Pharmacokinetics

Api137 dissolved in 0.9% saline was injected iv or sc in mice (18–26 g, total = 51) and rats (204–248 g, total = 12) at a dose volume of 10 and 5 mL/kg BW, respectively, according to the dosing scheme (Supplementary Figures 1, 2). Three mice per dose and time point were euthanized by carbon dioxide inhalation and blood was collected via cardiac puncture into K₂-EDTA Microtainer^® tubes (Becton Dickinson and Company, BD, Franklin Lakes, NJ, United States). Additionally, blood was collected via saphenous vein puncture from three rats into identic tubes.

After priming, as described above, pumps were implanted under isoflurane anesthesia into a pocket of the subcutaneous connective tissue between the scapula of the mice. The pump was inserted with the flow moderator pointing away from the incision to the tail. Three animals per dose were bled twice via submandibular puncture and euthanized at the third time point by carbon dioxide inhalation and cardiac puncture (Supplementary Table 1). EDTA-blood samples were inverted for 10 min and centrifuged (3000 × g, 5 min, 4°C). Plasma was transferred to polypropylene tubes and stored at −80°C prior to analysis.