FEV1 is a lung function measure defined as the maximal amount of air that can be forcefully exhaled in one second. Mean FEV1 from the 3 acceptable spirometric efforts were recorded after at pre-dose, 0.5 h, 1 h, 2 h, 8 h and 12 h post-dose on Day 1. Baseline was defined as the mean of the 3 pre-dose measurements on Day 1 of each treatment period. The change from Baseline was calculated by subtracting the Baseline value (Day 1, pre-dose) from the individual post-Baseline (0.5 h, 1 h, 2 h, 8 h and 12 h post-dose) values on Day 1. Adjusted mean was reported as LS mean.

FEV1 is a lung function measure defined as the maximal amount of air that can be forcefully exhaled in one second. Mean FEV1 from the 3 acceptable spirometric efforts were recorded after at pre-dose, 0.5 h, 1 h, 2 h, 8 h and 12 h post-dose on Day 1. Baseline was defined as the mean of the 3 pre-dose measurements on Day 1 of each treatment period. The change from Baseline was calculated by subtracting the Baseline value (Day 1, pre-dose) from the individual post-Baseline (0.5 h, 1 h, 2 h, 8 h and 12 h post-dose) values on Day 14. Adjusted mean was reported as LS mean.

PEF is a measure of lung function and measures how fast a person can breathe out. It was measured using a peak flow meter by the participants and recorded on diary cards during Day 1 to Day 14 pre-dose am and pm of each treatment period. The efforts were recorded after withholding salbutamol used as bronchodilator for at least 6 hours. The averaged PEFR over the 14-day treatment period was calculated as the mean of the 14 pre-dose observations during each treatment period. Baseline was defined as the assessment done during Screening (the average of am and average of pm measurements over the last 7 days pre-dose prior to Day 1 of treatment period 1). The change from Baseline was calculated by subtracting the Baseline value (average of am and average of pm screening values) from the individual post-Baseline (average of am and average of pm values over 14 days) values. Adjusted mean was reported as LS mean.

The exhaled nitric oxide assessment is used as a marker for airway inflammation. The exhaled nitric oxide was assessed using breath test on Day 1 (pre-dose and 1 h post-dose on Day 1), Day 7 (pre-dose) and Day 14 (pre-dose, 1 h and 24 h post-dose). Baseline was defined as the assessment done pre-dose on Day 1. The change from Baseline was calculated by subtracting the Baseline value (Day 1, pre-dose) from the individual post-Baseline (1 h post-dose Day 1, pre-dose Day 7 and Day 14 at pre-dose, 1 h post-dose and 24 h post-dose) values. Data was analyzed using repeated measures mixed effects model for the ratio of change from Baseline values, derived from dividing doses of GSK256066 by placebo, adjusted for period, treatment, Day (1, 7 or 14), time (pre-dose, 1 or 24 h), period-level Baseline of exhaled nitric oxide and participant-level Baseline exhaled nitric oxide as fixed effects and participant as a random effect. Adjusted geometric mean is reported as LS geometric mean.

The PC20 was assessed using the methacholine challenge. This is a physiological measurement derived from repeated breathing maneuvers which measures airway reactivity. Participants were exposed to higher and higher concentrations of an airway irritant (in this case methacholine), and between each dose spirometry was performed. The test was stopped after the FEV1 fell 20% below the Baseline. The concentration of methacholine at which this occurred was called the PC20. The methacholine challenge was performed at 24 h post-dose on Day 14 (morning of Day 15) after exhaled nitric oxide assessment. Data was analyzed using mixed effect model, where the model was adjusted for period and treatment as fixed effects and participant as a random effect. Adjusted geometric mean is reported as LS geometric mean.

Blood samples were collected on Day 1 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h and 12 h) and Day 14 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h). The AUC (0-t) was determined using the linear trapezoidal rule for increasing concentrations and the logarithmic trapezoidal rule for decreasing concentrations. AUC (0-infinity) was planned to be calculated, where data permitted, as the sum of AUC (0-t) and Ct/z, where Ct was the observed plasma concentration obtained from the log-linear regression analysis of the last quantifiable time-point and z is the terminal phase rate constant. Data for AUC (0-infinity) was not derived due to non quantifiable data.

