Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and Obese Subjects
Phase: NA
Conditions: Overweight, Obese
Sponsor: Tufts University

Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Version Date: 3/2/2018
Principal Investigator: Mohsen Meydani
Co-Investigator: Susan Roberts
Study Physician: Lisa Ceglia
Study Title: Body Fat Reducing Efect and Safety of Enzymatically Modifed 
Isoquercitrin in Overweight and Obese Subjects
I.
Aim and Hypotheses
Specifc Aim 1: To compare the body fat reducing efect of 360mg/day 
enzymatically modifed isoquercitrin (EMIQ) with that of a placebo.
Hypothesis 1: Compared to a placebo, participants receiving EMIQ 
supplementation will have a greater reduction in total body fat as evidenced by 
measures of whole body DXA scan, skinfold thickness, waist to hip 
circumference ratio, resting metabolic rate, plasma adiponectin levels, physical 
activity, and dietary intake.
Specifc Aim 2: To determine the safety of long term intake of EMIQ.
Hypothesis 2: Long-term intake of EMIQ has no ill efect on participant 
health as evidenced by measures of heart, liver, kidney function, and 
psychological state.
II.
Background and Rationale
Dietary quercetin is a natural bioactive polyphenol with potential health benefts
and the capacity to prevent development of various diseases including those of 
the cardiovascular system, central nervous system and others [ 1-3
 
 ]. It is widely 
distributed in various plant foods such as onions, broccoli, concord grapes, 
apples and more [4]. In plants, quercetin is present as conjugated with diferent 
forms of glucosides [5], which afect their bioavailability. The bioavailability of 
diferent forms of quercetin has been the subject of several investigations [6]. 
Enzymatically-modifed isoquercitrin (EMIQ) is a mixture of quercetin 
monoglucoside and its -oligoglucosides, which mainly consist of quercetin-3-O-
-glucoside (Q3G) and -glucosyl derivatives with 1-7 of additional linear 
glucose moieties [7, 8]. Supplementing the diet of apoE-defcient mice with 
EMIQ increased quercetin levels and suppressed atherogenesis in this mouse 
model of human atherosclerosis[9]. In human trials, the bioavailability of EMIQ 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
has been shown to be 17-fold higher than quercetin aglycone [10]. After 
efficient absorption of EMIQ, quercetin is released, and it exerts its biological 
activity such as reducing body fat, improving cardiovascular health and 
reducing blood pressure. 
EMIQ has been shown to be efective in reducing body fat in Japanese people 
who have diferent dietary habits and lifestyle from that of a person in the 
United States. Thus, this study is designed to investigate if EMIQ is also 
efective in reducing body fat in subjects with a western lifestyle. 
Quercetin, according to Seo et al [11] in addition to being a very strong 
antioxidant, prevents adipogenesis by regulating two sets of enzymes, a) by 
upregulating adipose triglyceride lipase (ATGL) and hormone sensitive lipase 
(HSL) expression and, b) by downregulating fatty acid synthesis (FAS), 
lipoprotein lipase (LPL) and adipocyte fatty acid-binding protein (aP2) 
expression, through expression of transcription factors such as C/EBPα, PPARγ, 
SREBP-1. These observations suggest that quercetin has therapeutic potential 
for body fat reduction through regulating the expression of transcriptional 
factors and enzymes associated with adipogenesis. Dong et al [12] also 
reported that dietary quercetin reduces high fat diet-induced body weight gain, 
fat deposition, and insulin resistance and suppresses glucose uptake by adipose
tissue. They reported that these efects of dietary quercetin is mediated through
quercetin activating of AMPK/ACC signaling pathway which promotes fatty acid 
-oxidation and suppresses fatty acid esterifcation into triglyceride in 
adipocyte.  Administration of 275 mg/d for 12 weeks reduced total body fat, 
visceral fat and subcutaneous fat in Japanese subjects.  EMIQ is classifed by the
Food and Drug Administration (FDA) as Generally Recognized As Safe (GRAS). 
Thus, it should be considered a safe compound in order to investigate its 
potential biological activities in humans.   
III.
Research Plan
A. Experimental Design
This is a randomized, double-blind, placebo controlled study.
B. Sample Size and Statistical Analyses
50 subjects will be required to complete this 12 week study (though 150 will be 
screened and 64 will be enrolled to ensure completion of 50 subjects). Subjects 
will be randomized 1:1 into either the treatment (EMIQ) or placebo group.  The 
study physician, Dr. Lisa Ceglia, will hold the code to the randomization which 
will be revealed upon completion of the study or if adverse event occurs. 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
To calculate number of subjects needed per group and provide 80% power at 
=0.05 we used mean diferences and standard deviation (EMIQ: 19224.62 and
Placebo: 1212.87) of whole abdominal fat loss from an early published study 
conducted in Japan, where the Japanese volunteers consumed 275mg EMIQ per 
day for 12 weeks and showed a positive impact of EMIQ on reduction of fat 
depots. We plugged in data from that study into this formula N=1+[16(xS²/d²)] 
to calculate sample size. Using this formula the number of subjects in each 
group were calculated to be n=18. However, we selected to use 25 subjects per
group which increases power of study to >85%.  25 subjects per group is a 
conservative sample size, which make us to expect a larger diference in the 
means with a higher dose of EMIQ intake.
The primary objective of this study is to demonstrate reduction of total body fat 
following 12 weeks of EMIQ intervention. Therefore, we will determine 
statistically if EMIQ is efective to reduce percent body weight, total abdominal 
fat, subcutaneous and visceral fat after 12 weeks of intervention compared to 
baseline values. We plan to compare the two group medians with the Mann-
Whithey U test.  After reviewing the data at the completion of the study, we can
assume normality, then we will compare the two group means with parametric 
t-test.  
 
C. Subject Characteristics
Inclusion Criteria:

Male or female age 18-65 years.

Premenopausal female with regular menstrual cycle or post-menopausal female
with cessation of menstrual cycle for a minimum of 6 months.

BMI in the range of 26.0-37.0  

Beck Depression Inventory Score less than 20.

Fluency in spoken or written English.

Willingness to be randomized to take EMIQ or placebo.

Must weigh at least 164lbs
Exclusion Criteria:

Any major illness or condition that may interfere with study outcomes at the 
discretion of the study physician.

Diabetes Type I & Type II, or use of any pharmacological treatment for diabetes.

Use of medications that interfere with energy metabolism 

Receiving hormone therapy growth hormone, testosterone, or estrogen with the
exception of hormone contraceptives. 

GI diseases, conditions or medications known to infuence GI absorption 
including active peptic ulcer disease or infammatory bowel disease such as 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
ulcerative colitis, Crohn’s disease, celiac disease, chronic diarrhea or 
constipation.

Undergone gastric bypass or other bariatric weight loss procedure.

Lipid lowering medications such as: bile acid sequestrants (Cholestyramine, 
Colestipol, Colesevelam, etc.), cholesterol absorption inhibitors (Ezetimibe-
Zetia), and fbrates (Gemfbrozil, Clofbrate, Fenofbrate, Triclor). Exceptions to 
this exclusion are HMG-CoA reductase (Statins). 

On or planning to participate in a weight loss program.

Weight loss or weight gain greater than 10 lbs in the past 6 months. 

History of eating disorders, anorexia, bulimia, or binge-eating in the past 5 
years.

Participation in any regular endurance exercise: running, biking, aerobics 
(except walking) or resistance training greater than 2.5 hours per week.

Regular use of acid lowering medications (greater than 3 times per week) such 
as antacids, PPIs (proton pump inhibitors), or H2 blockers.

Unstable thyroid disease.

Psychiatric disorders including schizophrenia, bipolar, major depression or 
psychosis.

Antidepressant medication: SSRIs (selective serotonin reuptake inhibitors), 
SNRIs (serotonin norepinephrine reuptake inhibitors), MAO inhibitors, or TCAs. 

