Xanthohumol Metabolism and Signature (XMaS) in Healthy Adults
Phase: PHASE1
Conditions: Healthy
Sponsor: National University of Natural Medicine

Study Product Guidelines and Considerations 
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1 June 2018 
Clinical Study 
Protocol 
 
 

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1 June 2018 
FULL PROTOCOL TITLE 
Xanthohumol Microbiome and Signature  (XMaS) in Healthy Adults 
Randomized placebo-controlled trial of xanthohumol (24 mg/day for 8 weeks) 
to determine safety and biological signature in healthy adults.  
  
Study Chairman or Principal Investigator:   
Co-Principal Investigator: Ryan Bradley ND, MPH, Director, Helfgott 
Research Institute, National University of Natural Medicine 
Co-Principal Investigator: Fred Stevens, PhD, Director, Linus Pauling 
Institute, Oregon State University 
 
Supported by:  
The National Center for Complementary and Integrative Health  
Grant Number: 1RO1AT010271-01 
 
Study Intervention Provided by:  
Metagenics Inc., Gig Harbor, WA 
Hopsteiner – S.S. Steiner, Inc., New York, NY 
 
 
 
 
Sponsor of IND (IDE):  
Fred Stevens, PhD, Oregon State University, IND#140626 

Study Product Guidelines and Considerations 
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Version 1.1 
 
 
1 June 2018 
Tool Revision History 
Version Number: 1.0 
Version Date: 1-29-2019 
Summary of Revisions Made: N/A 
Version Number:2.0 
Version Date:2-20-2019 
Summary of Revisions Made: Removal of Phase 2 content re: Crohn’s disease 
Version Number:3.0 
Version Date:5-20-2019 
Summary of Revisions Made: Specified time “window” for study visit scheduling, moved first 
product dispensation to coincide with first sample return, added PROMIS-29 as additional 
measure of tolerability 
Version Number: 4.0 
Version Date: 10-1-2019 
Summary of Revisions Made: Clarification of acceptable laboratory values and re-screening 
protocols for accessing candidate eligibility; clarification of stabilization periods for dietary 
supplements, changes in diet and exercise, alcohol reductions and discontinuation of Cannabis 
products

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TABLE OF CONTENTS 
Page 
Clinical Intervention Study Protocol Template ........................................................... 1 
FULL PROTOCOL TITLE ............................................................................................... 2 
Tool Revision History ................................................................................................... 3 
TABLE OF CONTENTS .................................................................................................. 4 
STUDY TEAM ROSTER .............................................................................................. 8 
PARTICIPATING STUDY SITES ................................................................................. 8 
PRÉCIS ....................................................................................................................... 8 
1. STUDY OBJECTIVES ................................................................................................ 8 
1.1 
Primary Objective .............................................................................................. 8 
1.2 
Secondary Objectives ....................................................................................... 8 
1.3 
Tertiary Objectives ............................................................................................ 8 
2. BACKGROUND AND RATIONALE ........................................................................... 8 
2.1 
Background on Condition, Disease, or Other Primary Study Focus .................. 9 
2.2 
Study Rationale ................................................................................................. 9 
3. STUDY DESIGN ....................................................................................................... 10 
4. SELECTION AND ENROLLMENT OF PARTICIPANTS .......................................... 10 
4.1 
Inclusion Criteria ............................................................................................. 10 
4.2 
Exclusion Criteria ............................................................................................ 11 
4.3 
Study Enrollment Procedures ......................................................................... 11 
5. STUDY INTERVENTIONS ........................................................................................ 13 
5.1 
Interventions, Administration, and Duration .................................................... 13 
5.2 
Handling of Study Interventions ...................................................................... 13 
5.3 
Concomitant Interventions .............................................................................. 14 
5.3.1 Allowed Interventions .................................................................................. 14 
5.3.2 Required Interventions ................................................................................ 14 
5.3.3 Prohibited Interventions ............................................................................... 14 
5.4 
Adherence Assessment .................................................................................. 14 
6. STUDY PROCEDURES ............................................................................................ 15 
6.1 
Schedule of Evaluations.................................................................................. 15 

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6.2 
Description of Evaluations............................................................................... 15 
6.2.1 Screening Evaluation................................................................................... 16 
6.2.2 Enrollment, Baseline, and/or Randomization ............................................... 17 
6.2.3 Blinding........................................................................................................ 18 
6.2.4 Followup Visits ............................................................................................ 18 
6.2.5 Completion/Final Evaluation ........................................................................ 18 
7. SAFETY ASSESSMENTS ........................................................................................ 20 
7.1 
Specification of Safety Parameters ................................................................. 20 
7.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters ................................................................................................................ 20 
7.3 
Adverse Events and Serious Adverse Events ................................................. 22 
7.4 
Reporting Procedures ..................................................................................... 23 
7.5 
Followup for Adverse Events .......................................................................... 24 
7.6 
Safety Monitoring ............................................................................................ 24 
8. INTERVENTION DISCONTINUATION ..................................................................... 25 
9. STATISTICAL CONSIDERATIONS .......................................................................... 26 
9.1 
General Design Issues .................................................................................... 26 
9.2 
Sample Size and Randomization .................................................................... 26 
Treatment Assignment Procedures ................................................................ 26 
9.3  
Definition of Populations ................................................................................. 26 
9.4 
Interim Analyses and Stopping Rules ............................................................. 26 
9.5 
Outcomes........................................................................................................ 26 
9.5.1 Primary Outcome ........................................................................................ 27 
9.5.2 Secondary Outcomes .................................................................................. 27 
9.5.3 Tertiary Outcomes ....................................................................................... 27 
9.6 
Data Analyses ................................................................................................. 28 
10. DATA COLLECTION AND QUALITY ASSURANCE ............................................. 32 
10.1 
Data Collection Forms .................................................................................... 32 
10.2 
Data Management .......................................................................................... 32 
10.3 
Quality Assurance ........................................................................................... 32 
10.3.1 
Training .................................................................................................... 32 
10.3.2 
Metrics ..................................................................................................... 32 
10.3.3 
Protocol Deviations .................................................................................. 32 
10.3.4 
Monitoring ................................................................................................ 32 
11. PARTICIPANT RIGHTS AND CONFIDENTIALITY ................................................ 33 

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11.1 
Institutional Review Board (IRB) Review......................................................... 33 
11.2 
Informed Consent Forms ................................................................................ 33 
11.3 
Participant Confidentiality................................................................................ 33 
11.4 
Study Discontinuation ..................................................................................... 34 
12. COMMITTEES ........................................................................................................ 34 
13. PUBLICATION OF RESEARCH FINDINGS ........................................................... 34 
14. REFERENCES ........................................................................................................ 34 
15. SUPPLEMENTS/APPENDICES ............................................................................. 38 
I. 
IRB-approved Informed Consent for Screening 
II. 
IRB-approved Informed Consent for Trial Participation 

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STUDY TEAM ROSTER  
Principal Investigator: Ryan Bradley ND, MPH (rbradley@nunm.edu), (503) 552-1862 
Co-Investigator: Jennifer Ryan, ND, MS (jryan@nunm.edu), (503) 552-1744 
Co-Investigator: John Phipps PhD, NUNM (jphipps@nunm.edu), (503) 552-1763 
Study Coordinator: Trina Soileau, NUNM (tsoileau@nunm.edu), (503) 552-1746 
Study Coordinator: Emily Stack, NUNM (estack@nunm.edu), (503) 552-1777 
Biostatistician: Doug Hanes PhD, NUNM (dhanes@nunm.edu), (503) 552-1741 
For all above, the fax number is (503) 227-3750 and the address is: 
 
National University of Natural Medicine 
Helfgott Research Institute 
2220 SW First Ave.  
Portland, OR 97201 
 
PARTICIPATING STUDY SITES 
Dr. Ryan Bradley (see above), is the principal investigator for all clinical activities. 
Additional laboratory procedures will be overseen by the following site investigators: 
 
Dr. Fred Stevens,  
307 Linus Pauling Science Center 
2900 SW Campus Way 
Corvallis, OR 97331 
Email: fred.stevens@oregonstate.edu, Phone: (541) 737-9534, Fax: (541) 737-5077  
 
Dr. Tom Metz 
Pacific Northwest National Laboratory 
PO Box 999 
MSIN: K8-98 
Richland, WA 99352 
Email: Thomas.Metz@pnnl.gov, Phone: (509) 371-6581, Fax: (509) 371-6555 
 
 
 
 

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PRÉCIS  
Study Title  
Xanthohumol Microbiome and Signature (XMaS) in Healthy Adults Trial  
Objectives  
The overarching objectives are to: 1. Determine safety and tolerability of xanthohumol (XN, 
24 mg/day for 8 weeks) and 2. Collect data on the metabolism and biological signature of 
XN as a baseline for possible future studies.  
Design and Outcomes   
This is an 8-week triple-masked, placebo controlled, randomized clinical trial of 
xanthohumol (XN), which is a constituent of hops (Humulus lupulus) in healthy volunteers 
aged 21 to 50 years. Participants will complete a series of bi-weekly clinical visits and 
provide samples of blood, urine, and stool, plus be interviewed for adverse events.  
Primary outcome measures include routine clinical toxicology measures (e.g. 
comprehensive metabolic panel and complete blood counts). Secondary measures include: 
XN metabolites, bile acid metabolism, biomarkers of inflammation and gut integrity, and 
composition of intestinal microbiome species.  
Interventions and Duration  
The intervention is xanthohumol (XN, 24 mg/day plus 288 mg of rice protein) or placebo 
(rice protein 288 mg/day), encapsulated in gelatin capsules, and taken by mouth once daily 
for 8 weeks.  
Sample Size and Population  
The target sample will be 32 healthy adults free of acute and chronic diseases aged 21-50 
years, divided equally between XN and placebo groups. 
1. STUDY OBJECTIVES 
1.1 
Primary Objective 
The primary objective is to determine the safety and tolerability of xanthohumol in healthy 
adults over 8 weeks in a Phase 1 trial. The primary hypothesis is that in the XN arm, the 
number of participants with an adverse event or abnormal lab value will not be more than 
50% higher than the number in the placebo group.  
1.2 
Secondary Objectives 
The secondary objective is to collect comparison data for a subsequent Phase 2 trial in adults 
with Crohn’s disease (if an acceptable safety/tolerability profile is established by the Phase 
1 trial). Our primary outcome measure for this secondary objective is TNF-alpha as a 
biomarker of systemic inflammation. Because we are enrolling healthy adults, we 
hypothesize there will not be a difference in changes in TNF-alpha between XN and placebo 
in Phase 1. Yet the sample size provides >80% power, at a threshold of alpha=0.05, for the 
detection of a 25% difference in TNF-alpha changes in response to XN between the healthy 
adults in this trial and participants in future trials.  

