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Abstract：
BACKGROUND: Cancer cells exhibit selective metabolic reprogramming to promote proliferation, invasiveness, and metastasis. Sphingolipids such as sphingosine and sphinganine have been reported to modulate cell death processes in cancer cells. However, the potential of extracellular sphinganine and its mimetic compounds as inducers of cancer cell death has not been thoroughly investigated.
METHODS: We obtained extracellular conditioned medium from HCT-116 cells treated with the previously reported anticancer composition, goat urine DMSO fraction (GUDF). The extracellular metabolites were purified using a novel and in-house developed vertical tube gel electrophoresis (VTGE) technique and identified through LC-HRMS. Extracellular metabolites such as sphinganine, sphingosine, C16 sphinganine, and phytosphingosine were screened for their inhibitory role against intracellular kinases using molecular docking. Molecular dynamics (MD) simulations were performed to study the inhibitory potential of a novel designed modified mimetic sphinganine (MMS) (Pubchem CID: 162625115) upon c-Src kinase. Furthermore, inhibitory potential and ADME profile of MMS was compared with luteolin, a known c-Src kinase inhibitor.
RESULTS: Data showed accumulation of sphinganine and other sphingolipids such as C16 sphinganine, phytosphingosine, and ceramide (d18:1/14:0) in the extracellular compartment of GUDF-treated HCT-116 cells. Molecular docking projected c-Src kinase as an inhibitory target of sphinganine. MD simulations projected MMS with strong (-7.1 kcal/mol) and specific (MET341, ASP404) binding to the inhibitory pocket of c-Src kinase. The projected MMS showed comparable inhibitory role and acceptable ADME profile over known inhibitors.
CONCLUSIONS: In summary, our findings highlight the significance of extracellular sphinganine and other sphingolipids, including C16 sphinganine, phytosphingosine, and ceramide (d18:1/14:0), in the context of drug-induced cell death in HCT-116 cancer cells. Furthermore, we demonstrated the importance of extracellular sphinganine and its modified mimetic sphinganine (MMS) as a potential inhibitor of c-Src kinase. These findings suggest that MMS holds promise for future applications in targeted and combinatorial anticancer therapy.
METHODS: We obtained extracellular conditioned medium from HCT-116 cells treated with the previously reported anticancer composition, goat urine DMSO fraction (GUDF). The extracellular metabolites were purified using a novel and in-house developed vertical tube gel electrophoresis (VTGE) technique and identified through LC-HRMS. Extracellular metabolites such as sphinganine, sphingosine, C16 sphinganine, and phytosphingosine were screened for their inhibitory role against intracellular kinases using molecular docking. Molecular dynamics (MD) simulations were performed to study the inhibitory potential of a novel designed modified mimetic sphinganine (MMS) (Pubchem CID: 162625115) upon c-Src kinase. Furthermore, inhibitory potential and ADME profile of MMS was compared with luteolin, a known c-Src kinase inhibitor.
RESULTS: Data showed accumulation of sphinganine and other sphingolipids such as C16 sphinganine, phytosphingosine, and ceramide (d18:1/14:0) in the extracellular compartment of GUDF-treated HCT-116 cells. Molecular docking projected c-Src kinase as an inhibitory target of sphinganine. MD simulations projected MMS with strong (-7.1 kcal/mol) and specific (MET341, ASP404) binding to the inhibitory pocket of c-Src kinase. The projected MMS showed comparable inhibitory role and acceptable ADME profile over known inhibitors.
CONCLUSIONS: In summary, our findings highlight the significance of extracellular sphinganine and other sphingolipids, including C16 sphinganine, phytosphingosine, and ceramide (d18:1/14:0), in the context of drug-induced cell death in HCT-116 cancer cells. Furthermore, we demonstrated the importance of extracellular sphinganine and its modified mimetic sphinganine (MMS) as a potential inhibitor of c-Src kinase. These findings suggest that MMS holds promise for future applications in targeted and combinatorial anticancer therapy.
