emonstrated a linear response range of 0.9 to 100 units/mL of pSTAT3 ( R 2 = 0.9996), where 1 unit is equivalent to 20 pg of pSTAT3 protein. The intra-assay precision ranged from 14.9% to 29.8% for ruxolitinib concentrations of 0 to 3.33  and was 85.9% at 10.0  of ruxolitinib concentration (this concentration is above the IC 90 of pSTAT3 inhibition by Linifanib ruxolitinib and hence corresponds to the lower limit of pSTAT3 quantity). The interassay precision ranged from 6.9% to 29.1% for ruxolitinib concentrations of 0 to 10.0 . Duplicate PD analyses were performed, and the average value was reported for each sample analyzed. For each participant, the percent inhibition of phosphorylated STAT3 was calculated by comparing predose values obtained before the first dose with values obtained at different times after dosing.

Pharmacokinetic and Pharmacodynamic Analysis Standard noncompartmental (model-independent) pharmacokinetic methods were used to analyze the plasma concentration data using actual blood sampling times. The maximum plasma drug concentration (C max ) and tysical examinations, including vital signs, electrocardiogram, and clinical laboratory Ridaforolimus test results. Women of childbearing potential enrolled in these studies were determined to be in a nongravid state and had agreed to take appropriate precautions (with at least 99% certainty) to avoid pregnancy from screening through follow-up. Applicants with hemo- globin levels and white blood and platelet counts below the lower reference limit for the parameters were excluded. Nonstudy medications were not allowed for 7, 14, and 30 days, for over-the-counter (including vitamins, minerals, and herbal/plant-derived prepara- tions), prescription, and investigational drugs, respec- tively, prior to the first dose of study medication until the completion of study, unless deemed necessary and acceptable by the investigator.

Use of any medica- tions known to affect cytochrome P450 enzymes or P-glycoprotein buy Agomelatine activity was prohibited within 15 days or 5 half-lives (whichever was longer) prior to the study start and throughout the study. Study Design Two clinical studies were conducted in full accor- dance with the Declaration of Helsinki, principles of good clinical practices (GCP), and local laws regard- ing the protection of the rights and welfare of human participants in biomedical research. The protocols were approved by an independent institutional review board (IRB), and informed consents for all participants were obtained prior to screening. Both the studies were single-center, open-label, 2-treatment, and 1-way crossover drug-drug interaction studies conducted in healthy male and female adult volunteers. Study A, employing 2 independent cohorts, evalu- ated the polymath effects of multiple doses of ketoconazole (cohort 1, n = 16) and erythromycin (cohort 2, n = 14), a potent and moderate CYP3A4 inhibitor, respectively, on the single-dose PK and PD of ruxolitinib. Study B enrolled a single cohort of 12 participants and evalu- ated the effect of multiple doses of rifampin, a potent CYP3A4 inducer, on the single-dose PK and PD of ruxolitinib. An overview for the study designs is pro- vided in Figure 1.

All doses of ruxolitinib phosphate were adminis- tered as tablets with 240 mL of water after an overnight fast of at least 10 hours, and participants continued to fast from food for at purchase Agomelatine least 1 hour postdose. For both the studies, blood samples for determination of rux- olitinib concentrations were collected at and 24 hours postdose, and blood samples for determination of ruxolitinib ex vivo PD activity were collected  24 hours postdose. Analytical Methods Pharmacokinetic plasma samples were assayed for ruxolitinib concentrations by a validated, good labora- tory practice (GLP), liquid chromatography/tandem mass spectrometry (LC/MS/MS) method at Incyte Corporation.