留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

肝移植术后受者依维莫司血药浓度检测体系的建立

陶斯湄 黄际薇 李海波 张英才 杨扬

陶斯湄, 黄际薇, 李海波, 等. 肝移植术后受者依维莫司血药浓度检测体系的建立[J]. 器官移植, 2021, 12(5): 595-600. doi: 10.3969/j.issn.1674-7445.2021.05.014
引用本文: 陶斯湄, 黄际薇, 李海波, 等. 肝移植术后受者依维莫司血药浓度检测体系的建立[J]. 器官移植, 2021, 12(5): 595-600. doi: 10.3969/j.issn.1674-7445.2021.05.014
Tao Simei, Huang Jiwei, Li Haibo, et al. Establishment of a blood concentration detection system for everolimus in recipients after liver transplantation[J]. ORGAN TRANSPLANTATION, 2021, 12(5): 595-600. doi: 10.3969/j.issn.1674-7445.2021.05.014
Citation: Tao Simei, Huang Jiwei, Li Haibo, et al. Establishment of a blood concentration detection system for everolimus in recipients after liver transplantation[J]. ORGAN TRANSPLANTATION, 2021, 12(5): 595-600. doi: 10.3969/j.issn.1674-7445.2021.05.014

肝移植术后受者依维莫司血药浓度检测体系的建立

doi: 10.3969/j.issn.1674-7445.2021.05.014
基金项目: 

广东省重点领域研发计划项目 2019B020236003

详细信息
    作者简介:

    陶斯湄,女,1986年生,主管药师,研究方向为药理学,Email: taosm3@mail2.sysu.edu.cn

    通讯作者:

    黄际薇,女,1969年生,副主任药师,研究方向为临床药学与医院药学, E-mail: 947543512@qq.com

    杨扬,男,1971年生,博士,教授、主任医师,研究方向为肝移植, E-mail: yysysu@163.com

  • 中图分类号: R617, R969

Establishment of a blood concentration detection system for everolimus in recipients after liver transplantation

More Information
  • 摘要:   目的  建立测定肝移植受者全血中依维莫司浓度的超高效液相色谱-串联质谱法(UPLC-MS/MS)检测体系。  方法  用甲醇和硫酸锌沉淀样本蛋白质,以依维莫司-D4为内标物。采用Phenomenex Kinetex PFP色谱柱;流动相A:水(含有2 mmol/L甲酸铵和0.1%甲酸),流动相B:甲醇(含有2 mmol/L甲酸铵和0.1%甲酸),梯度洗脱,流速1 mL/min,柱温50 ℃,进样量1 μL。在正离子模式下,以多反应监测方式进行定量测定。该UPLC-MS/MS检测体系只需100 μL全血,无需复杂的样品制备即可达到足够的定量下限,总运行时间4.5 min内。采用依维莫司峰面积/依维莫司-D4峰面积(y)和依维莫司浓度/依维莫司-D4浓度(x)进行线性回归(1/ x2)分析,计算校准函数,分析其准确度和线性关系。并应用UPLC-MS/MS检测5例肝移植术后受者血样本的依维莫司血药谷浓度。  结果  依维莫司在血药浓度1~100 ng /mL范围内的质控准确度都在15%以内,线性关系良好(R2 > 0.990)。测定的5例肝移植受者血样本的依维莫司血药谷浓度为3.77~9.27 ng/mL。  结论  本研究建立的UPLC-MS/MS检测体系测定肝移植受者全血中依维莫司血药浓度准确度高、样品处理方法简便、检测时间短,适用于肝移植术后受者血样本中依维莫司的药物浓度监测。

     

  • 图  1  依维莫司的标准曲线

    Figure  1.  Standard curve of everolimus

    表  1  依维莫司标准曲线质控的准确度结果

    Table  1.   Accuracy results of quality control of everolimus standard curve

    样本序号 待测物峰面积 理论浓度(ng/mL) 内标物峰面积 实测浓度(ng/mL) 准确度(%)
    STD1 5.13×103 1.00 1.01×105 1.00 100.0
    STD2 2.33×104 5.00 8.63×104 4.85 97.0
    STD3 5.05×104 10.00 9.21×104 9.75 97.5
    STD4 1.39×105 25.00 8.88×104 27.60 110.4
    STD5 2.62×105 50.00 9.09×104 50.80 101.6
    STD6 5.00×105 100.00 9.46×104 93.20 93.2
    L-QC 1.68×104 3.31 8.91×104 3.43 103.6
    M-QC 7.81×104 13.00 1.07×105 13.00 100.0
    H-QC 2.18×105 41.50 9.19×104 41.80 100.7
    下载: 导出CSV

