Alsibaee Aishah M, Aljohar Haya I, Attwa Mohamed W, Abdelhameed Ali Saber, Kadi Adnan A
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
Heliyon. 2023 Jun 9;9(6):e17058. doi: 10.1016/j.heliyon.2023.e17058. eCollection 2023 Jun.
Spebrutinib is a new Bruton tyrosine kinase inhibitor developed by Avila Therapeutics and Celgene. Spebrutinib (SPB) is currently in phase Ib clinical trials for the treatment of lymphoma in the United States. Preliminary studies were first performed to predict susceptible sites of metabolism, reactivity pathways and structural alerts for toxicities by StarDrop WhichP450™ module, Xenosite web predictor tool and DEREK software; respectively. SPB metabolites and adducts were characterized from rat liver microsomes (RLM) using LC-MS/MS. Formation of reactive intermediates was investigated using potassium cyanide (KCN), glutathione (GSH) and methoxylamine as trapping nucleophiles for the unstable and reactive iminium, iminoquinone and aldehyde intermediates, respectively, with the aim to produce stable adducts that can be detected and characterized using mass spectrometry. Fourteen phase I metabolites, four cyanide adducts, six GSH adducts and three methoxylamine adducts of SPB were identified and characterized. The proposed metabolic pathways involved in generation of phase I metabolites of SPB are oxidation, hydroxylation, -dealkylation, epoxidation, defluorination and reduction. Several reactive intermediates were identified and characterized, the formation of which can aid in explaining the adverse drug reactions of SPB. Several iminium, 2-iminopyrimidin-5()-one and aldehyde intermediates of SPB were revealed. Acrylamide is identified as a structural alert for toxicity by DEREK report and was found to be involved in the formation of several glycidamide and aldehyde reactive intermediates.
司布替尼是阿维拉治疗公司和新基公司联合开发的一种新型布鲁顿酪氨酸激酶抑制剂。司布替尼（SPB）目前正在美国进行Ib期淋巴瘤治疗的临床试验。最初分别通过StarDrop WhichP450™模块、Xenosite网络预测工具和DEREK软件进行初步研究，以预测代谢易感位点、反应途径和毒性结构警示。利用液相色谱-串联质谱法（LC-MS/MS）对大鼠肝微粒体（RLM）中的SPB代谢物和加合物进行了表征。分别使用氰化钾（KCN）、谷胱甘肽（GSH）和甲氧基胺作为捕获亲核试剂，研究不稳定且具有反应活性的亚胺鎓、亚氨基醌和醛中间体的反应性中间体形成情况，目的是生成可通过质谱检测和表征的稳定加合物。鉴定并表征了SPB的14种I相代谢物、4种氰化物加合物、6种GSH加合物和3种甲氧基胺加合物。SPB的I相代谢物生成所涉及的推测代谢途径包括氧化、羟基化、脱烷基化、环氧化、脱氟和还原。鉴定并表征了几种反应性中间体，其形成有助于解释SPB的药物不良反应。揭示了SPB的几种亚胺鎓、2-亚氨基嘧啶-5（）-酮和醛中间体。DEREK报告将丙烯酰胺鉴定为毒性结构警示，并发现其参与了几种缩水甘油酰胺和醛反应性中间体的形成。
