Stephen J. Lane

Efficacy and safety of casopitant mesylate, a neurokinin 1 (NK1)-receptor antagonist, in prevention of chemotherapy-induced nausea and vomiting in patients receiving cisplatin-based highly emetogenic chemotherapy: a randomised, double-blind, placebo-controlled trial

BACKGROUND Chemotherapy-induced nausea and vomiting (CINV) remains a clinical management problem after treatment with highly emetogenic chemotherapy (HEC). We therefore designed and carried out a multicentre, randomised, double-blind, placebo-controlled trial to assess whether a three-drug antiemetic regimen of ondansetron, dexamethasone, and the neurokinin-1-receptor antagonist casopitant mesylate was able to prevent acute and delayed CINV events in patients naive to chemotherapy with a malignant solid tumour who were scheduled to receive cisplatin-based HEC regimens. METHODS The study was done between Nov 6, 2006, and Oct 9, 2007, in 77 participating centres in 22 countries. All 810 patients enrolled in the trial received dexamethasone and ondansetron. Patients were randomly assigned to also receive placebo (n=269), single oral dose of casopitant mesylate (150 mg oral, n=271), or 3-day intravenous plus oral casopitant mesylate (90 mg intravenous on day 1 plus 50 mg oral on days 2 and 3, n=270). Randomisation was done using a central telephone system at the study level, because some centres were expected to recruit only a few patients during the study period. The primary endpoint was the proportion of patients achieving complete response (no vomiting, retching, or use of rescue medications) in the first 120 h after receiving HEC. Efficacy analysis was done on the modified intention-to-treat population (n=800), which included all patients who received placebo or study drug and HEC (n=265 control, n=266 single-dose oral casopitant mesylate, n=269 3-day intravenous and oral casopitant mesylate). Safety was reported in 802 patients who received either placebo or study medication. This study is registered with ClinicalTrials.gov, NCT00431236. FINDINGS Significantly more patients in each casopitant group achieved complete response in cycle 1 of HEC treatment than did those in the control group (175 [66%] patients in the control group, 228 [86%] in the single-dose oral casopitant mesylate group [p<0.0001 vs control], and 214 [80%] in the 3-day intravenous plus oral casopitant mesylate group (p=0.0004 vs control]). This improvement was sustained over multiple cycles of HEC. Adverse events occurred in 205 (77%) patients in the single-dose oral casopitant mesylate group and 203 (75%) patients in the 3-day intravenous and oral casopitant mesylate group compared with 194 (73%) of patients in the control group. The most common serious adverse events were neutropenia (n=5 [3%] in the control group, n=3 [1%] in the single-dose oral casopitant mesylate group, and n=11 [4%] in the 3-day intravenous plus oral casopitant mesylate group), febrile neutropenia (n=1 [<1%] in the control group, n=4 [1%] in the single-dose oral casopitant mesylate group, and n=6 [2%] in the 3-day intravenous plus oral casopitant mesylate group), and dehydration (n=4 [2%] in the control group, n=2 [<1%] in the single-dose oral casopitant mesylate group, and n=1 [<1%] in the 3-day intravenous plus oral casopitant mesylate group). INTERPRETATION A three-drug regimen including a single oral dose or 3-day intravenous plus oral regimen of casopitant mesylate plus dexamethasone and ondansetron significantly reduced CINV events in patients receiving HEC compared with a two-drug regimen of dexamethasone and ondansetron. FUNDING GlaxoSmithKline.

Capillary electrochromatography/mass spectrometry — a comparison of the sensitivity of nanospray and microspray ionization techniques

Capillary electrochromatography/mass spectrometry (CEC/MS) has so far only been performed using electrospray or microspray ionization. CEC has, to date, not been reported coupled online to nanospray ionization, due to practical difficulties in coupling the CEC column to the nanospray emitter. However this combination is ideally suited, as the flow rates for CEC (100nL/min, approximately) are directly compatible with larger orifice (10μm) nanospray emitters. A CEC unit has been designed and engineered in-house, to enable the use of short columns and high field strengths, facilitating fast electro-osmotic flow and short retention times. This device was initially coupled via a 50 μm × 37 cm C18/SCX column, and then by a C6/SCX 50 μm × 45 cm column, to microspray and nanospray interfaces, also designed and built in-house. CEC/microspray-MS and CEC/nanospray-MS were successfully evaluated on an ion-trap mass spectrometer using five corticosteriods with the addition of thiourea as a flow marker. Limits of detection (S/N = 3) were found to be 500 fg (1–2 mol) for CEC/microspray-MS and initial studies using the nanospray interface have predicted detection limits in the region of 50 fg (∼100 amol).