    表  2  5例肝移植受者的依维莫司血药谷浓度检测结果

    Table  2.   Test results of everolimus trough blood concentration in 5 liver transplant recipients

    样本序号 待测物峰面积 内标物峰面积 实测浓度(ng/mL)
    Sample-1 4.06×104 9.11×104 8.05
    Sample-2 3.28×104 8.99×104 6.60
    Sample-3 2.43×104 1.17×105 3.77
    Sample-4 4.07×104 9.59×104 7.67
    Sample-5 4.96×104 9.66×104 9.27
    下载: 导出CSV
  • [1] KIM WR, LAKE JR, SMITH JM, et al. OPTN/SRTR 2013 annual data report: liver[J]. Am J Transplant, 2015, 15(Suppl 2): 1-28. DOI: 10.1111/ajt.13197.
    [2] 沈中阳, 谷川, 郑虹, 等. 临床肝脏移植20年回顾[J]. 中华危重病急救医学, 2019, 31(3): 269-280. DOI: 10.3760/cma.j.issn.2095-4352.2019.03.004.

    SHEN ZY, GU C, ZHENG H, et al. A twenty-year review of clinical liver transplantation[J]. Chin Crit Care Med, 2019, 31(3): 269-280. DOI: 10.3760/cma.j.issn.2095-4352.2019.03.004.
    [3] 赵东, 夏强. 肝移植相关领域的研究进展[J]. 国际消化病杂志, 2020, 40(2): 71-74. DOI: 10.3969/j.issn.1673-534X.2020.02.001.

    ZHAO D, XAI Q. Research progress in related fields of liver transplantation[J]. Int J Dig Dis, 2020, 40(2): 71-74. DOI: 10.3969/j.issn.1673-534X.2020.02.001.
    [4] LEMAITRE F, VETHE NT, D'AVOLIO A, et al. Measuring intracellular concentrations of calcineurin inhibitors: expert consensus from the International Association of Therapeutic Drug Monitoring and Clinical Toxicology Expert Panel[J]. Ther Drug Monit, 2020, 42(5): 665-670. DOI: 10.1097/FTD.0000000000000780.
    [5] HOŠKOVÁ L, MÁLEK I, KOPKAN L, et al. Pathophysiological mechanisms of calcineurin inhibitor-induced nephrotoxicity and arterial hypertension[J]. Physiol Res, 2017, 66(2): 167-180. DOI: 10.33549/physiolres.933332.
    [6] FAROUK SS, REIN JL. The many faces of calcineurin inhibitor toxicity-what the FK?[J]. Adv Chronic Kidney Dis, 2020, 27(1): 56-66. DOI: 10.1053/j.ackd.2019.08.006.
    [7] 中华医学会器官移植学分会. 器官移植免疫抑制剂临床应用技术规范(2019版)[J]. 器官移植, 2019, 10(3): 213-226. DOI: 10.3969/j.issn.1674-7445.2019.03.001.

