西罗莫司治疗肾移植后排斥反应的疗效、不良反应与其血药浓度的相关性分析

目的:观察西罗莫司治疗肾移植后排斥反应时,其疗效、不良反应与血药浓度的相关性。方法:选择52名肾移植术后应用西罗莫司进行免疫抑制治疗的患者为研究对象,其中男性43人,女性9人。采用高效液相色谱(HPLC)法测定西罗莫司血药浓度,同时测定患者的肌酐清除率、肝功能指标、血常规等。结果:共收集西罗莫司血药浓度数据112份。西罗莫司的血药浓度与给药剂量/体重比呈正相关,与体重呈负相关,女性患者血药浓度明显高于男性患者(P<0.05),未发现西罗莫司血药浓度与年龄有明显相关性;西罗莫司的主要不良反应表现为肝功能损伤,将血药浓度分为<8ng·mL-1和>8ng·mL-1组,不同血药浓度组的肝功能差异有统计学意义(P<0.05);西罗莫司的临床疗效确切,将西罗莫司稳态谷浓度分成<4ng·mL-1、4～8ng·mL-1和>8ng·mL-1组,不同血药浓度组其肌酐清除率和肾功能恢复情况差异有统计学意义(P<0.05)。结论:西罗莫司副作用主要表现为肝功能异常,其血药浓度与临床疗效和不良反应均有相关性,西罗莫司稳态谷浓度宜控制在4～8ng·mL-1范围内。     

西罗莫司在全血中的稳定性

目的:考察西罗莫司在全血中的稳定性.方法:配制浓度分别为2.5,7.5,22.5μg·L-1的西罗莫司血样,在3种温度下(25℃,4℃,-20℃)避光储存0～8 d和在-40℃存放后经历3个冻融周期,考察西罗莫司在全血中的稳定性.结果:在3种温度下储存8d和经历3个冻融周期后,西罗莫司血药浓度无显著变化.结论:西罗莫司血样可在冰冻或室温条件保存8d,经历3个冻融周期后西罗莫司在全血中仍能保持稳定.     

新型大环内酯类免疫抑制剂-西罗莫司

目的:综述新型大环内酯类免疫抑制剂西罗莫司的相关药学资料,为进一步了解西罗莫司提供参考.方法:通过查阅相关文献,并在此基础上对西罗莫司的化学结构、理化性质、免疫抑制作用机理、药代动力学、药效学、临床疗效、用法用量和血药浓度监测以及不良反应等方面进行综述.结论:西罗莫司为毒性较小的有潜力的新型强效免疫抑制剂,为移植者提供了一种新的治疗替代方法.     

药物相互作用致三酰甘油升高1例

目的 报道并分析西罗莫司致高三酰甘油血症1例,提高临床重视西罗莫司对血脂代谢的影响。方法 1例肝移植病人使用西罗莫司后引起高三酰甘油血症的关联性评价及查阅文献印证。结果 住院第7日查血生化示:总胆固醇2.80 mmol/L,三酰甘油7.74 mmol/L;西罗莫司血药浓度为20 ng/mL;立即停用西罗莫司。住院第14日西罗莫司血药浓度为7 ng/mL,住院第21日复查血生化示:总胆固醇2.75 mmol/L,三酰甘油4.77 mmol/L。结论 该肝移植术后病人并发高三酰甘油血症很可能是西罗莫司引起。在应用西罗莫司时,应严密监测病人血脂情况,一旦出现三酰甘油值增加,应及时调整西罗莫司给药方案以期维持其血药浓度在基线范围内,从而降低其对血脂代谢的影响。     

西罗莫司不同血药浓度对肾移植患者肝肾功能的影响

目的探讨西罗莫司不同血药浓度对肾移植患者肝肾功能的影响。方法 45例肾移植术后服用西罗莫司治疗的患者,采用高效液相色谱法(HPLC法)检测西罗莫司的血液浓度,比较分析不同血药浓度下肝肾指标的变化情况。结果西罗莫司血药浓度≤8 ng/ml组患者的天门冬氨酸肌转酶(AST)、丙氨酸氨基转移酶(ALT)、直接胆红素(DBIL)分别为(65.26±12.68)U/L、(70.19±13.66)U/L、(11.05±9.64)μmol/L,显著低于>8 ng/ml组,差异均具有统计学意义(P<0.05);西罗莫司血药浓度为4~8 ng/ml组患者的内生肌酐清除率(Ccr)为(82.64±17.32)ml/(min·70 kg),显著高于<4 ng/ml组、>8 ng/ml组,差异具有统计学意义(P<0.05)。结论肾移植术后采用西罗莫司行免疫抑制治疗时,血药浓度控制在4~8 ng/ml最佳。     

