Effects of Colchicine in Non-Diabetic Adults With Metabolic Syndrome

Obesity affects one-third of the adult U.S. population and is a major risk factor for the
development of type 2 diabetes and cardiovascular disease. Mouse models and human data
suggest that obesity-induced chronic inflammation is one mechanism promoting
obesity-associated comorbid conditions. In obesity, innate immunity is activated by
circulating molecules such as fatty acids and cholesterol crystals bind to nucleotide-binding
oligomerization (NOD)-like receptor family, pyrin domain containing 3 (NLRP3) receptors in
adipocyte tissue macrophages (ATMs). This binding stimulates NLRP3 oligomerization,
inflammasome formation, and proinflammatory cytokine activation. The resultant inflammatory
cascade leads to insulin resistance and decreased pancreatic beta-cell reserve. It has been
proposed that the suppression of this chronic low-level inflammatory state may impede the
onset of diabetes and cardiovascular disease.

Recent studies have shown colchicine, a potent microtubule inhibitor commonly used for the
treatment of gout and some rare inflammatory conditions, disrupts intracellular localization
of NLRP3, thereby blocking inflammasome assembly. As there are limited medical therapies
proven effective to improve obesity-related metabolic dysregulation, we propose to determine
the efficacy of colchicine 0.6 mg twice daily in non-diabetic obese adults with metabolic
syndrome. We will conduct a randomized, double-blinded, placebo-controlled pilot trial of
colchicine in forty subjects. We will study changes in insulin resistance, beta-cell reserve,
and systemic inflammation. Using adipose tissue obtained from biopsies, we will also study
colchicine s local effects on inflammation and insulin resistance. Should results prove
promising, this pilot study will allow determination of the sample size needed for an
adequately powered study of the effects of colchicine in obese adults with metabolic
syndrome.

Seven patients with diet-controlled type 2 diabetes will be given open-label colchicine and
followed as described above. We also plan to perform baseline evaluations on 40 subjects who
are not eligible for the treatment protocol. This group will consist of non-obese adults,
obese adults who are not insulin-resistant, and adults with diet-controlled type 2 diabetes.

- INCLUSION CRITERIA FOR SUBJECTS RANDOMIZED TO COLCHICINE or PLACEBO::

Subjects will qualify for randomization to colchicine or placebo if they meet the following
criteria:

- Good general health. In general subjects should take no medications. However,
individuals taking medications for obesity-related co-morbid conditions, who have not
had changes in dosage for more than 3 months, may be included, at the discretion of
the principal investigator.

- Obesity, defined as a body mass index (BMI) greater than or equal to 30 kg/m^2, but
weight less than 450 lbs in order for subjects to undergo Dual-Energy X-ray
Absorptiometry (DXA) scanning.

- Age greater than or equal to 18 to 100 years.

- Metabolic Syndrome

--(Any 3 of the following 5):

- FPG greater than or equal to 100 mg/dl, or Impaired Glucose Tolerance (Glucose
greater than or equal to 140 mg/dl at 2 hours of OGTT)

- Triglycerides greater than or equal to 150 mg/dl, or on treatment

- Waist Circumference: Men greater than or equal to 40 in (greater than or equal to
102 cm); Women greater than or equal to 35 in (greater than or equal to 88 cm)

- Reduced HDL-C: Men < 40 mg/dl; Women < 50 mg/dl, or on treatment

- Hypertension: greater than or equal to 130 mmHg systolic, or greater than or
equal to 85 mmHg diastolic, or on treatment

- HOMA-IR greater than or equal to 2.6. Our goal is to enroll participants who have
pre-existing insulin resistance.

- hsCRP greater than or equal to 2.0 mg/L. We aim to recruit participants with increased
baseline level of inflammation. Individuals with hsCRP above 2.0 mg/L have been shown
to have an increased risk for cardiovascular events.

EXCLUSION CRITERIA FOR SUBJECTS RANDOMIZED TO COLCHICINE OR PLACEBO:

- Type 2 diabetes mellitus, as determined by either having:

- Clear clinical diagnosis of diabetes, such as a patient in a hyperglycemic crisis
or classic symptoms of hyperglycemia and a random plasma glucose greater than or
equal to 200 mg/dL

- Two of the following three:

- Fasting plasma glucose greater than or equal to 126 mg/dL

- Hemoglobin A1c greater than or equal to 6.5%

- An oral glucose tolerance test glucose concentration of greater than or
equal to 200 mg/dL at 2 hours.

