An Open-label Study to Evaluate the Use of the CapScan Intestinal Collection Device to Measure the Regional Metabolism of Sulfasalazine in the Digestive Tracts of Healthy Volunteers
Phase: NA
Conditions: Healthy
Sponsor: Envivo Bio Inc

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 1 of 40 
 
ENVIVO BIO 
CLINICAL TRIAL PROTOCOL 
Protocol Number: EB-03 
NCT05749068 
An open-label study to evaluate the use of the CapScan intestinal collection device to measure 
the regional metabolism of sulfasalazine in the digestive tracts of healthy volunteers. 
 
Sponsor Contact: 
Dari Shalon Ph.D. 
Envivo Bio Inc. 
733 Industrial Rd 
San Carlos, CA 94070 
 
 
 
Study Medical Monitor: 
 
 
 
 
Project Manager 
 
Shai Friedland MD 
900 Blake Wilbur Drive 
Palo Alto , CA 94304 
 
 
Dari Shalon Ph.D. 
Envivo Bio Inc. 
733 Industrial Rd 
San Carlos, CA 94070 
 
 
Biostatistician / Data Management: 
Pete Shabe 
Advanced Research Associates 
2350 Mission College Boulevard 
Suite 825 
Santa Clara, CA  95054 
 
Date of Protocol 
Version: 1.0 Dated: 5 May 2022 
 
Confidentiality Statement
This protocol is a confidential communication document of Envivo Bio Inc. The recipient of this document agrees 
not to disclose the information contained herein to others without prior written authorization of Envivo Bio Inc. 
except that this document may be disclosed to appropriate Institutional Review Boards, or duly authorized 
representatives of Regulatory authorities such as relevant national authorities, or the US Food and Drug 
Administration under the condition that they maintain confidentiality. 
Revision
Date
Description of change
Rev 1.0
5 May 2022 
First Issue

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 2 of 40 
 
 
APPROVAL AND SIGNATURE PAGE                 
 
 
SPONSOR’S APPROVAL: 
 
 
 
 
 
_____________________ 
         
 
 
5/5/2022                                                                                        
Dr. Dari Shalon                                     
 
Date 
Envivo Bio Inc. 
Project Manager 
 
 
 
 
 
_____________________ 
         
 
 
5/5/2022                                                                                        
Pete Shabe                                    
 
 
Date 
Biostatistics / Data Management 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 3 of 40 
 
 
 
 
 
 
 
Version 1.0 – Dated: 5 May 2022 
 
 
 
INVESTIGATOR’S STATEMENT 
 
 
I agree to conduct the study as outlined in the protocol entitled: An open-label study to 
evaluate the use of the CapScan intestinal collection device to measure the regional 
metabolism of sulfasalazine in the digestive tracts of healthy volunteers in accordance with all 
applicable guidelines and government regulations.  
 
I have read and understand all sections of the protocol. 
 
George Triadafilopoulos, M.D. 
Silicon Valley Gastroenterology 
2490 Hospital Dr, Suite 211 
Mountain View, CA 94040 
 
 
________________________________________    5/5/2022 
Principal Investigator’s Signature                                DATE 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 4 of 40 
 
 
 
 
 
 
Version 1.0 - Dated: 5 May 2022 
PROTOCOL SYNOPSIS 
Title 
An open-label study to evaluate the use of the CapScan® intestinal collection 
device to measure the regional metabolism of sulfasalazine in the digestive 
tracts of healthy volunteers. 
Sponsor 
Envivo Bio Inc. 
Protocol No. 
EB-03 
Clinical 
Phase 
Feasibility 
Device 
CapScan® intestinal sampling device 
Indication 
Healthy volunteers 
Study Period 
Total per-subject participation is up to 4 weeks 
Study 
Objective 
The objective is to evaluate the effectiveness of the CapScan intestinal collection 
device in characterizing the regional distribution of sulfasalazine, the metabolic 
breakdown products of sulfasalazine, and the gut microbiota in the digestive 
tracts of healthy volunteers.  
Study Design 
Prospective, open-label, single-arm study designed to provide effectiveness 
information on the CapScan intestinal collection device in healthy volunteers. 
Planned 
Sample Size 
This study will be conducted in up to 10 healthy subjects. 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 5 of 40 
 
Inclusion 
Criteria 
 
Subjects must meet all of the following criteria to be included in the study: 
Males or females 18 years of age or older and 70 years of age or younger at the 
time of the first Screening Visit. 
ASA Classification 1 or 2. 
For women of childbearing potential, negative urine pregnancy test within 7 days 
of Screening Visit and agreement to remain abstinent (refrain from heterosexual 
intercourse) or use adequate contraception during the treatment period and for 
14 days after the final dose of sulfasalazine.  
Subject is fluent in English and understands the study protocol and informed 
consent and is willing and able to comply with study requirements and sign the 
informed consent form. 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 6 of 40 
 
Exclusion 
Criteria 
Subjects with any of the following conditions or characteristics must be 
excluded from the study: 
 Known or suspected gastrointestinal obstructions, strictures or fistula 
 Known or suspected moderate to severe dysmotility, by the judgement 
of the principal investigator 
 Gastroparesis 
 A swallowing disorder 
 Known intolerance, reaction or hypersensitivity to sulfasalazine, its 
metabolites, sulfonamides, or salicylates 
 
Urinary obstruction 
 Porphyria 
 
G6PD deficiency 
 Known or suspected infection with SARS-CoV2, HIV, HBV or HCV  
 Pregnancy or planned pregnancy within 30 days from Screening Visit, or 
breast-feeding. 
 Use of concomitant medications, such as aspirin, antibiotics, laxatives 
and proton pump inhibitors that may interfere with the study in the 
judgement of the investigator.  
 Any form of active substance abuse or dependence (including drug or 
alcohol abuse), unstable medical or psychiatric disorder, or any chronic 
condition susceptible, in the opinion of the principal investigator, to 
interfere with the conduct of the study. 
 
A clinical condition that, in the judgment of the principal investigator, 
could potentially pose a health risk to the subject while involved in the 
study. 
Device 
Description 
The CapScan intestinal collection device is a single-use passive device that 
collects fluids from the gastrointestinal (“GI”) tract for analysis outside the body. 
The CapScan Device contains no medicinal product. The CapScan Device is 
strictly a collection device.  
Study 
Treatment 
At the Screening Visit, study eligibility is confirmed and the subject signs the 
informed consent document after all questions have been appropriately 
answered. 
The subject is given 4 CapScan Devices and one 1,000 mg dose of sulfasalazine. 
The subject is provided with a snack bar to consume before swallowing the 4 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 7 of 40 
 
CapScan Devices. The subject is also given a retrieval wand and stool collection 
containers to snare the expelled CapScan Devices from the stool. The subject is 
also given 2 Tasso+ blood collection devices for home collection of blood 
samples, saliva and stool collection tubes for analysis of saliva and stool 
microbiome and metabolites. The blood, saliva and stool samples will be 
compared to the intestinal samples collected by the CapScan Device. The 
subject is given Daily Diaries to document the time the CapScan Devices were 
swallowed, what the subject ate, and time of bowel movements. 
On Day 1, the subject swallows 2 CapScan Devices and collects stool and saliva 
for the baseline set of measurements. 
On Day 2, the subject takes a single dose of 1,000 mg of sulfasalazine and 
swallows 2 CapScan Devices and collects 2 blood samples, plus stool and saliva. 
The timing of the dose of sulfasalazine, sampling of blood, and the ingestion of 
the CapScan Devices on each day will be at the discretion of the PI. 
The subject continues to collect all CapScan Devices expelled in the stool and 
stool samples until all ingested CapScan Devices have been retrieved, or until 
Day 8. In the event of missing CapScan Devices that were not successfully 
retrieved by the end of Day 8, and at the discretion of the PI, an abdominal x-
ray will be performed on the subject to confirm no CapScan Devices are still in 
the body.  
In the event that a subject was unable to retrieve the 2 CapScan Devices 
swallowed on Day 2, or if the devices did not produce usable samples, the PI 
may request that the full set of procedures from Day 2 onwards will be 
repeated starting any day between Day 12-19 to give sufficient time for any 
sulfasalazine to be cleared from the body, including a second dose of 1,000 mg 
of sulfasalazine and swallowing and retrieval of the 2 additional CapScan 
Devices. The End of Study is defined when the last ingested CapScan Device is 
collected from the stool, or after determination by the PI that no CapScan 
Devices are in the subject’s body.  
If a subject complains of any Adverse Event (AE) within 5 days of the End of Study 
that the investigator deems related to the device, the event will be documented 
and the subject will be evaluated by the principal investigator within 72 hours of 
the investigator becoming aware of the AE. 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 8 of 40 
 
Endpoint 
The endpoint of this trial is the regional measurement of sulfasalazine in the 
intestinal lumen, the metabolic breakdown products of sulfasalazine and the gut 
microbiota in the digestive tracts of healthy volunteers.  
Safety 
Assessments 
Safety assessments will include the following: 
 Subject-reported adverse events, as communicated to the study staff. 
A medical evaluation, if necessary, to evaluate an adverse event. 
Statistical 
methods 
All measured variables and derived parameters will be listed individually and, if 
appropriate, tabulated by descriptive statistics. 
 
