Whole-blood sampling was conducted before dose 1 and at the following timepoints before and after dose 3: pre-dose (0 h; immediately before start of infusion), 2 (end of infusion), 2.25, 2.5, 3, 4, 6 and 8 h in K2EDTA-containing vacutainers (Becton Dickinson and Company, Franklin Lakes, NJ, USA). Samples were centrifuged at 1500 × g for 10 min at 4°C and separated plasma was stored at −80°C until concentration determination.
All subjects were randomly assigned to undergo a single bronchoscopy with bronchoalveolar lavage (BAL) at 2, 4, 6 or 8 h after start of the third drug infusion (five subjects per timepoint). Subjects fasted for at least 6 h prior to the procedure and were then prepared for bronchoscopy with aerosolized lidocaine in the nares and oropharynx and 2% lidocaine jelly in the nasal passageway within 30 min of the procedure. The subjects underwent conscious sedation with IV injection of midazolam and fentanyl (per SurgiCenter standard of care) as needed. Each BAL was conducted with a fibre-optic bronchoscope (Olympus BF-Q190, Olympus America Inc., Center Valley, PA, USA) into the right middle lobe and utilized four aliquots of sterile 0.9% saline for instillation and aspiration as previously described.15–18 (link) The initial aliquot (50 mL) was discarded and the subsequent three aliquots (50 mL each) were stored on ice immediately after aspiration. The three aliquots were pooled (total volume recorded) and an aliquot obtained for complete cell count and differential. The remaining volume of pooled BAL was immediately centrifuged at 400 × g for 10 min, and the supernatant and cell pellet were separated. Supernatant aliquots were obtained to determine drug and urea concentrations. A blood sample was collected at the time of bronchoscopy to determine the plasma drug and urea concentrations. Water:formic acid (100:2, v/v) was added at a 1:1 ratio to all non-plasma samples to ensure taniborbactam drug stability prior to storage at −80°C until concentration determination.
All subjects were randomly assigned to undergo a single bronchoscopy with bronchoalveolar lavage (BAL) at 2, 4, 6 or 8 h after start of the third drug infusion (five subjects per timepoint). Subjects fasted for at least 6 h prior to the procedure and were then prepared for bronchoscopy with aerosolized lidocaine in the nares and oropharynx and 2% lidocaine jelly in the nasal passageway within 30 min of the procedure. The subjects underwent conscious sedation with IV injection of midazolam and fentanyl (per SurgiCenter standard of care) as needed. Each BAL was conducted with a fibre-optic bronchoscope (Olympus BF-Q190, Olympus America Inc., Center Valley, PA, USA) into the right middle lobe and utilized four aliquots of sterile 0.9% saline for instillation and aspiration as previously described.15–18 (link) The initial aliquot (50 mL) was discarded and the subsequent three aliquots (50 mL each) were stored on ice immediately after aspiration. The three aliquots were pooled (total volume recorded) and an aliquot obtained for complete cell count and differential. The remaining volume of pooled BAL was immediately centrifuged at 400 × g for 10 min, and the supernatant and cell pellet were separated. Supernatant aliquots were obtained to determine drug and urea concentrations. A blood sample was collected at the time of bronchoscopy to determine the plasma drug and urea concentrations. Water:formic acid (100:2, v/v) was added at a 1:1 ratio to all non-plasma samples to ensure taniborbactam drug stability prior to storage at −80°C until concentration determination.
