A Clinical Study to Assess the Safety, Tolerability, and Activity of Oral SRT2104 Capsules Administered for 28 Days to Subjects With Type 2 Diabetes Mellitus

The areas under the glucose concentration time curve from time 0 (dosing) to 2 hours post-dose (AUC 0-2h, glucose) obtained during an oral glucose tolerance test (OGTT) on days 28 and 42.

AUC 0-2h, glucose was obtained pre-dose (Day -1), at the end of the dosing period (Day 28), and at end-of-study (Day 42). Blood samples were obtained prior to administration of 75 g of oral glucose which was administered as standard glucose beverage after an overnight fast, within 15 minutes. A peripheral blood sample was drawn from a peripheral arm vein prior to the participant consuming the beverage. Additional blood samples were drawn at 30, 60, 90, and 120 minutes after the participant had consumed the glucose beverage for the measurement of glucose, insulin and C-peptide concentrations. Blood glucose concentrations were determined with a glucose analyzer. On Day 28, on which the participant received also investigational product, the OGTT had to be performed prior to consumption of standard meal and administration of investigational product.

Time frame: Day 28 and Day 42

The areas under the serum insulin concentration time curve from time 0 (dosing) to 2 hours post-dose (AUC 0-2h, insulin) obtained during OGTT, on days 28 and 42.

AUC 0-2h, insulin was obtained pre-dose (Day -1), at the end of the dosing period (Day 28), and at end-of-study (Day 42). Blood samples were obtained prior to administration of 75 g of oral glucose which was administered as standard glucose beverage after an overnight fast, within 15 minutes. A peripheral blood sample was drawn from a peripheral arm vein prior to the participant consuming the beverage. Additional blood samples were drawn at 30, 60, 90, and 120 minutes after the participant had consumed the glucose beverage for the measurement of glucose, insulin and C-peptide concentrations. Blood glucose concentrations were determined with a glucose analyzer. On Day 28, on which the participant received also investigational product, the OGTT had to be performed prior to consumption of standard meal and administration of investigational product.unit of measurement was microunits×hours per milliliter

Time frame: Day 28 and 42

Measurement of insulin sensitivity consisting of the insulin sensitivity index (ISIest) obtained during OGTT.

Stumvoll Index: ISIest was obtained pre-dose (Day -1), at the end of the dosing period (Day 28), and at end-of-study (Day 42). Blood samples were obtained prior to administration of 75 g of oral glucose which was administered as standard glucose beverage after an overnight fast, within 15 minutes. ISI is based on insulin and glucose levels in a fasting state during OGTT and is calculated as: ISI (Matsuda) = 10000/√ G0 X I0 X Gmean X Imean where G0 - fasting plasma glucose (mg/dL) I0 - fasting plasma insulin (milliinternational units (miU)/L) Gmean - mean plasma glucose during OGTT (mg/dL) Imean - mean plasma insulin during OGTT (mIU/L). Unit of measure was (Deciliter × liter)(mg × milliinternational units)

Time frame: Up to Day 42

Measurement of insulin sensitivity consisting of the composite whole body sensitivity index (ISIcomposite) obtained during OGTT

ISI is based on insulin and glucose levels in a fasting state during an oral glucose tolerance test (OGTT) and is calculated as: ISI (Matsuda) = 10000/√ G0 X I0 X Gmean X Imean where G0 - fasting plasma glucose (mg/dL) I0 - fasting plasma insulin (mIU/L) Gmean - mean plasma glucose during OGTT (mg/dL) Imean - mean plasma insulin during OGTT (mIU/L). ISIcomposite was obtained pre-dose (Day -1), at the end of the dosing period (Day 28), and at end-of-study (Day 42).

Time frame: Up to Day 42

Measurement of insulin sensitivity consisting of the hepatic insulin sensitivity (HIRI) obtained during OGTT

HIRI was obtained pre-dose (Day -1), at the end of the dosing period (Day 28), and at end-of-study (Day 42). Blood samples were obtained prior to administration of 75 g of oral glucose which was administered as standard glucose beverage after an overnight fast, within 15 minutes. A peripheral blood sample was drawn from a peripheral arm vein prior to the participant consuming the beverage. Additional blood samples were drawn at 30, 60, 90, and 120 minutes after the participant had consumed the glucose beverage for the measurement of glucose, insulin and C-peptide concentrations. Blood glucose concentrations were determined with a glucose analyzer. On Day 28, on which the participant received also investigational product, the OGTT had to be performed prior to consumption of standard meal and administration of investigational product. Unit of measure was millimole × hour per liter)×(milliunits × hour per liter \[mmol\*h/L)\*(mU\*h/L\]

Time frame: Up to Day 42

Measurement of insulin sensitivity consisting of the muscular insulin sensitivity (MISI) obtained during OGTT.