 
 
 
 
 
 
 
 
 
 
 
 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 9 of 40 
 
 
List of Abbreviations and Terms 
 
Abbreviation/Term 
ADE 
Definitions 
Adverse device effect 
AE 
Adverse event 
ASA 
American Society of Anesthesiologists 
BMI 
Body mass index 
BSLN 
Baseline 
CE 
Conformité Européenne 
EC 
Ethics committee 
Cm 
Centimeter 
CRF 
Case report form 
CRO  
Contract research organization 
GCP 
Good Clinical Practice 
ICF 
Informed consent form 
ICH 
International Conference for Harmonisation 
ISO 
International Standard Organization 
Kg 
Kilogram 
M 
Meter 
MedDRA 
Medical Dictionary for Regulatory Activities 
Mm 
Millimeter 
l 
Microliter 
N 
Number of subjects 
PI 
Principal investigator 
SADE 
Serious adverse device effect 
SAE 
Serious adverse event 
SAP 
Statistical analysis plan 
SEM 
Standard error of the mean 
WHO 
World Health Organization 
USP 
United States Pharmacopoeia  
RFID 
Radiofrequency Identification Device 
 
 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 10 of 40 
 
Table of Contents 
1. 
INTRODUCTION .............................................................................................................................13 
1.1 
CURRENT APPROACHES.................................................................................................................. 13 
2. 
INVESTIGATIONAL DEVICE: CAPSCAN COLLECTION CAPSULE .........................................................14 
2.1 
DEVICE DESCRIPTION ..................................................................................................................... 14 
2.2 
TREATMENT PROCEDURES ............................................................................................................. 14 
2.3 
DEVICE INTENDED USE ................................................................................................................... 15 
2.4 
RISK BENEFIT ANALYSIS .................................................................................................................. 16 
2.4.1 
Possible Risks ........................................................................................................................ 16 
2.4.2 
Possible Benefits ................................................................................................................... 17 
2.4.4 
Safety Measures Taken ......................................................................................................... 17 
3. 
STUDY RATIONALE .........................................................................................................................17 
4. 
STUDY OBJECTIVES ........................................................................................................................18 
5. 
STUDY DESIGN ...............................................................................................................................18 
6. 
STUDY POPULATION ......................................................................................................................20 
6.3 SUBJECT IDENTIFICATION .................................................................................................................. 21 
6.4 
REMOVAL, REPLACEMENT, OR EARLY WITHDRAWAL OF SUBJECTS FROM THERAPY OR ASSESSMENT
 
21 
6.5 
HANDLING OF WITHDRAWALS ....................................................................................................... 22 
6.6 
SPONSOR'S TERMINATION OF STUDY ............................................................................................ 22 
7. 
STUDY PROCEDURES ......................................................................................................................22 
7.1 
STUDY PROCEDURES CHART .......................................................................................................... 22 
7.2 
SCREENING VISIT ............................................................................................................................ 24 
7.3          CAPSULE USE PHASE .................................................................................................................... 25 
7.3 
PROTOCOL AMENDMENTS, REVISION AND/OR DEVIATIONS ........................................................ 25 
7.4 
MANUFACTURING AND SUPPLY OF INVESTIGATIONAL DEVICE ..................................................... 26 
7.5 
STORAGE AND ACCOUNTABILITY OF INVESTIGATIONAL DEVICE ................................................... 26 
7.6 
RETURN OF INVESTIGATIONAL DEVICE .......................................................................................... 26 
7.7 
CONTRA-INDICATION ..................................................................................................................... 26 
8. 
STUDY ASSESSMENT METHODS .....................................................................................................26 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 11 of 40 
 
8.1 
SAFETY ASSESSMENTS .................................................................................................................... 26 
8.1.1 
Adverse Events ...................................................................................................................... 26 
8.1.2 
General Health Assessment .................................................................................................. 27 
9. 
STUDY ENDPOINTS ........................................................................................................................27 
10. 
SAFETY ...........................................................................................................................................27 
10.1 
DEFINITION OF ADVERSE EVENT ........................................................................................................ 27 
10.2 
DEFINITION OF SERIOUS ADVERSE EVENT ................................................................................. 28 
10.3 
DEFINITION OF RELATIONSHIP TO INVESTIGATIONAL DEVICE AND DEVICE DELIVERY AND REMOVAL 
PROCEDURES ........................................................................................................................................... 28 
10.4 
DEFINITION OF SEVERITY OF ADVERSE EVENTS ........................................................................ 29 
10.5 
REPORTING OF ADVERSE EVENTS.............................................................................................. 29 
10.6 
DOCUMENTATION OF AES ......................................................................................................... 30 
10.7 
PREGNANCY ............................................................................................................................... 30 
11. 
STATISTICAL ANALYSIS PLAN .........................................................................................................30 
12. 
SAMPLE SIZE CONSIDERATION .......................................................................................................31 
13. 
ETHICS ...........................................................................................................................................31 
12.1 
INSTITUTIONAL REVIEW BOARD ................................................................................................ 32 
12.2 
ETHICAL CONDUCT OF THE STUDY ............................................................................................ 32 
12.3 
SUBJECT INFORMATION AND CONSENT .................................................................................... 32 
12.4 
SUBJECT INSURANCE ................................................................................................................. 33 
12.5 
INFORMING THE GENERAL PRACTITIONER ............................................................................... 33 
12.6 
PERSONAL DATA PROTECTION .................................................................................................. 33 
14. 
QUALITY CONTROL AND QUALITY ASSURANCE..............................................................................33 
13.1 
STUDY MONITORING ................................................................................................................. 34 
13.2 
SOURCE DOCUMENT ................................................................................................................. 34 
13.3 
CASE REPORT FORMS (CRFS) ..................................................................................................... 35 
13.4 
DATA MANAGEMENT ................................................................................................................ 35 
15. 
STUDY ADMINISTRATION ..............................................................................................................35 
14.1 
STUDY PERSONNEL .................................................................................................................... 35 
14.1.1 
Site Staff ........................................................................................................................... 35 
14.1.2 
Sponsor............................................................................................................................. 35 
14.2 
REQUIRED DOCUMENTS PRIOR TO STUDY INITIATION ............................................................. 36 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 12 of 40 
 
14.3 
INVESTIGATOR'S STUDY FILE ..................................................................................................... 36 
14.4 
CLINICAL TRIAL SUPPLIES ........................................................................................................... 37 
14.5 
STUDY REPORTING REQUIREMENT ........................................................................................... 37 
14.6 
STUDY COMPLETION ................................................................................................................. 37 
14.7 
FINAL REPORT ............................................................................................................................ 38 
14.8 
RETENTION OF STUDY RECORDS ............................................................................................... 38 
14.9 
CONFIDENTIALITY AND PUBLICATION OF STUDY RESULTS ....................................................... 38 
 
References 
APPENDIX A – DAILY DIARY  
 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 13 of 40 
 
1. 
INTRODUCTION 
It has recently been recognized that microbes, viruses, and micro-eukaryotes in the human 
gastrointestinal (“GI”) tract perform many vital physiological functions that benefit their host 
organism, contributing to digestion, metabolizing drugs, producing essential amino acids and 
vitamins, regulating the immune system, providing resistance to disease, and even modifying 
appetite, and behavior.i Indeed it is estimated that the majority of drugs given orally today are 
metabolized or modified in some manner by the gut microbiome.ii Yet we know very little about 
the hundreds or thousands of microbial, viral, and micro-eukaryote species in the human GI tract 
(“microbiome”).iii The variety of microbes in a single individual at different regions of the GI tract 
is staggering. Due to the complexity of this microbial ecology in a single individual and the 
variability among individuals, it is necessary to routinely sample and analyze the microbial 
community living along with its associated metabolites in all regions of the GI tract and measure 
their interactions with orally-delivered drugs. 
 
The CapScan intestinal collection device (“CapScan Device”) is a non-invasive sampling device 
that collects gastrointestinal samples along the GI tract that are then analyzed outside the body. 
One specific utility for this type of sampling is that the CapScan Device may enable the 
characterization and the performance of various drugs by detecting the presence of the drug and 
its metabolic breakdown products in the digestive lumen. Samples collected by the CapScan 
Device will undergo DNA sequencing and metabolite analysis. The goal will be to determine the 
identity and function of sulfasalazine, the metabolic breakdown products of sulfasalazine, and 
the gut microbes in the different regions of the GI tract, for comparison to similar analyses 
conducted on concomitantly collected saliva, blood and stool samples.  
 