Cancer of any type (except for non-melanoma skin) in the past 5 years.

Use of chemotherapeutic agents. 

Pregnancy, planning to become pregnant during the study period, or 
breastfeeding.

Unwilling to use efective birth control during study.

History of bilateral mastectomy with nodal dissection.

Alcohol use, on average, greater than 3 servings per day, 20 servings per week 
(Serving size: 12oz beer, 4oz wine, 2oz hard liquor), or self-reported binge 
drinking.

Uncontrolled hypertension (HTN) determined at the discretion of the study MD 
or RN. HTN medications allowed in the study: angiotensin converting enzyme 
(ACE) inhibitors, calcium channel blockers, and diuretics.

Medications for chronic obstructive pulmonary disease (COPD) or kidney 
disease.

α-adrenergic or β-adrenergic blockers (oral or ocular) and diuretics.

Renal or chronic kidney disease due to any condition, renovascular disease, or 
dialysis.

Chronic liver disease such as hepatitis B, hepatitis C, or cirrhosis.

Cardiovascular disease including: myocardial infarction, cerebrovascular 
disease (CVA), coronary artery bypass graft, stenosis greater than 50%, angina, 
coronary artery disease (CAD), congestive heart failure (CHF),peripheral 
vascular disease (PVD) or dysautonomia.

History of autoimmune diseases such as rheumatoid arthritis, lupus, multiple 
sclerosis, vitiligo, or psoriasis.

HIV or AIDS based on self-report.

Steroid use with the exception of over-the-counter topical and nasal steroids 
such as Flonase.

Allergy medication or regular antihistamine use.
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018

Seizure disorders. Acceptable if managed with medication and free of seizure 
activity for 5 years.

Smoking or the use of nicotine replacement products in the past 6 months.

Use of dietary supplements containing vitamins (except Calcium and Vitamin 
D), minerals, herbal or plant-based preparations, fsh oil or homeopathic 
remedies during study participation unless willing to discontinue prior to 
enrollment.

Current participation in another research study.

Non-English speaking.

No social security number.

Participation in another research study.

Withdrawal/Termination Criteria: Low compliance including taking less 
than 80% of study supplements, or missing study visits. Subjects who 
manifest an allergic reaction to the study supplement or placebo. 
Negative health efects as determined by the study physician. In the 
event of a withdrawal/termination, study subjects will be asked to 
discontinue their study supplement, and, if necessary due to negative 
health efects, will be referred to their primary care physician.
*Note: Screening lab values that would interfere with study outcomes are 
determined at the discretion of the study physician.
D. Risk/Beneft Assessment: We do not anticipate any major risks or hazards to 
participants in this study. 
There are no benefts to participation in this study. EMIQ has been 
independently evaluated by an expert panel as safe and has been categorized 
as GRAS (generally recognized as safe) according to the Food and Drug 
Administration (FDA)(see attachment “5. FDA GRAS EMIQ”). Other investigators 
have conducted similar clinical studies with EMIQ at doses comparable to those 
in this study and reported no adverse efect.
While all measures to ensure confdentiality will be taken, it is an ever present 
possibility that it may be lost. Stored samples will be labeled with a unique 
number (code) that will not contain any private information such as name or 
date of birth to ensure the samples cannot be identifed.  There is no intention 
for allowing anyone other than the principal investigator to have access to these
codes.  If Dr. Mohsen Meydani turns this study over to a new principal 
investigator, he will give the key code to the new principal investigator.
During blood draws there may be a small amount of bruising and discomfort, 
and rarely infection.  Every efort will be made to minimize discomfort to 
volunteers. There may be discomfort associated with 12-hour fasting. Some 
subjects may experience dizziness during prolonged fasting.
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
There is minimal risk of psychological or cognitive dysfunction due to the 
intervention. Participants may experience claustrophobia during the DXA and 
RMR measurements.  As such, a research technician will be present at all times 
during these procedures. If a volunteer indicates discomfort during the 
measurement, the procedure will be terminated at that time.
DXA scans expose participants to minimal radiation equivalent to 0.73 days 
total of background or natural radiation. This is equivalent to the amount of 
radiation exposure experienced by spending about 18 hours outdoors in a given
day.
E. Subject Participation
I.
Overview
This study will be a double-blind and placebo-controlled study. Total of 50 
overweight and obese subjects with BMI: 26-37 will be randomly assigned into 
two groups: 25 subjects to placebo control group and 25 subjects into treatment
group. Subjects in both groups will be asked to consume their regular diet 
during the 12 weeks of the study period. Subjects in the treatment group will 
also be asked to swallow two capsules of 180mg each, totaling 360mg of EMIQ 
(enzymatically-modifed isoquercitrin) every day, one capsule at breakfast and 
one at dinner, for a period of 12 weeks. The subjects in the placebo group will 
also consume their regular diet and will swallow two identical placebo capsules, 
one at breakfast and one at dinner, for 12 weeks.  
According to San-Ei Gen, the producer of EMIQ, the No Observed Efect Level 
(NOAEL) of EMIQ is 489 mg/kg BW/day, and the Acceptable Daily Intake (ADI) is 
4.89mg/kg BW/day [13, 14]. This data was generated from several studies 
including: acute oral toxicity testing [15], 4 week oral toxicity testing [16], 13 
week oral toxicity testing [17-19], mutagenicity testing [20], and 2 year 
carcinogenic testing [13].
The goal of this study is to test the efect of EMIQ on body fat reduction in 
overweight and obese individuals. Thus we have selected a BMI range of 26-37 
kg/m2, assuming an average height of 1.72m (5’ 8”) and an average weight of 
94kg (207.2 lbs). 
To calculate the dose of 360 mg EMIQ/day that participants will receive, we 
used the dose administered in the previous human studies from Japan as a 
benchmark. We wanted the dose in our study to refect the greater average 
predicted weight of our participants compared to a person in the Japanese study
due to an increase in BMI range for our study.  The BMI range for the study 
completed in Japan was 24-31, while the range for this study is 26-37. Japanese 
study subjects weighed approximately 72kg while our prediction is that 
participants will average 94kg based on our database records. This % diference
(31%) is refected in the increase from 275mg/day to 360mg/day. 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Our rationale for setting a minimum weight requirement is that a subject may 
be within the BMI range outlined in the study, but still not weigh enough to 
meet the ADI of 4.89mg/kg BW/day. A minimum bodyweight of 74kg should 
place participants in a safe range below the specifed ADI. We believe that a 
minimum body weight of 74kg should be a requirement for study participation.  
This rationale is demonstrated in the following calculation:
4.89mg/day *(x)kg=360mg 
(x)kg=360/4.89
(x)kg=73.62 (162.3lbs)
This study will be carried out after review and approval of the Institutional 
Review Board (IRB) of Tufts University. Following IRB approval, a total of 50 
male and female volunteers will be recruited and admitted to the study. The 
scope and plan of the study will be described to the admitted volunteers and a 
written consent will be obtained before enrolling the subjects into the study. 
The study is planned to start in 2016 (i.e., begin enrollment) and volunteer 
activity completed within one to one and a half years. 
The admitted volunteers will be scheduled and asked to come to the Metabolic 
Research Unit (MRU) at Jean Mayer USDA Human Nutrition Research Center on 
Aging (HNRCA) at Tufts University at the beginning of the study for enrollment 
(visit 1), baseline (visit 2), 4 weeks after baseline (visit 3), a visit 8 weeks after 
baseline (visit 4), and the fnal visit after 12 weeks (visit 5) for a total of 5 visits. 
Overnight fasted blood samples will be collected at screening and visits 2, 3, 4 
and 5.  
In order to monitor adherence and safety of EMIQ intake over the 12 week of 
the study, volunteers will undergo electrocardiogram (EKG) testing as well as 
several hematological tests, liver functions tests and other biochemical tests 
(See below) carried out on the collected blood samples. In addition, volunteers 
will be asked to provide a spot urine sample during screening and visits 2 and 5 
to measure total protein, glucose, urobilinogen, bilirubin, ketones, pH and occult
blood in urine using urine dip stick test.  Volunteers will complete three (3) 24-
hour dietary recalls over the phone at three time points: between visit 1 and 
visit 2; between visit 3 and visit 4 (around week 6 from beginning the 
supplement); and between visit 4 and visit 5 (around weeks 11-12) to estimate 
total caloric intake and intake of three major nutrient groups:  protein, 
carbohydrates, and fat. Volunteers will be asked to bring back unused EMIQ 
capsules and a compliance calendar during each visit to determine adherence 
to the supplementation schedule.  Volunteers will be asked not to change their 
regular physical activity, and will fll out a 7-day physical activity record at visits
2, 3, 4 and 5 to monitor this. To assess changes in adiposity following EMIQ 
intervention, the volunteers will complete whole body and segmental Dual X-ray
Absoprtiometry (DXA), skinfold thickness and hip to waist circumferences 
measurements at visit 2, i.e., before the intervention, and after 12 weeks of 
intervention, at visit 5, for total and segmental changes in body fat and fat free 
7

Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
mass. DXA data will be further subjected to computational analysis to obtain 
changes in abdominal, visceral and subcutaneous fats.   
Per policy number 9.20.18.30 of Tufts Department of Pharmacy Policy and 
Procedure Manual, any supplement returned from study subjects, or leftover 
upon completion of the study, will be destroyed “on-site” by placing them in a 
chemotherapy waste container for disposal using the Hospital’s system.
The following is a summary of the 6 total visits required by volunteers 
in this study:
Screening: Volunteers that were pre-screened by telephone will be scheduled 
and asked to come to the MRU at the HNRCA. Volunteers will be asked to arrive 
fasted for 12 hours. When they arrive, volunteers will be given a Screening 
Consent Form along with an explanation of all procedures that will be 
completed at screening. Volunteers will have an opportunity to read the consent
form and have any of their questions answered to their satisfaction. Once their 
consent is given we will begin by measuring volunteer’s vital signs, height, body
weight wearing a pre-weighted pair of scrubs, and BMI.  Blood samples will be 
collected for blood chemistry analysis. Women of childbearing potential will be 
subjected to a blood pregnancy test. The Beck Depression Inventory will be 
administered to assess for depression.  Additionally, study inclusion and 
exclusion criteria and health history will be reviewed. Breakfast and a $25 
screening stipend will be provided once screening procedures are complete.    
All subjects will be mailed a copy of screening labs within 1-2 weeks following 
the screening visit. Volunteer’s enrollment eligibility will be determined based 
on meeting inclusion and exclusion criteria.  Subjects with abnormal 
hematologic parameters, as defned by the HNRCA Nutrition Evaluation 
Laboratory (NEL) reference ranges, which are determined by the study 
physician to interfere with study outcomes, will be excluded from study 
participation. At the discretion of the study physician, subjects will be alerted to 
abnormal results that warrant follow-up with their primary care physician. 
Subjects with abnormal hematologic values outside the NEL reference range 
that do not afect study outcomes will be included at the discretion of the study 
physician. Note that data generated from blood draws at this visit will be used 
as baseline values in lieu of drawing additional blood at visit 2 for subjects that 
continue the study after screening.
Enrollment, Visit 1:  Volunteers that meet the study inclusion and exclusion 
criteria will be invited to participate in the main study. Those interested will be 
asked to return to the HNRCA. Volunteers will be given a Main Study Consent 
Form as well as an Optional Tissue Banking Consent Form. Volunteers will have 
an opportunity to read the consent forms and have any questions answered to 
their satisfaction. Once consent is obtained, MRU staf will assess any volunteer 
health, medication or eligibility status changes. The Dietary Assessment Unit 
(DAU) will dispense instructions for completing 24-hour dietary recalls by phone 
and will complete 3 dietary recalls by phone between visit 1 and visit 2. A 
baseline visit will be scheduled. Subjects will be mailed a $50 check for 
completion of Visit 1.
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Baseline, Visit 2:  Subjects will be asked to arrive at the MRU fasted for 12 
hours. Vital signs, and EKG measure of heart rhythm, scrub weight and height 
will be obtained. Changes to volunteer health, medication and eligibility status 
will be assessed. Blood samples will be collected and archived pending tissue 
banking consent.  Several baseline measurements will be performed during this 
visit including: lipid profle, insulin test, adiponectin, a urine dip stick, skinfold 
thickness and waist/hip circumference, resting metabolic rate (RMR), and DXA 
measure of total body fat. To perform RMR analysis, subjects will be asked to 
rest for 30 min prior to the test conducted by study staf.  Systolic and diastolic 
blood pressure and RMR will be measured after 30 min rest.  Subjects will be 
escorted to The Body Composition Analysis Center (BCAC) of the Tufts 
University Gerald J. and Dorothy R. Friedman School of Nutrition Science and 
Policy for a whole body DXA scan to measure total and segmental body fats.  A 
three factor eating questionnaire, food craving questionnaire, and the Stanford 
7-Day Physical Activity Recall (PAR) will be administered to assess volunteer’s 
dietary inhibition, restraint, hunger, cravings and physical activity level. EMIQ 
capsules, compliance calendar, and instructions will be dispensed to 
participants by nursing staf in the form of 180mg capsules contained in blister 
packages at the quantity to last for 1½ months. A continental breakfast meal 
will be provided to the subjects. Subjects will be mailed a $150 check for 
completion of Visit 2. Subjects will be instructed by MRU staf to store the study 
supplement at room temperature and away from heat and moisture. These 
instructions will also be attached to the study supplement in the form of labels.  
 
Week 4, Visit 3:
 
  Subjects will be asked to return to the MRU 4 weeks (21 
week) from visit 2. This visit is required to monitor volunteer health and safety 
during EMIQ supplementation of 360mg/day for 4 weeks. Subjects will be asked 
to arrive fasted for 12 h and bring their unused supplements as well their 
compliance calendar. Vital signs, scrub weight and changes to volunteer health,
medication and eligibility status will be assessed. Blood samples will be 
collected for liver and kidney function. In addition, a second EKG will be 
performed to monitor any changes the supplement may have on heart function.
The PAR will be administered. Compliance with supplement intake will be 
determined by counting the unused capsules remaining in blister pack and 
review of compliance calendars. The second 1 to 1½ month supply of EMIQ 
capsules will be dispensed to the subject.  A continental breakfast meal will be 
provided to the subjects. Subjects will be mailed a $50 check for completion of 
Visit 3.   After visit 3 (around week 6), subjects will complete 3 dietary recalls by
phone.
Week 8, Visit 4:
 
  Subjects will be asked to come to the MRU 8 weeks after visit 
2 or 4 weeks (21 week) after their previous visit. This visit is also required to 
monitor and ensure safety of EMIQ supplement intake at the dose of 360 
mg/day for 8 weeks. Subjects will be asked to arrive fasted for 12 h and bring 
their unused supplements as well their compliance calendar. Vital signs, scrub 
weight and changes to volunteer health, medication and eligibility status will be 
assessed. Blood samples will be collected to monitor liver and kidney function. 
The PAR will be administered. Compliance with supplement intake will be 
9

Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
determined by counting the unused capsules remaining in blister pack and 
reviewing the compliance calendar. The fnal 1 to 1½ month supply of EMIQ 
capsules will be dispensed to subjects.  A continental breakfast meal will be 
provided to the subjects. Subjects will be mailed a $50 check for completion of 
Visit 4.  After visit 4 (around week 11-12), subjects will complete 3 dietary 
recalls by phone.
 
