Clinical trial NCT03047629
Conditions: Parkinson's Disease Constipation
Keywords: non-motor symptoms
Sponsor: Enterin Inc.
Start date: 2017-05-11; Completion date: 2018-06-14
Enrollment: 50 participants; Allocation: RANDOMIZED; Intervention model: SEQUENTIAL

Brief summary:
This is a Phase 1/2a study to assess the safety, tolerability, pharmacokinetics, and pharmacodynamics of an orally-administered medication to relieve symptoms of constipation associated with Parkinson's Disease. Ten patients will be enrolled in Phase 1, and will be studied over an 8-12 week period. Forty patients will be enrolled in Phase 2, and will be studied over an 8-10 week period. All subjects will receive the study drug during one of the observational periods of the study.

Detailed description:
Phase 1 will enroll 10 patients to assess the safety, tolerability, and pharmacokinetics of single escalating doses over a 30-60 day period. The dose-escalation period will be preceded by a 2-week run in period and followed by a 2-week wash-out period.

Phase 2 follow the safe completion of Phase 1. It will enroll 40 patients and is composed of 4 periods of study: 1) a 2-week run-in period, 2) a 3-5 week escalating dose period to identify a prokinetic dose in the initial set of 10 patients, 3) a 1-week period of randomized dosing (placebo versus the previously identified pro-kinetic dose), and 4) a 2-week wash-out period. Pharmacodynamics will be assessed along with safety and tolerability. Relative outcomes will be compared within each patient and across groups for the randomized dosing period.

Frequency of bowel movements and other non-motor symptoms of Parkinson's Disease will be collected over the course of both phases.

Arms: arm 1: ENT-01 at a to-be-determined dose taken by mouth every day upon awakening. arm 2: Placebo to be taken by mouth every day upon awakening
Interventions: intervention 1: ENT-01 intervention 2: Placebo
Intervention descriptions: intervention 1: Daily dosing with ENT-01. ENT-01 is an orally administered proprietary substance formulated as a small 25mg coated tablet. Dosing will range from 25-200mg, and the dose will be taken upon awakening on an empty stomach with 8oz water simultaneous to dopamine. intervention 2: Daily dosing with a placebo

Protocol documents:
Prot_000.pdf: 
 
 
 
 
 
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PROTOCOL TITLE 
A Multicenter, Single-Dose, Multiple-Dose, Randomized, Double-Blind, Placebo-Controlled 
Study to Evaluate Safety, Tolerability, Pharmacokinetics and Pharmacodynamics of Orally 
Administered ENT-01 for the Treatment of Parkinson’s Disease related Constipation.  IND# 
130770 
 
PROTOCOL NUMBER 
ENT-01 
 
STUDY PHASE 
Phase 1/2a 
 
 
 
 
 
 
 
 
 
Date issued: 
10 April 2017  
Date Amended: 
 27 July 2017 


 
 
 
 
 
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PROTOCOL / AMENDMENT APPROVAL PAGES(S) 
 
 
Authored by: 
Co-Authored by: 
 
 
Denise Barbut MD FRCP 
 
Michael Zasloff MD PhD 
 
Printed Name 
Printed Name 
 
 
 
 
 
 
Signature 
Signature 
9179751377 
 
4844337807 
 
Telephone Number 
Telephone Number 
dbarbut@gmail.com 
 
Maz5@georgetown.edu 
 
E-mail Address 
E-mail Address 
 
 
 
 
Date 
Date 
 
 
 
 
 
 
 
IMPORTANT CONTACTS AT ENTERIN 
 
Person responsible for this study 
Telephone  
E-mail address 
Denise Barbut 
917-975-1377 
dbarbut@gmail.com 
Michael Zasloff 
484-433-7807 
Maz5@georgetown.edu 
Matt Lowery 
727-317-6639 
m.lowery@enterininc.com  
Tom Malloy 
215-740-0699 
t.mallow@enterininc.com  
 


 
 
 
 
 
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STUDY SYNOPSIS 
 
Study Title 
Safety, Tolerability, Pharmacokinetic and Pharmacodynamic 
characteristics of single or multiple dose oral ENT-01 for 
Constipation in Parkinson’s Disease. 
Objectives 
Stage 1: 
Primary Objective:  
Safety and Tolerability of single oral doses of ENT-01 in patients 
with Parkinson’s Disease related constipation. 
Secondary objective: 
To evaluate the Pharmacokinetic (PK) characteristics of single 
oral doses of ENT-01 in patients with Parkinson’s Disease and 
constipation. 
 
Stage 2: 
Primary objective: 
To determine the Safety, Tolerability, PK and Pharmacodynamic 
(PD) characteristics of repeated oral doses of ENT-01 in patients 
with Parkinson’s Disease related constipation. 
 