The ability to combine information about the identities and diversity of microbial community 
members obtained from these measurements could provide an important understanding of the 
microbiome’s role in human physiology. Furthermore, understanding how to better 
characterize metabolic breakdown of drugs within the digestive tract may potentially improve 
the development and optimization of oral or enterally administered treatments for various 
diseases. 
1.1 CURRENT APPROACHES 
STOOL SAMPLING 
The least invasive way to assess the microbiome is analyzing stool samples. However, stool 
samples do not capture the microbial activity at proximal sites of the intestine or in microbes 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 14 of 40 
 
living close to the gut mucosa.iv Identification of microbes from stool reflects mainly the 
accumulated remnants of microbes that were more proximally active in the GI tract. 
Furthermore, many of the metabolites and proteins that are active in the small and large 
intestine are no longer present in the stool in their active form. Therefore, it is necessary to 
sample the microbiome directly from the stomach, small intestine and colon in order to 
accurately capture the full biochemical and immunological activity of the microbiome and its 
host interactions, and in order to better characterize the metabolic activity of drugs. 
ENDOSCOPIC BIOPSIES 
Endoscopic mucosal biopsies display greater microbial diversity than stool samples.v However, 
mucosal biopsies are associated with the invasiveness and risk of endoscopy, as well as the 
biopsy procedure that alters the mucosal surface of the GI tract. 
To the best of our knowledge, no device exists to safely, routinely and non-invasively collect 
fluid samples from the various regions of the GI tract to provide highly relevant samples for 
microbiome and pharmocological function analysis. 
BLOOD SAMPLES 
Blood samples show how much of the drug is in systemic circulation. However, for drugs such 
as sulfasalazine that work in the gut lumen, circulating levels of the drug in blood may not be 
informative. 
2. 
INVESTIGATIONAL DEVICE: CAPSCAN COLLECTION DEVICE 
2.1 DEVICE DESCRIPTION  
The CapScan Device is a single-use passive device that collects fluids from the gastrointestinal 
(“GI”) tract for analysis outside the body.  
The CapScan Device contains no medicinal product. The CapScan Device is strictly a collection 
device. 
  
2.2 TREATMENT PROCEDURES  
The subject is given 4 CapScan Devices and one 1,000 mg dose of sulfasalazine. The subject is 
provided with snack bars to consume before swallowing the CapScan Devices. The subject is 
given a retrieval wand to snare the expelled CapScan Devices from the toilet bowl and stool 
collection containers if they prefer to retrieve the device from stool in the container as opposed 
to from the toilet. The subject is also given 2 Tasso+ home-use blood collection devices, saliva 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 15 of 40 
 
and stool collection tubes for analysis of blood, saliva and stool microbiome and metabolites. 
The blood, saliva and stool samples will be compared to the intestinal samples collected by the 
CapScan Device. The subject is given Daily Diaries to document the time the CapScan Devices 
were swallowed, blood and saliva samples collected, what the subject ate, and time of bowel 
movements. 
On Day 1, after consuming a snack bar, the subject swallows 2 CapScan Devices and collects 
stool and saliva for the baseline set of measurements. 
On Day 2, after consuming a snack bar, the subject takes a single dose of 1,000 mg of 
sulfasalazine and swallows 2 CapScan Devices and collects blood, stool and saliva.  
The timing of the dose of sulfasalazine, sampling of blood, stool and saliva and the ingestion of 
the CapScan Devices on each day will be at the discretion of the PI. 
The subject continues to collect all CapScan Devices expelled in the stool until all ingested 
CapScan Devices have been retrieved, or until Day 8. In the event of missing CapScan Devices 
that weren’t successfully retrieved by the end of Day 8, and at the discretion of the PI, an 
abdominal x-ray will be performed on the subject to confirm no CapScan Devices are still in the 
body.  
In the event that a subject was unable to retrieve the two CapScan Devices swallowed on Day 2 
or if the devices did not produce usable samples, the PI may request that the full set of 
procedures from Day 2 will be repeated on one day between Day 12-19 to give sufficient time 
for any sulfasalazine to be cleared from the body, including a second dose of 1,000 mg of 
sulfasalazine and swallowing and retrieval of 2 additional CapScan Devices. The End of Study is 
defined when the last ingested CapScan Device is collected from the stool, or after 
determination by the PI that no CapScan Devices are in the subject’s body.  
If a subject complains of any Adverse Event (AE) within 5 days of the End of Study that the 
investigator deems related to the device, the event will be documented and the subject will be 
evaluated by the principal investigator or sub investigator within 72 hours of either investigator 
becoming aware of the AE. 
2.3         DEVICE INTENDED USE 
The CapScan Device is intended to collect liquid samples from the small intestine and colon.  

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 16 of 40 
 
2.4 RISK BENEFIT ANALYSIS 
2.4.1 Possible Risks 
The potential risks associated with the use of CapScan Device include transient discomfort when 
swallowing a CapScan Device. Additional risks include aspiration and intestinal blockage.  
The risks above are considered reasonable given the following safeguards: 
Physically, the size and shape of the CapScan Device is similar to a standard size 00 medicinal 
capsule.  As such, the swallowing of the CapScan Device does not pose any greater risk to the 
health, safety, or welfare of the patient than the swallowing of a standard medicinal capsule. 
The passage of the CapScan Device through the GI tract does not present any greater risk to the 
health, safety, or welfare of the patient than a Medtronic Pillcam®, which is routinely swallowed 
for diagnostic purposes and eliminated via defecation. The Pillcam® is hard and inflexible and is 
12 mm in diameter, versus the CapScan Device which is soft and flexible and only 6 mm in 
diameter.  A radio-opaque marker incorporated into the CapScan Device enables definitive 
confirmation that the device has been eliminated from the body, if necessary. 
Generally, healthy volunteers with any underlying intestinal stricture that would prevent the 
passage of items 6 mm in diameter through the GI tract would be symptomatic even before the 
study. Examples of symptoms of pre-existing intestinal blockage could include pain and bloating 
after eating corn or other vegetables, for example.  
In the unexpected event that the CapScan Device is retained in the intestines, the CapScan Device 
can be removed via motility enhancers (i.e. laxatives) or via laparoscopic surgery in a worst case 
scenario. In the event of a retained CapScan Device, there is a potential benefit for the subject to 
discover and treat the underlying cause of any pre-existing intestinal strictures inadvertently 
discovered as part of this study. 
All of the materials in the CapScan Device have been well characterized and understood to be 
biocompatible and used in a variety of cleared medical products and pharmaceuticals.  
Sulfasalazine is an FDA-approved drug indicated for treatment of ulcerative colitis and 
rheumatoid arthitis. In clinical practice, sulfasalazine is most commonly associated with the 
following side effects: gastric distress, headache, nausea, oligospermia (temporary reduced 
sperm count), vomiting, and anorexia. Less frequent side effects include pruritis (itching), 
urticaria (hives), rash, fever, Heinz body anemia, hemolytic anemia, and cyanosis.  
Research and clinical experience suggests that side effects are more frequent in individuals taking 
daily doses of 4,000 mg or higher and for prolonged periods of time (months to years). Due to 
the relatively low single dose of 1,000 mg sulfasalazine administered in this study, side effects 
are expected to be rare and less serious. Regardless, subjects will be instructed to report any 
experienced side effects to the study investigator. 
Finally, the proposed clinical investigation does not involve any anticipated surgical procedures, 
nor do any of the other clinical procedures proposed for use during the study present a potential 
for serious risk to the health, safety, or welfare of a subject.  

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 17 of 40 
 
2.4.2 Possible Benefits 
The possible benefits offered by the CapScan Device for future patients might be the 
development of better predictive models on the effectiveness of sulfasalazine and/or other 
drug therapies, disease diagnosis, patient stratification, and potentially the development of 
transformative treatments for various diseases.  
 