MISI was obtained pre-dose (Day -1), at the end of the dosing period (Day 28), and at end-of-study (Day 42). Blood samples were obtained prior to administration of 75 g of oral glucose which was administered as standard glucose beverage after an overnight fast, within 15 minutes. A peripheral blood sample was drawn from a peripheral arm vein prior to the participant consuming the beverage. Additional blood samples were drawn at 30, 60, 90, and 120 minutes after the participant had consumed the glucose beverage for the measurement of glucose, insulin and C-peptide concentrations. Blood glucose concentrations were determined with a glucose analyzer. On Day 28, on which the participant received also investigational product, the OGTT had to be performed prior to consumption of standard meal and administration of investigational product. Unit of measure was milligram per deciliter per minute per milliunits per liter (mg/dL/min/mU/L).

Time frame: Up to Day 42

Measurement of insulin sensitivity consisting of the insulin resistance index obtained during OGTT

Measurement of insulin sensitivity consisting of the insulin resistance index obtained during OGTT was not collected.

Time frame: Up to Da y 42

Measurement of insulin sensitivity consisting of the beta cell function based on homeostasis model assessment :HOMA-IR, obtained during OGTT

HOMA-IR was obtained pre-dose (Day -1), at the end of the dosing period (Day 28), and at end-of-study (Day 42). Blood samples were obtained prior to administration of 75 g of oral glucose which was administered as standard glucose beverage after an overnight fast, within 15 minutes. A peripheral blood sample was drawn from a peripheral arm vein prior to the participant consuming the beverage. Additional blood samples were drawn at 30, 60, 90, and 120 minutes after the participant had consumed the glucose beverage for the measurement of glucose, insulin and C-peptide concentrations. Blood glucose concentrations were determined with a glucose analyzer. On Day 28, on which the participant received also investigational product, the OGTT had to be performed prior to consumption of standard meal and administration of investigational product.

Time frame: Up to Day 42

Measurement of insulin sensitivity consisting of the beta cell function based on homeostasis model assessment : HOMA-%B, obtained during OGTT.

HOMA-%B was obtained pre-dose (Day -1), at the end of the dosing period (Day 28), and at end-of-study (Day 42). Blood samples were obtained prior to administration of 75 g of oral glucose which was administered as standard glucose beverage after an overnight fast, within 15 minutes. A peripheral blood sample was drawn from a peripheral arm vein prior to the participant consuming the beverage. Additional blood samples were drawn at 30, 60, 90, and 120 minutes after the participant had consumed the glucose beverage for the measurement of glucose, insulin and C-peptide concentrations. Blood glucose concentrations were determined with a glucose analyzer. On Day 28, on which the participant received also investigational product, the OGTT had to be performed prior to consumption of standard meal and administration of investigational product.

Time frame: Up to Day 42

Measurement of insulin sensitivity consisting of the quantitative insulin sensitivity check index (QUICKI) obtained during OGTT.

QUICKI was obtained pre-dose (Day -1), at the end of the dosing period (Day 28), and at end-of-study (Day 42). Blood samples were obtained prior to administration of 75 g of oral glucose which was administered as standard glucose beverage after an overnight fast, within 15 minutes. A peripheral blood sample was drawn from a peripheral arm vein prior to the participant consuming the beverage. Additional blood samples were drawn at 30, 60, 90, and 120 minutes after the participant had consumed the glucose beverage for the measurement of glucose, insulin and C-peptide concentrations. Blood glucose concentrations were determined with a glucose analyzer. On Day 28, on which the participant received also investigational product, the OGTT had to be performed prior to consumption of standard meal and administration of investigational product.

Time frame: Up to Day 42

Determination of sensitivity and metabolic clearance consisting of the insulin sensitive index (SI), Obtained during an hyperinsulinemic euglycemic glucose clamp (HEGC)

A HEGC was carried out at Day 0, 29 and 43. HEGC was performed in participants who developed coagulation abnormalities during the study. Prior to the start of the HEGC procedure, participants were instructed to refrain from alcohol and caffeine for at least 48 hours before each inpatient visit and from exercise at least 24 hours before each inpatient visit. In the evening of the day prior to the HEGC procedure, the participants received a standard dinner at approximately 19:00 hours. Afterwards, the participants remained fasting until the end of the HEGC procedure at approximately 16:00 hours the next day. The HEGC procedure consisted of three phases: Blood glucose adjustment, Insulin infusion step 1, Insulin infusion step 2. The last 60 min of the two insulin infusion steps were defined as "steady state period 1" (SS1) and "steady state period 2" (SS2). unit of measure was Milliliter per minute permeters squared per micro units (mL/min/m2/mcU/mL).