    Branch of Organ Transplantation of Chinese Medical Association. Technical specifcation for clinical application of immunosuppressive agents in organ transplantation (2019 edition)[J]. Organ Transplant, 2019, 10(3): 213-226. DOI: 10.3969/j.issn.1674-7445.2019.03.001.
    [8] HASSKARL J. Everolimus[J]. Recent Results Cancer Res, 2018, 211: 101-123. DOI: 10.1007/978-3-319-91442-8_8.
    [9] ZOU Y, LI W, ZHOU J, et al. ERK inhibitor enhances everolimus efficacy through the attenuation of dNTP pools in renal cell carcinoma[J]. Mol Ther Nucleic Acids, 2019, 14: 550-561. DOI: 10.1016/j.omtn.2019.01.001.
    [10] CUCCHIARI D, RÍOS J, MOLINA-ANDUJAR A, et al. Combination of calcineurin and mTOR inhibitors in kidney transplantation: a propensity score analysis based on current clinical practice[J]. J Nephrol, 2020, 33(3): 601-610. DOI: 10.1007/s40620-019-00675-2.
    [11] GEDALY R, DE STEFANO F, TURCIOS L, et al. mTOR inhibitor everolimus in regulatory T cell expansion for clinical application in transplantation[J]. Transplantation, 2019, 103(4): 705-715. DOI: 10.1097/TP.0000000000002495.
    [12] BENDTSEN MAF, GRIMM D, BAUER J, et al. Hypertension caused by lenvatinib and everolimus in the treatment of metastatic renal cell carcinoma[J]. Int J Mol Sci, 2017, 18(8): 1736. DOI: 10.3390/ijms18081736.
    [13] TAN PS, MUTHIAH MD, KOH T, et al. Asian Liver Transplant Network clinical guidelines on immunosuppressionin liver transplantation[J]. Transplantation, 2019, 103(3): 470-480. DOI: 10.1097/TP.0000000000002532.
    [14] CILLO U, DE CARLIS L, DEL GAUDIO M, et al. Immunosuppressive regimens for adult liver transplant recipients in real-life practice: consensus recommendations from an Italian Working Group[J]. Hepatol Int, 2020, 14(6): 930-943. DOI: 10.1007/s12072-020-10091-5.
    [15] RUBÍN SUÁREZ A, BILBAO AGUIRRE I, FERNÁNDEZ-CASTROAGUDIN J, et al. Recommendations of everolimususe in liver transplant[J]. Gastroenterol Hepatol, 2017, 40(9): 629-640. DOI: 10.1016/j.gastrohep.2017.05.008.
    [16] LIN M, MITTAL S, SAHEBJAM F, et al. Everolimus with early withdrawal or reduced-dose calcineurin inhibitors improves renal function in liver transplant recipients: a systematic review and Meta-analysis[J]. Clin Transplant, 2017, 31(2): e12872. DOI: 10.1111/ctr.12872.
    [17] YU L, CHEN X, WANG L, et al. The sweet trap in tumors: aerobic glycolysis and potential targets for therapy[J]. Oncotarget, 2016, 7(25): 38908-38926. DOI: 10.18632/oncotarget.7676.
    [18] FERRÍN G, GUERRERO M, AMADO V, et al. Activation of mTOR signaling pathway in hepatocellular carcinoma[J]. Int J Mol Sci, 2020, 21(4): 1266. DOI: 10.3390/ijms21041266.
    [19] VAN GELDER T, FISCHER L, SHIHAB F, et al. Optimizing everolimus exposure when combined with calcineurin inhibitors in solid organ transplantation[J]. Transplant Rev (Orlando), 2017, 31(3): 151-157. DOI: 10.1016/j.trre.2017.02.007.
    [20] MABASA VH, ENSOM MH. The role of therapeutic monitoring of everolimus in solid organ transplantation[J]. Ther Drug Monit, 2005, 27(5): 666-676. DOI: 10.1097/01.ftd.0000175911.70172.2e.
    [21] FALKOWSKI S, WOILLARD JB. Therapeutic drug monitoring of everolimus in oncology: evidences and perspectives[J]. Ther Drug Monit, 2019, 41(5): 568-574. DOI: 10.1097/FTD.0000000000000628.
    [22] STROBBE G, PANNIER D, SAKJI I, et al. Advantages of everolimus therapeutic drug monitoring in oncology when drug-drug interaction is suspected: a case report[J]. J Oncol Pharm Pract, 2020, 26(7): 1743-1749. DOI: 10.1177/1078155220904761.
    [23] GOIRAND F, ROYER B, HULIN A, et al. Level of evidence for therapeutic drug monitoring of everolimus[J]. Therapie, 2011, 66(1): 57-61. DOI: 10.2515/therapie/2010025.
    [24] VENTURA-AGUIAR P, CAMPISTOL JM, DIEKMANN F. Safety of mTOR inhibitors in adult solid organ transplantation[J]. Expert Opin Drug Saf, 2016, 15(3): 303-319. DOI: 10.1517/14740338.2016.1132698.
    [25] ARENA C, BIZZOCA ME, CAPONIO VCA, et al. Everolimus therapy and side-effects: a systematic review and Meta-analysis[J]. Int J Oncol, 2021, 59(1): 54. DOI: 10.3892/ijo.2021.5234.
    [26] YEE ML, TAN HH. Use of everolimus in liver transplantation[J]. World J Hepatol, 2017, 9(23): 990-1000. DOI: 10.4254/wjh.v9.i23.990.
    [27] NOGUERAS LÓPEZ F, ABELLAN ALFOCEA P, ORTEGA SUAZO EJ, et al. Impact of everolimus-based immunosuppression on renal function in liver transplant recipients[J]. Transplant Proc, 2020, 52(2): 556-558. DOI: 10.1016/j.transproceed.2019.12.012.
    [28] KOVARIK JM, TEDESCO H, PASCUAL J, et al. Everolimus therapeutic concentration range defined from a prospective trial with reduced-exposure cyclosporine in de novo kidney transplantation[J]. Ther Drug Monit, 2004, 26(5): 499-505. DOI: 10.1097/00007691-200410000-00007.
    [29] SHIPKOVA M, HESSELINK DA, HOLT DW, et al. Therapeutic drug monitoring of everolimus: a consensus report[J]. Ther Drug Monit, 2016, 38(2): 143-169. DOI: 10.1097/FTD.0000000000000260.
    [30] KLAWITTER J, NASHAN B, CHRISTIANS U. Everolimus and sirolimus in transplantation-related but different[J]. Expert Opin Drug Saf, 2015, 14(7): 1055-1070. DOI: 10.1517/14740338.2015.1040388.
    [31] JASIAK NM, PARK JM. Immunosuppression in solid-organ transplantation: essentials and practical tips[J]. Crit Care Nurs Q, 2016, 39(3): 227-240. DOI: 10.1097/CNQ.0000000000000117.
    [32] SATOH S, MIURA M. Therapeutic drug monitoring of immunosuppressive drugs[J]. Rinsho Byori, 2016, 64(12): 1381-1389.
    [33] MILLÁN O, WIELAND E, MARQUET P, et al. Pharmacodynamic monitoring of mTOR inhibitors[J]. Ther Drug Monit, 2019, 41(2): 160-167. DOI: 10.1097/FTD.0000000000000616.
    [34] ZHANG M, TAJIMA S, SHIGEMATSU T, et al. Development and validation of an LC-MS/MS Method to simultaneously measure tacrolimus and everolimus concentrations in kidney allograft biopsies after kidney transplantation[J]. Ther Drug Monit, 2021, DOI: 10.1097/FTD.0000000000000912[Epubaheadofprint].
    [35] ANTUNES NJ, KIPPER K, COUCHMAN L, et al. Simultaneous quantification of cyclosporin, tacrolimus, sirolimus and everolimus in whole blood by UHPLC-MS/MS for therapeutic drug monitoring[J]. Biomed Chromatogr, 2021, 35(6): e5071. DOI: 10.1002/bmc.5071.
    [36] 洪顺福, 吴国兰, 郑运亮, 等. 在线固相萃取LC-MS/MS法测定健康人体全血中依维莫司浓度及在药代动力学研究中的应用[J]. 药物分析杂志, 2016, 36(10): 1778-1784. DOI: 10.16155/j.0254-1793.2016.10.11.