2.4.3 Alternative Treatments Summary 
 
Alternative approaches for collecting samples from the GI tract and looking for drug 
pharmacokinetics, include measuring the drug levels in the blood and/or stool samples, which 
are not representative of the actual luminal drug levels or and only offer limited insight into 
effects of the gut microbiome on drug metabolism. 
2.4.4 Safety Measures Taken 
Physically, the size and shape of the CapScan Device is similar to a standard size 00 medicinal 
capsule.  As such, the swallowing of the CapScan Device does not present any greater risk to the 
health, safety, or welfare of the patient than the swallowing of a standard medicinal capsule.  
The passage of the CapScan Device through the GI tract does not present any greater risk to the 
health, safety, or welfare of the patient than a Given Imaging Pillcam®, which is routinely 
swallowed and eliminated via defecation and is significantly harder, larger and more 
voluminous than the CapScan Device. The radio-opaque marker incorporated into the CapScan 
Device enables definitive confirmation that the device has been eliminated from the body if 
necessary. 
All of the materials in the CapScan Device are well characterized and understood to be 
biocompatible and used in a variety of cleared medical products and pharmaceuticals.  
The considerations mentioned above have been taken during the design and manufacturing of 
the CapScan Device to ensure its safe and reliable performance. 
The selection of and dosage of sulfasalazine is intended to minimize observed side effects related 
to the drug used for metabolic analysis. 
3. 
STUDY RATIONALE 
Performing drug metabolism and microbiome analyses on stool samples does not fully represent 
the diversity of the microbial, viral and micro-eukaryotic species, metabolism and host 
interactions occurring in the intestines. Obtaining samples via endoscopic biopsy is invasive and 
not convenient. Understanding drug levels in the lumen of the gut will provide insight into drug 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 18 of 40 
 
metabolism and pharmacokinetics as well as enable an understanding of systemic versus 
intestinal lumen drug concentrations.  
The CapScan Device is designed to be a safe and convenient method for obtaining gastrointestinal 
samples from key regions of the GI tract for microbiome analysis. This study is focused on 
examining the safety and efficiency of sample collection of the CapScan Device in humans, as well 
as its suitability for adequate sample collection process aimed at subsequent DNA, metabolite 
and drug pharmacokinetics and metabolism analyses. Sulfasalazine was selected as the drug for 
metabolic characterization based on its safety profile and low likelihood of causing harm to 
healthy subjects when ingested for analytical purposes. Furthermore, sulfasalazine is degraded 
by gut microbes into known metabolites, although to the best of our knowledge, this process has 
not been observed directly in the human gut before. 
The trial sample size is considered the minimal size required to collect sample collection efficiency 
data.  
4. 
STUDY OBJECTIVES 
The objective is to evaluate the effectiveness of the CapScan Device for characterizing the 
regional distribution of sulfasalazine, the metabolic breakdown products of sulfasalazine and the 
gut microbiota in the digestive tracts of healthy volunteers.  
5. 
STUDY DESIGN 
This will be an open-label feasibility study conducted in up to 10 subjects at a single center. 
All subjects will undergo similar pre-study screening and evaluation and study procedures.  
SCREENING VISIT: 
During the Screening Visit, study eligibility is confirmed, the informed consent process is properly 
conducted and documented and instructions are given for use of the CapScan Device, saliva and 
stool collection, ingestion of sulfasalazine, and completion of the Daily Diaries.  
The CapScan Devices are provided in individually-sealed containers.  
DAY 1: 
2 CapScan Devices are swallowed on Day 1. The subject will eat a study-provided snack bar and 
swallow the CapScan Devices at times of the day that are at the discretion of the PI. The subject 
also collects saliva and stool samples using the collection kits provided to them. 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 19 of 40 
 
The subject uses a retrieval wand to snare any CapScan Devices that are expelled together with 
the stool. 
The retrieved CapScan Devices, saliva and stool samples are stored in closed containers inside a 
study-supplied refrigerator until collected by the study staff.  
The subject will complete a daily diary to document the time the CapScan Devices were 
swallowed, what they ate, when they sampled saliva, when they had bowel movements and the 
time the CapScan Devices were recovered, if any were recovered. 
DAY 2: 
Two CapScan Devices are swallowed on Day 2. The subject will also eat a study-provided snack 
bar and take the physician-directed dose of 1,000 mg of sulfasalazine. The subject also collects 
up to two samples of blood using the Tasso+ device, saliva and stool samples using the 
collection kits provided to them. The timing of the above procedures will be at the discretion of 
the PI. 
The subject will retrieve the CapScan Devices from the stool using the retrieval wand provided 
by the study sponsor.  
The subject will complete a daily diary to document the time the CapScan Devices were 
swallowed, what they ate, when they swallowed the sulfasalazine, sampled blood and saliva, 
had bowel movements and the time the CapScan Devices were recovered, if any were 
recovered. 
DAYS 3-8: 
The subject will continue to retrieve the CapScan Devices, collect stool and complete the daily 
diaries until the last device is retrieved. If not all devices have been retrieved by Day 8, the PI 
may elect to perform an abdominal x-ray to confirm that no CapScan Devices are still in the 
body. The End of Study is defined when the last ingested CapScan Device is collected from the 
stool, or after determination by the PI that no CapScan Devices are in the subject’s body.  
DAYS 12-19, if needed: 
In the event that a subject was unable to retrieve the two CapScan Devices swallowed on Day 2 
or if the devices did not produce usable samples, the PI may request that the full set of 
procedures from Day 2-8 will be repeated starting on any day between Day 12-19 to give 
sufficient time for any sulfasalazine to be cleared from the body, including a second dose of 
1,000 mg of sulfasalazine and swallowing and retrieval of 2 additional CapScan Devices. The end 
of the study is defined when the last swallowed CapScan Device is retrieved from the stool or 
after determination by the PI that no CapScan Devices are in the subject’s body. At this point, a 
study representative will go to the subject’s house to collect all unused CapScan Devices, 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 20 of 40 
 
retrieved devices, blood, saliva and stool samples, refrigerator  and the Daily Diaries from the 
subject.   
6. 
STUDY POPULATION 
Study subjects will be healthy volunteers (ASA classes 1 and 2).    
6.1 INCLUSION CRITERIA 
Subjects must meet all of the following criteria to be included in the study: 
 Males or females 18 years of age or older and 70 years of age or younger at the time of 
the first Screening Visit. 
 ASA Classification 1 or 2. 
 For women of childbearing potential, negative urine pregnancy test within 7 days of 
Screening Visit and agreement to remain abstinent (refrain from heterosexual 
intercourse) or use adequate contraception during the treatment period and for 14 days 
after the final dose of sulfasalazine.  
 Subject is fluent in English and understands the study protocol and informed consent and 
is willing and able to comply with study requirements and sign the informed consent form. 
 
6.2 EXCLUSION CRITERIA 
Subjects with any of the following conditions or characteristics must be excluded from the 
study: 
 
 Known or suspected gastrointestinal obstructions, strictures or fistula 
 Known or suspected moderate to severe dysmotility, by the judgement of the principal 
investigator 
 Gastroparesis 
 A swallowing disorder 
 Known intolerance, reaction or hypersensitivity to sulfasalazine, its metabolites, 
sulfonamides, or salicylates 
 
Urinary obstruction 
 Porphyria 
 
G6PD deficiency 
 Known or suspected infection with SARS-CoV2, HIV, HBV or HCV  
 Pregnancy or planned pregnancy within 30 days from Screening Visit, or breast-feeding. 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 21 of 40 
 
 Use of concomitant medications, such as aspirin, antibiotics, laxatives and proton pump 
inhibitors that may interfere with the study in the judgement of the investigator.  
 Any form of active substance abuse or dependence (including drug or alcohol abuse), 
unstable medical or psychiatric disorder, or any chronic condition susceptible, in the 
opinion of the principal investigator, to interfere with the conduct of the study. 
 A clinical condition that, in the judgment of the principal investigator, could potentially 
pose a health risk to the subject while involved in the study. 
6.3 SUBJECT IDENTIFICATION  
A unique subject identifying number will be assigned when an individual subject is qualified for 
enrollment. Before formal enrollment into the study, subjects will be uniquely identified by their 
initials and a subject identification number only. 
6.4 REMOVAL, REPLACEMENT, OR EARLY WITHDRAWAL OF SUBJECTS FROM THERAPY OR 
ASSESSMENT 
Subjects are free to discontinue their participation in the study at any time and without prejudice 
to further treatment, up until the time of publication of the study results, at which point 
withdrawal from the study is no longer possible. The Principal Investigator (PI) must withdraw 
any subject from the study, if that subject requests to be withdrawn. If a subject is withdrawn 
from the study or fails to return either at his or her request or at the PI’s sole discretion, every 
effort should be made to determine the reason.  This information will be recorded in the subject’s 
CRF. 
When a subject is removed from the study for a Serious Adverse Event, a final medical evaluation 
must be performed. Patients removed from the study because of an adverse event will be 
followed-up until the adverse event has resolved or stabilized. 
If a subject has discontinued study participation during the CapScan Device Use Phase, the 
subject will return all collected and unused CapScan Devices, collected saliva, blood and stool 
samples, the completed Daily Diaries and the refrigerator. 
Subjects withdrawn from the study will be replaced, regardless of the reason for withdrawal.  
The subject's participation in this study may be discontinued due to any of the following reasons: 
 Failure to follow the instructions of the PI and study staff 
 The PI decides that continuing participation could be harmful to the subject 
 Pregnancy  
 The study is cancelled 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 22 of 40 
 
 Other administrative reasons 
 Unanticipated circumstances 
6.5 HANDLING OF WITHDRAWALS 
If a subject is withdrawn from the study or fails to return either at his or her request or at the PI’s 
sole discretion, every effort should be made to determine the reason. This information will be 
recorded on the subject’s case report form (CRF).  
All subjects who withdraw from the study prematurely, regardless of cause, should undergo all 
early termination assessments; if relevant. It is vital to obtain follow-up data for any subject 
withdrawn due to an AE following device use.  
6.6 SPONSOR'S TERMINATION OF STUDY 
Envivo Bio Inc. reserves the unilateral right to discontinue the study at any time for any reason. 
Envivo Bio Inc. may also discontinue the study for inadequate site performance or compliance. 
Such a termination must be implemented by the PI, if instructed to do so by Envivo Bio Inc., in a 
time frame that is compatible with the subjects’ well-being. 
7. 
STUDY PROCEDURES 
7.1 STUDY PROCEDURES CHART 
A schedule of study procedures for the investigator and study subject are shown in Table 1 and 
Table 2 respectively. No protocol related procedures will be performed before subjects review 
and sign a written informed consent form.  
 