 
Exploratory 
To determine the effectiveness of study drug in improving 
colonic motility. 
To collect information on the effect of study drug on Quality of 
Life Measures 
Study Description 
Multicenter, Single dose, Multiple dose safety, tolerability and 
dose-finding study leading into a Randomized, Double-blind, 
Placebo-controlled Study 
Study Population 
Parkinson’s disease with constipation  
Study Drug 
Administration 
Oral 
Study Procedures 
Colonic motility will be determined by Sitzmarks method before 
and during treatment 


 
 
 
 
 
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Study Endpoints 
Stage 1: 
Safety Endpoint:  
Adverse events as evaluated with patient report, vital signs, 
chemical chemistry and EKG 
Tolerability Endpoint: 
• Recurrent vomiting 
• Recurrent diarrhea 
• Abdominal pain 
• Postural hypotension/dizziness 
 
Stage 2: 
Primary Efficacy Endpoint:  
• Bowel function as determined by stool frequency  
 
 
Exploratory Endpoints 
• Colonic motility as determined by Sitzmarks test 
• Sleep diary 
• Skin temperature (I-Button)-assessed sleep parameters 
• Mood assessed with Beck depression inventory (BDI) 
• QOL, non-motor symptoms questionnaires 
Initial Date 
March 2017 
Completion Date 
December 2017 
 
 
 


 
 
 
 
 
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TABLE OF CONTENTS 
1 
INTRODUCTION....................................................................................................... 10 
1.1 
Background ...................................................................................................... 10 
1.2 
Rationale for the Study ................................................................................... 12 
1.3 
ENT-01 Dosing Rationale ............................................................................... 15 
2 
OBJECTIVES ............................................................................................................. 17 
2.1 
Stage 1 ............................................................................................................... 18 
2.2 
Stage 2 ............................................................................................................... 19 
3 
STUDY DESCRIPTION ............................................................................................ 22 
3.1 
Periods .............................................................................................................. 23 
4 
STUDY POPULATION ............................................................................................. 38 
4.1 
Number of Patients .......................................................................................... 38 
4.2 
Selection Criteria ............................................................................................. 38 
4.3 
Inclusion Criteria ............................................................................................ 38 
4.4 
Exclusion Criteria ............................................................................................ 40 
4.5 
Discontinuation Criteria ................................................................................. 41 
4.6 
Concomitant Medication ................................................................................ 42 
5 
STRATIFICATION AND RANDOMIZATION (STAGE 2) ................................. 43 
6 
UNBLINDING............................................................................................................. 43 
7 
MATERIALS .............................................................................................................. 43 
7.1 
Study Drug ....................................................................................................... 43 
7.2 
Packaging and Labeling .................................................................................. 43 
7.3 
Storage, Dispensing and Reconciliation of Study Drug ............................... 43 
8 
WARNINGS AND PRECAUTIONS ........................................................................ 44 
9 
STUDY PROCEDURES ............................................................................................ 44 
9.1 
Sitzmarks assessment of colonic motility in Stage 2 ..................................... 44 
9.2 
Observations and Measurements ................................................................... 45 
9.3 
Informed Consent ............................................................................................ 45 
9.4 
Instructions to Patients ................................................................................... 45 
9.5 
Clinical Laboratory Procedures ..................................................................... 46 
9.6 
Other Procedures ............................................................................................ 46 
9.7 
CTCAE Definitions of Dose Limiting Adverse Events ................................ 47 


 
 
 
 
 
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9.8 
Criteria for DLT Endpoints: .......................................................................... 48 
9.9 
Pre-Existing Medical Conditions ................................................................... 48 
9.10 
Treatment Emergent Adverse events ............................................................ 48 
9.11 
Laboratory Abnormalities .............................................................................. 49 
9.12 
AE Assessment and Recording ....................................................................... 49 
9.13 
Reporting Requirements ................................................................................. 51 
9.14 
Serious Adverse Events ................................................................................... 52 
9.15 
Notification of Serious Adverse Events ......................................................... 53 
9.16 
Reporting a Serious Adverse Event ............................................................... 53 
9.17 
Serious Adverse Experiences: After Study Participation ............................ 55 
9.18 
Departure from Protocol for Emergency or Adverse Event ....................... 55 
9.19 
Data Safety and Monitoring Board ............................................................... 56 
9.20 
Review of Stage 1 Data by Data Safety Monitoring Board ......................... 56 
9.21 
Stopping Rules ................................................................................................. 56 
9.22 
Follow-Up and Final Reports ......................................................................... 56 
9.23 
Regulatory Aspects .......................................................................................... 56 
10 
DATA MANAGEMENT AND STATISTICS .......................................................... 57 
10.1 
Sample Size/Power Considerations ................................................................ 57 
10.1.1 
Stage 1 ............................................................................................................... 57 
10.1.2 
Stage 2 ............................................................................................................... 58 
10.2 
Data Collection and Case Report Form Monitoring .................................... 59 
10.3 
Endpoint definitions and measurements ....................................................... 59 
10.3.1 
Assessment of Safety ....................................................................................... 59 
10.3.2 
Efficacy Endpoints .......................................................................................... 61 
10.4 
Analysis Plan .................................................................................................... 62 
10.5 
Stage 2 ............................................................................................................... 62 
11 
ESTIMATED DURATION OF THE STUDY ......................................................... 64 
12 
ADMINISTRATIVE PROCEDURES AND ETHICAL CONSIDERATIONS ... 64 
12.1 
Medical Care .................................................................................................... 64 
12.2 
Patient Information and Informed Consent ................................................. 64 
12.3 
Study Monitoring ............................................................................................ 64 
12.4 
Audits and Inspections .................................................................................... 65 


