The assay for bendamustine, M3, and M4 used a Synergi™ Hydro-RP column, and the assay for HP2 used a Synergi™ Polar-RP column (Phenomenex, Inc.; Torrance, CA, USA). On both columns, gradient elution was performed with 5 mM ammonium formate with 0.1% formic acid in water and methanol. The quantifiable ranges for bendamustine, M3, and M4 were 0.5–500 ng/mL in plasma and 0.5–50 μg/mL Selleckchem Vorinostat in urine, and for HP2 were 1–500 ng/mL in plasma and 0.1–50 μg/mL in urine. Quality control samples were prepared and analyzed together with the study samples, and acceptance criteria
for bioanalytic data during routine drug analysis, as described in the US Food and Drug Androgen Receptor Antagonist research buy Administration (FDA) guidelines , were applied. 2.7 Pharmacokinetic Analysis Pharmacokinetic parameters for bendamustine, M3, M4, HP2, and TRA were estimated by noncompartmental analysis AG-881 research buy using WinNonlin™ software (version 4.1.a; Pharsight Corporation; Mountain View, CA, USA). Parameters that were determined for
all analytes included the maximum observed plasma concentration (Cmax), the elimination half-life (t½), and the area under the plasma concentration–time curve from time zero to infinity (AUC∞). Additionally, the plasma clearance (CL) and the apparent volume of distribution at steady state (Vss) were determined for bendamustine and estimated for TRA, and the renal clearance (CLR) was determined for bendamustine. 2.8 Safety Assessments The safety of bendamustine was assessed by evaluating AEs according to Common Terminology Criteria for AEs v3.0; serum chemistry, hematology, and urinalysis test results; vital signs; 12-lead electrocardiograms (ECGs); body weight; physical examinations; and concomitant medication. ECGs were performed prior BCKDHA to study drug administration and at multiple time points on day 1 of cycle 1. No formal statistical analysis was applied in this study; descriptive statistics were used when appropriate. 3 Results 3.1 Patients Six patients with confirmed relapsed or refractory
malignancy were enrolled (Table 1). They had a median age of 66 years (range 48–75), a mean weight of 72.7 kg (range 59–94), a mean height of 173.2 cm (range 155–181), and a mean body surface area of 1.9 m2 (range 1.6–2.2). All patients had a history of cancer drug therapy and anticancer surgery. At the time of enrollment, four patients (67%) had a WHO performance status of 0 and two (33%) had a status of 1. Table 1 Patient characteristics Characteristic Value Median age (years [range]) 66 [48–75] Sex (n [%]) Male 3  Female 3  Race (n [%]) White 6  Ethnicity (n [%]) Non-Hispanic and non-Latino 6  Mean weight (kg [range]) 72.7 [59–94] Mean height (cm [range]) 173.2 [155–181] Mean body surface area (m2 [range]) 1.9 [1.6–2.2] Mean time since cancer diagnosis (years [range]) 4.