Freyer Nora, Knöspel Fanny, Damm Georg, Greuel Selina, Schneider Christin, Seehofer Daniel, Stöhr Thomas, Petersen Karl-Uwe, Zeilinger Katrin
Berlin Brandenburg Center for Regenerative Therapies (BCRT), Charité - Universitätsmedizin Berlin, Berlin 13353, Germany,
Department of Hepatobiliary Surgery and Visceral Transplantation, University of Leipzig, Leipzig 04103, Germany.
Drug Des Devel Ther. 2019 Apr 2;13:1033-1047. doi: 10.2147/DDDT.S186759. eCollection 2019.
Remimazolam is an ultra-short acting benzodiazepine under development for procedural sedation and general anesthesia. It is hydrolyzed by CES1 to an inactive metabolite (CNS7054).
In this study, the effect of continuous remimazolam exposure on its metabolism and on expression was investigated in a dynamic 3-D bioreactor culture model inoculated with primary human hepatocytes.
Remimazolam was continuously infused into bioreactors for 5 days at a final concentration of 3,000 ng/ml (6.8 µM). In parallel, 2-D cultures were run with cells from the same donors, but with discontinuous exposure to remimazolam.
Daily measurement of clinical chemistry parameters (glucose, lactate, urea, ammonia, and liver enzymes) in culture supernatants indicated no noxious effect of remimazolam on hepatocyte integrity as compared to untreated controls. Concentrations of remimazolam reached steady-state values of around 250 ng/ml within 8 hours in 3-D bioreactors whereas in 2-D cultures remimazolam concentrations declined to almost zero within the same time frame. Levels of CNS7054 showed an inverse time-course reaching average values of 1,350 ng/ml in perfused 3-D bioreactors resp. 2,800 ng/ml in static 2-D cultures. Analysis of mRNA expression levels of indicated no changes in gene expression over the culture period.
The results indicated a stable metabolism of remimazolam during 5 days of continuous exposure to clinically relevant concentrations of the drug. Moreover, there was no evidence for a harmful effect of remimazolam exposure on the integrity and metabolic activity of in vitro cultivated primary human hepatocytes.
瑞马唑仑是一种正在研发用于手术镇静和全身麻醉的超短效苯二氮䓬类药物。它被羧酸酯酶1（CES1）水解为无活性代谢物（CNS7054）。
在接种原代人肝细胞的动态三维生物反应器培养模型中，研究瑞马唑仑持续暴露对其代谢及表达的影响。
将瑞马唑仑以终浓度3000 ng/ml（6.8 μM）持续输注到生物反应器中5天。同时，对来自相同供体的细胞进行二维培养，但瑞马唑仑暴露为间断性。
培养上清液中临床化学参数（葡萄糖、乳酸、尿素、氨和肝酶）的每日测量结果表明，与未处理的对照相比，瑞马唑仑对肝细胞完整性无有害影响。在三维生物反应器中，瑞马唑仑浓度在8小时内达到约250 ng/ml的稳态值，而在二维培养中，瑞马唑仑浓度在同一时间范围内下降至几乎为零。CNS7054水平呈现相反的时间进程，在灌注的三维生物反应器中平均值为1350 ng/ml，在静态二维培养中为2800 ng/ml。对……的mRNA表达水平分析表明，培养期间基因表达无变化。
结果表明，在持续暴露于临床相关浓度药物的5天内，瑞马唑仑代谢稳定。此外，没有证据表明瑞马唑仑暴露对体外培养的原代人肝细胞的完整性和代谢活性有有害影响。