中国肾移植患者中不同的CYP3A5*3基因型与西罗莫司血药浓度的关系

回顾性收集了服用西罗莫司的80例肾移植患者病例,建立了四引物法检测其CYP3A5基因型,并以聚合酶链反应-限制性片段长度多态性(PCR-RFLP)法进行对照.采用微粒子化学发光免疫法测定患者西罗莫司的谷浓度.80位患者的CYP3A5*3基因频率为79%.*1/*1基因型患者有3人(3.75%),*1/*3基因型有28人(35%),*3/*3基因型有49人(61.25%).三种基因型患者西罗莫司平均血药谷浓度分别为(4.17±0.90)、(5.96±2.43)和(6.39±2.86)ng/ml.表明不同CYP3A5基因型患者西罗莫司血药浓度差异显著,含有CYP3A5*1等位基因患者的西罗莫司血药浓度明显低于*3/*3组(P<0.05).     

游动放线菌ATCC 12065生物转化强效免疫抑制剂西罗莫司

游动放线菌ATCC 12065生物转化强效免疫抑制剂西罗莫司后产生与西罗莫司不同的4个未知组分(未知组分1,2,3,4),HPLC保留时间分别为25.41min,34.13min,48.46min和52.24 min(西罗莫司的保留时间为26.62min)。经UV、HPLC及LC-MS分析表明,转化产物为西罗莫司衍生物。未知组分1和未知组分2的分子量为916,比西罗莫司的分子量多2;未知组分3和未知组分4的分子量900,比西罗莫司的分子量少14。     

西罗莫司血药浓度测定方法的建立及在肝移植患者中的应用

目的建立高效液相色谱法测定西罗莫司血药浓度,测定该药在肝移植患者中的药代动力学参数。方法以沉淀-萃取法处理全血样品,采用Hypersil ODS C18(250 mm×4.6 mm,5μm)柱,柱温为50℃,乙腈-甲醇-水(7.5:62.5:30)为流动相,流速1.2 mL/min,以32-去甲基雷帕霉素为内标,在276 nm波长处,检测西罗莫司血药浓度,并应用此方法测定5例肝移植患者的西罗莫司血药浓度。结果全血中西罗莫司浓度在2.0～50.0ng/mL范围内线性良好(r=0.999 7),平均相对回收率为99.1%(n=5)。5例肝移植患者的Cmax为(12.66±4.1)ng/mL,t1/2为(19.2±11.2)h,tmax为(1.8±0.3)h。结论本方法快速、简便、准确,灵敏度高,重现性好,可用于肝移植患者的西罗莫司临床药物监测。     

肾移植受者应用西罗莫司治疗窗的临床研究

目的:探讨西罗莫司应用于国内肾移植受者的治疗窗范围。方法:采用多中心、开放性临床研究,来自国内4家移植中心的首次肾移植病人共100例。免疫抑制方案为西罗莫司联合环孢素和皮质激素的三联疗法。移植后48h内开始服用西罗莫司,首次负荷剂量为6mg·d-1,维持剂量为2mg·d-1,采用高效液相色谱法测定西罗莫司浓度。结果:100例病人西罗莫司总的全血谷浓度为(6.6±s2.8)μg·L-1,10及90百分位数浓度分别为3.2μg·L-1和10.26μg·L-1。肾移植后6mo内急性排斥发生率为10%(8/84),此8例病人急性排斥时的西罗莫司浓度明显低于非排斥时浓度(P<0.01)。主要不良反应为肝功能损害和高脂血症,三酰甘油浓度与西罗莫司浓度相关(r=0.276,P<0.01)。结论:西罗莫司浓度维持在4～8μg·L-1范围内较为合适,定期监测血药浓度,合理调整用量,可增加西罗莫司应用的有效性及安全性。     

HPLC法检测肾移植病人全血中西罗莫司的药物浓度

目的 :建立测定全血中西罗莫司含量的方法。 方法 :采用HPLC法 ,色谱柱为 :YMC PackODS A(15 0mm× 4 .6mm ,5 μm) ,预柱为AlltimaC18(7.5mm× 4 .6mm ,5 μm)。流动相为乙腈∶四氢呋喃∶水 =5 5∶5∶4 0 ;检测波长为 2 78nm ,流速为 1.5ml/min ,柱温 :5 0℃。 32 去甲氧基西罗莫司为内标。 结果 :西罗莫司及内标 32 去甲氧基西罗莫司的保留时间分别为 10 .1、12 .1min ,全血定量线性范围 :2 .4 9～ 76 .36ng/ml,最低检测浓度为 1.98ng/ml,方法回收率为 99.93%～ 10 5 .90 % ,日内、日间RSD <7.4 0 %。 结论 :本方法准确、可靠 ,适用于临床对西罗莫司的血药浓度监测。     