 
 
 
 
 
 
 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 23 of 40 
 
 
Table 1: SCHEDULE OF INVESTIGATOR STUDY PROCEDURES 
 
 PROCEDURES 
SCREENING 
CAPSCAN 
DEVICE USE 
 
CAPSCAN 
DEVICE 
RETRIEVAL 
1 Day 
2 days 
Up to 8 days 
Informed Consent 
X 
  
 
Inclusion/Exclusion 
X 
 
 
Demographics 
X 
 
 
Medical History 
X 
 
 
Concomitant 
Medications
X
 
General Health 
Assessment (ASA 
classification) 
X 
  
 
Urine Pregnancy 
X 
 
 
AE/SAE 
X  
X 
 
 
  

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 24 of 40 
 
 
Table 2: SCHEDULE OF SUBJECT STUDY PROCEDURES 
 
 PROCEDURES 
SCREENING 
CAPSCAN 
DEVICE USE 
 
CAPSCAN 
DEVICE 
RETRIEVAL 
Day 1 
Day 2 
Days 3-8 
Informed Consent 
X 
  
 
Swallows 2 CapScan 
Devices 
 
X 
X 
 
Collect saliva samples 
when swallowing 
CapScan Devices 
X 
X 
 
Collect blood using 
Tasso+ home-use tests 
 
X 
 
Retrieve CapScan 
Devices from stool 
using retrieval wand 
             X 
              X 
 
 
X 
Ingests 1000mg of 
sulfasalazine
X
 
Complete Daily Diaries 
to document the time 
the CapScan devices 
were swallowed, what 
was eaten, and time 
of bowel movements 
    X 
X 
 
 
 
 
 
X 
 
 
 
 
 
7.2 SCREENING VISIT 
The purpose and procedures of the study will be fully explained to potential subjects. Those 
individuals wishing to enroll in the study will sign a written informed consent form prior to 
initiating any evaluations or procedures. The Screening Visit includes: 
 Review and sign Informed Consent by subject and PI or PI designee 
 Assess suitability of subject to enroll in the study according to the Inclusion and 
Exclusion Criteria 
 Obtain subjects medical history (including clinically significant abnormalities of all 
body systems), demographic data and concomitant medications 
 Conduct health assessment of subject to include ASA classification  
 Provide instruction for and dispense Daily Diaries to complete at home. 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 25 of 40 
 
 Provide instruction for use and dispense, or have the Sponsor dispense, 4 CapScan 
Devices. 
 Provide instructions for use and dispense, or have the Sponsor dispense, blood 
collection devices, retrieval wands, stool and saliva collection containers for 
collecting CapScan Devices expelled in stool.  
 Provide instruction for and dispense, or have the Sponsor dispense, saliva and 
blood collection kits. 
 Provide instructions for use and dispense, or have the Sponsor dispense, 1,000 mg 
of sulfasalazine. 
 Assign subject a unique study number. 
 
7.3          CAPSCAN DEVICE USE PHASE 
 The CapScan Devices are provided in a sealed containers.  
 The subject uses a device retrieval wand to look for and collect the expelled 
devices in the stool after all bowel movements for up to 8 days after swallowing 
a CapScan Device.  
 The end of the study is defined when the last swallowed CapScan Device is 
retrieved from the stool. The study representative will collect all of the retrieved 
samples and the Daily Diaries from home of the subject.  
7.3 PROTOCOL AMENDMENTS, REVISION AND/OR DEVIATIONS 
The protocol must be read thoroughly and its instructions and guidelines must be followed 
exactly. Any changes in the protocol will require a formal amendment document. Such 
amendments will be agreed upon and approved in writing by the PI and Envivo Bio Inc. All 
modifications and revisions will be noted and summarized on page 1 of this protocol document. 
The Institutional Review Board (IRB) will be notified of all significant amendments to the protocol. 
Significant amendments to the protocol that may increase risk to the subject, and/or that may 
adversely affect the rights of the subject or the validity of the investigation will not be 
implemented until IRB committee written approval has been received.  
With the exception of emergency situations, no significant changes or deviations in the conduct 
of this protocol will be permitted without the prior documented approval of Envivo Bio Inc. The 
IRB that granted the original approval for the study must be notified of all material changes in 
the protocol, and must provide a written approval of any significant change or significant 
deviation that may increase risk to the subject, and/or that may adversely affect the rights of the 
subject or the validity of the investigation. 
In the event of an emergency, the PI will institute any medical procedures deemed appropriate. 
However, all such procedures must be promptly reported to Envivo Bio Inc. and the IRB. 
 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 26 of 40 
 
7.4 MANUFACTURING AND SUPPLY OF INVESTIGATIONAL DEVICE 
Envivo Bio Inc. will supply the CapScan Devices to the investigational site or subjects in 
appropriately labeled packaging. 
The CapScan Device will be manufactured and packaged by Envivo Bio Inc. and/or its designated 
sub-contractors in compliance with the relevant principles and guidelines applicable to 
investigational medical devices.  
 
7.5 STORAGE AND ACCOUNTABILITY OF INVESTIGATIONAL DEVICE 
The CapScan Devices will be kept in a secure, limited-access, room temperature storage area at 
the investigational site or dispensed directly to the subjects by the Sponsor.  Only authorized 
personnel will have access to the study devices. Complete traceability records will be kept of all 
devices during the study.  Specifically, a study accountability log will be completed to maintain 
accurate records of all CapScan Device inventory. 
7.6 RETURN OF INVESTIGATIONAL DEVICE 
Government regulations require that all investigational product materials not used in clinical 
trials be returned to the Sponsor at the completion of the study. The monitor will be responsible 
for the accountability of the returned of used and unused investigational devices. 
 
7.7 CONTRA-INDICATION  
No contraindications related to the CapScan Device are currently known.  
8. 
STUDY ASSESSMENT METHODS 
8.1 SAFETY ASSESSMENTS  
8.1.1 Adverse Events 
Adverse Events (AE’s) reported by subjects or observed by the PI or PI designee will be 
documented in the source document, assessed by the PI and will be individually listed on an 
adverse event page in the CRF. Recording of AE’s will begin immediately after the signing of the 
informed consent document and continue until the subject dies, withdraws consent, or 48 hours 
after the End of Study (EOS), regardless of the relationship of the AE to the CapScan Device.  If a 
subject complains of any AE within 5 days of the EOS that the PI deems definitely, probably, or 
possibly related to the device, the event will be documented and followed until all parameters 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 27 of 40 
 
(including laboratory) have returned to normal, stabilized, or are otherwise explained.  The 
subject will be evaluated by the PI within 72 hours of the PI becoming aware of the AE. 
Information to be collected includes type of event, date of onset, date of resolution, 
investigator-specified assessment of severity and relationship to device, seriousness, as well as 
any action taken.   
All conditions present before the Screening Visit will be documented as medical history. 
Specific procedures for recording and reporting serious adverse events are described in section 
10.5. 
8.1.2 General Health Assessment 
General health of the subject will be assessed at the Screening Visit utilizing the American 
Society of Anesthesiologists (ASA) Physical Status Classification System. Only subjects with an 
ASA classification of 1 and 2 will be eligible to participate in the trial.  
 ASA Physical Status 1 - A normal healthy patient 
 ASA Physical Status 2 - A patient with mild systemic disease 
 ASA Physical Status 3 - A patient with severe systemic disease 
 ASA Physical Status 4 - A patient with severe systemic disease that is a constant 
threat to life 
 ASA Physical Status 5 - A moribund patient who is not expected to survive without 
the operation 
 ASA Physical Status 6 - A declared brain-dead patient whose organs are being 
removed for donor purposes 
9. 
STUDY ENDPOINTS 
The study’s endpoint is the regional measurement of sulfasalazine, the metabolic breakdown 
products of sulfasalazine in the digestive tract and peripheral circulation, and the gut microbiota 
in the digestive tracts of healthy volunteers. 
Safety assessments will include the monitoring of subject-reported adverse events as 
communicated to the study staff. 
10. 
SAFETY 
10.1 Definition of Adverse Event 
An adverse event (AE) is any untoward, undesirable, unplanned clinical event in the form of signs, 
symptoms, or clinical significant deviation from baseline in laboratory or physiological 
observations occurring in a human being participating in a clinical study with a Sponsor study 
device regardless of a causal relationship to device or device delivery and removal procedures. 
This includes the following:  