Time frame: Up to Day 43

Determination of insulin sensitivity during first (M1) and second (M2) insulin infusion step.

M1 and M2 was carried out at Day 0, 29 and 43. HEGC was performed in participants who developed coagulation abnormalities during the study. Prior to the start of the HEGC procedure, participants were instructed to refrain from alcohol and caffeine for at least 48 hours before each inpatient visit and from exercise at least 24 hours before each inpatient visit. In the evening of the day prior to the HEGC procedure, the participants received a standard dinner at approximately 19:00 hours. Afterwards, the participants remained fasting until the end of the HEGC procedure at approximately 16:00 hours the next day. The HEGC procedure consisted of three phases: Blood glucose adjustment, Insulin infusion step 1, Insulin infusion step 2. The last 60 min of the two insulin infusion steps were defined as "steady state period 1" (SS1) and "steady state period 2" (SS2). Unit of measure was milligrams per minute per kilograms (mg/min/kg).

Time frame: Up to Day 43

Insulin sensitivity in relation to mean insulin infusion concentration (M/I1 and M/I2) ,Obtained during HEGC

M/I1 and M/I2 was carried out at Day 0, 29 and 43. HEGC was performed in participants who developed coagulation abnormalities during the study. Prior to the start of the HEGC procedure, participants were instructed to refrain from alcohol and caffeine for at least 48 hours before each inpatient visit and from exercise at least 24 hours before each inpatient visit. In the evening of the day prior to the HEGC procedure, the participants received a standard dinner at approximately 19:00 hours. Afterwards, the participants remained fasting until the end of the HEGC procedure at approximately 16:00 hours the next day. The HEGC procedure consisted of three phases: Blood glucose adjustment, Insulin infusion step 1, Insulin infusion step 2. The last 60 min of the two insulin infusion steps were defined as "steady state period 1" (SS1) and "steady state period 2" (SS2). milligrams per minute per kilograms per microunits per milliliter (mg/min/kg/μU/mL).

Time frame: Up to Day 43

Metabolic clearance rate during first and second insulin infusion step (MCR1, MCR2) on Day 29, and 43

MCR1, MCR2 was carried out at Day 0, 29 and 43. HEGC was performed in participants who developed coagulation abnormalities during the study. Prior to the start of the HEGC procedure, participants were instructed to refrain from alcohol and caffeine for at least 48 hours before each inpatient visit and from exercise at least 24 hours before each inpatient visit. In the evening of the day prior to the HEGC procedure, the participants received a standard dinner at approximately 19:00 hours. Afterwards, the participants remained fasting until the end of the HEGC procedure at approximately 16:00 hours the next day. The HEGC procedure consisted of three phases: Blood glucose adjustment, Insulin infusion step 1, Insulin infusion step 2. The last 60 min of the two insulin infusion steps were defined as "steady state period 1" (SS1) and "steady state period 2" (SS2). Unit of measure was kilograms per milliliters per kilo calories (kg/mL/kcal)

Time frame: Up to Day 43

Variables of metabolic clearance rate including M1ree and M2ree.

M1ree and M2ree was carried out at Day 0, 29 and 43. HEGC was performed in participants who developed coagulation abnormalities during the study. Prior to the start of the HEGC procedure, participants were instructed to refrain from alcohol and caffeine for at least 48 hours before each inpatient visit and from exercise at least 24 hours before each inpatient visit. In the evening of the day prior to the HEGC procedure, the participants received a standard dinner at approximately 19:00 hours. Afterwards, the participants remained fasting until the end of the HEGC procedure at approximately 16:00 hours the next day. The HEGC procedure consisted of three phases: Blood glucose adjustment, Insulin infusion step 1, Insulin infusion step 2. The last 60 min of the two insulin infusion steps were defined as "steady state period 1" (SS1) and "steady state period 2" (SS2).

Time frame: Up to Day 43

Variables of metabolic clearance rate including MCR1ree and MCR2ree .