Measurement of whole blood sirolimus by an HPLC assay using solid-phase extraction and UV detection

The authors developed an HPLC assay for determining blood sirolimus concentration using a relatively simple solid-phase extraction and UV detection. The retention times of sirolimus and the internal standard, 32-desmethoxyrapamycin, are 8.7 and 9.3 minutes, respectively. The assay possesses linearity up to 200 ng/mL, sensitivity to 2.0 ng/mL, and day-to-day reproducibility of 8.8, 9.8, 6.1, and 6.4% at sirolimus concentrations of 6, 10, 20, and 30 ng/mL, respectively. A patient correlation study using this HPLC method and an established LC/MS/MS assay revealed a slope of 0.982 and intercept of -0.021 ng/mL and a correlation coefficient of 0.99 (n = 37). Of the 31 different drugs tested none interfered with the measurement of the drug of interest, and a recovery study gave an overall mean recovery of 101.8%. The authors conclude that the method described here is suited for the therapeutic monitoring of blood sirolimus concentration.     

西罗莫司治疗儿童蓝色橡皮疱痣综合征5 例并文献复习

目的:探讨儿童蓝色橡皮疱痣综合征(BRBNS)的临床特征及应用西罗莫司治疗的临床疗效。方法:收集我院2019 年1 月 至2022 年4 月收治的5 例儿童BRBNS 的临床资料。通过PubMed、中国知网、万方数据库检索建库至2022 年4 月收录的文献, 分析BRBNS 的临床特征、诊断、治疗及预后情况。结果:5 例BRBNS 均有皮肤静脉畸形及胃肠道病变;伴随重度小细胞低色素 性贫血,表现为面色苍白、生长缓慢;内镜对BRBNS 诊断率达100%,病变分布在胃、十二指肠、空肠、回肠、结肠等。治疗上需多 次输血、口服普萘洛尔、局部介入或硬化处理病变。多数3~6 个月复发,改用西罗莫司后症状好转。西罗莫司起始日剂量为 0. 5~1. 0 mg/ m2 ,根据血药浓度调整剂量增加至每日2. 0 mg/ m2 ,达到目标浓度5. 0~10. 0 ng/ mL,维持血红蛋白至稳定水平。 结论:儿童BRBNS 临床症状较重且易反复,西罗莫司可作为安全有效的治疗药物。     

固相萃取-HPLC法测定西罗莫司的血药浓度及其临床应用研究

目的:建立以固相萃取-高效液相色谱法测定西罗莫司血药浓度的方法。方法:采用Waters OASIS固相萃取柱处理血样后进样分析,其中色谱柱为Supelcosil LC-18DB,流动相为甲醇-水(75∶25),流速1.5mL·min-1,检测波长276nm,柱温50℃。结果:西罗莫司血药浓度在1.25~70ng·mL-1范围内与峰面积比线性关系良好(r=0.999 7),最低检测浓度为0.5ng·mL-1;平均方法回收率为(101.7±2.7)%,萃取回收率≥82.8%,日内RSD≤4.0%,日间RSD≤5.4%。结论:本方法简便快速、灵敏准确,可用于西罗莫司血药浓度的快速监测。     

Evaluation of QMS everolimus assay using Hitachi 917 Analyzer: comparison with liquid chromatography/mass spectrometry

Everolimus is an immunosuppressant requiring routine monitoring in whole blood. We evaluated the analytical performance of a new immunoassay for everolimus, Quantitative Microsphere System (QMS) everolimus (Thermo Fisher Scientific), which is CE marked and currently under review by Food and Drug Administration of the United States by comparing results with values obtained by using liquid chromatography/mass spectrometry. The total coefficient of variations (CVs) were 8.3% for low control (mean: 3.8 ng/mL), 6.1% for the medium control (mean: 8.0 ng/mL), and 7.5% for the high control (mean: 14.4 ng/mL) (n = 80 for each control, run over 20 nonconsecutive days). The respective total CVs for patients' pool were 13.3% (mean: 4.0 ng/mL), 7.5% (mean: 8.2 ng/mL), and 8.7% (mean: 11.7 ng/mL) (n = 80 for each patient pool). The assay was linear from a whole-blood everolimus level between 1.5 and 20 ng/mL, and the limit of quantitation was 1.3 ng/mL. Comparison was carried out using 90 renal transplant patient samples, and we observed the following Passing and Bablok linear regression plot: y = 1.11, slope = -0.005 (R = 0.92). This assay was not affected by commonly used 70 drugs, but sirolimus, a drug structurally similar to everolimus, showed 46% cross-reactivity. We conclude that QMS everolimus immunoassay has adequate sensitivity and specificity for the determination of whole-blood everolimus and can be used for routine therapeutic drug monitoring.     