 
 
 
 
 
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12.5 
Ethics Committee Review ............................................................................... 66 
12.6 
Standards ......................................................................................................... 66 
12.7 
Confidentiality ................................................................................................. 66 
12.8 
Protocol Adherence ......................................................................................... 67 
12.9 
Amendments to the Protocol .......................................................................... 68 
12.10 
Protocol Deviations .......................................................................................... 68 
12.11 
Study Termination .......................................................................................... 68 
12.12 
Data handling and record keeping ................................................................. 69 
12.13 
Inspection of Records ...................................................................................... 69 
12.14 
Data Management ........................................................................................... 69 
12.15 
Data Capture and Management ..................................................................... 69 
12.16 
Liability and Insurance ................................................................................... 70 
12.17 
Retention of Records ....................................................................................... 70 
12.18 
Data Quality Assurance .................................................................................. 70 
13 
USE OF INFORMATION ......................................................................................... 71 
14 
INVESTIGATOR AGREEMENT ............................................................................ 71 
15 
REFERENCES ............................................................................................................ 72 
APPENDIX 1:  CLINICAL LABORATORY DETERMINATION .................................. 75 
APPENDIX 2: SCHEDULE OF EVENTS ........................................................................ 76 
APPENDIX 3: NUMERIC RATING SCALES FOR PAIN AND SWELLING ............ 1 
APPENDIX 4: ROME-IV CRITERIA FOR CONSTIPATION ...................................... 2 
APPENDIX 5: EASE OF EVACUATION SCALE ............................................................ 3 
APPENDIX 6: BRISTOL STOOL CHART ........................................................................ 4 
APPENDIX 7: SLEEP DIARY .............................................................................................. 5 
APPENDIX 8: I-BUTTON ....................................................................................................... 6 
APPENDIX 9: NON-MOTOR SYMPTOMS QUESTIONNAIRE (NMSQ) ...................... 8 
APPENDIX 10: BECK DEPRESSION INVENTORY (BDI) ........................................... 9 
APPENDIX 11: UNIFIED PARKINSON’S DISEASE RATING SCALE ................... 14 
APPENDIX 12: MINI MENTAL STATE EXAMINATION .......................................... 43 
APPENDIX 13: TRAIL MAKING A & B ............................................................................ 44 
APPENDIX 14: UM-PDHQ ................................................................................................. 47 
APPENDIX 15: PARKINSON’S DISEASE FATIGUE SCALE (PFS-16) ................... 49 


 
 
 
 
 
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APPENDIX 16: PATIENT ASSESSMENT OF CONSTIPATION SYMPTOMS ...... 51 
APPENDIX 17: PATIENT ASSESSMENT OF CONSTIPATION QUALITY OF LIFE
 ........................................................................................................................... 52 
APPENDIX 18: REM SLEEP BEHAVIOR DISORDER SCREENING 
QUESTIONNAIRE ........................................................................................ 55 
APPENDIX 19: PARKINSON’S DISEASE SLEEP SCALE ......................................... 56 


 
 
 
 
 
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LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
ADL 
Activities of Daily Living  
AE 
Adverse Event 
BDI 
Beck Depression Inventory 
CIOMS 
Council for International Organizations of Medical Sciences 
CFR 
Code of Federal Regulations 
CRF 
Case Report Form 
CRO 
Contract Research Organization 
CTCAE 
Common Terminology Criteria for Adverse Events 
DLT 
Dose Limiting Tolerability 
EMEA 
European Agency for the Evaluation of Medicinal Products 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practice 
IB 
Investigator’s Brochure 
ICF 
Informed Consent Form 
IEC 
Independent Ethics Committee 
IRB 
Institutional Review Board 
ITT 
Intent to Treat 
MMSE 
Mini Mental State Examination 
MTD 
Maximum Tolerable Dose 
NMSQ 
Non-Motor Symptom Questionnaire 
PAC-QOL 
Patient Assessment of Constipation - Quality of Life 
PAC-SYM 
Patient Assessment of Constipation Symptoms 
PDSS 
Parkinson’s Disease Sleep Scale 
PFS 
Parkinson's Disease Fatigue Scale 
RBDQ 
REM Sleep Behavior Disorder Questionnaire 
SAE 
Serious Adverse Event 
TEAE 
Treatment Emergent Adverse Event 
ULN 
Upper Limit of Normal 
UM-PDHQ 
University of Miami Parkinson’s Disease Hallucination Questionnaire 
UPDRS 
Unified Parkinson Disease Rating Scale 