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 28 of 40 
 
 Any clinically significant worsening of a pre-existing condition; 
 Any reoccurrence of a pre-existing condition; 
 An AE that has been associated with the premature removal of the device. 
An adverse device effect (ADE) is any untoward and unintended response to the investigational 
device or device delivery and removal procedures.  
10.2 
DEFINITION OF SERIOUS ADVERSE EVENT 
A serious adverse event (SAE) is any adverse event that suggest a significant hazard or side effect, 
regardless of the PI’s or Sponsor's opinion on the relationship to the investigational product and 
that results in, but may not be limited to, any of the following outcomes: 
 death (regardless of the cause) 
 a life-threatening event 
 hospitalization or prolongation of existing hospitalization (any inpatient hospital 
admission that includes a minimum of an overnight stay in a health care facility) 
 a persistent or significant disability/incapacity 
 a congenital anomaly or birth defect 
 significant medical events which requires medical intervention to prevent any of the 
above outcomes 
Significant medical events are those which may not be immediately life-threatening, but may 
jeopardize the subject and may require intervention to prevent one of the other serious 
outcomes listed above.  
Hospitalization for elective treatment of a pre-study condition that did not worsen while on study 
and hospitalizations for treatment of non-adverse events (e.g. cosmetic surgery) are not 
considered serious adverse events. 
An unexpected adverse medical device event is any adverse event, the specificity or severity of 
which is not consistent with information in the current Investigator's Brochure (IB) for an 
unapproved medical device. 
A serious adverse device effect (SADE) is any ADE that has resulted in any consequences 
characteristics of a serious adverse event or that might have led to any of these consequences if 
suitable action had not been taken or intervention had not been made or if circumstances had 
been less opportune. 
10.3 
DEFINITION OF RELATIONSHIP TO INVESTIGATIONAL DEVICE AND DEVICE DELIVERY 
AND REMOVAL PROCEDURES 
The PI will document his/her clinical judgment of the causal relationship of the investigational 
product to the AE. The PI should consider and evaluate alternative causes, such as underlying 
conditions or other risk factors, as well as the temporal relationship of the event and the study 
intervention in their assessment of causality. The AE relationship will be defined using the criteria 
outlined in Table 4.  

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 29 of 40 
 
 
Table 4: 
Definition of Adverse Event Relationship Criteria 
Relationship Criteria 
Definition 
PROBABLY  
The adverse event is most likely to be explained by the 
investigational device or device delivery procedure, rather 
than the subject’s clinical state or other agents/therapies; as 
the event follows a reasonable temporal sequence from the 
medical device delivery timepoint and it abates upon 
removal of the medical device. 
POSSIBLY  
The adverse event may be explained by the investigational 
device or device delivery procedure, or the subject’s clinical 
state or other agents/therapies. 
 UNRELATED 
The adverse event can be explained by the subject’s clinical 
state or other agents/therapies. 
10.4 
DEFINITION OF SEVERITY OF ADVERSE EVENTS  
Adverse events must be graded for severity. A severity category of mild, moderate,  severe or life 
threatening as defined below, must be entered on the AE CRF page.  
1. 
MILD – Awareness of signs or symptoms, but they are easily tolerated and no 
disruption of normal daily activity.  
2. 
MODERATE – Enough discomfort to cause interference with normal daily activity. 
No treatment or medical intervention is indicated.  
3. 
SEVERE – Inability to work or perform normal daily activity; medical intervention 
is indicated.  
4. 
LIFE THREATENING/DISABLING – An immediate threat to life or leading to a 
permanent mental or physical conditions that prevents work or performing 
normal daily activities. Treatment or medical intervention is required in order to 
maintain survival.  
10.5 
REPORTING OF ADVERSE EVENTS 
Any Serious adverse event (SAE) and device respectively procedure-related (Relatedness: 
definite, probably and possibly) Adverse Effect (SADE),  must be fully recorded utililzing the 
sponsor provided SAE form and reported in writing by the Principal Investigator to Envivo Bio Inc. 
within 24 hours of becoming aware of the event. 
The PI will complete the Serious Adverse Event Form and email a scanned copy to Envivo Bio Inc. 
sponsor and the medical monitor contacts as follows:  

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 30 of 40 
 
 
 
 
 
Additional information (follow-up) about any SAE or SADE unavailable at the initial reporting 
should be forwarded by the site to the Envivo Bio Inc. as soon as they become available (such as 
hospital discharge).  
Envivo Bio Inc. will review and assess with the PI any SAE’s and any serious device/procedure-
related adverse effects. Envivo Bio Inc. ensures that reporting requirements according to the local 
regulations are fulfilled.   
All SAE’s which are considered as device- or procedure-related will be reported to IRB by the PI.  
Subjects who have had an SAE during the Treatment Phase must be followed clinically until all 
parameters (including laboratory) have returned to normal, have stabilized, or are otherwise 
explained.  The study period for the purpose of SAE reporting is defined as the period from time 
of informed consent until 14 days after the End of Study.  
If the investigator detects a SAE in a trial subject after the end of the period of observation, and 
considers the event possibly related to the prior trial, he should contact the sponsor to determine 
how the AE should be documented and reported. 
10.6 
DOCUMENTATION OF AES 
All AEs reported by the subject or detected by the investigator will be documented on the 
appropriate pages of the case report form (CRF). AEs must also be fully documented in the 
subject’s source documents.  
If the adverse event is serious, the investigator must complete, in addition to the “Adverse Event” 
page, a “Serious Adverse Event” Report Form at the time the serious adverse event is detected. 
Every attempt should be made to describe the adverse event in terms of a diagnosis. If a clear 
diagnosis has been made, individual signs and symptoms will not be recorded unless they 
represent atypical or extreme manifestations of the diagnosis, in which case they should be 
reported as separate events. If a clear diagnosis cannot be established, each sign and symptom 
must be recorded individually. 
10.7 
PREGNANCY 
Female subjects should immediately inform the Investigator of any pregnancies during the study.  
The site should report all pregancies within 48 hours to the Envivo Bio Inc. Project Manager using 
the SAE form.  
 
11. 
STATISTICAL ANALYSIS PLAN 
Due to the small sample size and the study objectives, the statistics will be mainly descriptive by 
nature. For categorical variables, summary tables will be provided giving sample size, absolute 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 31 of 40 
 
and relative frequency. For continuous variables summary tables will be provided giving sample 
size, arithmetic mean, standard deviation, standard error of the mean (SEM), median, minimum 
and maximum, percentiles and 95% CI (Confidence Interval) for means of variables.  
 
 
Tables of descriptive statistics will be presented for the following variables:   
 Adverse events and device-related adverse events in the clinical documentation. 
 Effectiveness of the CapScan Device in collecting measurable quantities of metabolic 
breakdown products of sulfasalazine and for the identification of microbial enzyme 
genes. 
 
All safety data will be summarized across the cohort with descriptive statistics. The incidence of 
adverse events during the treatment period will be summarized.  
All adverse events will be classified by System Organ Class and preferred term according to the 
Medical Dictionary for Regulatory Activities (MedDRA). The latest version available at the time of 
coding will be used throughout the study. 
All adverse events will be listed together with information on onset, duration, frequency, 
severity, seriousness, relationship to the investigational product, outcome and action taken. 
Frequency tables for severity and relationship to the Envivo Bio Device device will be provided 
by System Organ Class and preferred term. 
The main analysis will not include events which started before the first use of CapScan Device 
number 1; only treatment emergent adverse events will be included in the summary tables. 
VAS scores and changes from baseline will be presented by appropriate summary tables. Changes 
from baseline will be analyzed using Paired T-test or Signed rank test for two means (paired 
observations, or as appropriate). 
Any deviation from the statistical analysis plan will be recorded together with an explanation of 
the deviation. 
The data will be analyzed using standard statistical software (e.g., SAS [Cary, NC] or R [Vienna, 
Austria]). 
12. 
SAMPLE SIZE CONSIDERATION 
10 healthy subjects will be included in this study. The planned sample size was considered 
adequate by the Sponsor for this feasibility study. The study is not expected to show statistical 
significance or statistical power, but only to demonstrate safety and a trend for effectiveness and 
suitability of the sample collection process.   
 