MCR1ree and MCR2ree was carried out at Day 0, 29 and 43. HEGC was performed in participants who developed coagulation abnormalities during the study. Prior to the start of the HEGC procedure, participants were instructed to refrain from alcohol and caffeine for at least 48 hours before each inpatient visit and from exercise at least 24 hours before each inpatient visit. In the evening of the day prior to the HEGC procedure, the participants received a standard dinner at approximately 19:00 hours. Afterwards, the participants remained fasting until the end of the HEGC procedure at approximately 16:00 hours the next day. The HEGC procedure consisted of three phases: Blood glucose adjustment, Insulin infusion step 1, Insulin infusion step 2. The last 60 min of the two insulin infusion steps were defined as "steady state period 1" (SS1) and "steady state period 2" (SS2).

Time frame: Up to Day 43

Variables of metabolic clearance rate including M/I1ree and M/I2ree.

M/I1ree and M/I2ree was carried out at Day 0, 29 and 43. HEGC was performed in participants who developed coagulation abnormalities during the study. Prior to the start of the HEGC procedure, participants were instructed to refrain from alcohol and caffeine for at least 48 hours before each inpatient visit and from exercise at least 24 hours before each inpatient visit. In the evening of the day prior to the HEGC procedure, the participants received a standard dinner at approximately 19:00 hours. Afterwards, the participants remained fasting until the end of the HEGC procedure at approximately 16:00 hours the next day. The HEGC procedure consisted of three phases: Blood glucose adjustment, Insulin infusion step 1, Insulin infusion step 2. The last 60 min of the two insulin infusion steps were defined as "steady state period 1" (SS1) and "steady state period 2" (SS2).

Time frame: Up to Day 43

Mean steady state concentration Free fatty acids (FFA), and glycerols

Mean steady state concentration Free fatty acids and glycerols was carried out at Day 0, 29 and 43. HEGC was performed in participants who developed coagulation abnormalities during the study. Prior to the start of the HEGC procedure, participants were instructed to refrain from alcohol and caffeine for at least 48 hours before each inpatient visit and from exercise at least 24 hours before each inpatient visit. In the evening of the day prior to the HEGC procedure, the participants received a standard dinner at approximately 19:00 hours. Afterwards, the participants remained fasting until the end of the HEGC procedure at approximately 16:00 hours the next day. The HEGC procedure consisted of three phases: Blood glucose adjustment, Insulin infusion step 1, Insulin infusion step 2. The last 60 min of the two insulin infusion steps were defined as "steady state period 1" (SS1) and "steady state period 2" (SS2).

Time frame: Up to Day 43

Mean C-peptide concentration (mean C-peptideSS1 and C-peptideSS2) obtained during HEGC

Mean C-peptideSS1 and C-peptideSS2 was carried out at Day 0, 29 and 43. HEGC was performed in participants who developed coagulation abnormalities during the study. Prior to the start of the HEGC procedure, participants were instructed to refrain from alcohol and caffeine for at least 48 hours before each inpatient visit and from exercise at least 24 hours before each inpatient visit. In the evening of the day prior to the HEGC procedure, the participants received a standard dinner at approximately 19:00 hours. Afterwards, the participants remained fasting until the end of the HEGC procedure at approximately 16:00 hours the next day. The HEGC procedure consisted of three phases: Blood glucose adjustment, Insulin infusion step 1, Insulin infusion step 2. The last 60 min of the two insulin infusion steps were defined as "steady state period 1" (SS1) and "steady state period 2" (SS2).

Time frame: Up to Day 43

Number of participants with all adverse events (AEs) and serious adverse events (SAEs)

Data for number of participants who presented one or more adverse events (serious or non serious) was reported. An AE was defined as any untoward medical occurrence (MO) in a participant temporally associated with the use of a medicinal product (MP), whether or not considered related to the MP and can therefore be any unfavorable and unintended sign (including an abnormal laboratory finding), symptom, or disease temporally associated with its use. The SAE was any untoward MO that, at any dose, results in death, life threatening, persistent or significant disability/incapacity, results in or prolongs inpatient hospitalization, congenital abnormality or birth defect, that may not be immediately life-threatening or result in death or hospitalization but may jeopardize the participant or may require medical or surgical intervention to prevent one of the other outcomes listed in this definition.

Time frame: Up to 28 days

Number of participants with hypoglycemic events

Symptomatic, symptomatic and Severe Symptomatic hypoglycemia definitions were used to analyze and report hypoglycemic events. Symptomatic hypoglycemic episodes were collected from Day 1 to Day 28 with the assistance of participants diaries. Symptomatic hypoglycemia and severe symptomatic hypoglycemia was recorded in CRFs as AEs in all cases.