Week 12, Visit 5: Subjects will be asked to return to the MRU approximately 
12 weeks from their baseline visit or 4 weeks (2 1 week) from their previous 
visit.  Subjects will be asked to arrive fasted for 12 hours and to bring back their
unused supplement and compliance calendar. Vital signs, scrub weight and 
changes to volunteer health, medication and eligibility status will be assessed. 
Blood samples will be collected and archived pending tissue banking consent.  
Final measurements will be taken during this visit including: routine health 
screening analysis, lipid profle, adiponectin, insulin test, a urine dip stick, liver 
and kidney function (SGOT, SGPT, BUN, creatinine, and calculated GFR), skin 
fold thickness and waist/hip circumference. Subjects will be asked to rest for 30 
min prior to measuring systolic and diastolic blood pressures and RMR analysis, 
which will be conducted by study staf.  After RMR and BP measurements, 
subjects will be escorted to the BCAC for a whole body DXA scan to measure 
total and segmental body fats.  A three factor eating questionnaire, food craving
questionnaire, and the PAR will be administered to assess changes in 
volunteer’s dietary inhibition, restraint, hunger, cravings and activity level 
throughout the study.  Compliance with supplement intake will be determined 
by counting the unused capsules remaining in blister pack and by reviewing the 
compliance calendar.  A continental breakfast meal will be provided to the 
subjects. Subjects will be mailed a $305 check for completion of the fnal study 
Visit 5.
Below is an outline of the visits mentioned above:
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Visit
Procedures
Length
Screening
Obtain Vital Signs
Height, Scrub Weight and BMI
10mL Blood Draw*
Urine Sample 
Review Inclusion/Exclusion Criteria
Review Health History
Complete Beck Depression Inventory
*Some women may be required to give an additional 
3mL blood for a pregnancy test
Visit 1
Assess Health/Medication/Eligibility Changes
Receive Instructions/Materials for Dietary Recalls
Between 
Visits 1 and 2
No visit required. Volunteers will be called 3x for 24-
hour dietary recalls.
Visit 2
Assess Health/Medication/Eligibility Changes
Obtain Vital Signs
Height, Scrub Weight and BMI
Undergo EKG Measurement
Urine Sample
30 Minutes Bed Rest Followed By RMR Measurement
Hip/Waist Circumference
Skinfold Thickness
17mL Blood Draw
DXA Measurement
Administer Questionnaires
Receive Supplement/Instructions/Compliance Calendar
2 Hours
1 Hour
5 Hours
Visit 3
Assess Health/Medication/Eligibility Changes
Obtain Vital Signs
Scrub Weight
Undergo EKG Measurement
5mL Blood Draw
Physical Activity Recall Questionnaire
Return Unused Supplement/Compliance Calendar
Receive Next Round Of Supplement
Between 
Visits 3 and 4
No visit required. Volunteers will be called 3x for 24-
hour dietary recalls.
Visit 4
Assess Health/Medication/Eligibility Changes
Obtain Vital Signs
Scrub Weight
5mL Blood Draw
Physical Activity Recall Questionnaire
Return Unused Supplement/Compliance Calendar
Receive Next Round Of Supplement
Between 
Visits 4 and 5
No visit required. Volunteers will be called 3x for 24-
hour dietary recalls.
Visit 5
Assess Health/Medication/Eligibility Changes
Obtain Vital Signs
Scrub Weight
Urine Sample
30 Minutes Bed Rest Followed By RMR Measurement
Hip/Waist Circumference
Skinfold Thickness
20mL Blood Draw
DXA Measurement
Administer Questionnaires
Complete Beck Depression Inventory
Return Unused Supplement/Compliance Calendar
5 Hours
2 Hours
2 Hours
11

Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
II.
Registration
This study will be registered on http://www.Clinical Trials.gov.
III.
Recruitment, Screening and Consent
Participants will be recruited from the general public using several 
recruitment strategies including local print and electronic media (such 
as advertisements in newspapers, craigslist, local newsletters, bulletin 
boards and diferent websites, [i.e., university jobsites at Tufts 
University, Boston University]), social media (i.e. Facebook, Twitter, 
LinkedIn)  as well as posting fiers in public places, with permission, 
such as local YMCAs, supermarkets, libraries, laundromats, local 
community organizations and health centers.  Additionally, the 
Metabolic Research Unit (MRU) resources include a roster of >20,000 
names of subjects from which potentially qualifying women and men 
can be identifed and contacted through the use of direct mailings.  
Only subjects in the MRU database who have agreed to be contacted 
for future studies will be contacted for this study.  Subjects who do not 
wish to be contacted are fagged in this database (Protocol Manager) 
with “DO NOT CONTACT.” Previous subjects with this fag in their 
record will not receive any unsolicited recruitment materials, i.e., the 
letter to previous volunteers. All advertisements, fiers, and letters will 
be pre-approved by the IRB.  
Although employees will not be targeted for recruitment and 
enrollment, employees of Tufts University and/or the HNRCA 
(employee-subjects) who voluntarily want to participate in the study 
will be eligible for screening and enrollment. In order to qualify for the 
study, employee-subjects must respond to IRB-approval advertisement
of their own accord and will not be directly approached by any person 
seeking to recruit them for participation in the study. Members of the 
research team, as direct-report subordinates of the PI and anyone who 
is direct-report subordinate to any of the research team members in 
any other capacity will not be eligible to participate in the study. If 
employee-subjects qualify to participate in the study, they cannot 
participate as volunteers during hours in which they are being 
compensated by Tufts University for their regular work.  This includes 
use of vacation, personal days and sick time.
Interested subjects will be pre-screened by telephone.  Potentially 
eligible subjects will be invited to screen in-person for the study in 
order to further determine eligibility.  During the screening visit, 
informed consent will be obtained by authorized MRU personnel  using 
the screening informed consent form. The consent interview will take 
place in a private area in the MRU at the HNRCA. Once all of the 
participant’s questions and concerns have been addressed, the 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
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PI: Mohsen Meydani
Study Protocol
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designated staf will obtain informed consent.  Subjects will then 
undergo routine blood and urine tests.  If subjects meet study criteria, 
they will be invited to participate in the main study.  Results of the 
blood and urine testing will be available at least 1-2 weeks after 
screening.  Subjects with lab values that do not meet eligibility 
requirements, assessed by the study physician or nurse, will be 
disqualifed and may be provided with a recommendation to see their 
physician.  Prior to study enrollment, subjects will be allowed to read 
the main study consent form. Once all of their questions and concerns 
are addressed, and they understand all procedures in the study, a 
designated staf member will obtain their informed consent using the 
main study informed consent form.
*Note: Non-English speaking individuals will not be enrolled in this 
study. The reasons being include the following: The HNRCA is not a 
health care facility, and does not have appropriate interpretive 
services to include non-English speakers. The standardized 
questionnaires have not been validated in other languages. This study 
will not directly beneft participants.
IV.
Study Assessments
Blood and Spot Urine Collection: The MRU will collect a total of 
60mL (12.2 teaspoons) of blood and three separate urine samples from
each subject to measure the following: 
Hematological measures: Complete blood count with auto-
diferential, white blood cell (WBC), red blood cell (RBC), hemoglobin 
(Hgb), hematocrit (Hct), platelet (Plt), Mean corpuscular volume (MCV),
mean corpuscular hemoglobin (MCH), and mean corpuscular 
hemoglobin concentration (MCHC). WBC/Diferential includes: 
neutrophils, lymphocytes, monocytes, eosinophils, and basophils.
 
