PK-PD, Toxicology and Formulation Division, CSIR-Indian Institute of Integrative Medicine, Jammu, Jammu and Kashmir, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Oct 1;1154:122283. doi: 10.1016/j.jchromb.2020.122283. Epub 2020 Jul 27.
Hydroxyurea (HU) is the first-ever approved drug by the United States Food and Drug Administration (USFDA) for the management of sickle cell anemia (SCA). However, its treatment is associated with severe liabilities like myelosuppression. Therefore, the aim of the present investigation was to identify phytotherapeutics through assessment of the pharmacokinetic interaction of HU with dietary bioflavonoids followed by elucidation of the same phytoconstituents for their ability to protect HU-induced toxicity in hematological profile. In this direction, we developed a sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to estimate HU in rat plasma at first and then validated as per USFDA guidelines as there is no such precedent in the literature. A simple plasma protein precipitation method was employed for plasma sample processing. The separation was achieved in gradient mode using Syncronis HILIC column (100 × 4.6 mm, 3 μm) with a mobile phase composition of water containing 0.1% (v/v) formic acid and acetonitrile. Ionization was carried out in positive heated-electrospray ionization (H-ESI) mode. Detection was done in selected reaction monitoring (SRM) mode with m/z 77.1 > 44.4 and m/z 75.1 > 58.2 for HU and methylurea (internal standard), respectively. All the validation parameters were within the acceptable criteria. This bioanalytical method was found to be useful in assessing the preclinical pharmacokinetic interaction of HU. Concomitant administration of chrysin or quercetin with HU in rats significantly enhanced the oral exposure of HU. Lowering of total red blood cells (RBC) and hemoglobin (Hb) level by HU in rats was significantly improved in the presence of chrysin, quercetin, and naringenin. Overall, both chrysin and quercetin showed potential to be a promising phytotherapeutics for concomitant therapy with HU to combat its dose-dependent side effects.
羟基脲 (HU) 是美国食品和药物管理局 (USFDA) 批准的第一种用于治疗镰状细胞贫血 (SCA) 的药物。然而，其治疗与严重的副作用有关，如骨髓抑制。因此，本研究旨在通过评估 HU 与膳食生物类黄酮的药代动力学相互作用来识别植物疗法，然后阐明相同的植物成分，以研究它们在血液学特征中保护 HU 诱导毒性的能力。为此，我们开发了一种灵敏的超高效液相色谱-串联质谱 (UPLC-MS/MS) 方法来首次估算大鼠血浆中的 HU，然后按照 USFDA 指南进行验证，因为文献中尚无此类先例。采用简单的血浆蛋白沉淀法处理血浆样品。采用梯度模式，使用 Syncronis HILIC 柱（100×4.6mm，3μm）和水相组成的流动相进行分离，其中含有 0.1%（v/v）甲酸和乙腈。离子化在正加热电喷雾电离 (H-ESI) 模式下进行。检测采用选择反应监测 (SRM) 模式，m/z 77.1>44.4 和 m/z 75.1>58.2 分别用于 HU 和甲基脲（内标）。所有验证参数均符合可接受标准。该生物分析方法可用于评估 HU 的临床前药代动力学相互作用。在大鼠中同时给予白杨素或槲皮素与 HU，可显著增强 HU 的口服暴露。在白杨素、槲皮素和柚皮素存在的情况下，HU 降低大鼠总红细胞 (RBC) 和血红蛋白 (Hb) 水平的作用明显改善。总的来说，白杨素和槲皮素均显示出作为 HU 伴随治疗的有前途的植物疗法的潜力，以对抗其剂量依赖性副作用。