Time frame: Up to Day 28

Number of participants with abnormal Physical examination findings

A complete physical examination, including an ophthalmological examination to rule out diabetic neuropathy, was performed at screening. A symptom-driven, directed physical examination was performed also pre dosing (Day-2), during the dosing period (Day 14 and 27) and after dosing (Day 41). Number of participants with abnormal physical findings was reported.

Time frame: Up to Day 41

Change from baseline in Vital signs-systolic blood pressure (SBP) and diastolic blood pressure (DBP).

SBP and DBP were investigated at screening, pre-dosing (Day -2), during the dosing period (Day 1, 14 and 27) and at end of dosing follow-up period/end-of-study (Day 41 and 43). At Day 1 the vital signs were collected pre-dose (baseline values) and 3 hours post-dose. Day 0 was Baseline and change from Baseline was a value at study day subtracted by value at baseline.

Time frame: Up to Day 43

Change from baseline in Vital signs-Heart rate

Heart rate was investigated at screening, pre-dosing (Day -2), during the dosing period (Day 1, 14 and 27) and at end of dosing follow-up period/end-of-study (Day 41 and 43). At Day 1 the vital signs were collected pre-dose (baseline values) and 3 hours post-dose. Day 0 was Baseline and change from Baseline was a value at study day subtracted by value at baseline.

Time frame: Up to Day 43

Change from baseline in Vital signs-Respiratory rate

Respiratory rate was investigated at screening, pre-dosing (Day -2), during the dosing period (Day 1, 14 and 27) and at end of dosing follow-up period/end-of-study (Day 41 and 43). At Day 1 the vital signs were collected pre-dose (baseline values) and 3 hours post-dose. Day 0 was Baseline and change from Baseline was a value at study day subtracted by value at baseline.

Time frame: Up to Day 43

Change from baseline in Vital signs-Temperature.

Time frame: Up to Day 43

Number of participants with abnormal electrocardiogram (ECG) recordings

A 12-lead ECG was recorded at screening, pre-dose (Day -2), at the beginning and end of the dosing period (Day 1 and 27) and at the end of the dosing follow-up period/end-of study (Day 41 and 43). Number of participants with abnormal ECG findings was reported.

Time frame: Up to Day 43

Mean hematology parameters including white blood cell (WBC) and platelets count

Blood samples for assessment of WBC and platelet count was collected at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean hematology parameters including WBC Differential count and Red cell distribution width (RDW)

Blood samples for assessment of WBC differential count and RDW was collected at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean hematology parameters including Hemoglobin (Hb) and Mean corpuscular hemoglobin concentration (MCHC)

Blood samples for assessment of Hb and MCHC was collected at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean hematology parameters including: hematocrit count

Blood samples for assessment of hematocrit count was collected at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean hematology parameters including corpuscular volume (MCV)

Blood samples for assessment of MCV was collected at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean hematology parameters including corpuscular hemoglobin (MCH)

Blood samples for assessment of MCH was collected at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean hematology parameters including Red Blood Cells (RBCs) count

Blood samples for assessment of RBC count was collected at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Summary of urinalysis data: PH

Samples for pH assessment were collected at screening, Day -1, 14, 28 and 42. Unscheduled and control assessments were not included in the analysis. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Number of participants with urinalysis

Data for urinalysis was collected at screening, Day -1, 14, 28 and 42. Data for participants with positive and negative urinalysis result was presented. Unscheduled and control assessments were not included in the analysis.

Time frame: Up to Day 42

Mean clinical chemistry parameters including calcium, chloride, magnesium, potassium, sodium, bicarbonate, phosphate, glucose, urea and blood urea

Clinical chemistry assessment for calcium, chloride, magnesium, potassium, sodium, bicarbonate, phosphate, glucose, urea ,blood urea was done at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean clinical chemistry parameters including direct bilirubin, indirect bilirubin, total bilirubin ,uric acid

Clinical chemistry assessment for direct bilirubin, indirect bilirubin, total bilirubin and uric acid was done at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean clinical chemistry parameters including alkaline phosphatase (ALP), Alanine aminotransferase (ALT), Aspartate aminotransaminase (AST), Lactate dehydrogenase (LDH), Creatine kinase

Clinical chemistry assessment for ALP, ALT, AST, LDH and creatine kinase was done at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean clinical chemistry parameters including albumin

Clinical chemistry assessment for albumin was done at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42

Mean clinical chemistry parameters including serum creatinine

Clinical chemistry assessment for serum creatinine was done at screening, Day -1, 14, 28 and 42. Data for mean and standard deviation was presented.

Time frame: Up to Day 42