Lipid profle: Triglycerides, Total Cholesterol, HDL-Cholesterol, LDL-
Cholesterol.
Metabolic and hormonal markers: Insulin, adiponectin in serum, 
and urine stick test (Includes: total protein, glucose, urobilinogen, 
bilirubin, ketone, pH, occult blood).
Liver  function:
  
Aspartate  aminotransferase  (AST),  alanin
aminotransferase  (ALT),  alkaline  phosphatase  (ALP),  gamma
glutamyltransferase  (GTP),  total
 bilirubin  (T-bill),  lactate
dehydrogenase (LDH), total protein (TP), and Albumin (Alb).
Muscle Function: Blood urea nitrogen (BUN), and creatine (Cre).
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Electrolytes: sodium (Na), potassium (K), chloride (Cl), calcium (Ca), 
magnesium (Mg).
Blood and Urine will be collected according to the following scheme:
Screening: 13mL (2.6 teaspoons) of blood will be drawn at this visit. 
5mL (1 teaspoon) will be drawn for complete blood count (CBC) with 
diferential, and another 5mL (1 teaspoon) for routine blood 
chemistries. CBC measures will be used as baseline values. 
Additionally, participants in the screening process will be required to 
provide a spot urine sample for urinalysis. A 3mL (0.6 teaspoons) blood
sample will be taken from women of childbearing potential at 
enrollment in order to conduct hCG testing. This test will determine 
whether the participant is pregnant, and thus ineligible for 
participation in this study.
Main Study: 
Visit 1, Enrollment: No blood will be drawn during this visit.
Visit 2, Baseline: A 10mL (2 teaspoons) blood sample will be drawn 
into red marble top tubes for liver and kidney function tests, insulin 
and adiponectin. An additional 7mL (1.4 teaspoons) of blood will be 
drawn into lavender top tubes for analysis of lipids. A spot urine 
sample will be provided for urinalysis. 
Visit 3, Week 4: A 5mL (1 teaspoon) blood sample will be drawn into 
a gold topped tube for liver/kidney function testing.
Visit 4, Week 8: A 5mL (1 teaspoon) blood sample will be drawn into 
a gold topped tube for liver/kidney function testing.
Visit 5, Final: A 3mL (0.6 teaspoons) blood sample will be drawn into 
a lavender topped tube for CBC diferential analysis. A 10mL (2 
teaspoons) blood sample will be drawn into red marble top tubes for 
liver and kidney function tests, insulin and adiponectin. An additional 
7mL (1.4 teaspoons) of blood will be drawn into lavender top tubes for 
analysis of lipids. A spot urine sample will be provided for urinalysis.
*Note: Visits 3 and 4 will be used to monitor participant’s liver and
kidney function during the study. If the study physician determines
that liver function appears to be in jeopardy, subjects will be asked to
discontinue  the  supplement  and  to  take  medical  action  at  the
discretion of the study physician.
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
*Note: Abnormal lab values found at screening will be provided to the
volunteers in writing. Abnormal values for safety labs (those falling
outside  of  the  study  physician’s  predetermined  range)  will  be
repeated.  The  study  physician  will  be  notifed  and  determine  any
follow-up action.
Nutritional Intake and Physical Activity:
Subjects will receive a Food Amount Booklet and information about the
24-hour recalls at visit 1. The Dietary Assessment Unit will collect three
(3) 24-hour recalls by phone (two weekdays and a weekend) between 
visits 1 and 2.  Three recalls will be collected by phone again between 
visits 3 and 4 (around  week 6,midway through the 12 week 
supplementation period) and between visits 4 and 5 (around weeks 11-
12).  Dietary intake data will be analyzed using Nutrition Data System 
for Research software version 2015, developed by the Nutrition 
Coordinating Center (NCC), University of Minnesota, Minneapolis, MN. 
Volunteers will be asked not to change their regular physical activity;
unless, their total physical activity per week amounts to greater than 
150 minutes, in which case they will be asked to refrain from exercise 
30 days prior to participating in the study and to continue refraining 
from exercise until the end of the study. In addition, volunteers will be 
subjected to four PAR questionnaires to assess physical activity at 
visits 2, 3, 4 and 5.   Subjects showing drastic changes in physical 
activity will be consulted, monitored, and if necessary, will be 
eliminated from the study.
To assess changes in dietary restraint, disinhibition and hunger, the 
Three Factor Eating questionnaire will be administered at both visits 2 
and 5. In addition to this, general susceptibility to food cravings will be 
assessed using the Food Cravings Questionnaire trait administered 
visits 2 and 5. 
Analysis of Whole Body Fat by DXA Scan: 
To obtain whole body and segmental changes in body fat and fat free 
mass including trunk, arms and legs, as well as visceral and 
subcutaneous fat, two (2) DXA scans will be performed, one at visit 2 
and another one at visit 5. 
Fasting Resting Metabolic Rate/Respiratory Quotient:  
Fasting RMR/RQ will be performed by a member of the study team at 
visits 2 and 5 to determine fat burning rate. 
 