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1.3    Tertiary Objectives 
The tertiary objective is to collect comparison data related to XN metabolism, gut 
permeability and impact on the microbiota for a subsequent Phase 2 trial in adults with 
Crohn’s disease. Thus our outcome measures for this objective include XN metabolites, bile 
acid metabolism, biomarkers of gut inflammation and gut integrity, and composition of 
intestinal microbiome species.  
2. BACKGROUND AND RATIONALE 
2.1 
Background on Condition, Disease, or Other Primary Study Focus 
The motivation for the current Phase 1 trial of xanthohumol (XN) in health adults is to serve 
as comparison data for a subsequent trial in adults with Crohn’s disease (CD). Mechanistic 
evidence supports the therapeutic potential of XN in CD, and thus Phase 1 safety data are 
required prior to moving on to Phase 2 trials in CD. CD is an important source of disease 
burden in the United States. The Centers for Disease Control and Prevention estimates 1.3% 
of U.S. adults (3 million) are diagnosed with Inflammatory Bowel Disease (IBD), and the 
prevalence of the disease is increasing.1 CD is the most prevalent form of IBD, characterized 
by predominantly small bowel inflammation (although up to 50% may also have colitis), 
including inflammation in the ileum. CD causes significant symptoms including diarrhea, 
abdominal pain, bleeding, micronutrient malabsorption, iron deficiency anemia, increased 
risk of colon cancer, and unintended weight loss. Medical therapy for CD is problematic and 
no treatment has consistently delivered success in the treatment of CD due to the episodic 
nature and variability in abnormality. Current treatments are costly, and associated with 
adverse outcomes including increased risk of lymphoma and infections.2 
2.2 
Study Rationale 
The pathophysiology of CD is multi-factorial including potential genetic, environmental and 
microbial factors that may serve as potential triggers, and perpetuators, of the disease 
process. One consequence of inflammation in the ileum so common in CD is reductions in 
the absorption of bile acids (BA).3,4 BAs are important agonists to the nuclear farnesoid X 
receptor (FXR) in the intestines, as well as in the liver. The FXR receptor is known to regulate 
intestinal permeability, as well as innate immunity, including production of pro-inflammatory 
cytokines IL-1𝛃, IL-2, IL-6, tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-
𝛄).4,5 FXR agonists have been tested in animal models and early clinical trials, and show 
promise for altering inflammatory signaling pathways.4,6 Definitive clinical trials in humans 
with CD have not been performed.  
 
Mechanistic evidence supports the therapeutic potential of XN in CD, and thus Phase 1 safety 
data are required prior to moving on to Phase 2 trials in CD. XN is a flavonoid constituent of 
hops (Humulus lupulus), with antioxidant and anti-inflammatory activities. XN acts as a 
prebiotic for intestinal microflora, and along with its bacterial metabolites, alters the gut 
microbiome.7 In silico models suggest XN and its metabolites are receptor agonists for the 
FXR receptor,8 and our preliminary in vitro data suggest XN activates FXR-regulated genes 

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(SREBP1, G6Pase, and PEPCK) at low concentrations, suggesting XN acts as a 
pharmacologic agonist of FXR (unpublished data). Additional preliminary data from our 
collaborators demonstrates the bioavailability of XN, including increased bioavailability of XN 
when combined in a rice protein matrix.9 Thus XN has the potential to be a novel therapeutic 
in CD, by acting as a FXR agonist, reducing inflammation, increasing BA re-absorption, and 
reducing gut permeability.  
 
 
3. STUDY DESIGN 
This study will be an 8-week randomized, triple-masked, placebo-controlled parallel groups 
trial, in healthy adults aged 21 to 50 years. 
 
The intervention is xanthohumol (XN, 24 mg/day plus 288 mg of rice protein) or placebo (rice 
protein 288 mg/day), encapsulated in gelatin capsules, and taken by mouth once daily for 8 
weeks.  
 
The primary objective is to determine the safety and tolerability of xanthohumol in healthy 
adults over 8 weeks in a Phase 1 trial. The primary hypothesis is that not more than 50% 
more participants receiving XN will report an adverse event or experience a laboratory 
abnormality compared to those receiving placebo.  
 
The secondary objective is to collect comparison data for a subsequent Phase 2 trial in adults 
with Crohn’s disease (if an acceptable safety/tolerability profile is established by the Phase 
1 trial). Our primary outcome measure for this secondary objective is TNF-alpha as a 
biomarker of active inflammation. Because we are enrolling healthy adults, we hypothesize 
there will not be a difference in changes in TNF-alpha between XN and placebo.  
 
The tertiary objective is to collect comparison data related to XN metabolism, gut permeability 
and impact on the microbiota for a subsequent Phase 2 trial in adults with Crohn’s disease. 
Thus our outcome measures for this objective include XN metabolites, bile acid metabolism, 
biomarkers of gut inflammation and gut integrity, and composition of intestinal microbiome 
species.  
4. SELECTION AND ENROLLMENT OF PARTICIPANTS 
4.1 
Inclusion Criteria 
Participants must meet all of the following inclusion criteria to participate in this study:  
• 
Men and women aged 21-50 years 
• 
Willing to take isolated xanthohumol as a dietary supplement for 8 weeks 
• 
Willing to have blood drawn bi-weekly and fast for 10-12 hours before blood draws 
 

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• 
Willing and able to collect bi-weekly stool samples at home 
• 
Willing and able to collect 24-hour urine specimens prior to bi-weekly visits 
• 
Able to speak, read, and understand English 
• 
Must be able to provide written informed consent 
• 
Non-smokers (including tobacco and Cannabis products, combusted or vaporized) 
• 
For women of child-bearing potential, willingness to use an IUD or two other concurrent 
forms of birth control (e.g., 2 of the following categories: condoms, spermicide-containing 
gels, films or sponges; and/or vaginal rings) to prevent pregnancy while enrolled.  
4.2 
Exclusion Criteria 
Any candidate meeting any of the following exclusion criteria at baseline will be excluded 
from study participation:  
• 
History of any chronic disease including, but not limited to: diabetes (type 1 or 2); 
uncontrolled hypertension; coronary artery disease resulting in angina; cardiovascular 
disease requiring percutaneous coronary intervention (PCI), bypass, or past myocardial 
infarction or stroke; blood disease including current anemia; cancer (except non-
melanoma skin cancer) within the last year or still requiring chemotherapy or hormonal 
therapy; chronic kidney disease; liver disease including viral hepatitis, non-alcoholic 
fatty liver disease, or alcoholic hepatitis/cirrhosis; any immunocompromising condition 
including human immunodeficiency virus/acquired immunodeficiency syndrome or 
organ transplant requiring anti-rejection medications; chronic osteoarthritis requiring 
joint replacement or daily use of NSAIDs; chronic endocrine condition including but not 
limited to: Cushing's, Addison's, Hashimoto's thyroiditis, Grave's disease, etc. 
• 
Body Mass Index (BMI) less than 20 (underweight) or greater than 30 (obese) 
• 
Consumption of more than 1 beer per day. Candidates will be given the option to 
stabilize intake to this level for 14 days and re-contact the study team. 
• 
Use of NSAIDs more than once per week for headaches, routine aches/pains, etc. 
• 
Use of any prescription drugs, including oral contraceptives (due to potential 
interference with mechanisms under investigation) 
• 
Use of prescription opioids for any reason within the past 3 months 
• 
Use of prescription corticosteroids for any reason within the past 3 months 
• 
Acute viral or bacterial infection, or recent infection within the last 14 days or still 
requiring prescription medication for treatment 
• 
Recent acute trauma occurring within the last 14 days 
• 
Currently or recently (within last 14 days) taking any dietary supplements containing 
xanthohumol flavonoids, or other known herbal "anti-inflammatories" including: 
curcumin, turmeric, fenugreek, hops, rosemary, ginger, white willow, Devil's claw, fish oil 
(doses>1 g/day), or quercetin. Candidates will be given the option to "wash out" for 14 
days and re-contact the study team. 