β-氧化代谢基因多态性与癫痫患儿丙戊酸肝毒性的相关性

目的:分析ACSM2A、CPT1A基因多态性与癫痫患儿丙戊酸(VPA)血药浓度及肝毒性之间的相关性.方法:采用聚合酶链式反应(PCR)和直接测序法检测110例服用丙戊酸单药治疗癫痫患儿外周血ACSM2A rs1133607、CPT1A rs597316基因多态性,并采用液相色谱-质谱联用技术(LC-MS)测定血清VPA...     

Sirolimus steady-state trough concentrations are not affected by bolus methylprednisolone therapy in renal allograft recipients

AIMS: To determine whether bolus doses of methylprednisolone affect the steady-state trough concentrations of sirolimus. METHODS: Fourteen renal transplant recipients received concentration-controlled sirolimus therapy in combination with azathioprine and steroids (n=8) or mycophenolate mofetil and steroids (n=6). Bolus doses of methylprednisolone (mean total dose over 1-5 days, 1694 mg; range, 500-3000 mg) were given for the treatment of acute rejection. For each patient, the sirolimus dose (mean, 24.1 mg; range, 3.3-52.5 mg) was the same before and during methylprednisolone therapy. RESULTS: Mean sirolimus whole blood trough concentrations before and after treatment with methylprednisolone were 28.8 ng ml-1 (range, 13.9-45.3 ng ml-1), and 28.5 ng ml-1 (range, 13.0-47.9 ng ml-1), respectively (P=0.85; 95% confidence interval on the difference -3.3, 4.0 ng ml-1). CONCLUSIONS: Bolus methylprednisolone treatment does not affect steady-state sirolimus trough concentrations.     

Determination of sirolimus blood concentration using high-performance liquid chromatography with ultraviolet detection

BACKGROUND: Different HPLC methods have been developed and used to determined sirolimus blood concentrations. These methods show different performance characteristics, mostly related to peak interference, recovery, assay sensitivity, and turnaround times. OBJECTIVE: We adapted, improved, and validated an HPLC method with UV detection for measurement of sirolimus in whole blood clinical samples. METHODS: The standards, quality controls, or patient samples (0.25 or 0.5 mL) and internal standard (desmethoxysirolimus) were extracted with 1-chlorobutane. After evaporation, the extract was reconstituted in a 70% acetonitrile/water mixture and analyzed onto a reverse-phase C18 column at 50 degrees C under a flow rate of 1.0 mL/min in the HPLC system. Ultraviolet detection was performed at 278 nm, with sensitivity setting of 0.010 AUFS. Identification of peaks of interest was by retention time; quantification of sirolimus was based on a peak area ratio. RESULTS: Analytic recovery ranging from 96 to 120% (CV = 3.7 to 16.8%; bias = -4.2 to 16.7%) was observed throughout the assay's linear range (2.5-150.0 ng/mL). The lower limit of quantification for both sample volumes (0.25 or 0.5 mL) was 2.5 ng/mL (CV = 12 and 15%, bias = -1.2 and 4%, respectively). The intra- and interassay imprecision ranged from 6.2 to 14.4% and from 9.1 to 18.6%, with bias ranging from 1.3 to 12.9% and -1.8% to 7.1, for quality control levels of 3, 10, and 20 ng/mL. Whole blood and extracted samples are stable at room temperature and at 4 and -20 degrees C for 1 week and 3 days, respectively. Chromatograms showed good separation free of interfering peaks. A set of 45 samples can be extracted in 2 h, allowing results within 24 h. CONCLUSION: This HPLC-UV method shows good and reproducible performance, satisfying all requirements of an assay designated to be applied in therapeutic drug monitoring strategies after organ transplantation.     

Development and validation of a high-throughput assay for quantification of the proliferation inhibitor ABT-578 using LC/LC-MS/MS in blood and tissue samples

We report here a specific, automated LC/LC-MS/MS assay for the quantification of ABT-578 in human and rabbit blood and rabbit tissues for drug-eluting stent development. After protein precipitation, samples were injected into the HPLC system and extracted online using a high flow of 5 mL/min. The extracts were then backflushed onto the analytic column. The [M+Na] of ABT-578 (m/z 988.6-->369.4) and its internal standard sirolimus (m/z 936.5-->409.3) were monitored. Extraction and analysis took 4 minutes. The assay was validated following the US Food & Drug Administration guidelines. Linearity was 0.025-25 ng/mL for most matrices. In human blood, interday accuracies were 81.8% (at 0.025 ng/mL), 91.0% (1 ng/mL), and 99.5% (50 ng/mL), and interday precisions were 10.7% (0.025 ng/mL), 3.0% (1 ng/mL), and 1.8% (50 ng/mL).