Pharmacokinetic (PK) analyses
cancer
BLOOD, CANCER
Blood samples for PK analysis were obtained at pre-dose prior to the first infusion dose on cycle 1 day 1 (C1D1), end-of-infusion (EOI), then at 1 h (hr) post EOI, 4 h post EOI, 24 h post EOI, 72 h post EOI, 7 days post EOI and 14 days post EOI. Spare sampling (pre/trough and end of infusion) was performed on cycle 1 day 15 (C1D15), cycle 2 day 1 (C2D1), cycle 2 day 15 (C2D15), cycle 3 day 1 (C3D1) and samples were used to assess accumulation with biweekly dosing. Assessment into saturable elimination was also made. Blood samples were collected into Serum Separator Tubes (SST®), left to sit at room temperature for 30 min to allow clotting, then centrifuged to obtain serum. The serum was aliquoted into cryovials and frozen at -80 °C until bioanalysis. The first dose (starting from cycle 1, day 1 [C1D1]) exposure metrics of the maximum serum concentration (Cmax), minimum serum concentration (Cmin), area under the serum concentration vs time curve (AUC), half-life, clearance and volume of distribution through dense PK sampling were assessed.NEO-201 concentration in serum of cancer patients was determined by a fully-validated enzyme-linked immunosorbent assay (ELISA) method with a linear range of 0.25 – 2.0 mg/L (250 – 2000 ng/mL) performed by the NCI Clinical Pharmacology Program. NEO-201 was stable through 2 freeze/thaw cycles in serum at -80 °C, allowing for sample re-analysis. All data on serum NEO-201 concentrations met FDA guidelines for bioanalytical testing.Specifics of the ELISA and PK analysis are described in Supplementary materials and methods.