Blood samples were collected on Day 1 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h and 12 h) and Day 14 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h). The first occurrence of the Cmax was determined directly from the raw concentration-time data.

Blood samples were collected on Day 1 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h and 12 h) and Day 14 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h). The time at which Cmax was observed was determined directly from the raw concentration-time data. The t1/2 was planned to be obtained as the ratio of ln2/λz, where λz is the terminal phase rate constant estimated by linear regression analysis of the log transformed concentration-time data. The data for t1/2 was not derived due to non quantifiable data.

GSK614917 is a metabolite of GSK256066. Blood samples for PK analysis were collected on Day 1 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h and 12 h) and Day 14 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h). The AUC 0-t was planned to be determined using the linear trapezoidal rule for increasing concentrations and the logarithmic trapezoidal rule for decreasing concentrations. AUC (0-infinity) was planned to be calculated, where data permitted, as the sum of AUC (0-t) and Ct/z, where Ct was the observed plasma concentration obtained from the log-linear regression analysis of the last quantifiable time-point and z is the terminal phase rate constant. Data for this outcome measure was not derived due to non quantifiable data.

GSK614917 is a metabolite of GSK256066. Blood samples for PK analysis were collected on Day 1 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h and 12 h) and Day 14 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h). The first occurrence of the Cmax was planned to be determined directly from the raw concentration-time data. Data for this outcome measure was not derived due to non quantifiable data.

Blood samples were collected on Day 1 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h and 12 h) and Day 14 (pre-dose, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h). The time at which Cmax was observed was planned to be determined directly from the raw concentration-time data. The t1/2 was planned to be obtained as the ratio of ln2/λz, where λz is the terminal phase rate constant estimated by linear regression analysis of the log transformed concentration-time data. Data for this outcome measure was not derived due to non quantifiable data.

An AE is defined as any untoward medical occurrence in a participant, temporally associated with the use of a medicinal product, whether or not considered related to the medicinal product. An SAE is defined as any untoward medical occurrence that, at any dose, results in death, is life-threatening, requires hospitalization or prolongation of existing hospitalization, results in disability/incapacity, or is a congenital anomaly/birth defect, may jeopardize the participant or may require medical or surgical intervention to prevent one of the other outcomes listed in this definition.

The ECG measurements were made with the participant in a supine position having rested in this position for at least 10 minutes before each time-point. 12-lead ECG assessments were obtained on Day 1 (pre-dose triplicate assessment, 0.5 h, 1 h, 2 h, 8 h and 12 h), Day 7 (pre-dose) and Day 14 (pre-dose, 0.5 h, 1 h, 2 h, 8 h, 12 h and 24 h) using a machine that automatically calculated PR, QRS, QT and QTcB. Abnormal ECG findings (CNS and CS) were categorized. The abnormal PCI range included QTc interval of <300 and >500 milliseconds (msec) and increase from baseline QTc interval of >60 msec, PR interval <110 and >220 msec and QRS interval <75 and >110 msec. Baseline was Day 1 (mean of triplicate assessments done at pre dose).

The assessments were done at pre-dose on Day 1, Day 7 and Day 14 of each treatment period. The following laboratory parameters of clinical haematology were analyzed: platelet count, red blood cells (RBC) count, absolute white blood cells (WBC) count, reticulocyte count, hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), total neutrophils, lymphocytes, monocytes, eosinophils and basophils. Only those parameters for which at least one value of PCI was reported are summarized.

The assessments were done at pre-dose on Day 1, Day 7 and Day 14 of each treatment period. The following laboratory parameters of clinical chemistry were analyzed: blood urea nitrogen (BUN), creatinine, total cholesterol, sodium, potassium, chloride, total bicarbonate, calcium, C-reactive protein (CRP), aspartate aminotransferase (AST), alanine amino transferase (ALT), gamma glutamyltransferase (GGT), alkaline phosphatase, creatine kinase (CK), lactate dehydrogenase (LDH), total and direct bilirubin, uric acid, albumin and total protein. Only those parameters for which at least one value of PCI was reported are summarized.

