单甲基澳瑞他汀E（本妥昔单抗（Adcetris）中的有效载荷）对人肝细胞中细胞色素P450的抑制作用与微管破坏有关。
CYP Suppression in Human Hepatocytes by Monomethyl Auristatin E, the Payload in Brentuximab Vedotin (Adcetris), is Associated with Microtubule Disruption.
作者信息
Wolenski Francis S, Xia Cindy Q, Ma Bingli, Han Tae H, Shyu Wen C, Balani Suresh K
机构信息
Drug Safety Research and Evaluation, Takeda Pharmaceuticals International Co., Cambridge, MA, USA.
Drug Metabolism and Pharmacokinetics, Takeda Pharmaceuticals International Co., 35 Landsdowne Street, Cambridge, MA, USA.
出版信息
Eur J Drug Metab Pharmacokinet. 2018 Jun;43(3):347-354. doi: 10.1007/s13318-017-0455-5.
BACKGROUND AND OBJECTIVES
Monomethyl auristatin E (MMAE), the toxin linked to CD30-specific monoclonal antibody of Adcetris (brentuximab vedotin), is a potent anti-microtubule agent. Brentuximab vedotin has been approved for the treatment of relapsed or refractory Hodgkin lymphoma and anaplastic large cell lymphoma. Cytochrome P450 (CYP) induction assessment of MMAE was conducted in human hepatocytes to assess DDI potentials and its translation to clinic.
METHODS
MMAE was incubated at 1-1000 nM with cultured primary human hepatocytes for 72 h, and CYP1A2, CYP2B6, and CYP3A4 mRNA expression was assessed by quantitative reverse transcription-polymerase chain reaction and CYP-specific probe substrate by liquid chromatography coupled with mass spectrometry, along with microtubule disruption by immunofluorescence staining using anti-β-tubulin antibody and imaging.
RESULTS
MMAE up to 10 nM had no significant effect on CYP1A2, CYP2B6, and CYP3A4 mRNA expression and activity, whereas at higher concentrations of 100- and 1000-nM MMAE, the CYP mRNA expression and activity were diminished substantially. Further investigation showed that the degree of CYP suppression was paralleled by that of microtubule disruption by MMAE, as measured by increase in the number of β-tubulin-positive aggregates. At the clinical dose, the concentration of MMAE was 7 nM which did not show any significant CYP suppression or microtubule disruption in hepatocytes.
CONCLUSIONS
MMAE was not a CYP inducer in human hepatocytes. However, it caused a concentration-dependent CYP mRNA suppression and activity. The CYP suppression was associated with microtubule disruption, supporting the reports that intact microtubule architecture is required for CYP regulations. The absence of CYP suppression and microtubule disruption in vitro at the clinical plasma concentrations of MMAE (< 10 nM) explains the lack of pharmacokinetic drug interaction between brentuximab vedotin and midazolam, a sensitive CYP3A substrate, reported in patients.
背景与目的
单甲基澳瑞他汀E（MMAE）是与Adcetris（本妥昔单抗）的CD30特异性单克隆抗体相连的毒素，是一种有效的抗微管药物。本妥昔单抗已被批准用于治疗复发或难治性霍奇金淋巴瘤和间变性大细胞淋巴瘤。在人肝细胞中进行了MMAE的细胞色素P450（CYP）诱导评估，以评估药物相互作用的可能性及其临床转化情况。
方法
将MMAE以1 - 1000 nM的浓度与培养的原代人肝细胞孵育72小时，通过定量逆转录-聚合酶链反应评估CYP1A2、CYP2B6和CYP3A4 mRNA的表达，并通过液相色谱-质谱联用测定CYP特异性探针底物，同时使用抗β-微管蛋白抗体通过免疫荧光染色和成像评估微管破坏情况。
结果
高达10 nM的MMAE对CYP1A2、CYP2B6和CYP3A4 mRNA的表达及活性无显著影响，而在100 nM和1000 nM的较高浓度下，CYP mRNA的表达及活性显著降低。进一步研究表明，MMAE对CYP的抑制程度与微管破坏程度平行，这通过β-微管蛋白阳性聚集体数量的增加来衡量。在临床剂量下，MMAE的浓度为7 nM，在肝细胞中未显示出任何显著的CYP抑制或微管破坏。
结论
MMAE在人肝细胞中不是CYP诱导剂。然而，它会导致浓度依赖性的CYP mRNA抑制和活性降低。CYP抑制与微管破坏有关，这支持了完整的微管结构是CYP调节所必需的这一报道。在MMAE的临床血浆浓度（< 10 nM）下，体外未出现CYP抑制和微管破坏，这解释了患者中报道的本妥昔单抗与咪达唑仑（一种敏感的CYP3A底物）之间缺乏药代动力学药物相互作用的原因。
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