Soluble Epoxide Hydrolase Inhibition and Insulin Resistance

Subjects are randomized to treatment with the sEH inhibitor GSK2256294 (10mg/day) or matching
placebo for one week. On the seventh day of drug treatment, subjects will report to the CRC
in the morning after an overnight fast to undergo a hyperinsulinemic-euglycemic clamp with
adipose tissue biopsies.

During the Hyperinsulinemic-euglycemic clamp, insulin will be infused for 2 hours at low dose
(20 mU/m2/min) and 2 hours at high dose (80 mU/m2/min) to assess insulin sensitivity. The
Glucose Infusion Rate (GIR) will be adjusted to maintain glucose near 95 mg/dL. The average
GIR during the final 30 minutes of the high dose period will be used as the measure of
insulin sensitivity.

After completion of the study day, subjects will undergo a seven-week washout from study drug
and then receive the opposite drug for one week. On the seventh day of treatment they will
report to the CRC after an overnight fast and repeat the study day protocol.

Inclusion Criteria:

1. Men and women,

2. Age 21 to 50 years, and

3. Pre-diabetes as defined by

1. Fasting plasma glucose 100-125 mg/dL, or

2. Two-hour plasma glucose 140-199 mg/dL, or

3. HbA1c 5.7-6.4%

4. BMI ≥ 30 kg/m2, inclusive

5. For female subjects, the following conditions must be met:

1. Postmenopausal status for at least one year, or

2. Status-post surgical sterilization, or

3. If of childbearing potential, utilization of adequate birth control and
willingness to undergo serum β-hcg testing prior to drug treatment and on every
study day.

Exclusion Criteria:

1. Diabetes type 1 or type 2, as defined by a fasting plasma glucose of 126 mg/dL or
greater, a two-hour plasma glucose of 200 mg/dL or greater, a HbA1c >6.4%, or the use
of anti-diabetic medication

2. Subjects who have participated in a weight-reduction program during the last six month
or whose weight has increased or decreased more than two kg over the preceding six
months

3. Resistant hypertension, defined as hypertension requiring the administration of more
than three anti-hypertensive agents including a diuretic to achieve control

4. Use of spironolactone

5. Pregnancy or breast-feeding

6. Any history of smoking

7. Any history of cancer including skin cancer, any history of a precancerous lesion,
abnormal PSA, or lack of screening adherent to American Cancer Society Guidelines for
the Early Detection of Cancer

8. Cardiovascular disease such as myocardial infarction within six months prior to
enrollment, presence of angina pectoris, significant arrhythmia, congestive heart
failure (left ventricular hypertrophy acceptable), deep-vein thrombosis, pulmonary
embolism, second- or third-degree heart block, mitral valve stenosis, aortic stenosis,
or hypertrophic cardiomyopathy

9. Abnormal corrected QT interval on screening ECG (QTc).

10. Treatment with anticoagulants

11. History of serious neurologic disease such as cerebral hemorrhage, stroke, or
transient ischemic attack

12. History or presence of immunological or hematological disorders

13. Diagnosis of asthma requiring regular inhaler use

14. Clinically significant gastrointestinal impairment that could interfere with drug
absorption

15. Impaired hepatic function (aspartate amino transaminase [AST] and/or alanine amino
transaminase [ALT] >3.0 x upper limit of normal range)

16. History of gastrointestinal bleed

17. Estimated glomerular filtration rate (eGFR)<60 mL/min/1.73 m2 or with an
albumin-to-creatinine ratio (UACR) >300µg/mg, where eGFR is determined by the
four-variable Modification of Diet in Renal Disease (MDRD) equation, where serum
creatinine is expressed in mg/dL and age in years: eGFR (mL/min/1.73m2)=186 •
Scr-1.154 • age-0.203 • (1.212 if black) • (0.742 if female)

18. Hematocrit <35%

19. Any underlying or acute disease requiring regular medication which could possibly pose
a threat to the subject or make implementation of the protocol or interpretation of
the study results difficult

20. Treatment with chronic systemic glucocorticoid therapy

21. Treatment with lithium salts

22. History of alcohol or drug abuse

23. Treatment with any investigational drug in the month preceding the study

24. Mental conditions rendering a subject unable to understand the nature, scope, and
possible consequences of the study

25. Inability to comply with the protocol, e.g., uncooperative attitude, inability to
return for follow-up visits, and unlikelihood of completing the study