Electrocardiogram (EKG): 
To ensure the safety and suitability of study participants, as well as of 
the supplement, two separate EKGs will be performed at visits 2 and 3.
EKGs will be performed to assess heart activity and any changes that 
may occur throughout the study.
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Vital signs and anthropometric measures:
The following measurements will be completed at selected study visits:
Body weight: screening, visits 2, 3, 4 and 5
Body height: screening, visits 2, 3, 4 and 5
Waist and hip circumferences:  visits 2 and 5
Skinfold thickness: visits 2 and 5
Systolic blood pressure: screening, visits 2, 3, 4 and 5
Diastolic blood pressure: screening, visits 2, 3, 4 and 5
Heart rate: screening, visits 2, 3, 4 and 5
Subjects with weight loss of more than 10 lbs between visits will be 
evaluated by the study MD, and further action will be determined at his
discretion.
Compliance: 
Study participants will be contacted by a member of the study team 
each week either by phone or by email to monitor adherence to the 
intervention and to monitor self-reported health changes. Study team 
members will fll out a weekly log to ensure that volunteers have been 
contacted. If any abnormalities are reported, the study physician will 
be informed. The study physician will advise the participant to follow 
up with their primary care physician and also decide if the participant 
can continue with the intervention.
Participants will receive a compliance calendar used to track their 
supplement intake at visit 2.
Participants will be required to return any unused supplements and 
compliance calendar at visits 3, 4 and 5. Participants that miss a visit 
beyond the allowed interval (4 weeks 21 week from previous visit) will 
be instructed to discontinue their supplement, if they have not done so
already, and they will be removed from the study. 
V.
Location
This study will be performed at the Metabolic Research Unit and 
Vascular Biology Laboratory at the Jean Mayer USDA Human Nutrition 
Research Center on Aging at Tufts University, located at 711 
Washington St, Boston, MA 02111.
DXA scans will be performed at the Body Composition Analysis Center 
at Tufts University Friedman School of Nutrition and Science Policy, 
located at 150 Harrison Ave, Boston, MA 02111.
VI.
Transportation
Volunteers will be required to provide their own transportation to the 
HNRCA. The building is accessible by public transportation. However, if 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
participants should choose to drive, they can obtain parking validation 
at a reduced cost. 
VII.
Personnel who will conduct the study
a. Present During Study Procedures:
Study Nurse
Study Coordinators
Principal Investigator
b. Primary Responsibility for the Following Activities
i.
Obtaining Informed Consent
Authorized MRU staf
Principal Investigator
Study Coordinators
ii.
Providing On-Going Information to the Study Sponsor 
and the IRB
Principal Investigator
Co-Investigator
Study Physician
iii.
Maintaining Participant’s Research Records
Authorized MRU staf
Principal Investigator
Study Coordinators
VIII.
Subject Payment
Screening Stipend: $25
Visit 1 Stipend: $50
Visit 2 Stipend: $150
Visit 3 Stipend: $50
Visit 4 Stipend: $50
Visit 5 Stipend: $305
Subjects will only be paid for each completed visit, and will not be paid 
if they withdraw or are withdrawn from the study.
IX.
Confdentiality
The identity of the volunteers, medical records and data relating to this
research will be kept confdential, except as required by law and 
except for inspections by the USDA or the Tufts Medical Center 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Institutional Review Board.  The medical records will be fled in the 
medical record room only available to designated research staf.  Upon
entry into the research study, the volunteer will be assigned a unique 
code number.  For storage, blood and urine samples will be identifed 
with this unique code number (not related to medical record number), 
date, laboratory generated login number and sample description (i.e. 
plasma, serum, urine).
All samples and data generated from the research study will be labeled
and/or recorded with a unique code number and no other identifer. 
The information that will allow the unique code number to be linked to 
the volunteer’s name will be stored as a password-protected computer 
fle.  Samples will be stored until all analyses are complete.  If blood 
samples collected during the course of this research study are sent 
outside the HNRCA for analysis, they will be labeled in a manner that 
contains no information that can make it possible to trace the sample 
back to the volunteer as an individual.  At the conclusion of the study, 
the information fle that would allow linking of the volunteer’s 
identifcation information with the sample labels will be destroyed.  
After the research study is completed data and samples will be stored 
in a manner such that the identity will not be traceable.
Volunteers’ records will be fled in the Metabolic Research Unit’s 
Medical Records File Room located on the 11th Floor of the Human 
Nutrition Research Center.  Research records, including volunteer 
records, will be retained for a period of 7 years after completion of the 
study.
Dr. Mohsen Meydani will have access to the key to identity codes. Dr. 
Meydani will have access to the de-identifed data. Laboratory 
personnel responsible for analyzing the samples will have access only 
to the de-identifed data. The study statistician will have access only to
the de-identifed data.
The study physician and nurses, the PI and Co-I, and Research 
Coordinators will have access to the research records.
X.
Alternatives
Participation in this study is entirely the participant’s choice. 
Participants may withdraw from the study at any time for any reason. 
Versions of the study supplement are available for purchase from 
vendors outside of the study. 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
XI.
Payment
All stipends will be mailed to participants in the form of checks 2-3 
weeks after they complete each visit. Participants who are removed 
from the study or choose to drop out will be paid for their completed 
visits only, and will not be paid if they are removed or choose to drop 
out before the completion of any study visit. In the event of a 
government shutdown or any other emergency situation, stipend will 
be delayed until the shutdown or emergency has ended.  
F. Specifc Procedures Used Throughout this Study
Distribution and Storage of Dietary Supplement
Supplements and placebo will be labeled in a binary (e.g., A or B) 
fashion without study staf’s knowledge of which is which. Both 
supplement and placebo will be stored in locked cabinets at the MRU. 
The two will be distributed to participants, in accordance with their 
randomization scheme, starting at visit 2. Participants will receive an 
extra week’s worth of supplements at each visit wherein supplement is
distributed in anticipation of events that may keep volunteers from 
attending their next visit after 4 weeks. 
Weight and Height:
Body weight will be measured during each visit to the HNRCA, aside 
from visit 1. Participants will be asked to change into scrubs provided 
by MRU during weight measurement to ensure the most accurate 
measurement.
Height will be measured at each visit to the HNRCA except enrollment. 
Shoes and head dressings or ornaments will be removed. 
Measurements will be taken according to standard procedure.
Waist and Hip Circumference:
Waist and hip circumference will be measured at visits 2 and 5. 
Measurements will be taken against the skin. Waist circumference will 
be measured at the approximate midpoint between the lower margin 
of the last palpable rib and the top of the iliac crest. Hip circumference 
will be measured around the widest portion of the buttocks with the 
tape parallel to the foor. These measures will be performed in 
duplicate and averaged. 
Resting Metabolic Rate (RMR):
RMR measures will be performed only at the visits 2 and 5. Subjects 
will arrive at the HNRCA in a fasted state. Subjects will undergo bed 
rest for 30 min prior to measurement. Systolic and diastolic blood 
pressure will be measured.  Measures of VO2 and VCO2 will be made 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
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using an indirect calorimeter calibrated using test gases of known 
concentration. Subjects will be instructed to relax and avoid fdgeting 
and sleeping during the measurements. Measures will be made under 
thermo-neutral conditions for 40 minutes. Rates of energy expenditure 
will be calculated from VO2 and VCO2 using Weir’s equation. Fasting 
oral temperature will be measured on days in which RMR is measured.
Dual-Energy X-Ray Absorptiometry (DXA):
DXA measurement will be performed only at the visits 2 and 5. Once 
subjects have completed measures at the HNRCA, they will be 
escorted to the Body Composition Analysis Center. The subject should 
be asked about and examined for metal that could be in the scan path.
Typical items to look for are earrings, eyeglasses, wristwatches, coins, 
rings, buttons, buckles, zippers, and support braces. The subject must 
remove shoes, and it may be necessary to remove skirts, slacks, 
brassieres, etc. If clothes are removed, a medical cotton gown will be 
provided during the scan. Subjects will be asked to lie down on the 
scan table without allowing their hands and legs to overlap while the 
scan is performed.  The analysis of each whole body scan will generate
values for total body lean, total BMC, total body fat, total body mass, 
trunk fat, trunk lean, total bone mineral density (BMD), lumbar BMD, 
upper extremity fat, upper extremity lean, lower extremity fat and 
lower extremity lean. To minimize error, each patient will be scanned 
on the same machine from baseline to fnal. Scans will be analyzed by 
the Body Composition Analysis Lab.
Electrocardiogram (EKG):
An EKG will be performed by MRU staf at visits 2 and 3. Areas of 
participant’s chest, arms and legs will be rubbed with alcohol and 12 
electrodes will be attached to the skin. Participants will be required to 
lie still on their backs in bed while electrical activity is recorded by 
these electrodes. Electrodes will be removed after the test, which 
takes approximately 20 minutes to administer.
Skinfold Thickness: 
To obtain more specifc subcutaneous fat measurements we will 
perform skinfold thickness tests. Tests will be performed at visit 2 and 
visit 5. Participants will be asked to remove their shirts in order to 
obtain the measurements. Skinfold thickness will be measured at four 
sites: subscapular, suprailliac, the biceps and the triceps. All measures 
will be performed on the participant’s right side unless it is not possible
to do so.  Sites will be identifed through anatomical landmarks and 
measurements. The skinfold site will be marked. The participants skin 
will be pinched approximately 1cm above the mark using the thumb 
and index fnger, and simultaneously pulled away from the underlying 
muscle to form the skinfold. While the pinch is maintained the caliper 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
jaws will be opened and positioned on the mark approximately 1cm 
below the thumb and index fnger. The caliper jaws will be allowed to 
close around the skinfold. Two to four seconds after the caliper 
pressure is released the reading will be taken. This process will be 
repeated until subsequent measurements are within 1-2mm of each 
other. The subscapular measure will be taken from the bottom point of 
the shoulder blade. Suprailliac measure will be taken just above the 
top of the hip bone. Triceps will be measured at the posterior midline 
of the upper arm, and biceps at the anterior midline of the upper arm.
Questionnaires:
The following questionnaires will be administered:
Beck Depression Inventory:  this questionnaire will be administered 
by MRU staf at screening and visit 5 to assess volunteer’s 
psychological state. If subjects score ˃20, they will be informed of 
the result and a recommendation will be made to follow up with 
their physician. 
Three Factor Eating Questionnaire: This questionnaire will be 
administered by study team staf at visits 2 and 5.
Food Cravings Questionnaire-trait: This questionnaire will be 
administered by study team staf at visits 2 and 5. 
Stanford 7-Day Physical Activity Recall: This questionnaire will be 
administered by study team staf at visits 2, 3, 4 and 5.
Volunteer Consent and Safety:
The protocol will be reviewed and approved by the Tufts Institutional 
Review Board (IRB), and written informed consent will be obtained 
from all participants. Study volunteers will be given the opportunity to 
read consent forms in a quiet area free of noise and distraction. 
Consent forms will be thoroughly reviewed with the volunteer and all of
their questions will be answered to the volunteer’s satisfaction. 
Comprehension of study information will be assessed by asking 
volunteers to restate their understanding of presented information in 
their own words. Participants will be told they can decline participation 
at any time for any reason. Participant identifers will be kept in a 
separate fle from data. Data collection forms will not have participant 
identifers. The participant’s identity and records relating to the 
prescreening and screening procedures will be kept confdential in 
locked cabinets at all times at the Volunteer Recruitment Services, and
at the MRU, respectively, whether or not the participant qualifes for 
the study. The information kept may be used to invite the individual to 
participate in future studies at the HNRCA. If a participant chooses not 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
to have their contact or other information kept in our records for this or
other reasons, the participant need only to contact the Volunteer 
Recruitment Services Department by telephone or email to make this 
request, and all of their records will be destroyed. All study records for 
all participants will be kept confdential, and only the study P.I., Co-P.I.,
study physician, and study coordinators will have access to participant 
identifying information, if needed; otherwise, only de-identifed data 
will be transferred to co-investigators. All data will be kept in secure 
fles and locked data cabinets. Identifers will not be destroyed at any 
point.
Participants and nursing staf will be able to reach the study physician, 
Dr. Lisa Ceglia, or a covering physician 24 hours/day. Specifc 
participant complaints will be addressed on an individual basis and 
appropriate referrals to primary care providers or other health care 
providers will be made. Participants’ health status will be assessed 
weekly through phone or email and will be updated by the members of
the study team.  While there is no evidence to suggest so, there might 
be some adverse efects from EMIQ’s efect on fat loss, or EMIQ itself. If
these problems are reported by participants, the condition will be 
assessed to determine if these adverse events are potentially study-
related. Further, in the case of these and any other adverse event, the 
study physician will advise participants on the appropriate assistance 
or action to take, and then counseled to seek further appropriate 
assistance. 
Compensation will not be available for any research-related injury, and 
treatment may incur charges for the subject. The Nursing staf at the 
MRU is basic BLS (CPR) certifed and trained in the use of an automatic 
external defbrillator (AED). In all emergency situations, the MRU 
Nursing staf will provide immediate basic life support (BLS) response, 
call 911 and continue to provide emergency supportive care as needed
(e.g., monitoring, oxygen, airway maintenance) until emergency 
medical service (EMS) arrives. Their philosophy is to provide early 
access to the most appropriate and advanced care for treatment of 
study subjects. Treatment by the MRU Nursing staf and/or the staf 
physician is provided at no cost to the participants. All additional 
treatment including the cost of an ambulance is charged to the study 
subject. There are no funds available for these costs to be paid by the 
HNRCA or by study funds. This information is provided to participants 
prior to their signing an informed consent form.
A participant may be dropped from the study should a reported 
adverse reaction to the study supplement be considered severe.
G. Subject Safety and Data and Safety Monitoring Plan
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Classifcation and Reporting of Adverse Events and Unanticipated Problems for
This Study
An Unanticipated Problem is defned as: An incident, experience, or 
outcome that meets all of the following criteria: 
1. The nature, severity, or frequency is unexpected for the subject population 
or research activities as described in the current IRB approved protocol, 
supporting documents, and the ICF(s). 
2. It is related or possibly related to participation in the research. 
3. It suggests the research may place the subject or others at a greater risk of 
harm then was previously recognized.
An Adverse Event (AE) is defned as: Any untoward or unfavorable medical 
occurrence in a human subject, including any abnormal physical exam or 
laboratory fnding, symptom, or disease, temporally associated with a 
subject’s participation in the research.
AEs are classifed by three categories:
1.
Severity:
 