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• 
Currently receiving intravenous nutrition support therapy (or within the last 30 days) 
• 
Currently taking anti-coagulant or anti-platelet prescription medications (or they were 
taken within the last 30 days) 
• 
Currently taking antibiotic, antiparasitic, or antifungal medications orally or intravenously 
(or they were taken within the last 30 days) 
• 
Initiation of or changes to supplements or medications within 14 days prior to screening. 
Candidates will be given the option to stabilize their allowable dietary supplements and 
medications for 14 days and re-contact the study team. 
• 
Initiation of or changes to an exercise regimen within 14 days prior to screening. 
Candidates will be given the option to stabilize their exercise for 14 days and re-contact 
the study team. 
• 
Initiation of or changes to a food plan within 14 days prior to screening. Candidates will 
be given the option to stabilize their diet for 14 days and re-contact the study team. 
• 
Current involvement or within 30 days prior to screening of a significant diet or weight 
loss program, such as NutriSystem, Jenny Craig, Atkin's or other low-carb diet 
programs, or very low-calorie liquid diet programs (such as optifast, medifast, and/or 
HMR) 
• 
Hospitalization (for any reason other than an elective medical procedure) within 3 
months prior to screening 
• 
Gastrointestinal surgery within 3 months prior to screening 
• 
Undergoing UV therapy (e.g. treatment for skin conditions such as psoriasis). 
• 
Engaging in vigorous exercise (e.g. activity at or above 80% of maximum perceived rate 
of exertion) more than 6 hours per week.  
• 
Women who are lactating, pregnant or planning pregnancy within the next four months 
• 
Typical intake of more than 2 alcohol-containing beverages per day, more than 14 per 
week, or more than 4 in any single day within the past 28 days. Candidates will be given 
the option to stabilize intake to these levels for 14 days and re-contact the study team. 
• 
Smoking tobacco or nicotine products (combusted or vaporized) 
• 
Use of illicit drugs/substances (such as but not limited to cocaine, phencyclidine (PCP), 
and methamphetamine) within 30 days of screening 
• 
Use of inhaled or ingested Cannabis products, including CBD. Candidates will be given 
the option to discontinue intake for 14 days and re-contact the study team. 
• 
Currently participating in another interventional research study or participated in another 
interventional study within 30 days of screening 
4.3 
Study Enrollment Procedures  
Participants will be recruited from the general community via posted fliers, newspaper 
advertisements, online recruitment outlines, and targeted social media posts, plus at the 
NUNM clinic via posted fliers and IRB-approved mailings to current patients. Retention of 
study participants will be achieved through proactive and timely communication, and 
through detailed attention to professionalism and safety. Additional retention strategies 

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include the use of stipends dispensed for clinical screening ($50) and each clinical visit 
completed ($100 each) following enrollment for up to a total of $550. Ineligible candidates 
will be notified of reasons for ineligibility, and this information will be recorded in a screening 
log. All participants must read, attest understanding of, and agree to all items on the 
informed consent form.   
 
Randomization will occur in up to 8 blocks of 4 based on biological sex. Initial randomization 
series will be generated using readily available random sequence generators designed to 
do so such as Randomization.com, which allows for block randomization.  
 
5. STUDY INTERVENTIONS  
5.1 
Interventions, Administration, and Duration  
The intervention is xanthohumol (XN, 24 mg/day plus 288 mg of rice protein) or placebo (rice 
protein 288 mg/day), encapsulated in gelatin capsules, and taken by mouth once daily for 8 
weeks.  
5.2 
Handling of Study Interventions  
The study product and visually-identical placebo will be delivered by Metagenics, Inc. to the 
NUNM via two separate shipments, i.e., one intervention and one placebo. Products will be 
stored at room temperature in separately locked cabinets in a secured storage location in the 
NUNM clinical research facility. Each locker will contain a Dispensing Log which will maintain 
a chain-of-custody record from receipt of the product to dispensing. Dispensation will require 
a Study Coordinator’s signature, data of dispensation, and number of bottles removed.   
 
Masking will be accomplished by using trained staff not involved in trial operations to:  
 
1. Produce the “code” (e.g., A=XN and B=Placebo);  
 
2. Label XN and placebo bottles according to the assigned code;  
 
3. Generating the blocked randomization sequence;  
 
4. Create 32 sequentially-numbered, sealed envelopes that contain the random group 
assignments. Envelopes will be opaque and signed across the seal. The randomization 
“code” will be kept in a sealed envelope with a signature across the label and dated the day 
of creation. This envelope will be kept in a dedicated study folder in the office of the Research 
Integrity Officer at NUNM. 
 
Upon confirmation of eligibility and interest, Study Coordinators involved with trial operations 
will then open the next sequential envelop in the stack at the time of randomization, the 
envelop number will be recorded, and allocation disclosed to both participant and coordinator 
at that time as simply “A” or “B”,  which will correspond to a separately locked cabinets within 
a secure storage location from which the designated bottle(s) will be dispensed. 
 

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Upon return of the unused study agent at Study Visit 4 (4-week) and Study Visit 6 (8-week) 
unused capsules will be counted and noted on the corresponding CRF (see CRFs and 
SOPs for the relevant visits).  Immediately following the completion of the study visit, the 
bottle containing unused capsules will be placed in a locked cabinet in the secure medical 
records facility in Helfgott Research Institute, where it will await a site visit by Westat 
personnel, at which time it may be disposed of in the dumpster.  The study agent is a 
natural product extracted from the hops plant, which is native to North America, South 
America, and Eurasia.  As such, it is widely distributed in the environment and does not 
represent a health or ecological hazard. 
5.3 
Concomitant Interventions  
5.3.1 Allowed Interventions 
• Use of over-the-counter medications as indicated on the label for less than 2 days per 
week 
• Multi-vitamin and/mineral  
• Dietary supplements except as detailed below  
 
5.3.2 Required Interventions 
N/A 
5.3.3 Prohibited Interventions 
• Dietary supplements containing xanthohumol 
• Dietary supplements containing flavonoids 
• Other known herbal “anti-inflammatories” including: curcumin, turmeric, fenugreek, 
hops, rosemary, ginger, willow, fish oil (doses>1 g/day), or quercetin. 
• Anti-coagulant or anti-platelet prescription medications 
• Antibiotic, antiparasitic, or antifungal medications orally or intravenously 
• Biologic therapies (e.g., infliximab, adalimumab, certolizumab pegol, natalizumab, 
vedolizumab, ustekinumab) 
• Glucocorticoids (e.g., prednisone, budesonide) (except in emergency management of 
flares) 
5.4 
Adherence Assessment  
Participant adherence to the intervention will be determined by asking participants if they 
have missed any capsules during regularly scheduled bi-weekly study visits, and by counting 
unused capsules returned at week 4 and week 8 visits. A participant will be counted as 
adherent to the intervention if they consume at least 80% of the scheduled daily doses. 

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6. STUDY PROCEDURES  
6.1 
Schedule of Evaluations 
Assessment 
Screening
Visit  
Baseline, 
Enrollment, 
Randomization 
Visit 1 (Week 0) 
Visit 2 
(Week 
2) 
Visit 3 
(Week 
4) 
 Visit 4 
(Week 
6) 
Closeout 
Visit 5 
(Week 8) 
Informed Consent Form   
X 
X 
 
 
 
 
Demographics 
X 
 
 
 
 
 
Inclusion/Exclusion 
Criteria  
X 
X 
 
 
 
 
Medical History  
X 
X 
 
 
 
 
Current Medications 
X 
X 
 X 
X 
X 
X 
Anthropometrics/Vitals 
X 
X 
X 
X 
X 
X 
Blood Chemistries 
X 
X 
X 
X 
X 
X 
Hematology 
X 
X 
X 
X 
X 
X 
Enrollment/Randomization 
 
X 
 
 
 
 
Inflammatory markers 
(blood) 
 
X 
X 
X 
X 
X 
Urine Analysis 
 
X 
X 
X 
X 
X 
Stool Analysis 
 
X 
X 
X 
X 
X 
Adverse Events  
 
X 
X 
X 
X 
X 
Tolerability: PROMIS-29 
 
X 
X 
X 
X 
X 
 
 
6.2 
Description of Evaluations  
All biomarkers will be collected as closely to the indicated timepoint as possible, allowing a window of +/- 3 
days at each time point, e.g., 2 weeks +/- 3 days is allowable.  
 
 
Measures to meet Primary Objective: Clinical Toxicology  
 
Clinical safety will be determined based on subjective reports of AEs, as well as, objective monitoring of changes 
in clinical laboratory measures. PROMIS-29 will also be administered as an additional indication of tolerability. 
The lab measures of safety in this Phase 1 trial include measures of hematological, renal and liver safety as: 
mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV), and hematocrit (derived from a 
complete blood count); estimated glomerular filtration rate; blood urea nitrogen to creatinine ratio; aspartate 
aminotransferase (AST); alanine aminotransferase (ALT); gamma-glutamyl transferase (GGT). All parameters 
will be measured at Baseline/Enrollment and after 2, 4, 6 and 8 weeks of the intervention. Results for all 
participants will be summarized and the following reported: % abnormal, % new abnormals, and mean change 
from Baseline/Enrollment. 
 

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Measures to meet Secondary Objective: Biologic Signature 
• Change in levels of metabolic byproducts of xanthohumol: Xanthohumol and xanthohumol 
metabolites in blood, urine, and stool will be measured by ultra-high performance liquid 
chromatography-quadrupole time-of-flight mass spectrometry, and expressed as change over 
time from baseline. Metabolites will be measured at Baseline/Enrollment and after 2, 4, 6 and 8 
weeks of the intervention. 
 
• Bile acid profile: Bile acid concentrations in stool samples will be measured by ultra-high 
performance liquid chromatography-quadrupole time-of-flight mass spectrometry, and 
expressed as mean change over time from baseline. All parameters will be measured at 
Baseline/Enrollment and after 2, 4, 6 and 8 weeks of the intervention. 
 
 
• Gut inflammation: Fecal calprotectin, a protein associated with gut inflammation and irritable 
gut syndrome, will be measured by enzyme-linked immunosorbent assay, and expressed as 
mean change over time from baseline. All parameters will be measured at Baseline/Enrollment 
and after 2, 4, 6 and 8 weeks of the intervention. 
 