- One of the above three criteria (bi.-biii.) meeting the T2DM cutoff on two
different days. If only one of the above three criteria (bi.-biii.) meet the T2DM
threshold during the Screening Visit, that test will be repeated on another day
to determine if the subject has T2DM or not. As per ADA guidelines, The diagnosis
[of T2DM] is made on the basis of the confirmed test.Moreover, because HbA1c has
been shown to be higher in African Americans (AA) as compared to other races for
the same glycemia, non-diabetic AA may be unfairly excluded by their HbA1c alone.
Therefore, for AA subjects, if their 2 hour OGTT and fasting glucoses are in the
non-diabetic range, and the HbA1c is < 7.0%, we will consider them non-diabetic.

- Presence of a significant active or chronic illness likely to limit life span and/or
increase risk of intervention, including renal (GFR less than or equal to 60
ml/min/1.73m2), cardiovascular, hepatic (other than obesity-related steatosis),
gastrointestinal, immunologic, endocrinologic (e.g. Cushing syndrome), pulmonary
(other than either asthma not requiring continuous medication or sleep apnea-related
disorders), or other disorders at the discretion of the investigators.

- Recent use of colchicine or anorexiant medications in the last 3 months.

- Known allergy to colchicine.

- Previous history of agranulocytosis, gout, or significant myositis.

- Females who are pregnant, planning to become pregnant, currently nursing an infant, or
have irregular menses, defined as cycles less than 21 days or greater than 45 days.

- Individuals who have current substance abuse or a psychiatric disorder or any other
condition that in the opinion of the investigators would impede competence,
compliance, or participation in the study.

- Subjects who regularly use prescription medications unrelated to the complications of
obesity, especially those known to affect enzymes involved in colchicine metabolism,
such as CYP3A4 or P-glycoprotein (P-gp) . Oral contraceptive use will be permitted,
provided the contraceptive has been used for at least two months before starting study
medication. The use of over-the-counter and prescription medications will be reviewed
on a case-by-case basis; depending on the medication, subjects who have continued to
take prescription medication or have stopped taking an exclusionary medication for at
least 3 months prior to study entry may be eligible .

- Participation in a formal weight loss program (e.g. Weight Watchers) or recent weight
change of more than 3% of body weight in the past two months.

- Use of anti-inflammatory medications (e.g. prednisone, NSAIDs) chronically or in the
last 7 days prior to fat biopsy.

- History of keloid formation.

- Current users of tobacco or nicotine products (e.g. nicotine patch, e-cigarettes).

INCLUSION CRITERIA FOR SUBJECTS WHO ARE EVALUATED BUT NOT ELIGIBLE FOR RANDOMIZATION:

Subjects will qualify for the Evaluation-only arm if they meet the following criteria:

- Good general health. In general subjects should take no medications. The use of
over-the-counter and prescription medications will be reviewed on a case-by-case
basis; depending on the medication, subjects who have continued to take prescription
medication or have stopped taking an exclusionary medication for at least 3 months
prior to study entry may be eligible.

- Weight less than 450 lbs in order for subjects to undergo Dual-Energy X-ray
Absorptiometry (DXA) scanning.

- Age 18 years to 100 years.

EXCLUSION CRITERIA FOR SUBJECTS WHO ARE EVALUATED BUT NOT ELIGIBLE FOR RANDOMIZATION:

- Type 2 diabetes mellitus that is not well controlled with diet alone: subjects taking
an antidiabetic medication (e.g. metformin, insulin, sulfonylureas, etc.) or having a
Hemoglobin A1c > 9.0%

- Presence of a significant active or chronic illness likely to limit life span and/or
increase risk of intervention, including renal (GFR less than or equal to 60
ml/min/1.73m2), cardiovascular, hepatic (other than obesity-related steatosis),
gastrointestinal, immunologic, endocrinologic (e.g. Cushing syndrome), pulmonary
(other than either asthma not requiring continuous medication or sleep apnea-related
disorders), or other disorders at the discretion of the investigators.

- Recent use of colchicine or anorexiant medications in the last 3 months.

- Females who are pregnant, planning to become pregnant, or are currently nursing an
infant.

- Individuals who have current substance abuse or a psychiatric disorder or any other
condition that in the opinion of the investigators would impede competence,
compliance, or participation in the study.

- Participation in a formal weight loss program (e.g. Weight Watchers) or recent weight
change of more than 3% of body weight in the past two months.