  Non-serious or Serious
2.
Relation to Participation in Research:
 
  Related, Possibly Related, or Unrelated
The classifcation of the potential relationship to the intervention will be 
determined based on the following scheme:
Defnite
Temporal pattern and known or expected adverse event response 
pattern, confrmed by stopping the intervention and reappearance of 
adverse event on re-challenge.
Probabl
e
Temporal pattern and known or expected adverse event response 
pattern, confrmed by stopping the intervention and which could not 
be explained by the participant’s clinical state. 
Possible
Temporal pattern and known or expected adverse event response 
pattern, confrmed by stopping the intervention and which could have 
been produced by a number of other factors.
Unknow
n
Relationship for which no evaluation can be made. 
Not 
Related
Adverse event for which sufficient information exists to indicate that 
the cause is unrelated to the study intervention.
3. Expectation:
 
  Expected or Unexpected (determination is based on the 
known risks associated with the study supplement and procedures, 
natural progression of an underlying illness, and health characteristics of 
the study population)
4. Location:
 
  Internal (occurring at the HNRCA) or External (occurring at a site
that is not under the jurisdiction of the Tufts MC/TUHS IRB)
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
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Version Date 3/2/2018
Further, a Serious Adverse Event (SAE) is an AE that meets the following 
criteria:
1. Results in death, or
2. Is life-threatening, or
3. Results in hospitalization or prolongation of existing hospitalization, or
4. Results in a persistent or signifcant disability/incapacitation, or
5. Results in congenital anomaly/birth defect, or
6. May jeopardize the subject’s health and may require medical or surgical 
intervention to prevent one of the other outcomes listed above.
AEs are Unanticipated Problems only when they meet all three criteria in 
the defnition of an unanticipated problem.
In the event of an Unanticipated Problem, or adverse event, staf will 
immediately notify the PI and study physician. Once notifed, the PI, Dr. 
Mohsen Meydani, will take immediate action to eliminate or minimize risk to 
enrolled subjects. The study physician, Dr. Lisa Ceglia, will assess the severity
of the unanticipated problem, and determine its relation to the study 
protocol. Each unanticipated problem will be reported, with all supporting 
documentation, according  to the following IRB guidelines within 5 business 
days:
Information that does not fall under any of the categories does not require reporting
to the IRB.
1.
Harm experienced by a subject or other individual, which in the opinion of the
investigator is unexpected andrelated/possibly related to the research 
procedures.
A.
A harm is “unexpected” when its specifcity and/or severity 
is inconsistent with risk information previously reviewed and approved 
by the IRB in terms of nature, severity, frequency, and characteristics of 
the study population.
B.
A harm “possibly related” when there is a reasonable likelihood that 
the incident, experience, or outcome may have been caused by the 
procedures involved in the research.
b.
Information that indicates a new or increased risk, or a new safety issue. For 
example:
A.
New information (e.g., an interim analysis, safety monitoring report, 
publication in the literature, sponsor report, or investigator fnding) indicates 
an increase in the frequency or magnitude of a previously known risk, or 
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Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
uncovers a new risk or greater risk of harm (including physical, psychological, 
economic, or social harm) than was previously known or recognized.
B.
An investigator brochure, package insert, or device labeling is revised 
to indicate an increase in the frequency or magnitude of a previously 
known risk, or describe a new risk
C.
Withdrawal, restriction, or modifcation of a marketed approval of a 
drug, device, or biologic used in a research protocol
D.
Complaint of a subject that indicates subjects or others might be at 
increased risk of harm or at risk of a new harm
E.
Any changes or new information adversely afecting the conduct of the
research (that indicates a new or increased risk, or a new safety issue)
b.
Non-compliance with the federal regulations governing human research or 
with the requirements or determinations of the IRB, or an allegation of such 
non-compliance
c.
Audit, inspection, or inquiry by a federal agency and any resulting reports 
(e.g. FDA Form 483)
d.
Any site monitoring or other audit report fnding that directly and materially 
afects subject safety or their willingness to continue participation
e.
Failure to follow the protocol that harmed a subject or placed subject at risk 
of harm (for example, missed study test or visit (or out of window) that could 
afect subject safety, enrollment of a subject who did not meet all eligibility 
criteria, failure to follow safety monitoring plan. These examples are intended 
as a guide and are not all-inclusive.)
f.
Breach of confdentiality
g.
Change to the protocol taken without prior IRB review to eliminate an 
apparent immediate hazard to a subject
h.
Incarceration of a subject in a study not approved by the IRB to involve 
prisoners
i.
Complaint of a subject that indicates that the rights, welfare, or safety of the 
subject have been adversely afected or a subject complaint that cannot be 
resolved by the research team
j.
Premature suspension or termination of the protocol by the sponsor, 
investigator, or institution
k.
State medical board actions against any member of the research team
l.
Unanticipated adverse device efect
 
  (any serious adverse efect on health or 
safety or any life-threatening problem or death caused by, or associated with, 
a device, if that efect, problem, or death was not previously identifed in 
nature, severity, or degree of incidence in the investigational plan or 
application (including a supplementary plan or application), or any other 
unanticipated serious problem associated with a device that relates to the 
rights, safety, or welfare of subjects.)
m.
Information that, per the sponsor, requires prompt reporting to the IRB
25

Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Data and Safety Monitoring Plan:
As this study is low-risk, it does not have a Data and Safety Monitoring Board 
(DSMB). The PI, along with the IRB, will ensure that volunteer safety and data 
integrity are maintained through our Data and Safety Monitoring Plan. This 
Data and Safety Monitoring Plan (DSMP) outlines the PI’s, Dr. Mohsen 
Meydani, role as follows:

Dr. Meydani will regularly monitor potential risks and procedures in order to 
protect participants from risk.