 
6.2.1 Screening Evaluation 
Consenting Procedure 
Upon initial determination of eligibility over the phone, candidates will be invited to an in person 
Clinical Screening Visit conducted by the Study Coordinator. Administration of a written informed 
consent form will be the first activity of this visit. This first consent applies to drawing a blood 
sample for confirmation of eligibility to participate in the full study.  
 
Informed consent materials will be written in English at an eighth-grade reading level, and study 
personnel will be available to answer any questions the participant may have during the process. 
Candidates will have adequate time to review the written informed consent document (or have 
it read to them, according to their preference) which clearly indicates the voluntary nature of their 
participation and provides recourse should they feel their safety or privacy was violated in any 
way. Candidates will have an opportunity to ask any questions of the Coordinator or the site PI. 
 
Upon confirmation of eligibility to participate in the trial following review of laboratory results, a 
second informed consent form will be administered during the Baseline/Enrollment visit which 
follows the same procedures above but will apply to participation in the full trial.  
 
The informed consent form for full trial participation also includes a statement of consent to re-
use the biologic samples collected for these trials for subsequent analyses related to the Aims 
of the proposed trials, should they become necessary. Participants will have the autonomy to 
provide, or refuse, additional consent for this purpose. A small volume (<2 mL) of blood and 
urine will be collected and frozen at -70C for the purposes of potential future analyses. 
 

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Any revisions of the informed consent documents deemed necessary will be approved by the 
NUNM IRB prior to implementation.  
 
Signed forms will be retained for the duration of the study in a secure, dedicated records room 
with restricted access. 
 
Screening 
The screening visit will allow for: 
• Administration of a written informed consent  
• Review of eligibility criteria 
• Measurement of anthropometrics and vitals 
• Collection of blood to rule out anemia, hepatitis, and renal disease 
• Collection of urine for determination of pregnancy status (where appropriate) 
• Oral swab for HIV screening 
 
Lab results will be reviewed within 48 hours of receipt and, if eligible, the candidate will be 
required to return for a Baseline/Enrollment visit within 30 days of screening, or else will require 
re-assessment of laboratory measures to re-assess eligibility.  
Upon review and confirmation of eligibility, candidate participants will be contacted to schedule 
a Baseline/Enrollment visit, during which they will be given stool and urine collection kits and 
instructed on their proper use. The day following their Baseline/Enrollment visit, participants will 
return the baseline stool and urine specimens, and will receive their first bottle of study product.  
6.2.2 Enrollment, Baseline, and/or Randomization 
Enrollment 
Enrollment in the study will occur at the baseline visit, immediately prior to randomization.  
Baseline Assessments 
• Demographics 
• Anthropometrics (BMI, waist circumference) and vitals (BP and pulse rate) 
• Health-related quality of life (PROMIS-29) 
• Symptoms/Adverse-event questionnaire for Baseline anchoring 
• Collection of blood for: 
§ Complete blood count  
§ Electrolytes 
§ Estimated glomerular filtration rate 
§ Blood urea nitrogen to creatinine ratio  
§ Aspartate aminotransferase (AST)  
§ Alanine aminotransferase (ALT)  
§ gamma-Glutamyl transferase (GGT) 

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§ Alkaline phosphatase  
§ Bilirubin  
• Collection of blood for Baseline measurement of: 
§ Metabolic byproducts of xanthohumol  
• Collection of a urine specimen for Baseline measurement of: 
§ Metabolic byproducts of xanthohumol  
• Collection of a stool specimen for Baseline measurement of: 
§ Microbiota sequencing  
§ Fecal calprotectin 
§ Gut permeability biomarkers  
§ Metabolic byproducts of xanthohumol 
 
Randomization 
Randomization will occur within 30 days of screening during the Baseline/Enrollment visit, 
following confirmation of eligibility and signing of the informed consent form. Study 
Coordinators involved with trial operations will assign the participant to a treatment arm by 
opening the next sequential envelop in a previously created stack of randomized, sealed 
envelopes (see detailed description in section 5.2). The envelope number will be recorded, 
and allocation disclosed to both participant and coordinator at that time as simply “A” or “B”, 
indicating which bottles will be dispensed. Group allocation will be recorded in the baseline 
visit case report form. On the day following this visit (i.e. at the time of baseline sample 
return), study product will be provided to the participant to take home, but the container will 
only indicate “A” or “B”. 
6.2.3 Blinding/Masking/Allocation Concealment 
Masking procedures are detailed in section 5.2 above. This intervention has a low potential 
of producing effects that would compromise the masking of group allocation. 
Unmasking is permissible if the Data Safety Monitoring Board requires it upon review and in 
the case of a Serious Adverse Event (SAE). If the DSMB identifies a need to break the 
allocation masking, the Chair of the DSMB will consult with relevant officials at NCCIH and 
FDA prior to unmasking.  
6.2.4 Follow-up Visits 
Bi-weekly follow-up visits will be conducted for 8 weeks total. Each follow-up visit must occur 
within a 72 hours window of each planned visit, e.g., the 4-week follow-up visit must occur 
within 4 weeks +/- 3 days. Follow-up visits for the total duration of the trial will be scheduled 
upon Enrollment whenever possible. Each follow-up visit will end with the dispensation of 
urine and stool collection kits for the next suitable follow-up visit, with the necessary 
instructions on sample collection. The following will be included in each follow-up visit: 
• Anthropometrics (BMI, waist circumference) and vitals (BP and pulse rate) 
• Health-related quality of life (PROMIS-29) 
• Adverse events interview/questionnaire 
• Collection of blood for: 
§ Complete blood count  

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§ Electrolytes 
§ Estimated glomerular filtration rate 
§ Blood urea nitrogen to creatinine ratio  
§ Aspartate aminotransferase (AST)  
§ Alanine aminotransferase (ALT)  
§ gamma-Glutamyl transferase (GGT)  
§ Alkaline phosphatase  
§ Bilirubin 
• Collection of blood for Baseline measurement of: 
§ Metabolic byproducts of xanthohumol  
• Collection of a urine specimen for Baseline measurement of: 
§ Metabolic byproducts of xanthohumol 
• Collection of a stool specimen for Baseline measurement of: 
§ Microbiota sequencing  
§ Fecal calprotectin 
§ Gut permeability biomarkers  
§ Metabolic byproducts of xanthohumol 
6.2.5 
Completion/Final Evaluation 
• Anthropometrics (BMI, waist circumference) and vitals (BP and pulse rate) 
• Health-related quality of life (PROMIS-29) 
• Adverse events interview/questionnaire 
• Collection of blood for: 
§ Complete blood count  
§ Electrolytes 
§ Estimated glomerular filtration rate 
§ Blood urea nitrogen to creatinine ratio  
§ Aspartate aminotransferase (AST)  
§ Alanine aminotransferase (ALT)  
§ gamma-Glutamyl transferase (GGT)  
§ Alkaline phosphatase  
§ Bilirubin 
• Collection of blood for Baseline measurement of: 
§ Metabolic byproducts of xanthohumol  
• Collection of a urine specimen for Baseline measurement of: 
§ Metabolic byproducts of xanthohumol  
• Collection of a stool specimen for Baseline measurement of: 
§ Microbiota sequencing  
§ Fecal calprotectin 
§ Gut permeability biomarkers  
§ Metabolic byproducts of xanthohumol 
 
For participants who withdraw from the trial prior to completing all scheduled visits, every 
attempt will be made to contact them and determine their reason for their withdrawal. 
Common reasons include inconvenience, geographical challenges, unexpected life events, 
and adverse effects. If adverse effects are noted as the reason, all reported adverse effects 

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will be graded and reported according to specifications noted below. All withdrawn 
participants will be queried for any/all adverse effects, and all reported events will be graded 
and included in summary reports.  
7. SAFETY ASSESSMENTS  
Safety will be assessed at every visit through the measurement of clinical toxicology 
laboratory measures (blood counts, liver and renal function tests as detailed above) and 
multi-system self-reported adverse events (per NCI definitions).  
Previous clinical trials of XN have not resulted in any acute or chronic adverse events. 
However, potential adverse events are anticipated as possible: 
• Allergy to study product 
• Blood draw discomfort/bruising 
• Gastrointestinal symptoms (described below) 
Risks associated with any dietary supplement include gastrointestinal symptoms such as 
gas, bloating, digestive upset and change in bowel movement frequency or consistency. Also 
there is a chance of identifying a previously unknown allergy, and therefore a risk of rash, 
breathing difficulty and serious allergic reaction. 
7.1 
Specification of Safety Parameters 
Standard clinical Complete Blood Counts and toxicology panels including liver and renal 
function tests will be measured at each bi-weekly clinical visit. Values will be interpreted 
based on: new abnormal value, change since baseline, new emergency or alert values, 
and/or the emergence of new diagnostic concerns. All participants’ labs will be reviewed by 
the clinical investigator. 
7.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety Parameters 
In light of the benign nature of XN, historical experience with its consumption as a common 
food ingredient, and the availability of preliminary human studies, the FDA has approved 
extending the duration of the proposed intervention to 8 weeks. Based on the available 
human data, it is expected that the study participants will not develop adverse effects at the 
proposed dose during the 8-week exposure period. However, a primary purpose of this study 
is to provide a clearer picture of the safety and physiological effects of XN. Therefore, blood 
samples will be taken at baseline and at each bi-weekly visit for complete blood counts and 
comprehensive metabolic panels, including liver and renal function tests, accompanied by 
determination of weight, blood pressure, and pulse rate. Additionally, adverse events will be 
solicited by interview at each visit, and participants will be encouraged to report any concerns 
as needed between visits.  
All assessments, together with reported adverse events and unanticipated problems, will be 
collected and reviewed regularly, and per occurrence where appropriate (see Monitoring 
table in section 7.6) by the PI and specified monitoring bodies. As specified in section 7.1, 
values will be interpreted based on: new abnormal value, change since baseline, new 

