Verification of the Bühlmann fCAL turbo faecal calprotectin assay on the Binding Site Optilite benchtop analyser.

Objectives: Inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS) are increasingly prevalent disorders. Faecal calprotectin is useful in the differential diagnosis of IBD from IBS and monitoring IBD activity. We verified the Bühlmann fCAL turbo faecal calprotectin assay on the Binding Site, Optilite benchtop analyser. Design: Accuracy, precision, lower limit of quantitation (LLoQ), and linearity of the Bühlmann fCAL turbo faecal calprotectin assay on the Binding Site, Optilite benchtop analyser were ascertained. Comparison with the Bühlmann Quantum Blue fCAL extended and DiaSorin, Liaison calprotectin assays were also undertaken. Difference between assays was evaluated using the Wilcoxon signed-rank test and method comparison was undertaken using Spearman's rank correlation (rs), difference plots and Passing-Bablok regression analyses. Results: The fCAL turbo assay was linear between 25 and 10,000 µg/g, and the LLoQ was 25 µg/g. Intra-, and inter-assay imprecision was <5%. There was a good agreement (rs = 0.96) and no significant bias (3%, p = 0.10) present between the fCAL turbo and Quantum Blue extended assays. Between the fCAL turbo and DiaSorin, liaison assays there was a good agreement (rs = 0.97), but a significant bias (53%, p = <0.01) was present. Conclusions: The fCAL turbo assay performs well on the Binding Site, Optilite benchtop analyser. Calprotectin results are commutable between with Bühlmann fCAL turbo and Quantum Blue fCAL extended assays, but not between Bühlmann and DiaSorin calprotectin assays.

Phase I first-in-human study of TAK-285, a novel investigational dual HER2/EGFR inhibitor, in cancer patients.

BACKGROUND: This phase I first-in-human study was conducted in Japanese patients to investigate the safety, pharmacokinetics (PKs), and determine the maximum tolerated dose (MTD) of oral TAK-285, a novel dual erbB protein kinase inhibitor that specifically targets human epidermal growth factor receptor (EGFR) and HER2. METHODS: The TAK-285 dose was escalated until MTD was determined. A second patient cohort received TAK-285 at the MTD for at least 4 weeks. RESULTS: In all, 26 patients received TAK-285 at doses ranging from 50 to 400 mg once daily (q.d.) or twice daily (b.i.d.); 20 patients made up the dose escalation cohort and the remaining 6 patients were the repeated administration cohort. TAK-285 was well tolerated. Dose-limiting toxicities noted in two patients who received 400 mg b.i.d. were grade 3 increases in aminotransferases and grade 3 decreased appetite. Consequently, the MTD was determined to be 300 mg b.i.d. Absorption of TAK-285 was rapid after oral dosing, and plasma exposure at steady-state increased in a dose-proportional fashion for doses ranging from 50 to 300 mg b.i.d. A partial response was observed for one patient with parotid cancer who received 300 mg b.i.d. CONCLUSION: The toxicity profile and PK properties of oral TAK-285 warrant further evaluation.

Application of liquid chromatography-turbo ion spray tandem mass spectrometry for quantitative analysis of a potent motilin receptor agonist, EM574, and its metabolites in human plasma.

A liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the simultaneous determination of a new potent motilin receptor agonist as erythromycin derivative, EM574 (erythromycin derivative), and its three metabolites, M-IV, M-V and M-VI, in human plasma was developed. The internal standards (I.S.s) used were deuterated EM574, M-IV and M-V. For the quantitation of M-VI, deuterated M-V was used. The analytes and I.S. were extracted from plasma samples with diethyl ether at neutral pH. A turbo ion spray interface was used as the ion source of LC-MS-MS, and the analysis was performed in the selected reaction monitoring mode. The lower quantitation limits for all the analytes were 0.05 ng/ml when 0.2 ml of plasma was used, and the standard curves were linear in the range 0.05 to 20 ng/ml. The method was precise; the intra- and inter-day precisions of the method were not more than 19.8% for all the analytes. The accuracy of the method was good with the deviations between added and calculated concentrations of each analyte being typically within +/- 11.2%.

Moment analysis of stereoselective biliary excretion and chiral inversion of ketoprofen enantiomers in perfused rat liver.

The stereoselective local disposition of ketoprofen was evaluated by the single-pass perfusion experiment following a bolus injection of R(-)- or S(+)-ketoprofen into the liver from the portal vein. The elution time profiles of enantiomers into the hepatic vein and the excretion time profiles into the bile were kinetically assessed by local moment analysis. The hepatic recovery ratios (FH) of both enantiomers were < 1%, and the mean hepatic transit times (tH) were approximately 7 s. After the injection of S-ketoprofen into the liver, the biliary excretion ratio (Fb) of total S-ketoprofen was 68% (15% S-ketoprofen and 53% glucuronide) and the mean biliary transit time (tb) of S-ketoprofen was 10 min. R-Ketoprofen inversion from S-ketoprofen was not observed in either the perfusate or in the bile. After the injection of R-ketoprofen, the Fb of total R-ketoprofen was 12% (3% R-ketoprofen and 9% glucuronide), and tb of R-ketoprofen was 8 min. The Fb of total S-ketoprofen inverted from R-ketoprofen was 24% (7% S-ketoprofen and 17% glucuronide), and the tb of inverted S-ketoprofen was 17 min. Forty-six percent of R-ketoprofen was inverted to S-ketoprofen during a single pass through the rat liver, and the mean inversion time was 7.5 min. It was concluded that the unidirectional chiral inversion of ketoprofen was stereospecific, and the hepatic uptake and biliary excretion were stereo-nonspecific.

Development of glycoside-bound radiopharmaceuticals: novel radioiodination method for digoxin.

We combined 2-hydroxy-3-methylbenzoylhydrazide (HMBH) with glycosides as a novel method for the radioiodination of physiologically active glycosides. This method was tested using digoxin, which is one of the cardiac glycosides. A digoxin-HMBH conjugate was synthesized by periodate cleavage of the third sugar ring, and was readily radiolabeled with Na[125I] by the chloramine-T method. 125I labelled digoxin-HMBH conjugate retained Na+, K(+)-ATPase binding in vivo and in vitro, and also retained immunoreactivity to an anti-digoxin antibody. Thus, this 125I labelled digoxin-HMBH conjugate represents a potential radiopharmaceutical for Na+, K(+)-ATPase imaging, as well as for the radioimmunoassay of digoxin.