Sustiva

Delamanid tablets were manufactured by Otsuka Pharmaceutical Co., Ltd. (Japan). Efavirenz was supplied as commercially available Sustiva manufactured by Bristol-Myers Squibb (USA). Subjects were randomly assigned to receive one of the following two treatments: group 1 received 600 mg of efavirenz (one 600-mg Sustiva tablet) once daily for 10 days and group 2 received 100 mg of delamanid (two 50-mg tablets) twice daily for 7 days, followed by 100 mg of delamanid twice daily plus 600 mg of efavirenz once daily for 10 days, for a total of 18 days of dosing. Consistent with labeling, the once-daily dose of efavirenz in both groups was administered in the evening at about 8 p.m. on an empty stomach (2 h after a standard meal and 2 h prior to a snack) (20 ). The first dose of delamanid was the evening dose administered on day 1, and the last delamanid dose was the evening dose administered on day 18. Each evening dose of delamanid was administered at about 6 p.m., and each morning dose of delamanid was administered at about 8 a.m.; all delamanid doses were given within 30 min of the start of a standard meal.

Lansoprazole was purchased by Sigma-Aldrich (Milan, Italy) and was dissolved at 20 mM in DMSO in absence of direct light and reconstituted immediately prior its use. Efavirenz was purified from commercially available Sustiva (Bristol-Myers Squibb) as described [3 (link)], dissolved in DMSO and added to the culture medium in the indicated concentrations and times. RPMI-1640 cell culture medium (BE12-702F), antibiotics (DE17-603E), phosphate buffer saline (PBS) (BE17-512F), trypsin/EDTA (BE17-171E) and fetal bovine serum (DE14-701F) were obtained from Lonza (Milan, Italy). Trypan blue was bought from Alexis Biochemicals (Florence, Italy) and Annexin V-FITC Apoptosis detection kit from Enzo Life Sciences (Lausen, Switzerland). 4-Nitrophenyl phosphate disodium salt hexahydrate tablets for proliferation assay was from Sigma-Aldrich.
The pH of all cell culture supernatants were estimated by the use of a pH 123 Microprocessor pH Meter (Hanna Instruments, Italy).