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emergency or alert values, and/or the emergence of new diagnostic concerns. If statistical 
analysis is required for interpretation, the NUNM study Biostatistician, Dr. Doug Hanes, will 
conduct the analysis.  
The general population is primarily exposed to XN through consumption of beer. A major 
brand beer contains about 0.5-1 mg prenylflavonoids (mainly XN and isoXN) per liter and a 
bitter microbrew ale may contain as much as 4 mg of total prenylflavonoids.15  
 
XN is not currently FDA approved for the treatment, cure, prevention or mitigation of disease. 
Based on suspected mechanisms of action, potential medical uses include: inflammatory 
bowel disease, mucous colitis, prostate cancer, breast cancer, ovarian cancer, menopausal 
symptoms, hyperlipidemia, tuberculosis, cystitis, intestinal cramps, and neuralgia. XN is likely 
safe when used orally in amounts commonly found in foods. Hops and hops oil have 
Generally Recognized as Safe (GRAS, Flavoring Extract Manufacturers’ Association records 
2578 and 2580 respectively) status in the US.10 The Stevens lab at Oregon State University 
has completed the first human pharmacokinetics study of XN in which 48 study participants 
ingested a single dose of XN of 20, 60, or 180 mg (16 men and women per dose group).11 
No treatment-related adverse effects were reported in this study (ClinicalTrials.gov Identifier: 
NCT01367431). Risks associated with any dietary supplement include gastrointestinal 
symptoms such as gas, bloating, digestive upset and change in bowel movement frequency 
or consistency.  
 
Xanthohumol has been extensively studied in animals. Gerhauser and colleagues studied 
the safety of XN administered orally to Sprague-Dawley rats for four weeks.12 At a dose of 
1000 mg XN/kg body weight, authors reported a reduction of liver weight, but found no 
macroscopic or histopathologic changes of the liver, kidney, lung, heart, stomach and spleen. 
The mammary glands of treated rats appeared less developed compared to the control 
animals. In a two-generation study of the teratogenic effects of XN, Gerhauser’s group found 
no differences in development of SD rats treated lifelong with a daily dose of 100 mg/kg body 
weight. 
 
In another safety study of XN, mice exposed to XN for four weeks via the drinking water at a 
concentration of 0.5 mM (=177 mg XN per liter) showed no changes in blood counts or in the 
integrity of the bone marrow, liver, exocrine pancreas, kidneys, muscles, thyroid, ovaries or 
the surrenal cortex.13 We estimate that the XN exposure in this study is equivalent to a human 
dose of 265 mg per day, which is much higher than the highest dose we propose in our 
clinical trial, i.e., 24 mg/day. In Zucker fa/fa rats study14, there was no difference in uterus 
weight (n=6, females) or testes weight (n=6, males) between dose groups and the zero dose 
control group (n=6, females/males). Animals were given daily oral doses of 1.86 mg/kg, 5.64 
mg/kg, or 16.9 mg/kg body weight for six weeks. These doses correspond to 20 mg, 60 mg 
and 180 mg XN in a 64 kg human by allometric interspecies scaling. 
Based on in vitro data, the trial measured endpoints related to protection of DNA (single cell 
gel electrophoresis, 8-oxo-guanosine and 8-oxo-7,8-dihydro-2′-deoxyguanosine levels in 
urine); oxidative stress (15F2t-isoprostanes in urine, and malondialdehyde, oxidized low-
density lipoprotein, oxygen radical absorbance capacity, and ferric reducing ability in 
plasma); energy metabolism (glucose, cholesterol, and lipoproteins); inflammation (C-

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reactive 
protein 
and 
urea); 
hormone-related 
parameters 
(17beta-estradiol 
and 
progesterone); and osteogenesis (alkaline phosphatase activity and osteocalcin). The 
observed differences between XN and placebo groups were limited to protective effects 
against DNA damage in the XN group (indicated through single cell gel electrophoresis in 
blood lymphocytes, and reduced urinary excretion of 8-oxo-7,8-dihydro-2′-deoxyguanosine 
and 8-oxo-guanosine). 
Additional research on XN has been performed in humans which support its safety and 
potentially beneficial mechanisms of action. Researchers at the Institute of Cancer Research 
at the University of Vienna, Austria, conducted a human study with the non-alcoholic 
beverage, XAN Wellness (TA-XAN AG Wiesbaden, Germany).16 The tested beverage 
contained 12 mg/L XN. The study was designed as a cross-over, double-masked, placebo-
controlled study with 22 subjects (11 men and 11 women, ages 19-37, nonsmokers). The 
subjects consumed one liter of the beverage per day, for 14 days. After a washout period of 
14 days, subject crossed over to the alternate treatment (placebo or XN treatment). The daily 
dose was 12 mg XN.  
Stevens and co-workers investigated the possible protective effects of XN on oxidative DNA 
damage and oxidative stress in a crossover, double-blinded, placebo-controlled human 
intervention study at three dose levels of XN: 6, 12 and 24 mg/day for 3 weeks 
(ClinicalTrials.gov Identifier: NCT02432651; IND #122633). Healthy human subjects 
consumed a non-alcoholic beverage without (placebo control) and with XN for 3 weeks, with 
a washout period of 3 weeks. Urine and blood were collected at the start and at the end of 
both 3-week intervention periods. Oxidative DNA damage was evaluated in peripheral white 
blood cells using single cell gel electrophoresis assay and by the measurement of urinary 8-
oxo-7,8-dihydro-2´-deoxyguanosine (8-oxo-dG). 8-Oxo-7,8-dihydroguanosine (8-oxo-Guo), 
a marker for RNA oxidation, was also measured in urine samples. The results showed that 
consumption of XN at 12 mg/day for 3 weeks protected DNA in white blood cells from 
oxidative damage as measured by the single cell gel electrophoresis, but did not influence 
the urinary excretion of 8-oxo-dG and 8-oxo-Guo. The blood levels of oxidized glutathione 
were significantly reduced by XN taken at a dose of 6 mg/day for 3 weeks. These findings 
suggest that XN administered in the form of a fortified non-alcoholic beverage is protective 
against DNA damage and oxidative stress in healthy individuals. No treatment-related 
adverse effects were reported or observed in this study. Thus the dose chosen for this trial, 
24mg/day, aligns with the highest dose for which we have preliminary evidence of safety, 
tolerability and biologic effects.  
The Van Breemen group evaluated the interaction of a hop-derived botanical supplement 
with drug metabolism in women.17 Low doses of an oral cocktail of caffeine, tolbutamide, 
dextromethorphan, and alprazolam (probes for metabolism by CYP1A2, CYP2C9, CYP2D6, 
and CYP3A4, respectively) were administered to 16 peri- and post-menopausal women at 
baseline and then again after consuming a hop dietary supplement (2 capsules, twice daily 
for 14 days). The capsules contained 8 mg XN, 0.8 mg isoXN, 0.25 mg 8-prenylnaringenin, 
and 1.3 mg 6-prenylnaringenin. The pharmacokinetics values for each probe substrate 
showed no clinically relevant changes due to consumption of the hop dietary supplement. 
They concluded that consumption of the hop dietary supplement did not interact with drug 
metabolism. 

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There are no known toxicities for human subjects taking oral supplements containing XN. 
The general population is primarily exposed to XN through consumption of beer. A major 
brand beer contains about 0.5-1 mg prenylflavonoids (mainly XN and iso-XN) per liter and a 
bitter microbrew ale may contain as much as 4 mg of total prenylflavonoids.15 In Europe, a 
beer/beverage, containing 4 mg of XN per liter, has been on the market for several years 
without adverse effects having been reported by the consumers. Based on the limited human 
data and the results from the above cited animal studies, we expect that the study participants 
will not develop adverse effects at the proposed dosage regimens during the 8-week 
exposure period. 
7.3 
Adverse Events and Serious Adverse Events  
An adverse event (AE) is generally defined as any unfavorable and unintended diagnosis, 
symptom, sign (including an abnormal laboratory finding), syndrome or disease which either 
occurs during the study, having been absent at baseline, or if present at baseline, appears 
to worsen. Adverse events are to be recorded regardless of their relationship to the study 
intervention.   
A serious adverse event (SAE) is generally defined as any untoward medical occurrence 
that results in death, is life threatening, requires inpatient hospitalization or prolongation of 
existing hospitalization, results in persistent or significant disability/incapacity, or is a 
congenital anomaly. 
Adverse event (AE) monitoring will occur using a standardized multi-system AE 
questionnaire based on the National Cancer Institute’s Common Terminology Criteria for 
Adverse Events v4.0 (2009). Adverse events of grades 4 and 5 will be considered Serious 
Adverse Events (SAE). 
Standard clinical Complete Blood Counts and toxicology panels including liver and renal 
function tests will be measured at each bi-weekly clinical visit. Values will be interpreted and 
summarized based on: new abnormal value, change since baseline, new emergency or alert 
values, and/or the emergence of new diagnostic concerns. All participants’ labs will be 
reviewed by the Clinical Investigator. 
7.4 
Reporting Procedures 
Serious AEs will be reported to the PI within the same business day; all other AEs within 48 
hours. Serious AEs, and any AE that in the discretion of the Clinical Investigator may require 
a modification to medications, will be reported to primary care doctors via phone contact 
between the Clinical Investigator and the PCP within the same business day, if possible, or 
the following. All other AEs will be reported to the PCPs in a summary report from each 
clinical visit within 1 week of scheduled visits. Adverse events will be dealt with directly, 
discussed with co-investigators and advisors, and adjudicated by the DSMB as related, 
possibly related, or unrelated to the study procedures. 
 