 
 
 
 
 
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WHO 
World Health Organization  
1 
INTRODUCTION 
Constipation is a common problem worldwide, affecting 2% to 27% of the population, with 
most estimates varying from 12% to 20% (Bouras and Tangalos, 2009). The prevalence of 
constipation increases to 30%-40% among people aged >65 years and women are 
disproportionately affected. In North America, 63M people meet the Rome IV criteria for 
constipation and in the US alone, constipation is responsible for over 2M physician visits annually. 
Laxatives are prescribed to 2-3M patients every year and furthermore, in most patients, the 
condition is chronic requiring life-long treatment (Bouras and Tangalos, 2009).   
Constipation is much more common among patients with Parkinson’s disease than in the 
general population. There are 1M people suffering from Parkinson’s disease  in the US, of which 
roughly 60%, or 600,000 suffer from chronic constipation and in most, the condition is chronic, 
severe and unresponsive to standard therapy (Ondo et al., 2012; Zangaglia et al., 2007). This 
represents an economic burden to the individual with Parkinson’s disease and to the healthcare 
system. According to the most recent Federal Supply Schedule (FSS; April 2016), the average 30-
day reimbursed price for a basket of orally administered drugs for constipation is approximately 
$260 or $3120 per year. This represents about $1.8B of prescription laxatives just for patients with 
Parkinson’s disease.  
Constipation not only constitutes a major economic burden but it also significantly affects 
the quality of life of the individual with Parkinson’s disease, contributing to social isolation and 
depression. Furthermore, the severity of the symptoms correlates negatively with patient reported 
quality of life.  
An effective pro-kinetic medication for individuals with Parkinson’s disease would be a 
useful addition to the currently available treatments for this disabling condition. 
1.1 
Background 
Constipation is common in Parkinson’s disease and often becomes symptomatic years 
before the onset of the motor dysfunction and the subsequent diagnosis of Parkinson’s disease (Lin 
et al., 2014; Savica et al., 2009). There is substantial evidence that the neurodegenerative process 


 
 
 
 
 
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associated with Parkinson’s disease, namely the accumulation of toxic aggregates of alpha-
synuclein, occurs within the enteric nervous system years before they appear within the brain 
(Braak et al., 2006). It is believed that the enteric nervous system (ENS), with its vast surface area, 
is subject to continuous insults from infectious agents and toxic substances. Although the function 
of alpha- synuclein is not known, inflammation within the nervous system leads to an increase in 
its intracellular levels (Allen Reish and Standaert, 2015).   In individuals with Parkinson’s disease 
the increase in alpha- synuclein leads to the formation of neurotoxic aggregates, perhaps because 
of a failure  by the neuron (due to genetic factors) to effectively dispose of them (Sahay et al., 
2016).  The aggregates of alpha- synuclein then traffic along the vagal nerve to the dorsal motor 
nucleus within the brainstem, and from there to more rostral structures (Greene, 2014).  
The individual with Parkinson’s disease suffers from a form of constipation that is believed 
to be caused principally by delayed transit through the colon. In addition, defecation is often 
impaired by dysfunction of the Parkinson’s disease patient’s anorectal reflex. For many individuals 
bowel issues represent a significant detriment to quality of life. Failure to effectively manage this 
problem can also lead to bowel obstruction, especially as the terminal phase of Parkinson’s disease 
approaches. A limited number of therapies have been subjected to clinical trials and they include 
agents that increase the fluid content of the stool, either by blocking fluid resorption or increasing 
the osmolar load within the intestine (Ondo et al., 2012; Zangaglia et al., 2007). 
Squalamine is a small molecule, originally discovered in the liver of the dogfish shark, 
where it is delivered following a meal via the biliary tract into the intestine (Moore et al., 1993). 
When squalamine is applied to the isolated colon and/or jejunum of a mouse, the compound 
stimulates organized peristaltic waves (Kunze et al, manuscript in preparation). This effect occurs 
through direct stimulation by squalamine of the intrinsic primary afferent neurons (IPANS) which 
are the most abundant type of neuron within the ENS. The IPANs, in turn, excite the ganglia of 
the myenteric plexus, and promote propulsive peristalsis. In addition, intense electrical signals are 
directed to the brainstem via the vagus following exposure of the ENS to squalamine which persist 
for some time. Of relevance to this clinical study, squalamine reverses the impaired motility of the 
colon from aged animals engineered to express a Parkinson’s disease phenotype.   