- Use of anti-inflammatory medications (e.g. prednisone, NSAIDs) chronically or in the
last 7 days prior to fat biopsy.

- History of keloid formation.

- Current users of tobacco or nicotine products (e.g. nicotine patch, e-cigarettes).

INCLUSION CRITERIA FOR SUBJECTS WITH DIET-CONTROLLED T2DM

Subjects will qualify for the Open Label arm if they meet the following criteria:

- A diagnosis of T2DM.

- Not on any diabetic/hypoglycemic agents

- Not having an alternate cause of hyperglycemia (e.g. T1DM, glucocorticoidinduced,
lipodystrophy, acromegaly, etc.)

- Hemoglobin A1c less than or equal to 9.0%

- Good general health. In general subjects should take no medications. The use of
over-thecounter and prescription medications will be reviewed on a case-by-case basis;
depending on the medication, subjects who have continued to take prescription
medication or have stopped taking an exclusionary medication for at least 3 months
prior to study entry may be eligible

- Age greater than or equal to 18 to 100 years.

- Obesity, defined as a body mass index (BMI) greater than or equal to 30 kg/m2, but
weight less than 450 lbs in order for subjects to undergo Dual-Energy X-ray
Absorptiometry (DXA) scanning.

- Metabolic Syndrome

(Any 3 of the following 5):

- FPG greater than or equal to100 mg/dl or Impaired Glucose Tolerance (Glucose greater
than or equal to 140 mg/dl at 2 hours of OGTT)

- Triglycerides greater than or equal to 150 mg/dl, or on treatment

- Waist Circumference: Men greater than or equal to 40 in (greater than or equal to 102
cm); Women greater than or equal to 35 in (greater than or equal to 88 cm)

- Reduced HDL-C: Men < 40 mg/dl; Women < 50 mg/dl, or on treatment

- Hypertension: greater than or equal to 130 mmHg systolic, or greater than or equal to
85 mmHg diastolic, or on treatment

- HOMA-IR greater than or equal to 2.6. Our goal is to enroll participants who have
pre-existing insulinresistance.

- hsCRP greater than or equal to 2.0 mg/L. We aim to recruit participants with
increased baseline level of inflammation. Individuals with hsCRP above 2.0 mg/L
have been shown to have an increased risk for cardiovascular events.

EXCLUSION CRITERIA FOR SUBJECTS WITH DIET-CONTROLLED T2DM

- T2DM that is not well controlled with diet alone: subjects will not be eligible if
they take an anti-diabetic medication (e.g. metformin, insulin, sulfonylurea, etc.),
or have HbA1c >9%.

- Presence of a significant active or chronic illness likely to limit life span and/or
increase risk of intervention, including renal (GFR less than or equal to 30
ml/min/1.73m2), cardiovascular, hepatic (other than obesity-related steatosis),
gastrointestinal, immunologic, endocrinologic (e.g. Cushing syndrome), pulmonary
(other than either asthma not requiring continuous medication or sleep apnea-related
disorders), or other disorders at the discretion of the investigators.

- Recent use of colchicine or anorexiant medications in the last 3 months.

- Known allergy to colchicine.

- Previous history of agranulocytosis, gout, or significant myositis.

- Females who are pregnant, planning to become pregnant, currently nursing an infant, or
have irregular menses, defined as cycles less than 21 days or greater than 45 days.

- Individuals who have current substance abuse or a psychiatric disorder or any other
condition that in the opinion of the investigators would impede competence,
compliance, or participation in the study.

- Subjects who regularly use prescription medications unrelated to the complications of
obesity, especially those known to affect enzymes involved in colchicine metabolism,
such as CYP3A4 or P-glycoprotein (P-gp). Oral contraceptive use will be permitted,
provided the contraceptive has been used for at least two months before starting study
medication. The use of over-the-counter and prescription medications will be reviewed
on a case-by-case basis; depending on the medication, subjects who have

continued to take prescription medication or have stopped taking an exclusionary medication
for at least 3 months prior to study entry may be eligible.

- Participation in a formal weight loss program (e.g. Weight Watchers) or recent weight
change of more than 3% of body weight in the past two months.

- Use of anti-inflammatory medications (e.g. prednisone, NSAIDs) chronically or in the
last 7 days prior to fat biopsy.

- History of keloid formation.

- Current users of tobacco or nicotine products (e.g. nicotine patch, e-cigarettes).