13. ETHICS 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 32 of 40 
 
12.1 
INSTITUTIONAL REVIEW BOARD 
Prior to initiation of the study, the PI will submit the study protocol and significant amendments, 
sample Informed Consent Form (ICF), and any other documents that may be requested to the 
Instituional Review Board (IRB) for review and approval. The PI will request that the IRB provide 
written approval of the study and will keep on file records of approval of all documents pertaining 
to this study. The Investigator will not begin the study until the protocol and ICF have been 
approved by the IRB. The PI must agree to make any required progress reports to the IRB, as well 
as reports of device-related SAEs, life-threatening problems, or death. The PI will provide a copy 
of the written IRB approval of the protocol and ICF to the Sponsor prior to the start of the study. 
12.2 
ETHICAL CONDUCT OF THE STUDY 
This trial will be conducted in compliance with this protocol, Sponsor instruction, the guidelines 
for good clinical practice (GCP), ISO 14155 parts 1 and 2 (2003), Medical Device Directive 
93/42/EEC, Declaration of Helsinki; and applicable federal and local regulatory requirements. 
All clinical work conducted under this protocol is subject to GCP rules. This includes an inspection 
by Envivo Bio Inc. or its designee, health authority, IRB representatives at any time. The PI must 
agree to the inspection of study-related records by health authority representatives and/or 
Envivo Bio Inc. or its designee. 
 
12.3 
SUBJECT INFORMATION AND CONSENT 
At the Screening Visit the subject will be informed in detail about the investigational CapScan  
Device, and the nature of the clinical investigation with known or foreseeable benefits, risks and 
discomforts that subject may experience.  The basic elements of informed consent as specified 
by ICH-GCP will be followed.  
Written informed consent will be obtained from each subject to be involved in the clinical trial 
by using the Informed Consent Form (ICF) prior to the conduct of any study-related activity. Each 
subject will be given a copy of the written ICF. The subject must understand that throughout the 
study his or her participation remains voluntary and protected by the Declaration of Helsinki. The 
investigator is responsible for obtaining written (or witnessed) informed consent from potential 
subjects prior to study entry.   
Subjects will be given time to read the informed consent and ask any questions before being 
asked to sign the form.  The subjects will also be instructed that they are free to withdraw their 
consent and discontinue their participation in the study at any time without prejudice. If a 
particpant withdraws during the Treatment Phase, the investigator will schedude an Early 
Termination visit.  
Prior to the start of the study, the PI or PI designee will provide Envivo Bio Inc. with a blank copy 
of the ICF version approved by the IRB. Each subject’s study file will include the signed ICF for 
study participation.  

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 33 of 40 
 
When the study Treatment Phase is completed and the CRF has been monitored, the ICF will be 
kept in the investigator’s central study file for the required period of time.  
12.4 
SUBJECT INSURANCE 
The sponsor has subscribed to an insurance policy covering, in its terms and provisions, its legal 
liability for injuries caused to participating persons and arising out of this research performed 
strictly in accordance with the scientific protocol as well as with applicable law and professional 
standards. The insurance was taken out with a maximum limit of $1,000,000 per participating 
person. 
Any impairment of health which might occur in consequence of trial participation must be 
notified to the insurance company. The study subject is responsible for notification. The insured 
person will agree to all appropriate measures intended to clarify the cause and the extent of 
damage as well as the reduction of damage. 
During the conduct of the trial, the study subject must not undergo other clinical investigational 
treatment except for cases of emergency. The study subject is bound to inform the investigator 
immediately about any AE’s and additionally drugs taken. The terms and conditions of the 
insurance should be delivered to the study subject. 
12.5 
INFORMING THE GENERAL PRACTITIONER 
The PI should inform the study subject’s regular physician of his/her participation in this study, 
by sending a letter to the physician, if the study subject agrees.  
12.6 
PERSONAL DATA PROTECTION 
Envivo Bio Inc. complies with the principle of subject's right to protection against invasion of 
privacy. Throughout this trial, all data will be identified only by an identification number and 
subject initials. The data will be blinded in all data analyses. The study subject must be informed 
and consent is required that authorised personnel of Envivo Bio Inc. (Study Monitor, Auditor, 
Investors and Consultants etc.) and relevant Health regulatory agency will have direct access to 
personal medical data to assure a high quality standard of the study. 
14. 
QUALITY CONTROL AND QUALITY ASSURANCE 
Envivo Bio Inc. maintains quality assurance to ensure that clinical trials are conducted and data 
are generated, documented and reported in compliance with the protocol, GCP and applicable 
regulatory requirments.  
This clinical trial may be audited by the Sponsor to determine whether the study is being 
conducted and monitored in compliance with the protocol as well as recognized GCP guidelines 
and local regulations. These audits will also increase the likelihood that the study data and all 
other study documentation can withstand a subsequent inspection by any regulatory authority. 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 34 of 40 
 
Such audits, if necessary, will be pre-arranged with the site and conducted within a reasonable 
period. 
The study may also be inspected by regulatory agencies. These inspections may take place at any 
time during or after the study and are based on the federal regulations, as well as ICH-GCP 
guidelines. 
13.1 
STUDY MONITORING 
Monitoring of the study is the responsibility of the Sponsor. The study monitor will advise the PI 
regarding the practical conduct of the study and maintaining compliance with the protocol, GCP 
and all applicable regulatory requirements. Throughout the course of the study, the Sponsor and 
the study monitor will oversee the conduct and the progress of the study by frequent contacts 
with the investigator. This will include telephone calls and on-site visits. During the on-site visits, 
the CRFs will be reviewed for completeness with corresponding source documents. As part of the 
routine data monitoring, source documents will be made available for review by the study 
monitor. The study monitor will also perform subject's validation checks of the study particpant’s 
questionnaires and may periodically request review of the Investigator's Study File to ensure 
completeness of documentation in all respects of clinical study conduct.  
Upon completion of the study, the study monitor will arrange for a final review of the study files 
after which the files should be secured for the appropriate time period. The PI or PI designee will 
receive the study monitor during these on-site visits, cooperate in providing the documents for 
inspection, and respond to inquiries. 
13.2 
SOURCE DOCUMENT 
The investigator(s)/Institution(s) will permit study-related monitoring, audits by or on behalf of 
the Sponsor, IRB review and regulatory inspections providing direct access to source data 
documents. Source documents are original records in which raw data are first recorded. These 
may be office / clinic / hospital records, charts, diaries, questionnaires and laboratory results, 
printouts, pharmacy records, care records, completed questionnaires for each study subject. 
Source documents should be kept in a secure and limited access area. All source documents must 
be accurate, clear, unambigous, permanent and capable of being audited. They should be made 
using a permanent form of recording (ink, typing, printing, optical disc etc). They should not be 
obscured by correcting fluid or have temporary attachements (such as removable self-stick 
notes). Source documents that are computer generated and stored electronically must be printed 
(for review by the study monitor), signed and dated by the investigator.  
Source data for subjects should indicate date informed consent was signed, participation in 
clinical protocol number EB-01 and title, evidence that inclusion/exclusion criteria have been met 
and study execution dates. 
 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 35 of 40 
 
13.3 
CASE REPORT FORMS (CRFS) 
CRFs will be provided to the study site by Envivo Bio Inc. The PI will be responsible for the 
timeliness, completeness, and accuracy of the information on the CRF. All entries must be legibly 
recorded. The entry to be corrected is to be crossed out with a single line so that the original 
entry remains legible. The correction then has to be made right next to the entry and confirmed 
by date and initials of the person making the correction. Corrections that cannot be made in this 
fashion have to be explained in a detailed statement (e.g. Data Clarification Form), reference to 
which has to be documented on the relevant CRF page(s). No one can erase, overwrite, or use 
correction fluid on the original. The CRF for each subject must be reviewed and signed by the PI. 
This should be done as soon as possible after each subject completes the study. The PI will make 
the CRF pages available for review and collection by the designated Envivo Bio Inc.'s 
representative at each scheduled monitoring visit. A clinical study monitor will review the CRFs 
for all subjects enrolled and will compare the content versus the source data. The CRF will be on 
colored paper (for example, yellow). At the end of the study, the CRF will be photocopied with 
the original colored CRF (yellow) retrieved by the monitor to the Sponsor and the photocopied 
CRF (white) will remain at the site.  
All clinical work conducted under this protocol is subject to GCP rules. This includes an inspection 
by Envivo Bio Inc. and/or health authority representatives at any time. The PI will agree to the 
inspection of study-related records by health authority representatives and/or Envivo Bio Inc. 
13.4 
DATA MANAGEMENT 
All protocol-specified data documented on the CRFs as well as the subjects’ questionnaires and 
diaries will be entered into an electronic database by a representative of the Sponsor. 
14.0 STUDY ADMINISTRATION 
14.1 
STUDY PERSONNEL 
14.1.1 Site Staff 
The Principal Investigator (PI) for this trial will be qualified by education and experience to 
conduct this clinical trial in accordance with the protocol, ICH Good Clinical Practice (GCP) 
guidelines and all state, local and federal regulations. The PI is responsible for the conduct of the 
clincial trial at a trial site.   
14.1.2 Sponsor 
Below is a list of key individuals and their roles from the Sponsor and designee that will contribute 
to this study: 
Sponsor:  
 
