INTRODUCTION To generate biomarkers of target engagement or predictive response for multi-target drugs is challenging. One such compound is the multi-AGC kinase inhibitor AT13148. Metabolic signatures of selective signal transduction inhibitors identified in preclinical models have previously been confirmed in early clinical studies. This study explores whether metabolic signatures could be used as biomarkers for the multi-AGC kinase inhibitor AT13148. OBJECTIVES To identify metabolomic changes of biomarkers of multi-AGC kinase inhibitor AT13148 in cells, xenograft / mouse models and in patients in a Phase I clinical study. METHODS HILIC LC-MS/MS methods and Biocrates AbsoluteIDQ™ p180 kit were used for targeted metabolomics; followed by multivariate data analysis in SIMCA and statistical analysis in Graphpad. Metaboanalyst and String were used for network analysis. RESULTS BT474 and PC3 cells treated with AT13148 affected metabolites which are in a gene protein metabolite network associated with Nitric oxide synthases (NOS). In mice bearing the human tumour xenografts BT474 and PC3, AT13148 treatment did not produce a common robust tumour specific metabolite change. However, AT13148 treatment of non-tumour bearing mice revealed 45 metabolites that were different from non-treated mice. These changes were also observed in patients at doses where biomarker modulation was observed. Further network analysis of these metabolites indicated enrichment for genes associated with the NOS pathway. The impact of AT13148 on the metabolite changes and the involvement of NOS-AT13148- Asymmetric dimethylarginine (ADMA) interaction were consistent with hypotension observed in patients in higher dose cohorts (160-300 mg). CONCLUSION AT13148 affects metabolites associated with NOS in cells, mice and patients which is consistent with the clinical dose-limiting hypotension.

中文翻译：

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细胞、小鼠和患者中多 (-AGC-) 激酶抑制剂 AT13148 的代谢组变化与 NOS 调节相关。


简介 生成多靶点药物的靶点参与或预测反应的生物标志物具有挑战性。其中一种化合物是多 AGC 激酶抑制剂 AT13148。临床前模型中鉴定的选择性信号转导抑制剂的代谢特征先前已在早期临床研究中得到证实。本研究探讨代谢特征是否可以用作多 AGC 激酶抑制剂 AT13148 的生物标志物。目的 在 I 期临床研究中确定细胞、异种移植/小鼠模型和患者中多 AGC 激酶抑制剂 AT13148 生物标志物的代谢组变化。方法 HILIC LC-MS/MS 方法和 Biocrates AbsoluteIDQ™ p180 试剂盒用于靶向代谢组学；接下来是 SIMCA 中的多变量数据分析和 Graphpad 中的统计分析。 Metaboanalyst 和 String 用于网络分析。结果 用 AT13148 处理的 BT474 和 PC3 细胞影响代谢物，这些代谢物位于与一氧化氮合酶 (NOS) 相关的基因蛋白代谢物网络中。在携带人类肿瘤异种移植物 BT474 和 PC3 的小鼠中，AT13148 治疗没有产生常见的强大的肿瘤特异性代谢变化。然而，AT13148 对非荷瘤小鼠的治疗显示出 45 种代谢物与未治疗的小鼠不同。在观察到生物标志物调节剂量的患者中也观察到了这些变化。对这些代谢物的进一步网络分析表明与 NOS 途径相关的基因富集。 AT13148 对代谢物变化的影响以及 NOS-AT13148-不对称二甲基精氨酸 (ADMA) 相互作用的影响与在较高剂量组 (160-300 mg) 患者中观察到的低血压一致。 结论 AT13148影响细胞、小鼠和患者中与NOS相关的代谢物，这与临床剂量限制性低血压一致。