摘要：
背景：癌细胞表现出选择性代谢重编程以促进增殖，侵入性,和转移。已经报道了鞘脂例如鞘氨醇和鞘氨醇调节癌细胞中的细胞死亡过程。然而,细胞外鞘氨醇及其模拟化合物作为癌细胞死亡诱导剂的潜力尚未得到彻底研究.
方法：我们从用先前报道的抗癌组合物处理的HCT-116细胞获得细胞外条件培养基，山羊尿DMSO部分（GUDF）。使用新颖且内部开发的垂直管凝胶电泳（VTGE）技术纯化细胞外代谢物，并通过LC-HRMS进行鉴定。细胞外代谢产物如鞘氨醇，鞘氨醇,C16鞘氨醇,并使用分子对接筛选了植物鞘氨醇对细胞内激酶的抑制作用。进行分子动力学（MD）模拟以研究新型设计的修饰的模拟鞘氨醇（MMS）（PubchemCID：162625115）对c-Src激酶的抑制潜力。此外，MMS的抑制潜力和ADME谱与木犀草素进行了比较，已知的c-Src激酶抑制剂。
结果：数据显示鞘氨醇和其他鞘脂类如C16鞘氨醇的积累，植物鞘氨醇，和神经酰胺(d18:1/14:0)在GUDF处理的HCT-116细胞的细胞外室中。分子对接投射c-Src激酶作为双鞘氨醇的抑制靶标。MD模拟预测MMS具有与c-Src激酶的抑制袋的强(〜7.1kcal/mol)和特异性(MET341,ASP404)结合。预计的MMS显示出与已知抑制剂相当的抑制作用和可接受的ADME谱。
结论：总之，我们的发现强调了细胞外鞘氨醇和其他鞘脂的重要性，包括C16鞘氨醇,植物鞘氨醇，和神经酰胺（d18：1/14：0），在HCT-116癌细胞中药物诱导的细胞死亡的背景下。此外，我们证明了细胞外鞘氨醇及其修饰的模拟鞘氨醇（MMS）作为c-Src激酶潜在抑制剂的重要性。这些发现表明MMS有望在靶向和组合抗癌治疗中的未来应用。
方法：我们从用先前报道的抗癌组合物处理的HCT-116细胞获得细胞外条件培养基，山羊尿DMSO部分（GUDF）。使用新颖且内部开发的垂直管凝胶电泳（VTGE）技术纯化细胞外代谢物，并通过LC-HRMS进行鉴定。细胞外代谢产物如鞘氨醇，鞘氨醇,C16鞘氨醇,并使用分子对接筛选了植物鞘氨醇对细胞内激酶的抑制作用。进行分子动力学（MD）模拟以研究新型设计的修饰的模拟鞘氨醇（MMS）（PubchemCID：162625115）对c-Src激酶的抑制潜力。此外，MMS的抑制潜力和ADME谱与木犀草素进行了比较，已知的c-Src激酶抑制剂。
结果：数据显示鞘氨醇和其他鞘脂类如C16鞘氨醇的积累，植物鞘氨醇，和神经酰胺(d18:1/14:0)在GUDF处理的HCT-116细胞的细胞外室中。分子对接投射c-Src激酶作为双鞘氨醇的抑制靶标。MD模拟预测MMS具有与c-Src激酶的抑制袋的强(〜7.1kcal/mol)和特异性(MET341,ASP404)结合。预计的MMS显示出与已知抑制剂相当的抑制作用和可接受的ADME谱。
结论：总之，我们的发现强调了细胞外鞘氨醇和其他鞘脂的重要性，包括C16鞘氨醇,植物鞘氨醇，和神经酰胺（d18：1/14：0），在HCT-116癌细胞中药物诱导的细胞死亡的背景下。此外，我们证明了细胞外鞘氨醇及其修饰的模拟鞘氨醇（MMS）作为c-Src激酶潜在抑制剂的重要性。这些发现表明MMS有望在靶向和组合抗癌治疗中的未来应用。