In accordance with NUNM IRB AE reporting guidelines, SAEs (grades 4-5) will be reported 
to the NUNM IRB within 24 hours of PI awareness. Unanticipated AEs of any grade that are 
both related to research, result in a change to risk/benefit, or require protocol and/or consent 
modifications will be reported to the NUNM IRB within 10 working days of PI awareness. All 

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Anticipated AEs regardless whether they are classified as severe or related to the study 
intervention will be reported at the time of annual continuing review.   
 
All adverse events will be recorded in the Adverse Event Log.  
 
The DSMB will meet quarterly to conduct continuous review of data and participant safety.  
 
The DSMB will review monthly safety reports, and adjudicate adverse events as required.  
 
The DSMB will recommend amendments to the protocol, changes in study procedures, 
changes to the data collection plan or study forms, or study termination due to safety or other 
issues as needed.   
 
The PI will have final responsibility for all reporting to the NUNM IRB, DSMB and/or NCCIH 
PO as required.   
7.5 
Follow-up for Adverse Events 
All adverse events, regardless of grade, will be recorded with start dates, graded severity, 
medical interventions as relevant, clinical monitoring plan as indicated, and date of resolution.  
If participants believe they have suffered an AE related to study procedures, but outside of 
routinely scheduled clinical follow-ups, they are welcome to contact us at any time and AEs 
will be adjudicated. At each study visit, the investigator will inquire about the occurrence of 
AE/SAEs since the last visit. Events will be followed for outcome information until resolution 
or stabilization for at least 30 days following discontinuation of any study-related 
interventions.  
 
7.6 
Safety Monitoring  
The NUNM IRB, DSMB, and NCCIH Program Officials will receive copies of all study 
monitoring/audit or inspection reports within 14 day of PI receipt. 
Safety Review Plan  
Study progress and safety will be reviewed monthly (and more frequently if needed). 
Progress reports, including participant recruitment, retention/attrition, and AEs will be 
provided to the DSMB. An Annual Report will be compiled to address (1) whether AE rates 
are consistent with pre-study assumptions; (2) reason for dropouts from the study; (3) 
whether all participants met entry criteria; (4) whether continuation of the study is justified on 
the basis that additional data are needed to accomplish the stated aims of the study; and (5) 
conditions whereby the study might be terminated prematurely. The Annual Report will be 
sent to the DSMB and will be forwarded to the IRB and NCCIH. The IRB and other applicable 
recipients will review progress of this study on an annual basis. 
 
 

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Monitoring 
Data type 
Frequency of review 
Reviewer 
Subject accrual 
(including compliance 
with protocol enrollment 
criteria) 
Weekly 
PI 
Monthly 
Study team 
Quarterly 
Independent Monitors 
Status of all enrolled 
subjects, as of date of 
reporting 
Weekly 
PI 
Quarterly 
Independent Monitors 
Data entry quality control 
checks on 15% of charts 
Monthly 
PI 
Adherence data 
regarding study visits 
and intervention 
Per visit 
Study team 
 
Monthly 
PI 
 
Quarterly 
Independent Monitors 
 
Annually 
NCCIH 
SAEs (unexpected and 
related) 
Per occurrence 
PI, Independent 
Monitors NIH/NCCIH 
SAEs (expected or 
unrelated) 
Per occurrence 
PI 
Annually 
Independent Monitors, 
NIH/NCCIH 
Unanticipated Problems 
Weekly 
PI 
Per Policy 
IRB 
 
8. INTERVENTION DISCONTINUATION  
Participants may discontinue the intervention at any time by their own choosing. However, 
the intervention may be discontinued by the study staff if they experience moderate or greater 
AEs requiring them to withdraw or new onset laboratory abnormalities (confirmed with repeat 
testing), including new elevation of liver function tests (AST, ALT, bilirubin or alkaline 
phosphatase), newly abnormal electrolytes, new onset anemia, newly abnormal creatinine 
or estimated glomerular filtration rate (eGFR).  
Of course, the intervention may also be discontinued if, for some reason, trial operations are 
suspended by the Institution, sponsor, or FDA requirement.  
Because this trial is a mechanistic, proof-of-concept trial focused on highly mechanistic 
outcome measures that require participants to consume the intervention (or placebo) 

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participants will not be maintained in the trial, or complete follow-up visits, if they discontinue 
the intervention (or placebo). The study staff will however follow up with them to query any/all 
adverse events as relevant and attempt to ascertain the reason for withdrawal.  
9. STATISTICAL CONSIDERATIONS  
9.1 
General Design Issues  
The primary hypothesis is that not more than 50% more participants receiving XN will report 
an adverse event or experience a laboratory abnormality compared to those receiving 
placebo. For our Secondary Objective, we hypothesize there will not be a difference in 
changes in TNF-alpha between XN and placebo. 
A placebo-controlled, parallel group trial is the only design conducive to determining if 
xanthohumol is the cause of any measured changes in safety parameters, inflammation and 
gut-related biomarkers and/or changes in microbiota composition or products in a timely 
manner. Season-, diet-, and stress-induced changes in the microbiota necessitate timely 
completion of the trial objectives, plus the unknown residual effects of XN supplementation 
preclude crossover or n-of-1 trial designs.  
9.2 
Sample Size and Randomization 
A sample size of 12 per group provides 80% power to detect a 50% difference in proportions 
of participants between groups by chi2 test experiencing a laboratory abnormality or adverse 
event at a significance threshold of α=0.05.  
 
Randomization and Treatment Assignment Procedures 
Allocation concealment and randomization procedures are described in Section 6.2.2 and 
6.2.3 above.  
9.3  
Definition of Populations 
Only per protocol analyses will be performed for this mechanistic, proof-of-concept study. 
The per protocol population will be those randomized to the intervention or placebo and 
who take 80% or more of the assigned product.  
9.4 
Interim Analyses and Stopping Rules 
Interim analyses will only include those analyses necessary for continuing safety 
assessments, including the accumulation of AEs by grade, the frequency of new onset 
laboratory abnormalities (confirmed with repeat testing), including new elevation of liver 
function tests (AST, ALT, bilirubin or alkaline phosphatase), newly abnormal electrolytes, 
new onset anemia, newly abnormal creatinine or estimated glomerular filtration rate (eGFR). 
The analyses will remain masked to group and will be prepared by the NUNM biostatistician 
periodically to determine accumulated frequencies of events.  
 
This study will be stopped prior to its completion if the intervention is associated with adverse 
effects that call into question the safety of the intervention, if any new information becomes 

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available during the trial that necessitates stopping the trial; or if other situations occur that 
might warrant stopping the trial. The ultimate decision to stop the trial may be made by the 
DSMB, NCCIH, and/or the FDA. 
The trial will be halted if >25% of the total sample experience moderate or greater AEs 
requiring them to withdraw or new onset laboratory abnormalities (confirmed with repeat 
testing), including new elevation of liver function tests (AST, ALT, bilirubin or alkaline 
phosphatase), newly abnormal electrolytes, new onset anemia, newly abnormal creatinine 
or estimated glomerular filtration rate (eGFR).  
 
The trial may be stopped if:  
1. SAEs attributable to the study intervention occur in 25% of the intervention group.  
2. New onset laboratory abnormalities (upon repeat testing) occur in 50% of the intervention 
group.  
3. The FDA, IRB, DSMB, and/or NCCIH recommend stopping the trial completing after 
review of the available data after a temporary suspension of the trial.  
 
The DSMB will provide a final recommendation to the PI to halt the study or amend the study 
protocol, and will develop a response plan, in consultation with the NUNM IRB and the 
NCCIH Program Officer.   
If immediate changes are required for participant safety, enrollment and other study activities 
will not continue until the modifications/amendment is approved by the IRB. 
9.5 
Outcomes  
In this study, measured outcomes will take the form of laboratory values. Consequently, we 
do not anticipate the need to adjudicate any of the assessments.  
9.5.1 Primary Outcome   
As the primary objective relates to safety and tolerability, the associated primary outcome 
measures relate to cumulative routine clinical toxicology measures including: complete blood 
count, electrolytes, estimated glomerular filtration rate, blood urea nitrogen to creatinine ratio, 
aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl 
transferase (GGT), alkaline phosphatase, and bilirubin, plus self-reported adverse events 
and health-related quality of life (PROMIS-29). 
 
Cumulative safety will be reported as the frequency of new onset laboratory abnormalities 
(confirmed with repeat testing), including: new elevation of liver function tests (AST, ALT, 
bilirubin or alkaline phosphatase), newly abnormal electrolytes, new onset anemia, newly 
abnormal creatinine or estimated glomerular filtration rate (eGFR).   
 
9.5.2 Secondary Outcomes   
The outcome measure for the secondary objective of assessing changes in inflammation is 
TNF-alpha as a biomarker of active inflammation.  
 
