Effect of SRT2104 on endotoxin-induced inflammation.

Rationale:
Activation of SIRT1 (silent information regulator transcript) results in inhibition of
inflammation. SRT2104 is a potent small molecule activator of SIRT1 that has been found to
inhibit systemic inflammation induced by intravenous injection of lipopolysaccharide (LPS) in
mice. SRT2104 may be a novel compound in the treatment of inflammatory disorders in man.

Primary objective:
To determine if (a) single or (b) 7 daily doses of SRT2104 attenuates
the inflammatory response in normal healthy male subjects after exposure to low-dose
endotoxin (LPS).

Secondary objectives:
(1) To determine PK of SRT2104 in normal healthy male subjects
exposed to low-dose endotoxin (LPS); (2) To determine the safety profile of SRT2104 in
healthy male subjects exposed to low-dose endotoxin (LPS).

Exploratory objectives:
To determine the effect of SRT2104 on other parameters following
low-dose endotoxin (LPS) exposure in humans e.g., lipid profile, serum amyloid
phospholipids, metabolic profiles and gene expression analysis etc.

Study design:
Double-blind, placebo-controlled intervention study.

Study population:
24 healthy male subjects (18-35 years).

Intervention:
This study consists of three treatment arms (N = 8 per arm). Subjects in arm
one will receive once daily doses of SRT2104 (2.0 g/day) for seven consecutive days.
Subjects in arm two will receive once daily doses of placebo for seven consecutive days.
Subjects in arm three will receive once daily doses of placebo on Days 1-6 and a single dose
of SRT2104 (2.0 g/day) on Day 7. Test material administration (SRT2104 or placebo) will
occur on all days during the study, i.e., from Day 1 to Day 7 inclusive. Subjects will take
SRT2104 or placebo approximately 15 minutes following consumption of a standardized
meal on all dosing days. Subjects must wait at least 1-2 hours after dosing before
consuming additional calories. On day 7, all subjects will be given an intravenous dose of
LPS (standardized LPS preparation provided by the National Institutes of Health (NIH),
Bethesda, USA; 4 ng/kg body weight). LPS will be given 3 hours after dosing.

Main study parameters/endpoints:
Primary study endpoints include clinical signs and
symptoms and laboratory parameters for inflammation (cytokines, activation of leukocytes,
coagulation and vascular endothelium). Secondary endpoints include pharmacokinetics and
safety recordings. Exploratory endpoints include lipid profiles, acute phase proteins,
metabolic profiles and gene expression analysis from white blood cells.

Nature and extent of the burden and risks associated with participation, benefit and
group relatedness:
The burden of this study involves a screening visit, two 2-nights
admissions to the clinical research unit, the ingestion of SRT2104 and the intravenous
injection of LPS. Intravenous LPS induces a transient influenza-like/inflammatory syndrome
in humans consisting of chills, fever, nausea, headache and muscle ache. SRT2104 has
been well-tolerated following both the single and multiple dose periods at all dose levels
investigated. In the current study the potential anti-inflammatory effects of SRT2104 will be
tested in the human endotoxemia model. The risks are low, whereas the study will generate
information regarding the anti-inflammatory activity of SRT2104. This knowledge may be of
future benefit to patients with inflammatory diseases.

Activation of SIRT1 (silent information regulator transcript) results in inhibition of
inflammation in humans.

1. Medication or placebo intake:
Day 1 – day 7, 7:00 AM.

2. Endotoxin (LPS, 4 ng/kg) administration:
Day 7 at 10 AM (3 hours after last medication or placebo intake).

3. Measurements ( Blood, urine, ECG, physical examination):
Day 1-2 and day 6-8.

This study consists of three treatment arms (N = 8 per arm). Subjects in arm one will receive once daily doses of SRT2104 (2.0 g/day) for seven consecutive days. Subjects in arm two will receive once daily doses of placebo for seven consecutive days. Subjects in arm three will receive once daily doses of placebo on Days 1-6 and a single dose of SRT2104 (2.0 g/day) on Day 7. Test material administration (SRT2104 or placebo) will occur on all days during the study, i.e., from Day 1 to Day 7 inclusive. Subjects will take SRT2104 or placebo approximately 15 minutes following consumption of a standardized meal on all dosing days. Subjects must wait at least 1-2 hours after dosing before consuming additional calories. On day 7, all subjects will be given an intravenous dose of LPS (standardized LPS preparation provided by the National Institutes of Health (NIH), Bethesda, USA; 4 ng/kg body weight). LPS will be given 3 hours after dosing.