Will discuss any adverse events with the study physician on a weekly basis.

Will report any adverse events associated with participation in the study 
protocol and fle all required annual paperwork with the IRB.

Will evaluate the progress of the study including periodic assessments of 
data quality and timeliness, recruitment, enrollment and retention, protocol 
violations, unanticipated events, volunteer risk vs. beneft, and anything else 
that may have an impact on the study outcome.

Maintain vigilance of developments outside of the study that could afect 
participant’s safety.

Perform, at minimum, two reviews of the data per year to ensure safety and 
efficacy of the study.

Responsibility for all procedures performed. 

Reviews all data as it may relate to volunteer safety. This includes factors like
participant retention and reason for drop-out.

Reporting all serious adverse events regardless of their relation to the study 
protocol.

Provide the IRB with summaries of all adverse events.

At the discretion of Dr. Meydani, the Co-I, Dr. Roberts, or Dr. Saltzman, the 
study may be terminated at any time if the risks to study subjects are found 
to outweigh the potential benefts.
Additionally, no personal results will be communicated to study participants. 
However, participants will be informed of any necessary new fndings 
developed during the course of this research study. Written results may be 
given directly, or mailed, to participants if they are still in the study, or 
mailed to them if they have completed their participation in the study.
H. Outcome
Our goal is to assess the safety and efficacy of EMIQ as it relates to body fat 
reduction. We predict that there will be no changes in health, and that there 
will be a signifcant diference between change in total body fat between the 
treatment and placebo groups.
I.
Tissue Banking Considerations
26

Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Optional participation in tissue banking is ofered during the informed 
consent process for participation in the main study.  All pertinent information 
regarding what will happen to the stored samples, how long they will be 
stored; their potential future use for research, etc. will be outlined clearly. 
Participants will be informed as to how they can withdraw their sample from 
the tissue bank if they choose to do so. Participants will be asked to take the 
time to think about their decision, and will be encouraged to ask questions. 
All questions and concerns will be addressed to the participant’s satisfaction 
to ensure the participant is giving informed consent.
J.
References
1.
Boots, A.W., G.R.M.M. Haenen, and A. Bast, Health effects of quercetin:
From antioxidant to nutraceutical. European Journal of Pharmacology, 2008. 
585(2-3): p. 325-337.
2.
Khurana, S., et al., Polyphenols: Benefits to the Cardiovascular System 
in Health and in Aging. Nutrients, 2013. 5(10): p. 3779-3827.
3.
Pandey, A.K., et al., Quercetin in hypoxia-induced oxidative stress: 
Novel target for neuroprotection. International Review of Neurobiology 2012. 
102: p. 107-146.
4.
Rothwell, J.A., et al., Phenol-Explorer 3.0: a major update of the Phenol-
Explorer database to incorporate data on the effects of food processing on 
polyphenol content. Database, 2013. 2013(0): p. bat070-bat070.
5.
Nabavi, S.F., et al., Role of quercetin as an alternative for obesity 
treatment: You are what you eat! Food Chemistry, 2015. 179: p. 305-310.
6.
Guo, Y. and R.S. Bruno, Endogenous and exogenous mediators of 
quercetin bioavailability. The Journal of Nutritional Biochemistry, 2015. 26(3):
p. 201-210.
7.
Akiyama, T., et al., Constituents of Enzymatically Modified Isoquercitrin
and Enzymatically Modified Rutin (Extract). Journal of the Food Hygienic 
Society of Japan, 2000. 41(1): p. 54-60.
8.
Makino, T., et al., Enzymatically Modified Isoquercitrin, a-Oligoglucosyl 
Quercetin 3-O-Glucoside, Is Absorbed More Easily than Other Quercetin 
27

Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
Glycosides or Aglycone after Oral Administration in Rats. Biological and 
Pharmaceutical Bulletin, 2009. 32(12): p. 2034-2040.
9.
Motoyama, K., et al., Atheroprotective and plaque-stabilizing effects of 
enzymatically modified isoquercitrin in atherogenic apoE-deficient mice. 
Nutrition, 2009. 25(4): p. 421-427.
10.
Murota, K., et al., α-Oligoglucosylation of a sugar moiety enhances the 
bioavailability of quercetin glucosides in humans. Archives of Biochemistry 
and Biophysics, 2010. 501(1): p. 91-97.
11.
Seo, Y.S., O.H. Kang, S.B. Kim, S.H. Mun, D.H. Kang, D.W. Yang, J.G. 
Choi, Y.M. Lee, D.K. Kang, H.S. Lee and D.Y. Kwon, Quercetin prevents 
adipogenesis by regulation of transcriptional factors and lipases in OP9 cells. 
Int J Mol Med, 2015. 35: 1779-85.
12.
Dong, J., X. Zhang, L. Zhang, H.X. Bian, N. Xu, B. Bao and J. Liu, 
Quercetin reduces obesity-associated ATM infltration and infammation in 
mice: a mechanism including AMPKalpha1/SIRT1. J Lipid Res, 2014. 55: 363-
74.
13.
Salim EI, Kaneko M, Wanibuchi H, Morimura K, Fukushima S. Lack of 
carcinogenicity of enzymatically modifed isoquercitrin in F344DuCrj rats. 
Food Chem Toxicol 2004; 42:1949-69.
14. 
January 5, 2006 letter from Professor Shoji Fukushima to Professor 
Joseph F. Borzelleca regarding ADI (See Supplementary Materials)
15. 
Acute oral toxicity study of SANMELIN 11130 in rats, San-Ei Gen F.F.I., 
January 10, 1987 (See Supplementary Materials)
16. 
Preliminary study for 90-day oral toxicity of enzymatically modifed 
isoquercitrin in F344DuCrj rats, San-Ei Gen F.F.I., Inc., January 28, 2003 (See 
Supplementary Materials)
17. 
Tamano S., Hatahara Y., Sano M., Hagiwara A., Nakamura M., Washino 
T. and Imaida K. (2001). 13-Week oral toxicity and 4-week recovery study of 
enzymatically modifed isoquercitrin in F344DuCrj rats. Jpn. J. Food Chem. 
8(3), 161-1 67
18. 
Results of peer review on the fnal report of “Ninety-Day Repeated Oral
Toxicity Study and 28-Day recovery study of enzymatically modifed 
isoquercitrin in rats,” reviewed by Dr. Michihito Takahashi, October 8, 2002 
(See Supplementary Materials)
28

Study Title: Body Fat Reducing Effect and Safety of Enzymatically Modified Isoquercitrin in Overweight and 
Obese Subjects
PI: Mohsen Meydani
Study Protocol
Version Date 3/2/2018
19. 
Answer for the peer review, Dr. Seiko Tamano, November 14, 2002 
(Attached)
20. 
Bacterial reverse mutation study of enzymatically modifed 
isoquercitrin (SANMELIN), San-Ei Gen F.F.I., Inc., December 19, 1986 (See 
Supplementary Materials)
29