 
 
 
 
 
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Furthermore, in a recent study, squalamine has been shown to prevent the formation of 
toxic alpha-synuclein aggregates ((Perni et al., 2017)). The molecule accomplishes this as a 
consequence of its mechanism of action and the mechanism by which alpha-synuclein aggregates 
to form toxic polymers. Squalamine enters a neuron and binds to the negatively charged 
phospholipids that are present on the cytoplasmic face of the neuron’s plasma membrane, 
effectively neutralizing the negative charge of the membrane surface. Alpha-synuclein normally 
binds to the cytoplasmic face of the membrane via the positive charge on its N-terminus. Thus, 
when squalamine binds to the cytoplasmic face of the plasma membrane it displaces alpha-
synuclein into the cytoplasm. As the concentration of alpha-synuclein on the membrane decreases, 
the likelihood that monomers will aggregate into higher order polymers decreases, thus reducing 
the rate of formation of aggregates and thereby rescuing neurons from certain death. At the same 
time, squalamine excites the neuron by displacing certain ion channels and transporters from the 
membrane.  Thus, squalamine both excites the neurons of the ENS and protects them from alpha-
synuclein induced cytotoxicity. It is for these reasons that we will evaluate squalamine as a 
therapeutic to treat the constipation associated with PD and why we will explore its effect on sleep 
architecture. 
1.2 
Rationale for the Study 
Constipation is a major clinical component of Parkinson’s disease and is reported to occur 
in greater than 60% of affected individuals. The pathophysiological basis of constipation in 
Parkinson’s disease is generally believed to be due to delayed transit through the colon (Edwards 
et al., 1991; Klingelhoefer and Reichmann, 2015). Several studies have demonstrated that transit 
of stool through the colon of an individual with Parkinson’s disease is about 50% that measured 
in age matched controls. As a consequence both stool frequency and stool consistency are 
abnormal in Parkinson’s disease. For many patients, as well as those caring for these individuals, 
constipation remains a significant morbidity associated with the condition (Salat-Foix and 
Suchowersky, 2012; Sung et al., 2014).  
 Few placebo controlled clinical trials have been conducted in the Parkinson’s disease 
population to assess the efficacy of therapeutics that could be of value. Addition of fiber to the 
diet, although increasing stool volume, is reported to have no effect on colon transit time (Ondo et 


 
 
 
 
 
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al., 2012; Zangaglia et al., 2007). An osmotic laxative, polyethylene glycol (Magrogol) has been 
studied in a small placebo controlled clinical trial of individuals with mild constipation, and shown 
to provide benefit with respect to stool frequency and consistency (Zangaglia et al., 2007). A short 
term placebo controlled trial of Lubiprostone, a chloride channel activator which increases 
intestinal fluid secretion, was only effective in about 50% of those treated, and resulted in passage 
of loose stools/diarrhea in place of constipation (Ondo et al., 2012). Furthermore, Lubiprostone 
delays gastric emptying (Camilleri et al., 2006), a function already compromised in Parkinson’s 
disease.   
The pathophysiology of the GI dysfunction in Parkinson’s disease involves deposition of 
alpha-synuclein within both the ENS as well as within the brainstem.  For reasons that remain 
unknown alpha-synuclein, which is a protein normally produced in neurons, forms neurotoxic 
intracellular aggregates in Parkinson’s disease. Numerous studies suggest that the alpha- synuclein 
aggregate formation begins in the ENS of the  Parkinson’s disease individual many years before 
the onset of the motor symptoms (Braak et al., 2006).  As a consequence of the normal retrograde 
neuronal trafficking that occurs within the vagus nerve, toxic aggregates are transported from the 
neurons of the ENS to the dorsal motor nucleus of the vagus, and then, gradually to sites within 
the  brain that are involved in  physical movement and balance (Greene, 2014; Visanji et al., 2014). 
Because the constipation is fundamentally of an acquired neurodegenerative nature, it differs, so 
far as we now understand, from other forms of this condition (Visanji et al., 2014). 
The mechanism of action of squalamine has been elucidated over the past decade. The 
molecule is structural related to a bile salt, but bears a polyamine, giving it a net positive charge. 
It is produced naturally in the liver of the dogfish shark and delivered along with bile salts to small 
intestine via the bile duct. Based on studies conducted ex vivo with isolated segments of mouse 
jejunum and colon, and our knowledge of squalamine’s mechanism of action at a cellular level, 
we can understand the prokinetic effects of squalamine on intestinal motility.  Once within the 
lumen of the intestine squalamine is transported from the epithelium into the lamina propria where 
it enters neurons of the ENS, including the intrinsic primary afferent neurons, the most abundant 
neuron of the ENS (Furness et al., 1999). Once within the neuron, the positively charged 
squalamine molecule is attracted to the inner face of the plasma membrane, which has a negative 


 
 
 
 
