Trace analysis of SN-38 in human plasma by high-performance liquid chromatography

A reversed-phase high-performance liquid chromatographic method with fluorescence detection was developed and validated for the quantitation of SN-38, the active metabolite of irinotecan (CPT-11), a new anticancer drug. This method uses solid-phase extraction with a C₁₈ column for sample clean-up and concentration following acidification of human plasma with two volumes of 0.1 M HCl. Using blank plasma spiked with SN-38, we found the assay to be linear over the concentration range of 10–500 pM (3.9–195 pg/ml) with acceptable total and within-day imprecision. The recovery of SN-38 ranged from 48.3% (10 pM) to 91.5% (500 pM) whereas that of the internal standard, 20-(S)-camptothecin, was 96.9% (500 pM). This method represents a sizeable increase in sensitivity over other published methods and is shown to be suitable for the measurement of ‘trough' concentrations of SN-38 during the treatment of patients with a weekly regimen of irinotecan.     

Separation methods for camptothecin and related compounds

This paper reviews working procedures for the analytical determination of camptothecin and analogues. We give an overview of aspects such as the chemistry, structure–activity relationships, stability and mechanism of action of these antitumor compounds. The main body of the review describes separation techniques. Sample treatment and factors influencing high-performance liquid chromatography development are delineated. Published high-performance liquid chromatographic methods are summarized to demonstrate the variability and versatility of separation techniques and a critical evaluation of separation efficiency, detection sensitivity and specificity of these methods is reported.     

替吉奥联合伊立替康治疗晚期结肠癌的临床研究

目的:比较替吉奥联合伊立替康与替吉奥单药治疗晚期结肠癌的临床疗效。方法:选取2011年3月~2012年3月我院收治的晚期结肠癌患者90例,按照随机数字表法将患者分为研究组和对照组,每组45例,研究组给予替吉奥联合伊立替康,对照组给予替吉奥单药治疗,比较两组近期疗效、远期疗效和不良反应。结果:研究组的总有效率51.1%(23/45)显著高于对照组的26.7%(12/45),两组比较差异具有统计学意义(X2=13.281,P=0.017);研究组中位进展时间和中位生存时间均显著长于对照组,两组比较差异具有统计学意义(t=9.029,10.412,P=0.021,0.019);两组中性粒细胞减少和腹泻比较差异具有统计学意义(X2=11.517,12.041;P=0.023,0.019)。结论:替吉奥联合伊立替康治疗晚期结肠癌较单用替吉奥具有更好的治疗效果,值得临床推广应用。     

地衣芽胞杆菌活菌制剂联合庆大霉素预防伊立替康所致腹泻

目的观察地衣芽胞杆菌活菌制剂（整肠生胶囊）联合庆大霉素预防伊立替康（irinoteean,CPT-11）所致腹泻的疗效。方法31例接受含伊立替康方案化疗的晚期癌症患者,采用自身随机交叉对照的方法,分别于2个治疗周期接受化疗（对照组）、化疗＋整肠生胶囊＋庆大霉素（预防组）,比较腹泻及其他不良反应的差异。结果31例患者在2个治疗周期中,预防组较对照组腹泻发生率有显著降低（腹泻发生率分别为48．4％VS61．3％,P〈0．05;3／4级腹泻发生率分别为9．7％vs16．1％,P〈0．05）,且减少合并用药;同时骨髓抑制、食欲减退、恶心呕吐的发生率也有所降低（P〈0．05）,而2组便秘、肝肾功能异常差异无显著性。结论地衣芽胞杆菌活菌制剂联合庆大霉素能有效预防伊立替康所致腹泻。     

Simple and rapid determination of irinotecan and its metabolite SN-38 in plasma by high-performance liquid-chromatography: application to clinical pharmacokinetic studies

