This was a open-label, single-dose study carried out at Pharma Bio-Research Group BV . The research was approved by a completely independent ethics committee Medische Ethische Toetsings Commissie van p Stichting RAF265 Beoordeling Bio-Medisch Onderzoek, Assen, Holland and carried out based on the concepts of excellent Clinical Practice and also the Promise of Helsinki (October 1996 version). Written informed consent was acquired all participants before study entry. After an overnight fast (a minimum of 10 h), subjects received just one dental 15 mg dose (calculated because the free base) of afatinib (equal to 22.2 mg of afatinib dimaleinate salt) solution that contains 2.25 MBq of [14C]-radiolabeled afatinib (Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany) within the sitting/standing position. The [14C]-afatinib powder was reconstituted with 50 mL of isotonic sodium chloride solution. This solution was given orally towards the volunteers. The empty vial was washed once again with another 50 mL of isotonic sodium chloride solution.

          that was then given towards the subjects. Subjects continued to be within the study center not less than 120 h for that assortment of bloodstream, urine and feces samples. When the radioactivity counts measured in urine and feces from day 5 let’s start continued to be over the termination limits (50 dpm/mL in urine MLN8054 and 75 dpm per 100 mg feces), the remain in the middle was extended to no more than ten days. After that, assortment of urine and/or feces was ongoing in your own home before the [14C]-radioactivity quick counts fell beneath the termination criteria. Sample collection All bloodstream samples were collected in potassium-EDTAcontaining tubes. Venous bloodstream samples for measurement of plasma amounts of afatinib and [14C]-radioactivity were acquired For pharmacokinetic checks, roughly 11 mL of bloodstream was collected each and every time point. A Couple-mL aliquot was taken for the determination of [14C]-radioactivity in whole blood and stored at -20C. The remaining 9 mL was centrifuged immediately at 2,000g (4C) for 10 min.

            Two aliquots of at least 1 mL each were used for the determination of [14C]- radioactivity in plasma, and 2 aliquots of at least 1 mL each were used for the analysis of the parent compound (afatinib) in plasma. Plasma aliquots were frozen immediately and stored at -20C until analysis. Additional blood samples were collected pre-dose and 1, 2 and 6 h after dosing for metabolic profiling (50 mL).AZD8931  Blood samples were centrifuged at 2,000g (4C) for 10 min. Each blood cell pellet was divided into two approximately equal parts and transferred to two suitable storage tubes. The blood cell samples were stored at -20C until shipment to the metabolic laboratory at Boehringer Ingelheim Pharma GmbH & Co. KG. Plasma was also transferred to separate tubes and stored at -20C until shipment to the same laboratory. Additional blood samples (10 mL) for the determination of ABT-263  protein binding were collected pre-dose and 1, 2 and 6 h after dosing. Blood samples were centrifuged at 2,000g for 10 min. Plasma was transferred to separate tubes and stored at -20C until shipment to the analytical laboratory at Pharma Bio-Research Group BV.