 
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surface charge, due to the presence of anionic phospholipids, neutralizing the negative charge 
(Zasloff et al., 2011).  As a consequence, many proteins that are bound to the inner face of the 
cytoplasmic face of the plasma membrane are displaced or functionally altered, including ion 
channels and transporters (Sumioka et al., 2009; Yeung et al., 2008). By this mechanism, 
squalamine activates the firing of the IPANs, which in turn, activate the myenteric plexus, resulting 
in propulsive migrating motor complexes (MMCs), and direct electrical activity rostrally to the 
brainstem via the vagus (Kunze et al, manuscript in preparation).  
Alpha-synuclein normally binds to membranes composed of a high proportion of anionic 
phospholipids via its positively charged N-terminus (Shi et al., 2015). When the intracellular 
concentration of alpha-synuclein increases, the concentration of alpha-synuclein molecules bound 
to the inner face of the cytoplasmic membrane increases proportionally, and as it does the 
probability that they will physically contact one another and aggregate increases. In individuals 
with Parkinson’s disease these aggregates grow in size and eventually overwhelm the cell’s 
capacity to digest the inclusions, whereas in those without Parkinson’s disease these aggregates 
are cleared by autophagy (Recchia et al., 2004).  In a recent study we show that squalamine 
effectively displaces alpha-synuclein from the surface of a negatively charged membrane surface, 
and completely prevents aggregate formation. These studies have been conducted in specialized 
in vitro systems as well as in a validated in vivo model of Parkinson’s disease, a genetically 
engineered strain of C. elegans expressing human alpha-synuclein. Oral treatment of C. elegans 
with squalamine prevents the aggregation of alpha-synuclein within the muscles of the animal and 
reverses the paralysis caused by these aggregates (Perni et al., 2017). 
Thus, squalamine has the remarkable property of stimulating peristaltic activity by directly 
activating the neurons of the ENS and via the same mechanism, inhibiting the formation of 
neurotoxic alpha-synuclein aggregates within the same neurons. In principle, squalamine could 
reverse the neurodegenerative process within the ENS of the individual with Parkinson’s disease. 
If indeed the pathophysiology of the CNS component of Parkinson’s disease occurs as a 
consequence of the trafficking of alpha-synuclein aggregates from the ENS to the brain via the 
connecting nerves, then squalamine could have an impact on the disease process occurring within 
the CNS both therapeutically and prophylactically.  


 
 
 
 
 
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1.3 
ENT-01 Dosing Rationale 
ENT-01 is the phosphate salt of squalamine. For this study it has been formulated as a 
small 25 mg coated tablet. Dosing will range from 25mg to 200mg. The dose will be taken upon 
awakening on an empty stomach along with 8 oz. of water simultaneously to dopamine. The patient 
will not be allowed to ingest any food for at least 60 minutes after study medication.    The 
compound is highly charged and will adsorb to foodstuffs, so it is being administered prior to 
feeding.  
The dose rationale for this study was based on the extensive clinical experience gathered 
in the setting of clinical trials involving systemic administration of squalamine, experience 
gathered from its consumption in nutraceutical products, and pre-clinical pharmacology studies.   
  
ENT-01, the phosphate salt of squalamine, is weakly soluble in water at neutral pH but 
readily dissolves at pH< 3.5 (the pH of gastric fluid).  Squalamine, as the highly water soluble 
dilactate salt has been extensively studied in over  3 Phase 1 and 8 Phase 2 human clinical trials as 
an intravenous agent for the treatment of cancer and diabetic retinopathy (Bhargava et al., 2001; 
Connolly et al., 2006; Hao et al., 2003; Herbst et al., 2003). The compound is well tolerated in 
single and repeat intravenous administration, alone or in combination with other agents, to doses 
of at least 300mg/m2). In the current clinical trial, ENT-01 will be administered orally to subjects 
with Parkinson’s disease who have long standing constipation. Although this trial will be the first 
in man oral dosing study of ENT-01, humans have long been exposed to low doses of squalamine 
(milligram to microgram) in the various commercial dogfish shark liver extracts available as 
nutraceuticals (e.g., Squalamax). In addition, following systemic administration squalamine is 
cleared by the liver and excreted as the intact molecule (in mice) into the duodenum through the 
biliary tract. Drug related GI toxicology has not been reported in published clinical trials involving 
systemic administration of squalamine. ENT-01 has limited bioavailability in rats and dogs. Based 
on measurement of portal blood concentrations following oral dosing of radioactive ENT-01 to 
rat’s absorption of ENT-01 from the intestine is low. As a consequence the principal focus of safety 
is on local effects on the gastrointestinal tract. However, ENT-01 appears to be well tolerated in 
both rats and dogs.   