Irinotecan (CPT-11) is an anticancer agent widely employed in the treatment of colorectal carcinoma. A simple, rapid and sensitive high-performance liquid chromatographic method for the simultaneous determination of CPT-11 and its metabolite SN-38 in plasma, and their preliminary clinical pharmacokinetics are described. Both deproteinisation of plasma specimens (100 μl) and addition of the internal standard, camptothecin (CPT), are achieved by incorporating to samples 100 μl of a solution of CPT (1 μg/ml) in acetonitrile–1 mM orthophosphoric acid (90:10); 200 μl of this acidified acetonitrile solution, drug-free, is also added to accomplish complete deproteinisation: this procedure reduces sample preparation time to a minimum. After deproteinisation, samples are treated with potassium dihydrogenphosphate (0.1 M) and injected into a Nucleosil C₁₈ (5 μm, 250×4.0 mm) column. Mobile phase consists of potassium dihydrogenphosphate (0.1 M)–acetonitrile (67:33), at a flow-rate of 1 ml/min. CPT-11, SN-38 and CPT are detected by fluorescence with excitation wavelength set at 228 nm and emission wavelengths of CPT-11, SN-38 and CPT fixed, respectively, at 450, 543 and 433 nm. The limits of quantitation for CPT-11 and SN-38 are 1.0 and 0.5 ng/ml, respectively. This method shows good precision: the within day relative standard deviation (RSD) for CPT-11 (1–10 000 ng/ml) is 5.17% (range 2.15–8.27%) and for SN-38 (0.5–400 ng/ml) is 4.33% (1.32–7.78%); the between-day RSDs for CPT-11 and SN-38, in the previously described ranges, are 6.82% (5.03–10.8%) and 4.94% (2.09–9.30%), respectively. Using this assay, plasma pharmacokinetics of CPT-11, SN-38 and its glucuronidated form, SN-38G, have been determined in one patient receiving 200 mg/m² of CPT-11 as a 90 min intravenous infusion. The peak plasma concentration of CPT-11 at the end of the infusion is 3800 ng/ml. Plasma decay is biphasic with a terminal half-life of 11.6 h. The volume of distribution at steady state (V_ss) is 203 l/m², and the total body clearance (Cl) is 14.8 l/h·m². The maximum concentrations of SN-38 and SN-38G reach 28.9 and 151 ng/ml, respectively.     

伊立替康联合阿帕替尼治疗术后转移性胃癌患者临床疗效和安全性分析

摘要 目的：探讨伊立替康联合阿帕替尼治疗术后转移性胃癌患者临床疗效和安全性。方法：选取我院2017年5月-2018年10月期间收治的术后一线化疗失败转移性胃癌患者105例,根据随机数字表法分为研究组（53例）和对照组（52例）,对照组患者给予伊立替康静脉滴注治疗,研究组在此基础上使用阿帕替尼进行联合治疗,4周为一个周期,连续治疗两个周期。比较两组患者疾病控制率、生存情况,并对治疗前后两组患者肿瘤标志物[癌胚抗原（CEA）、糖类抗原125（CA125）和糖类抗原199（CA199）]水平、基质金属蛋白酶-9（MMP-9）和血管内皮生长因子（VEGF）水平进行比较,观察治疗过程中两组患者不良反应发生情况。结果：治疗后,研究组疾病控制率、中位生存时间和中位进展时间均优于对照组（P<0.05）。治疗后,两组肿瘤标志物、MMP-9和VEGF水平均降低,且研究组低于对照组（P<0.05）。研究组不良反应发生率低于对照组（P<0.05）。结论：伊立替康联合阿帕替尼治疗术后转移性胃癌患者临床疗效确切,可延长患者生存时间,延缓疾病进展,且安全性较好,其作用机制可能与降低肿瘤标志物及MMP-9、VEGF水平有关。     

伊立替康联合替莫唑胺对儿童难治性实体瘤近期疗效的研究

目的：研究伊立替康（Irinotecan,CPT-11）联合替莫唑胺（Temozolomide,TMZ）对儿童难治性实体瘤的近期疗效;方法：选取了11名难治性儿童实体瘤患者并争取了家属同意,分别给予了伊立替康：60mg／m2／d,ivgtt,连用5天;替莫唑胺：75mg／m2／d,连用5天,在伊立替康前一小时口服;用药期间同时给予思密达或者头孢菌素类抗生素预防药物相关性腹泻,并定期复查血象以及肝肾功;28天一疗程,每疗程行复查CT或MRI进行疗效评定和药物安全性评价。结果：11例患者经过2-6个疗程的治疗：疾病控制率（DCR）达到54％;明显延长了患者的无疾病进展生存期,中位无疾病进展生存期3．3个月。用药期间不良事件主要包括0-Ⅲ级的药物相关性腹泻、呕吐、血小板减少、贫血等;结论：伊立替康联合替莫唑胺方案能明显延长儿童难治实体瘤患者未进展存活时间,且耐受性好、安全性高,可以作为儿童晚期实体瘤患者的首选治疗方案。     

