。

方法

生物碱是通过色谱分离方法从苦参中获得的。生物碱的结构通过光谱法和单晶X射线衍射确定。通过MTT法、western blotting、细胞迁移和侵袭试验、平板集落形成试验、管形成试验、免疫组化试验、苏木精和伊红染色评估细胞模型体外抗NSCLC的机制。在 NSCLC 异种移植模型中测试体内抗肿瘤功效。

结果

从苦参的根中分离出一种新型水溶性苦参碱衍生生物碱，该生物碱包含 6/8/6/6 四环系统，名为苦黄碱 A (SFA) 。与常见的苦参碱类生物碱相比，SFA 具有显着增强的细胞毒性，具有 IC ₅₀48 小时时，A549 中的值为 11.3 μM，H820 细胞中的值为 11.5 μM。从机制上讲，SFA 通过激活 NLRP3/caspase-1/GSDMD 信号通路诱导细胞焦亡来促进 NSCLC 细胞死亡，并通过阻断 PI3K/AKT/mTOR 信号通路增加 ROS 产生以激活自噬来抑制癌细胞增殖。此外，SFA还通过抑制EMT通路抑制NSCLC细胞迁移和侵袭，抑制癌细胞集落形成和人脐静脉内皮细胞血管生成。与上述结果一致，SFA 治疗阻止了携带 A549 细胞的原位小鼠模型中的肿瘤生长。

结论

该研究揭示了一种新型苦参碱衍生生物碱的潜在治疗机制，不仅为苦参的临床应用提供了合理解释，而且为 NSCLC 治疗提供了一种潜在的候选化合物。