The assessments were done at Day 1, Day 7 and Day 14 for urine glucose, protein, occult blood, ketones, nitrites, bilirubin, leukocytes esterase and urobilinogen by dipstick. The participants were categorized with results of 1 +, 2+, 3+, 4+, 7+, positive, trace, not rated (NR), 0, 0.2, 1, 16 and 3.2. Only those parameters for which at least one value of these categories reported are summarized.

The assessments were done at pre-dose on Day 1 (taken in triplicate at pre-dose, 0.5, 1, 2, 8 and 12 h post-dose), Day 7 (pre-dose) and Day 14 (24 h post-dose) of each treatment period. Supine vital signs measurements were made with the participant in a supine position having rested in that position for at least 5 minutes before the first reading at each time point. Vital signs assessment included heart rate (HR), systolic blood pressure (SBP) and diastolic blood pressure (DBP). Criteria for vital sign values meeting PCI included: SBP < 70 millimeters of mercury (mmHg) and > 160 mmHg; DBP < 45 mmHg and > 110 mmHg; < 35 beats per minute and > 160 beats per minute. Only those parameters for which at least one value of PCI was reported are summarized.

The assessment of troponin (I and or T) was done to ensure participants safety and further characterize potential renal and cardiac effects. It was monitored at pre dose on Day 1, Day 7 and Day 14. Data is categorized according to the troponin levels of <0.03 µg/L, >0.03 µg/L but <0.10 µg/L and negative. The PCI value for troponin T: low= 0 µg/L and high >0.4 µg/L.

This included the study of variability in drug response due to hereditary factors in different populations. There is increasing evidence that an individual's genetic composition (i.e., genotype) may impact the pharmacokinetics (absorption, distribution, metabolism, elimination), pharmacodynamics (relationship between concentrations and pharmacologic effects or the time course of pharmacologic effects) and/or clinical outcome (in terms of efficacy and/or safety and tolerability). A whole blood sample (10ml) was planned to be collected. The data for this outcome measure was not collected.

PDEs are a super family of enzymes that catalyze the breakdown of cyclic adenosine monophosphate (cAMP) and/or cyclic guanosine monophosphate (GMP) to their inactive form. PDE4 is the main selective cAMP-metabolizing enzyme in inflammatory and immune cells. Because PDE4 is highly expressed in leukocytes and other inflammatory cells involved in the pathogenesis of inflammatory lung diseases, such as asthma, inhibition of PDE4 had been predicted to have an anti inflammatory effect for them. PDE4 was assessed in sputum samples, collected 2-3 h post-dose on Day 7 by transcriptomic analysis on sputum ribonucleic acid (RNA) samples. Data is reported for PDE4B.

Sputum samples were collected 2-3 h post-dose on Day 7 to measure the differential cell counts which is used as an indicator of the anti-inflammatory activities. The cells include neutrophils, macrophages, lymphocytes, total leukocyte count and bronchial epithelial cells.

The anti-inflammatory activity was assessed by measuring novel candidate biomarkers of the biological response associated with the action of GSK256066, in sputum samples, collected 2-3 h post-dose on Day 7 by transcriptomic analysis on sputum ribonucleic acid (RNA) samples. Data is reported for the biomarkers of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), oncostatin M (OSM), cyclophilin, B actin, suppressor of cytokine signaling 3 (SOCS3), ETS proto-oncogene 2 transcription factor (ETS2), amphiregulin (AREG), cAMP responsive element modulator (CREM) and salt inducible kinase 1 (SNF1LK).

Cytokine sampling was planned of Day 7, 2-3 h post-dose to measure the IL10, IL-13 and TNFα for anti-inflammatory activity in sputum. However the data was not collected for this outcome measure.

Sputum samples were collected 2-3 h post-dose on Day 7 to measure the anti-inflammatory activities by SNF1LK nuclear intensity of staining and cytoplasmic intensity of staining of the cells.

Sputum samples were collected 2-3 h post-dose on Day 7 to measure the anti-inflammatory activities by the ratio of SNF1LK nuclear intensity of staining and cytoplasmic intensity of staining of the cells.