Esterification promotes the intracellular accumulation of roxadustat, an activator of hypoxia-inducible factors, to extend its effective duration
Esterification promotes the intracellular accumulation of roxadustat, an activator of hypoxia-inducible factors, to extend its effective duration
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DOI：
10.1016/j.bcp.2022.114939
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发表时间：
2022-02-03
影响因子：
5.8
通讯作者：
Suzuki，Norio
中科院分区：
文献类型：
--
作者：
Nakai，Taku;Saigusa，Daisuke;Suzuki，Norio
肾损伤通常由于肾脏中红细胞生长因子促红细胞生成素（EPO）的缺乏而导致贫血。Roxadustat是通过激活低氧诱导因子（HIF）诱导肾性贫血患者产生EPO的首批口服药物之一，HIF是EPO基因表达的激活剂。在这项研究中，开发roxadustat的前药，通过细胞膜的渗透性提高，我们研究了8种类型的酯化作用对roxadustat的药代动力学和生物活性的影响，使用HIF依赖性产生EPO的Hep3B肝癌细胞。用酯化的罗沙杜司他衍生物孵育的细胞的质谱分析显示，所设计的化合物在被细胞吸收后脱酯化，并且与原始化合物相比显示出低的细胞毒性。在短暂暴露于化合物的细胞中，酯化延长了roxadustat在EPO基因诱导和HIF激活方面的有效持续时间。在小鼠的肾脏和肝脏（两者都是EPO产生的独特位点）中，大部分甲基酯化的罗沙司他在给药后6小时内被脱酯化。给药后至少48小时，去酯化的罗沙度他衍生物在血浆和尿液中可持续检测，而给药后24小时，给药化合物变得不可检测。此外，我们证实甲基酯化的roxadustat通过诱导EpomRNA仅在肾间质细胞中表达来激活小鼠的红细胞生成，肾间质细胞具有内在的EPO产生潜力。这些数据表明，酯化可导致roxadustat前药的开发，其具有改善的细胞膜渗透性、有效持续时间和细胞毒性。
Kidney injury often causes anemia due to a lack of production of the erythroid growth factor erythropoietin (EPO) in the kidneys. Roxadustat is one of the first oral medicines inducing EPO production in patients with renal anemia by activating hypoxia-inducible factors (HIFs), which are activators ofEPOgene expression. In this study, to develop prodrugs of roxadustat with improved permeability through cell membrane, we investigated the effects of 8 types of esterification on the pharmacokinetics and bioactivity of roxadustat using Hep3B hepatoma cells that HIF-dependently produce EPO. Mass spectrometry of cells incubated with the esterified roxadustat derivatives revealed that the designed compounds were deesterified after being taken up by cells and showed low cytotoxicity compared to the original compound. Esterification prolonged the effective duration of roxadustat with respect toEPOgene induction and HIF activation in cells transiently exposed to the compounds. In the kidneys and livers of mice, both of which are unique sites of EPO production, a majority of the methyl-esterified roxadustat was deesterified within 6 h after drug administration. The deesterified roxadustat derivative was continuously detectable in plasma and urine for at least 48 h after administration, while the administered compound became undetectable 24 h after administration. Additionally, we confirmed that methyl-esterified roxadustat activated erythropoiesis in mice by inducingEpomRNA expression exclusively in renal interstitial cells, which have intrinsic EPO-producing potential. These data suggest that esterification could lead to the development of roxadustat prodrugs with improvements in cell membrane permeability, effective duration and cytotoxicity.