    HONG FS, WU GL, ZHENG YL, et al. Determination of everolimus in whole blood by online SPE LC-MS/MS system and its application in pharmacokinetic study[J]. Chin J Pharm Anal, 2016, 36(10): 1778-1784. DOI: 10.16155/j.0254-1793.2016.10.11.
    [37] 翟晓慧, 刘晓雪, 陆佳倩, 等. LC-MS/MS法同时分析血液中几种免疫抑制剂浓度[J]. 中国医院药学杂志, 2019, 39(8): 774-780. DOI: 10.13286/j.cnki.chinhosppharmacyj.2019.08.02.

    ZHAI XH, LIU XX, LU JQ, et al. Establishment of LC-MS/MS assay for the determination of blood immunosuppressive agents level simultaneously[J]. Chin J Hosp Pharm, 2019, 39(8): 774-780. DOI: 10.13286/j.cnki.chinhosppharmacyj.2019.08.02.
  • 加载中
图(2) / 表(2)
计量
  • 文章访问数:  464
  • HTML全文浏览量:  134
  • PDF下载量:  83
  • 被引次数: 0
出版历程
  • 收稿日期:  2021-08-26
  • 网络出版日期:  2021-09-15
  • 刊出日期:  2021-09-15

目录

    /

    返回文章
    返回