 
 
 
 
 
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Because local gastrointestinal intolerance is the primary adverse effect, safety margins 
based on mg/kg/day of ENT-01 are considered an appropriate safety metric based on allometric 
comparisons of gastrointestinal dimensions between species (Kararli, 1995; Sharma and McNeill, 
2009).  
The proposed starting dose in the Stage 1 segment of the planned trial is 25 mg (0.33 mg/kg 
for a 75 kg subject). The NOAEL of a single oral dose in the rat is ≥ 1000 mg/kg, and in the dog, 
≥200 mg/kg representing a relative safety margin for the proposed starting dose of about 3000x 
based on the rat and about 600x based on the dog. The proposed maximum single dose in Stage 1 
is 200 mg (2.7 mg/kg for a 75 kg subject) is covered by a safety margin of about 300x based on 
the rat, and about 70x based on the dog. 
The maximum proposed dose to be evaluated in Stage 2 of the planned trial is 175 
mg/kg/day (2.3 mg/kg/day for a 75 kg subject), and the total daily dosing exposure will last no 
longer than 25 days. The NOAEL of 25 daily dosing in the rat is ≥ 80 mg/kg/day and in the dog, 
≥ 20 mg/kg/day, representing a relative safety margin for the proposed maximum daily dose of at 
least 35x based on the rat, and at least 9x based on the dog. 
No gastrointestinal lesions, either macroscopic or microscopic, were associated with oral 
dosing of ENT-01 any of the non-clinical studies conducted. Systemic blood levels of ENT-01 will 
be determined in this clinical trial. The chemical structure of squalamine (highly water soluble, 
amphipathic cationic zwitterion of molecular weight 628) predicts poor oral bioavailability. 
Squalamine is well tolerated when administered by continuous intravenous dosing   at 200-300 
mg/m2 over days to months of 5 day continuous infusions.  At these dosing rates steady state 
plasma concentrations range between 5-10 µg/ml. Reversible elevation of hepatic enzymes is the 
major dose limiting toxicity, observed at dose rates above 300 mg/ m2, which is understood since 
squalamine is cleared by the liver.  
The daily dosing range in the planned clinical trial is from 25 mg (14.7 mg/m2) to 200 mg 
(114 mg/m2). Oral dosing of ENT-01, because of its low oral bioavailability, is not anticipated to 
reach significant plasma concentrations in human subjects, based on nonclinical studies. Based on 
an oral bioavailability of 0.1% observed in both dogs and rats and the PK data from prior Phase 1 
studies of IV administration, (where an IV dose of 12 mg/m2 yielded a Cmax of 0.8 µg/ml, and 90 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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mg/m2, a Cmax of 2 µg/ml)   the maximum plasma concentrations in man following a 25mg oral 
dose would be on the order of about 0.5ng/ml, and following a 200mg oral dose, about 2ng/ml.   
 The principal uncertainty related to potential toxicology on oral dosing in humans is the 
local effect of ENT-01 on the GI tract. However, in both rats and dogs, oral dosing was not 
associated with any evidence of local damage to the GI tract, including the gastric mucosa, which 
experienced the highest direct concentration of ENT-01.  
The principal adverse events we anticipate in human subjects will be those relating to the 
gastrointestinal tract, such as diarrhea, cramps, nausea, and vomiting. The trial is also designed to 
establish the pharmacodynamics of ENT-01 on the GI tract. These effects will be determined by 
correction of constipation, and by semi-quantitative Sitzmarks test of colonic motility. 
From preclinical electrophysiological and motility studies conducted in the ex vivo mouse 
jejunum and colon, squalamine concentrations of between 30-100M were required to stimulate 
the ENS and drive peristaltic contractions (Kunze et al, manuscript in preparation).  Based on the 
known fluid volume of the fasting stomach in normal subjects after ingesting an 8 oz. glass of 
water (about 240 ml) (Mudie et al., 2014), a 100 mg dose of Enterin would achieve  a concentration 
of about   500 M  in the stomach and would undergo progressive dilution as the bolus advanced 
rectally.  If we assume that 100mg (about 60 mg/m2) of ENT-01 is a prokinetic dose in Parkinson’s 
disease patients, that the bioavailability of squalamine (the active moiety) is no greater than  1% 
then blood levels would correspond roughly to no greater than that observed after a 0.6 mg/m2 i.v. 
dose (Bhargava et al., 2001; Hao et al., 2003; Herbst et al., 2003), which is at least 400-fold below 
the maximum tolerated  intravenous dose of about 300mg/ m2. 
We will monitor for potential systemic adverse events after oral dosing of ENT-01, with 
our principal focus on the local effects of the compound in the GI tract. 
 