 
 
 
 
Envivo Bio Inc. 
Project Manager: 
 
 
 
 
Dr. Dari Shalon 
Study Medical Monitor:  
 
 
 
Dr. Shai Friedland 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 36 of 40 
 
14.2 
REQUIRED DOCUMENTS PRIOR TO STUDY INITIATION 
Prior to the initiation of the study and enrollment of subjects, the following items must be 
received by the Sponsor from the investigational site: 
 All applicable country-specific regulatory requirements and forms. 
 Confidential Disclosure Agreement.  
 Signed protocol, amendment(s), Investigator's Brochure and notification(s) page(s).  
 Provisions for direct access to source/data documents if necessary for trial-related 
monitoring, audits, IRB review, and regulatory inspection. 
 Curriculum vitae (current < 2 years) and current medical licenses for the Principal 
Investigator and sub-investigators. 
 Signed Clinical Study Agreement and Financial Agreement. 
 Appropriate local health authority documentation properly signed and dated by the 
required investigators. 
 Letter of approval from the IRB and local regulatory agency for protocol (identified by 
protocol title and number), informed consent form (identified by protocol title and 
number), Subject Information Sheet (if applicable), advertisements (if applicable). 
 Copy of the IRB-approved written ICF to be used in the study (that has also been approved 
by the Sponsor). 
 IRB name, address and membership list.  
 Financial disclosure information for the principal investigator and sub investigators. 
 Name and location of all investigative sites. 
Upon satisfactory receipt of all required regulatory documents, Envivo Bio Inc. will arrange that 
all study material be delivered to the study site. Subject entry should not begin until after the 
required regulatory documents are confirmed as received and the Investigator Initiation visit has 
occurred. All personnel expected to be involved in the conduct of the study will undergo 
orientation to include review of GCP, study protocol, instructions for CRF completion, AE 
reporting, and overall responsibilities, including those for study file maintenance. 
 
14.3 
INVESTIGATOR'S STUDY FILE 
Envivo Bio Inc. will provide the site with an Investigator's Study File. This file should be used for 
all trial-related documents. The site study staff will be responsible for keeping the Investigator's 
file updated and ensuring that all required documents are filed appropriately. The file will be 
inspected during monitoring visits and may be audited by the Sponsor and/or regulatory 
agencies.  

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 37 of 40 
 
The Investigator's Study File will contain the protocol/amendments, Case Report and Query 
Forms, IRB and governmental approval with correspondence, sample informed consent, study 
staff curriculum vitae and authorization forms and other appropriate documents or 
correspondence etc. 
14.4 
CLINICAL TRIAL SUPPLIES 
Clinical trial supplies include, however, not limited to: CRFs, subjects' Daily Diary, saliva collection 
kit, stool collection kits, and Capscan  Devices.  
The PI, PI designee or Sponsor will be responsible for the dispensing, inventory, and 
accountability of all clinical trial supplies. An accurate and timely record of the disposition of all 
clinical supplies must be maintained. The supplies and inventory record must be made available 
for inspection by the designated Sponsor's representative upon request. Upon completion or 
termination of the study the Investigator will return all remaining clinical supplies to Envivo Bio 
Inc., or Sponsor designee along with a copy of the inventory record and a record of the clinical 
supplies returned. Under no circumstances will the PI allow the investigational medical device to 
be used other than as directed by this protocol. 
14.5 
STUDY REPORTING REQUIREMENT 
The Principal Investigator reporting requirements during the study include:  
1. Notifying the Envivo Bio Inc. immediately in case of any Serious Adverse Event 
and any device-related Adverse Effect. 
2. Inform the IRB about any Serious Adverse Device Effects (SADE’s) as per IRB 
reporting policy. 
3.  Reporting to the Sponsor within five (5) working days of a withdrawal of IRB 
approval at the site. 
4.  Notifying the IRB and the Sponsor within five (5) working days of the occurrence 
of any emergency protocol deviations necessary to protect study subjects. 
 
14.6 
STUDY COMPLETION 
This study is expected to end when all required subjects have been enrolled and the last subject 
has completed the study and the query resolution has been completed. 
A declaration of the end of the clinical trial will be made according to local regulations. 
Data and materials that are required before the study can be considered complete and/or 
terminated are: 
 All clinical data and all special test results from screening through the end of the 
follow-up period 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 38 of 40 
 
 CRFs (including correction forms) properly completed by appropriate study personnel 
and signed by the PI 
 Statement of outcome for each serious adverse event reported 
 Copies of approved protocol amendments by IRB committee as well as relevant health 
authority approval/notification (if applicable) 
14.7 
FINAL REPORT 
The PI will complete a report notifying the IRB of the conclusion of the clinical study.  
The final report sent to IRB will also be sent to the Sponsor and, along with the completed CRFs, 
will constitute the final summary to the Sponsor, thereby fulfilling the PI’s regulatory 
responsibility. 
14.8 
RETENTION OF STUDY RECORDS 
The PI will retain copies of the approved protocol, completed CRFs, informed consent documents, 
relevant source documents, and all other supporting documentation related to the project in a 
secured and safe facility for one of the following periods based on notification from the Sponsor: 
 A period of at least two years from last device registration and notification from the 
Sponsor 
 Or a period of at least two years after discontinuation of clinical development of the 
investigational product as confirmed by the Sponsor 
 Or longer if required by local regulations. 
These files must be made available for inspection upon reasonable request by authorized 
representatives of Envivo Bio Inc., or the corresponding regulatory agencies of the relevant 
countries. 
Envivo Bio Inc. will provide the PI with information concerning the current status of the 
investigational medical device as it relates to the investigator’s obligation for the retention of 
study records. The PI should contact Envivo Bio Inc. prior to disposing of any such records and to 
provide written notification to the Sponsor of any change in the location, disposition, or custody 
of the study files. Envivo Bio Inc. will arrange for continued storage of all records, if necessary. 
14.9 
CONFIDENTIALITY AND PUBLICATION OF STUDY RESULTS 
All unpublished information given to the PI by the Sponsor shall be kept confidential. Results of 
this study will not be submitted for presentation or publication or disclosed to a third party 
without the prior written permission of Envivo Bio Inc. The publication and communication rights 
are found in the Investigator agreement. The Sponsor requires that manuscripts or abstracts 
containing information about or data derived from this study be submitted to the Sponsor at 
least 30 days prior to submission to the journal or conference presentation, to safeguard against 
disclosure of confidential information.  

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 39 of 40 
 
When the Sponsor generates reports for presentations to regulatory agencies, an endorsement 
of the final report may be sought from the investigators who have contributed significantly to 
the study.  
No patent application based on the results of this study shall be made by the Investigator, and 
no assistance shall be given to any third party for such an application, without the written 
authorization of the Sponsor. 
 
 
REFERENCES 
 
 
i Shreiner AB et. al., The gut microbiome in health and in disease, Curr Opin Gastroenterol. 2015 January ; 31(1): 
69–75. 
 
ii Weersma RK, Zhernakova A, Fu J. Interaction between drugs and the gut microbiome, Gut, doi:10.1136/gutjnl-
2019-320204 
 
iii Blaser MJ., The microbiome revolution, J Clin Invest. 2014;124(10):4162–4165 
 
iv Wlodarska M. et. al. An integrative view of microbiome-host interactions in inflammatory bowel diseases, Cell 
Host Microbe. 2015 May 13; 17(5): 577–591. 
 
v Li G, Yang et al. Diversity of duodenal and rectal microbiota in biopsy tissues and luminal contents in healthy 
volunteers. J Micro Biotech. 2015;25(7):1136–45. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

PROTOCOL: EB-03 
ENVIVO BIO INC. 
CONFIDENTIAL 
 
 
 Page 40 of 40 
 
 
 
 
 
Appendix A 
Subject initials: _______     Subject no: ______     
 
 
Today’s Date:  ___________      
 
 
 
 
  
                                                
(Date notation 01 May 2022)               
ENVIVO BIO DAILY DIARY  
Instructions: Please complete all questions below. 
 
1) Circle CapScan Device number swallowed today and time of swallowing:  
Capsule 
number 
1 
2 
3 
4 
5 
6 
Time 
swallowed 
 
 
 
 
 
 
(Time notation 9:15 am, 5:30 pm) 
 
       2) Time sulfasalazine taken:________  Time blood sampled:____________ and _______________ 
 
 
3) What did you eat and drink today: 
Food  
 
 
 
 
 
 
 
 
 
  Time Finished Eating 
 
 
 
 
 
 
 
 
 
 
 
 
4) When did you have bowel movements today and did you retrieve a CapScan Device in the toilet: 
Bowel movement number 
1 
2 
3 
4 
5 
Time  
 
 
 
 
 
Retrieved device (Yes or No)