Effect of increasing concentration of ammonium acetate as an additive in supercritical fluid chromatography using CO2–methanol mobile phase

The effects of increasing concentrations of ammonium acetate additive in supercritical fluid chromatography were studied on silica, 2-ethyl-pyridine and endcapped 2-ethyl-pyridine stationary phases. The study involved the addition of increasing concentrations of the ammonium acetate either in the mobile phase modifier (methanol) or in the sample solvent. The effects of ammonium acetate on retention and peak shape of the analytes were evaluated. Compounds that exhibited satisfactory chromatographic behaviour in the absence of the additive were virtually unaffected by its presence in the mobile phase or sample solvent. Nevertheless, compounds that exhibited late elution and strongly tailing peak shapes when pure methanol was used showed dramatically improved chromatographic behaviour in the presence of the additive. Shorter retention was observed not only when the modifier was introduced in the mobile phase but also when it was in the sample solvent.

Defining and maintaining a high quality screening collection : the GSK experience

Understanding the quality of a screening collection is the first step to improving it and, as a result, the quality of the screening process. This article outlines how this issue was approached at GlaxoSmithKline and some of the hurdles that needed to be overcome to achieve success. The article focuses specifically on the necessary software and hardware infrastructure needed, and at some of the extra benefits of such a project in terms of data mining and data modelling.

An automated electro‐osmotic sample introduction/separation interface for automated electro‐osmotic flow flow‐injection mass spectrometry, infusion mass spectrometry and capillary electrochromatography mass spectrometry

A novel automated electro-osmotic flow (EOF) sample introduction and separation interface has been designed, developed and successfully coupled to an electrospray mass spectrometer. The system consists of a small-footprint automated capillary electrochromatography (CEC) injection and separation interface that has an integrated autosampler and connects to the outside world via a proprietary controller for automated CEC/MS, EOF infusion and flow-injection analysis (EOFFIA). The system utilizes a previously described CEC/MS/MS micro-electrospray interface that allows optimization of the spray conditions. Short CEC columns or open capillaries are immobilized in a hollow stainless steel needle that is located in a pincher bar through which injection and running high voltage is applied. The needle assembly is easily removed for ease of column installation and changeover. At the heart of the automated system is a microprocessor-based controller which allows both manual and automated control via a 12-way key pad. The controller provides automatic indexing of a 10 position sample carousel, control of kV supply so that injection and running voltages can be set independently for full flexibility and mass spectometer synchronizing signals. The design, although currently on a Finnigan TSQ 7000, has been made flexible in both mechanical and electronic design to be easiliy adaptable to other mass spectrometers. Examples of all functions of the system are presented here including rapid flow injection and controllable infusion experiments. Simple mixtures could be separated by CEC/MS with great efficiency and sensitivity in a short time. An example of multiple automated analytical runs performed continuously over a period of 10 hours are described. We believe this is the first report of a custom built CEC system with an integrated CEC/MS/MS interface that obviates the need for an expensive and unsuitable converted commercial CE system for sample introduction whilst allowing for a useful degree of automation. This system addresses many of the robustness and reduction-to-practice problems of CEC/MS and will catalyse the development of CEC/MS and allow the systematic optimization and evaluation of the technique.

Single bead and hard tag decoding using accurate isotopic difference target analysis-encoded combinatorial libraries.

A single fully automated generic microbore liquid chromatography electrospray time-of-flight protocol has been developed for the analysis of single beads, single beads with analytical constructs, and isotopically labelled dialkylamine hard tags. The protocol relies upon the incorporation of an isotopic signature into the linker and/or derivatising molecule to give a specific isotopic ratio together with an accurate isotopic difference. These properties facilitate highly specific and sensitive analysis of beads and hard tags using accurate isotopic difference analysis (AIDA) without prior knowledge of the molecular weight. Three open access methods with automated processing have been developed around a core generic approach.