 
 

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9.5.3 Tertiary Outcomes   
The tertiary objective is to collect comparison data related to XN metabolism, gut permeability 
and impact on the microbiota for a subsequent Phase 2 trial in adults with Crohn’s disease. 
Thus our outcome measures for this objective include XN metabolites in the blood, stool, and 
urine, assessment of bile acid metabolism, biomarkers of gut permeability, enterocyte and 
systemic inflammation; and microbiota population and expression.  
Specific biomarkers include: fecal calprotectin, soluble CD14, lipopolysaccharide-binding 
protein (LBP), cytokines (IL-8, IL-1β, IL-6, IL-10, and IL-12p70), 16s rRNA sequencing and 
gene product results measured by metagenomic DNA sequencing. 
9.6 
Data Analyses  
Analysis of laboratory toxicology measures: 
Laboratory measures including red blood cell count, white blood cell count, hematocrit (%), 
hemoglobin, mean corpuscular volume, AST, ALT, creatinine-estimated glomerular filtration 
rate (eGFR), blood urea nitrogen (BUN), and creatinine will be analyzed by first assessing 
descriptive statistics including mean, median and standard deviation. All distributions will be 
tested for skew and transformed (e.g., natural log transformed) as needed to reduce the 
influence of skew in subsequent analyses. All parameters will be evaluated by formally 
comparing means from baseline to each time point by unpaired, 2-sided t-tests, as well as 
for trends over time by applying linear mixed ANOVA models with time point as a repeated 
factor.  
As an element of the evaluation for safety of the intervention, the % laboratory newly 
abnormal values within the clinically normal range at baseline per clinical laboratory 
reference ranges for each lab parameter (per list above) will be reported at each 2-week time 
point and the % compared between XN and placebo groups by Fisher’s exact test.  
Assessment for changes in the distribution of each lab parameter will be performed by 
presenting means and confidence intervals (CI) for the change in each group, as well as the 
mean CI for the difference between groups; significance will be tested using 2-sided, 
unpaired t-tests (or, if inspection of distributions indicates strong non-normality, with a non-
parametric Wilcoxon signed rank test) of the mean values for XN compared to placebo.  
Assessment for overall increases or decreases in parameters of interest will be tested for 
linear trends of the means by linear mixed ANOVA with time point as a repeated factor, which 
may be suggestive of evidence for cumulative toxicity if trending in a clinical deleterious 
direction.  
 
Analysis of inflammatory cytokines, gut permeability biomarkers, and endotoxemia:  
Given the small sample size of these clinical trials, results related to changes in inflammatory 
cytokines (IL-1β, IL-2, IL-6, IL-8, IL-10, TNF-α, and IL-12p70), gut permeability biomarkers 
(CD14, intestinal fatty acid binding protein) and markers of endotoxemia (LPS, LPS-binding 
protein) are considered preliminary, and thus our principle assessment will be measurement 
of effect size as Cohen’s d statistic between XN- and placebo-treated groups at each 2-week 
interval compared to baseline, with the primary comparison being between baseline and 

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week 8. We will also perform formal statistical comparisons as unpaired, 2-sided t-tests (or, 
if inspection of distributions indicates strong non-normality, with a non-parametric Wilcoxon 
signed rank test) of the means at relevant time points. The following findings will be 
considered evidence of possible effects: 1. Clinically and statistically significant effects for 
changes in cytokines of interest and/or 2. Calculation of a Cohen’s d value for effect size 
estimation greater than 0.5 (with larger d indicating a larger potential effects size).  
Assessment for overall increases or decreases in parameters of interest will be tested for 
linear trends of the means by linear mixed ANOVA with time point as a repeated factor. 
Trends will be considered significant if p<0.05 for the model overall.  
 
Analysis of XN, its metabolites, and BAs to determine a small molecule signature of longer-
term oral treatment with XN:  
We will identify and quantify XN and XN-derived metabolites in plasma, stool, and urine. Bile 
acid profiles will be also be determined in stool samples. The metabolite profile data will then 
be used for correlation with the time-resolved gut microbiome analysis. This approach will 
allow us to determine links between metabolite profiles and gut microbiome compositions. 
Determining differential abundance between treatments: High-level exploratory analyses will 
be performed through metrics such as alpha and beta diversity and ordination techniques. 
Further, differential abundance analysis for 16S and metagenomics data will be conducted 
through negative binomial models, with specifications to deal with inflation of zero counts, 
such as those implemented in statistical R packages edgeR18 and DESeq219 or through a 
compositional Bayesian data analysis approach, such as the approach implemented in the 
R package ALDEx2.20 Exploratory data analysis will first be done to evaluate which modeling 
technique is appropriate for the data generated based on assumptions made by each 
modeling approach. For MS-based omics data, we use a suite of robust statistical 
approaches for: 1) processing the data to identify outliers,21–23 and 2) identifying statistically 
different proteins and metabolites between comparative samples. As a first step to statistical 
testing, we use the Statistical Procedure for the Analysis of molecular abundance 
Normalization Strategies (SPANS) approach to simultaneously evaluate several MS-based 
omics data set normalization strategies to determine the most appropriate normalization 
factor in the context of the experimental design.24 For individual data streams, there are 
several strategies for identifying molecular features that statistically classify patients, 
depending on the structure of the data and the phenotype data being used for classification. 
For the determination of statistically relevant features, if the features follow the standard 
normal distribution, then we utilize standard ANOVA methods; otherwise, a non-parametric 
version, such as Kruskal-Wallis or sum-rank test, can be utilized.25 To evaluate the statistical 
relevance of missing data, for example when the data is absent in the control group and 
present in the treatment group, we will utilize a G-test.23 However, with multiple comparisons 
comes the problem of increased likelihood of a significant result by random chance. To 
counter this problem, we use either the Dunn-Sidak or Bonferroni correction26 to adjust p-
values. If the data have a linear relationship, then we use partial least squares discriminant 
analysis (PLS-DA) 27,28 and rank the molecules by p-value in a pair-wise comparison to the 
control group.  
Longitudinal regression modeling:  

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We will apply systems biology tools to quantify how an individual’s inflammation state, as 
measured by calprotectin, changes over time in association with XN metabolite status and 
microbial taxa, their genes, gut metabolites, or BAs. To link the time dependence of 
inflammation to these various parameters, we will follow our prior work29 and construct 
compound Poisson generalized linear models (CPGLMs) that allow us to test for 
differences in temporal trends in the relative abundance of specific OTUs, genes, 
metabolites, or BAs as a function of various experimental parameters. CPGLMs include a 
point mass at zero as well as a weighted mixture of Poisson and gamma distributions. 
These features enable accurate modeling of the sparse (zero-inflated) but otherwise 
continuous and non-normal data usually produced during microbiome and metabolome 
investigations.29 CPGLMs also correctly account for multiple sources of variation30 due to 
different experimental parameters (e.g., XN metabolite status, inflammation state, time, and 
sex). We will construct CPGLMs that model how the temporal variation of each OTU, gene, 
or metabolite (i.e., response variables) relates to these experimental parameters. We will 
adopt a stepwise approach to model construction and optimal models will be identified. 
Models will include interaction terms that consider co-dependencies among the 
experimental parameters. Random intercepts and time slopes for each individual will be 
included in the model to account for within-individual correlation of a response variable’s 
abundance over time. Response variables that manifest as significantly different slopes as 
a function of XN metabolite status are those variables that putatively interact with XN 
metabolites. Related analyses of model slopes will resolve parameters that elicit 
dependencies with inflammation state, as well as parameters that vary in accordance with 
the XN metabolite-inflammation interaction. We will also test models that include a lag 
effect between response variables and these parameters to resolve, for example, taxa 
whose temporal change in abundance predicts subsequent inflammation changes. We 
have extensive experience using GLMs and related techniques for the study of 
microbiome-phenotype associations.19,31,32 Recognizing that the proposed CPGLMs are 
complex and may suffer from a lack of power, we may consider an alternative approach 
which avoids models that attempt to accommodate the data structure through zero-inflated 
distributional adjustments and instead consider a compositional data analysis approach, 
where the data is transformed and normalized using the Aitchison simplex, normalized 
through e.g. centered log ratio adjustment, and followed by fitting with traditional GLMs and 
accounting for other covariates and covariance structures. This approach would retain 
higher statistical power because there no longer would be an attempt to resolve the zero-
inflation problem. This approach has been shown to perform better in terms of differential 
expression with small sample/replicate sizes (https://doi.org/10.1186/1471-2105-14-91). An 
alternative, more well-powered approach to variable/feature selection would be to perform 
lasso or ridge regression via elastic net, which is known to operate well in small sample 
relative to large feature scenarios. 
 
Time-resolved alterations of the gut microbiome by 16S rRNA gene sequencing and 
metagenomics analysis:  
Microbiome data generation:  
We will profile the structure and functional capacity of the microbiome using 16S rRNA and 
shotgun metagenomic DNA sequencing. Following our prior work,24 we will extract whole 