伊立替康联合替莫唑胺对儿童难治性实体瘤近期疗效的研究

目的:研究伊立替康(Irinotecan,CPT-11)联合替莫唑胺(Temozolomide,TMZ)对儿童难治性实体瘤的近期疗效;方法:选取了11名难治性儿童实体瘤患者并争取了家属同意,分别给予了伊立替康:60mg/m~2/d,ivgtt,连用5天;替莫唑胺:75mg/m~2/d,连用5天,在伊立替康前一小时口服;用药期间同时给予思密达或者头孢菌素类抗生素预防药物相关性腹泻,并定期复查血象以及肝肾功;28天一疗程,每疗程行复查CT或MRI进行疗效评定和药物安全性评价。结果:11例患者经过2-6个疗程的治疗;疾病控制率(DCR)达到54%;明显延长了患者的无疾病进展生存期,中位无疾病进展生存期3.3个月。用药期间不良事件主要包括0-Ⅲ级的药物相关性腹泻、呕吐、血小板减少、贫血等;结论:伊立替康联合替莫唑胺方案能明显延长儿童难治实体瘤患者未进展存活时间,且耐受性好、安全性高,可以作为儿童晚期实体瘤患者的首选治疗方案。     

Sex and Dosing-Time Dependencies in Irinotecan-Induced Circadian Disruption

Circadian disruption accelerates malignant growth; thus, it should be avoided in anticancer therapy. The circadian disruptive effects of irinotecan, a topoisomerase I inhibitor, was investigated according to dosing time and sex. In previous work, irinotecan achieved best tolerability following dosing at zeitgeber time (ZT) 11 in male and ZT15 in female mice, whereas worst toxicity corresponded to treatment at ZT23 and ZT3 in male and female mice, respectively. Here, irinotecan (50?mg/kg intravenous [i.v.]) was delivered at the sex-specific optimal or worst circadian timing in male and female B6D2F1 mice. Circadian disruption was assessed with rest-activity, body temperature, plasma corticosterone, and liver mRNA expressions of clock genes Rev-erbα, Per2, and Bmal1. Baseline circadian rhythms in rest-activity, body temperature, and plasma corticosterone were more prominent in females as compared to males. Severe circadian disruption was documented for all physiology and molecular clock endpoints in female mice treated at the ZT of worst tolerability. Conversely, irinotecan administration at the ZT of best tolerability induced slight alteration of circadian physiology and clock-gene expression patterns in female mice. In male mice, irinotecan produced moderate alterations of circadian physiology and clock-gene expression patterns, irrespective of treatment ZT. However, the average expression of Rev-erbα, Per2, and Bmal1 were down-regulated 2- to 10-fold with irinotecan at the worst ZT, while being minimally or unaffected at the best ZT, irrespective of sex. Corticosterone secretion increased acutely within 2?h with a sex-specific response pattern, resulting in a ZT-dependent phase-advance or -delay in both sex. The mRNA expressions of irinotecan clock-controlled metabolism genes Ce2, Ugt1a1, and Top1 were unchanged or down-regulated according to irinotecan timing and sex. This study shows that the circadian timing system represents an important toxicity target of irinotecan in female mice, where circadian disruption persists after wrongly timed treatment. As a result, the mechanisms underling cancer chronotherapeutics are expectedly more susceptible to disruption in females as compared to males. Thus, the optimal circadian timing of chemotherapy requires precise determination according to sex, and should involve the noninvasive monitoring of circadian biomarkers. (Author correspondence: francis.levi@inserm.fr)     

High-performance liquid chromatographic assay with fluorescence detection for the simultaneous measurement of carboxylate and lactone forms of irinotecan and three metabolites in human plasma

Irinotecan (CPT-11), a camptothecin analog, is metabolized to SN-38, an active topoisomerase I inhibitor, and inactive metabolites, including APC and SN-38 glucuronide (SN-38G). A high-performance liquid chromatographic assay method to simultaneously measure the lactone and carboxylate forms of CPT-11, SN-38, SN-38G, and APC in human plasma was developed. Chromatography was accomplished with a reversed-phase C(8) column and fluorescence detection. A gradient mobile phase system was used. The buffer for mobile phase A consisted of 0.75 M ammonium acetate, 5 mM tetrabutylammonium phosphate (pH 6.0), and acetonitrile (86:14, v/v). The buffer for mobile phase B was identical to mobile phase A with the exception of the concentration (50:50, v/v). Precipitation of plasma proteins was performed with cold methanol. The linear range of detection of the lactone and carboxylate forms of SN-38, SN-38G, and APC was 2-25 ng/ml, and 5-300 ng/ml for CPT-11. The limit of quantitation for the analytes ranged from 0.5 to 5 ng/ml. Analysis of patients' plasma samples obtained before and after CPT-11 administration showed that the assay is suitable for measuring lactone and carboxylate forms of CPT-11, SN-38, SN-38G, and APC in clinical studies.