Ionisation in the absence of high voltage using supercritical fluid chromatography: a possible route to increased signal

Supercritical fluid chromatography (SFC) is fast becoming a technique of choice for the analysis of a wide range of compounds and has been found to be complementary to high-performance liquid chromatography (HPLC). The combination of SFC and mass spectrometry (MS) affords a very useful tool in the separation and analysis of compounds. In this study the ionisation of samples in the absence of an applied electrospray voltage has been observed when using SFC/MS, with some compounds showing increased sensitivity when all ionisation source high voltage (HV) is removed. In an attempt to understand the mechanism of ionisation, a series of test compounds were analysed using standard electrospray ionisation (ESI) and atmospheric pressure chemical ionisation (APCI) source configurations and also different API source designs. In both cases, data were acquired with the applied high voltage on (normal conditions) or with the high voltage off, i.e. no voltage spray (novo-spray). The standards were analysed with a range of pressure and modifier percentage conditions. To understand the nature of the ionisation process observed, this was compared with three established liquid-to-gas ionisation mechanisms. These were thermospray (TSP), charge residue model (CRM) of ESI and sonic spray ionisation (SSI). Experiments were undertaken in an attempt to explain this ionisation phenomenon and quantify any observed change in sensitivity. The most important point to note is that enhanced ionisation was observed under novo-spray conditions in a SFC/MS configuration, which in certain cases provides a lowering in the overall limit of detection (LOD).

On-Chip Post-Column Derivatisation Reactions in Capillary Liquid Chromatography — Mass Spectrometry

A micromixer chip used previously for organic reaction chemistries [1] is applied for post-column derivatisation in µ-HPLC hyphenated to a Time-of-Flight mass spectrometer (TOFMS).

Cardiac Safety Data for Casopitant, a Neurokinin-1 Receptor Antagonist, Given with Anthracycline.

Objectives: The NK-1 receptor antagonist casopitant is a promising agent for the prevention of chemotherapy-related and postoperative nausea and vomiting. However, findings suggestive of cardiac damage were noted in animal studies of long-term administration of casopitant at supratherapeutic doses. Cardiac evaluation of the concomitant use of casopitant with anthracyclines was therefore of particular interest due to myocyte loss associated with anthracycline use. Methods: Chemotherapy-naive patients receiving anthracyclines as part of a Phase III moderately emetogenic chemotherapy study regimen were randomized to receive ondansetron and dexamethasone plus one of the following casopitant regimens: single-dose 150-mg oral casopitant (ORAL1); 3-day IV/oral casopitant (90 mg IV/50 mg oral/50 mg oral) (IV/ORAL); 3-day oral casopitant (150/50/50 mg) (ORAL3); or placebo. Serial cardiac troponin (cTnI) levels were measured using the Beckman assay. Although this assay is generally considered reliable within the range of 0.04-0.5 ng/mL, levels below the limit of quantitation (LOQ) ( Results: In total, 611 patients were evaluated: 155 ORAL1, 145 IV/ORAL, 159 ORAL3, and 152 placebo; no risk factor variation between groups was observed. Adverse events were balanced across treatment groups. Cardiovascular adverse events occurred in 3% of patients in each of the placebo, ORAL1, and ORAL/IV groups and in 2% of the ORAL3 group. Three patients (0.5%) had cTnI >0.12 ng/mL (1 each IV/ORAL, ORAL3, and placebo); all LVEF determinations were normal at follow-up, with the exception of 1 patient with mitral regurgitation but normal ejection fraction. A total of 59 patients (9.7%) had median cTnI 0.04-0.12 ng/mL. Of the remaining patients with cTnI below LOQ, median baseline levels across groups were similar (ORAL1 and IV/ORAL 0.008 ng/mL; ORAL3 0.01 ng/mL; placebo 0.009 ng/mL). After 4 cycles of anthracycline, median cTnI increased to 0.02 ng/mL in all groups, with no difference noted between casopitant and placebo groups. Conclusions: Thorough monitoring of cardiac-related toxicities including cardiac (S)AEs, ECG, and ejection fraction data showed no evidence of increased risk of cardiac damage in the casopitant groups compared with control. Subclinical elevations in cTnI occurred in patients receiving anthracyclines but were not considered to be related to or accentuated by simultaneous administration of casopitant. cTnI elevations were similar across treatment groups and were well below rates reported in the literature for a patient population receiving higher doses of anthracycline. Use of appropriate cardiac biomarkers in ranges below those commonly used to detect infarction may be valuable to detect or exclude subclinical toxicity of potentially cardiotoxic agents used in combination. Additional analysis is ongoing. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1118.