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genomic DNA from each stool sample using a QIAGEN PowerFecal DNA kit. We will then 
PCR amplify the V4V5 region of the 16S gene33 and sequence amplicons on an Illumina 
MiSeq at OSU’s Center for Genome Research and Biocomputing. We will produce 300 bp, 
paired-end sequences and target a sequence depth of 50,000 reads per sample. We will also 
generate metagenomes from the same DNA extracts using NexteraXT library prepa- ration 
and the CGRB’s Illumina HiSeq 3000 (150 bp paired-end sequences). We will target a 
sequence depth of 15 million reads per sample, which our prior work showed was more than 
sufficient for resolving associations between IBD and microbiome taxonomy and functional 
capacity.29,34  
Microbiome data analysis:  
We will analyze 16S sequences to profile gut microbiome structure. The DADA235 software 
pipeline will quality filter sequences, assemble contigs between paired ends, resolve unique 
sequence variants (i.e., OTUs), and taxonomically annotate these sequences using the RDP 
database. The Phyloseq36 R package will quantify the alpha- and beta-diversity of 
microbiome samples. Non-parametric tests (e.g., PERMANOVA) will measure differences in 
α- and β-diversity and β-dispersion across patient groups and over time. We have experience 
developing and applying methods for the analysis of microbiome data.24,34,37–39 Metagenomic 
analysis will complement 16S assessments of taxonomy and quantify the functional capacity 
of microbiome samples. After quality controlling metagenomes using Shotcleaner, which 
follows Human Micro- biome Project40 guidelines to trim adapters, filter low-quality 
sequences, and informatically subtract host genomic DNA from metagenomes, we will apply 
several strategies for the analysis of metagenomes. First, we will assemble paired-end 
metagenomes using the Automatic Tool for Local Assembly Structures (ATLAS), an open-
source tool for NGS data analysis developed in-house.41 ATLAS transforms raw sequence 
data into functional and taxonomic data at the microbial population level and provides 
genome-centric resolution by integrating genome binning. The full ATLAS protocol is 
developed and maintained as open-source software on GitHub and is implemented in 
Snakemake for modular, flexible, and reproducible workflows. Individual protein sequence 
databases will be created based on a combined metagenome that will serve as references 
for paired metaproteome analysis. To complement these assemblies and produce as 
comprehensive an assessment of microbiome structure and functional capacity as possible, 
we will also conduct analyses at the level of individual, unassembled reads. Specifically, we 
will analyze quality controlled metagenomes using MetaPhlan242 to measure the abundance 
of specific microbial taxa in each metagenome. We will also use ShotMAP to assign reads 
into KEGG Orthology groups based on homology.34 Briefly, ShotMAP predicts coding 
sequences from unassembled metagenomic data using Prodigal and applies RAPsearch or 
DIAMOND to rapidly identify reads that show evidence of homology to KEGG Orthology 
groups. Our prior statistical simulations surrounding this technique highlight its accuracy34 
and our prior applications of these tools resolved taxa and microbiome metabolic modules 
that predict immune activation in mouse models of IBD29 and flare-ups in IBD patients.34  
 

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10. DATA COLLECTION AND QUALITY ASSURANCE 
10.1 Data Collection Forms  
All data will be collected on Case Report Forms and by direct data transmits from the 
clinical laboratory.  
10.2 Data Management  
The investigators are responsible for ensuring the accuracy, completeness, legibility, and 
timeliness of the data reported. All source documents should be completed in a neat, legible 
manner to ensure accurate interpretation of data. The investigators will maintain adequate 
case histories of study subjects, including accurate case report forms (CRFs), and source 
documentation. 
Data collection and accurate documentation are the responsibility of the study staff under the 
supervision of the investigator.  All source documents and laboratory reports must be 
reviewed by the study team and data entry staff, who will ensure that they are accurate and 
complete. Unanticipated problems and adverse events must be reviewed by the investigator 
or designee.   
This study will use a secure customized REDCap (Research Electronic Data Capture) 
database. REDCap is a secure, web-based application that supports electronic data capture 
for research studies. REDCap features include: 1) intuitive data entry features; 2) audit trails 
for tracking data manipulation and export; 3) user-based privileges that support HIPAA 
compliance, 4) seamless data export to common statistical packages; and 5) procedures for 
importing data from external sources. The REDCap database is secured with user login IDs 
and password protection. The database incorporates an electronic audit trail to show 
change(s) to data after original entry including the date/time and user making the change. 
Data will be stored as de-identified data; participants’ names will be linked to their unique 
study ID in a single password protected electronic file stored on a limited access server with 
select access limited to the Principal Investigator, NUNM Co-Investigators and Study 
Coordinator. Identifiers will be destroyed after completion of the study and analysis of the 
data. . Electronic communication with outside collaborators will involve only unidentifiable 
information.  
All data resulting from study visits will be collected on standardized case report forms (CRFs). 
Data will be transferred from CRFs to a secure REDCap database for data management and 
will be accessible only by the study team.  
All completed forms will be kept in locked files to which only project personnel have access. 
These files will also be in locked rooms. Data from paper forms will be manually entered into 
REDCap and kept as electronic files. Electronic files will be password protected with access 
given to study personnel only. All analyses will be conducted by study staff at the NUNM 
Helfgott Research Institute or the University of Washington.  The study statistician will only 
access de-identified data exported from REDCap to Excel or SPSS. Only de-identified data 
will be presented in any reports, presentations or manuscripts. 
All data will be managed using Research Electronic Data Capture (REDCap). 

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10.3 Quality Assurance  
10.3.1 Training 
All study investigators and staff complete Good Clinical Practices (GCPs), Human Subjects 
Research (HSR) and Responsible Conduct of Research (RCR) trainings from CITI, plus 
HIPAA and blood-borne pathogens training.  
 
All study coordinators have been training in the administration of informed consent, 
participant interviews for adverse effects, and standardized training for all visit components.  
10.3.2 Metrics 
Quality control metrics include documentation of training requirements and automated 
reminders for training updates as indicated. Training logs for study-specific trainings will be 
maintained in the study regulator binder.  
10.3.3 Protocol Deviations 
Protocol deviations are captured by interview for changes in eligibility, product adherence 
and adverse events at each scheduled clinical research visit. Protocol deviations such as 
lack of follow-up within acceptable timeframes for each visit, i.e., within “windows” are 
captured by regular audits of the visit schedule. Protocol deviations are reported to the IRB 
via periodic progress reports, as well as to the sponsor and the FDA.  
10.3.4 Monitoring 
Site monitoring will include internal and external audits. Internal audits of individual study 
participant records will occur monthly during trial operations by the Principal Investigator and 
will include reviews of eligibility confirmation, consent forms and all CRFs. Audit 
documentation will be maintained in the study binder for review by external auditors.  
External site monitoring will be at the discretion of the sponsor and the FDA.  
 
11. PARTICIPANT RIGHTS AND CONFIDENTIALITY  
11.1 Institutional Review Board (IRB) Review  
Any/all research activities involving human subjects are be reviewed and must approved by 
the NUNM IRB prior to implementation. All protocol amendments, other than minor 
administrative changes as defined by the NCCIH Guidance on Changes in Clinical Studies 
in Active Awards will be submitted in a prospective manner to NCCIH except when necessary 
to protect the safety, rights, or welfare of subjects. Prior to submission to NCCIH, the 
proposed changes will be reviewed and approved by the Independent Monitoring Committee. 
IRB approval will not be sought until after NCCIH approval of the protocol amendment has 
been obtained. 
11.2 Informed Consent Forms 
A signed consent form will be obtained from each participant by a trained study coordinator. 

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The consent form will describe the purpose of the study, the procedures to be followed, and 
the risks and benefits of participation. A copy will be given to each participant or legal 
guardian and this fact will be documented in the participant’s record. Non-English speakers 
and children will not be enrolled in this trial. Non-writing individuals will be able to sign with 
an “X”, stamp or personal mark, or assign a proxy for signing their consent form following 
witnessed verbal agreement by the candidate participant.  
11.3 Participant Confidentiality  
Throughout the study, measures to ensure the privacy of information on study participants 
will be maintained. All study investigators and staff are certified in Good Clinical Practices 
(GCPs), Human Subjects Research (HSR) and Responsible Conduct of Research (RCR) 
and have received training in HIPAA regulations. Participants and staff will be informed of 
the confidentiality of information and assured that data will be used only for statistical 
purposes and group analyses in which the individual cannot be identified. No data beyond 
what is stated in the Informed Consent will be sought without authorization from the 
participant. Information on illnesses and hospitalizations will not be sought from hospitals or 
doctors without a signed medical release from the subject. Conversely, no information on 
any individual will be released to anyone other than study personnel without a signed medical 
release from the participant, or where appropriate, the next of kin or a physician in case of a 
life-threatening emergency to the subject.   
Each participant will be assigned a unique alpha-numeric ID upon screening. All blood and 
urine aliquot tubes, stool collection vials and associated paperwork will be marked using the 
unique ID to protect patient confidentiality. All data resulting from study visits will be collected 
on standardized case report forms (CRFs). Data will be transferred from CRFs to a secure 
REDCap database for data management. The REDCap database will be accessible only by 
the study team. Data will be destroyed 3 years after completion of the study. Computer files 
will be deleted from the server and paper files will be destroyed using a professional 
document shredding service.    
All completed forms will be kept in locked files to which only project personnel have access. 
These files will also be in locked rooms. Data from paper forms will be manually entered into 
REDCap and kept as electronic files. Electronic files will be password protected with access 
given to study personnel only. 
11.4 Study Discontinuation  
This study will be stopped prior to its completion if the intervention is associated with adverse 
effects that call into question the safety of the intervention, if any new information becomes 
available during the trial that necessitates stopping the trial; or if other situations occur that 
might warrant stopping the trial. The ultimate decision to stop the trial may be made by the 
DSMB, NCCIH, and/or the FDA. 
The trial will be halted if >25% of the total sample experience moderate or greater AEs or 
laboratory abnormalities requiring them to withdraw.   
The DSMB will provide a final recommendation to the PI to halt the study or amend the study 
protocol, and will develop a response plan, in consultation with the NUNM IRB and the 
NCCIH Program Officer.   

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If immediate changes are required for participant safety, enrollment and other study activities 
will not continue until the modifications/amendment is approved by the IRB. 
12. COMMITTEES 
N/A 
13. PUBLICATION OF RESEARCH FINDINGS  
Authorship decisions on all resulting manuscripts will be made by the co-PIs of this trial. 
The principal manuscripts will be dependent on the safety and tolerability profile of XN in 
healthy adults and future comparisons made to adults with Crohn’s disease.   
All resulting manuscripts will be made available for review by the sponsor and the NCCIH 
prior to journal submission.  
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15. SUPPLEMENTS/APPENDICES 
Informed Consent for Screening (File: XN_Phase 1Screening_ICF_1.10.19) 
Informed Consent for Trial Participation (File: XN_Phase 1 Trial_ICF_1.10.19)

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