Department of Environmental and Occupational Health, The University of Arkansas for Medical Sciences, Little Rock, AR, 72205, USA.
The Winthrop P. Rockefeller Cancer Institute, The University of Arkansas for Medical Sciences, Little Rock, AR, 72205, USA.
Rapid Commun Mass Spectrom. 2019 Nov 15;33(21):1635-1642. doi: 10.1002/rcm.8509.
Treosulfan is a substance that is being studied as part of the conditioning regimen given prior to allogeneic stem cell transplantation in patients with hematological malignancies. It is known to decompose into 1,2:3,4-diepoxybutane (DEB) under physiologic conditions. In this study, we investigate whether N-terminal valine adducts can be utilized to monitor differences in DEB formation of patients receiving treosulfan as part of the conditioning regimen for transplantation.
Blood samples were collected from a group of 14 transplant recipients and analyzed for N,N-(2,3-dihydroxy-1,4-butadiyl)valine (pyr-Val) and 2,3,4-trihydroxybutylvaline (THB-Val) adducts as biomarkers for drug uptake and metabolism before treosulfan treatment and 6 days after treatment.
A new direct injection liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed and validated prior to clinical analysis. The assay precision was determined by 3 replicate analyses on 3 individual days using control globin spiked with known amounts of pyr-Val and THB-Val. The intra- and inter-day precision coefficients of variance (CVs) and accuracy were < 10% and 15%, respectively. In clinical specimens, the means ± SD of pyr-Val and THB-Val background were 0.29 ± 0.10 pmol/g HB and 5.17 ± 1.7 pmol/g HB, respectively.
These values are similar to those found previously. Treosulfan treatment leads to a significant increase in pyr-Val and THB-Val adducts in each patient (Student's t-test p <0.0001). The mean ± SD amounts of adduct formed were 245.3 ± 89.6 and 210 ± 78.5 pmol/g globin for pyr-Val and THB-Val, respectively. Importantly, these results show that this direct injection method can quantitate both background and treosulfan-induced pyr-Val and THB-Val N-terminal valine globin adducts in humans.
三嗪氟烷是一种物质，作为异体干细胞移植患者预处理方案的一部分，正在进行研究。已知其在生理条件下会分解为 1,2:3,4-二环氧丁烷（DEB）。在这项研究中，我们研究了在接受三嗪氟烷预处理方案的患者中，是否可以利用 N-末端缬氨酸加合物来监测 DEB 形成的差异。
采集了一组 14 名移植受者的血液样本，用于分析在接受三嗪氟烷治疗前和治疗后 6 天的药物摄取和代谢的生物标志物 N,N-(2,3-二羟-1,4-丁二基）缬氨酸（吡啶-Val）和 2,3,4-三羟基丁基缬氨酸（THB-Val）加合物。
在临床分析之前，开发并验证了一种新的直接进样液相色谱/串联质谱（LC/MS/MS）方法。使用含有已知量吡啶-Val 和 THB-Val 的球蛋白标准品进行 3 天 3 次重复分析，确定了分析精密度。日内和日间变异系数（CV）和准确度的精度分别小于 10%和 15%。在临床标本中，吡啶-Val 和 THB-Val 的背景平均值±标准差分别为 0.29±0.10 pmol/g HB 和 5.17±1.7 pmol/g HB。
这些值与以前的研究相似。三嗪氟烷治疗会导致每个患者的吡啶-Val 和 THB-Val 加合物显著增加（Student's t 检验 p<0.0001）。吡啶-Val 和 THB-Val 形成的平均加合物量分别为 245.3±89.6 和 210±78.5 pmol/g 球蛋白。重要的是，这些结果表明，这种直接进样方法可以定量分析人类中的背景和三嗪氟烷诱导的吡啶-Val 和 THB-Val N-末端缬氨酸球蛋白加合物。