 
2 
OBJECTIVES 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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The purpose of this study is to evaluate the safety, tolerability, pharmacokinetics and 
pharmacodynamics of orally administered ENT-01 at doses ranging from 25mg to 200mg in 
patients with constipation secondary to Parkinson’s disease. Dosing will be individually titrated to 
tolerability and effect.  A second objective of the study is to determine the effect of orally 
administered ENT-01 on gastric emptying and colonic motility. A third objective of the study is 
to collect information relating to sleep and other non-motor symptoms of PD. 
2.1 
Stage 1 
Primary objective:  
The primary objective of the Stage 1 study is to determine the safety and tolerability of a 
single oral dose of ENT-01 in patients with PD and constipation. The dose will be titrated to 
tolerability, starting at 25mg daily and increasing up to 200mg daily.  
10 patients will be enrolled in the study for 7-12 weeks.  
Safety end-point will be: 
Adverse events as measured by patient report, vital signs, clinical chemistry, and EKG 
Tolerability end-points (referred to as the dose-limiting tolerability (DLT)):  
• Recurrent vomiting defined as 3-5 episodes of vomiting within 24 hours of taking drug 
• Recurrent diarrhea defined as 4-6 episodes of diarrhea within 24 hours of taking drug 
• Abdominal pain defined as moderate pain that limits instrumental activities of daily living 
(ADL) (e.g., preparing meals, shopping for groceries or clothes, using the telephone, 
managing money, etc.) within 24 hours of taking drug  
• Postural hypotension defined as dizziness/lightheadedness or fainting on rising from lying 
to sitting or standing that limits instrumental ADL and severe enough to indicate non-
urgent medical intervention within 24 hours of taking drug. 
• A fall in systolic blood pressure to <80mm Hg upon rising from lying to sitting or standing  
• A fall in diastolic blood pressure to <40mm Hg upon rising from lying to sitting or standing  
• Elevation of LFTs > 3 times the upper limit of normal (ULN) 
• A reduction of body weight of 10% or more 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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Secondary objective:  
The secondary objective of the Stage 1 study is to evaluate the Pharmacokinetic (PK) 
characteristics of a single oral dose of ENT-01 in patients with PD and constipation.  
Pharmacokinetics:  
Animal studies have shown < 1% ENT-01 absorption following oral administration. 
Plasma samples will be obtained for pharmacokinetics before dose administration and at 1, 2, 4, 
8, and 24 hours after dose administration on day 1 of each dose level. A PK analysis will be 
conducted if plasma concentrations exceed the lower limit of quantitation of 5 ng/ml using a 
validated analytical method  (Li et al., 2004). The PK analysis of ENT-01 will be conducted by 
using model-independent methods as implemented in WinNonlin™.  The PK variables analyzed 
will include AUC0-24 (ss), Cmax (ss), Cmin (ss), Tmax (ss), CL/F, and z,.  
The PK variables are summarized as follows: 
Cmax (ss) 
Maximum or peak measured plasma concentration  
Cmin (ss) 
Minimum or trough measured plasma concentration  
Tmax (ss) 
Time of maximum or peak measured plasma concentration  
t½z 
Apparent terminal elimination half-life; calculated as ln(2)/ z.  The 
number of data points included in the regression will be determined by 
visual inspection, but a minimum of three data points in the terminal phase, 
excluding Cmax, will be required to estimate z. 
AUC0-24 (ss) 
Area under the concentration versus time curve over the 16-hour dosing 
interval; calculated using linear trapezoid rule 
CL/F 
Clearance defined as: Dose/AUC0-24 (ss). 
 
2.2 
Stage 2 
Primary objectives:  
The primary objective of the Stage 2 study is to determine the safety, tolerability, 
pharmacokinetic and pharmacodynamic characteristics of repeated oral doses of ENT-01 in 
patients with Parkinson’s disease and constipation. 
Animal studies have shown that orally administered ENT-01 has significant pro-kinetic 
effect on ex-vivo colonic preparations and on in-vivo rats and dogs. The effect of orally 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
20 
 
administered ENT-01 on bowel function in patients with PD and constipation will be determined 
clinically using stool frequency, consistency and ease of passage. Constipation will be defined 
according to history and Rome-IV criteria at baseline (See Inclusion Criteria). During the study, 
all participants will maintain a daily diary of stool frequency, consistency and ease of passage. 
Date and dose of study medication and of bowel movement, stool consistency according to the 
modified Bristol scale and ease of passage of each stool on a 7-point adjectival scale as well as use 
of manual aids and suppositories will be recorded.  
Exploratory Objectives:  
The first exploratory objective of the Stage 2 study is to determine the pro-kinetic effect of 
repeated oral doses of ENT-01 on colonic motility as determined by Sitzmarks capsule retention. 
The Sitzmarks test will be performed on days 1-5 of the two-week run-in period and again at the 
end of the randomized treatment period.  
The exploratory objectives will be to collect information on the effect of orally 
administered ENT-01 on sleep, temperature, memory, energy and mood in patients with 
Parkinson’s disease. During the study, all patients will maintain a daily sleep diary. They will 
record bed time, sleep time and morning waking, hours slept each night, number and duration of 
wakes, and leg twitching/thrashing during the night. In addition, we will evaluate sleep indirectly 
using an I-Button which measures skin temperature. Diurnal variations in temperature correlate 
well with the sleep-wake cycle (Sarabia et al., 2008; van Marken Lichtenbelt et al., 2006). The 
patients will be asked to wear the I-Button attached to a wrist-band for five consecutive 24 hour 
periods during the baseline period (Period 3.1), during the stable-dose period (Period 3.2), the 
randomized period (Period 4) and part of the wash-out period. They will remove it only during 
showering. Mood, fatigue, hallucinations, memory and executive function will all be assessed at 
baseline, during the fixed dose period and at the end of the randomization period using the Beck 
Depression Index (BDI), REM behavior disorder questionnaire (RBDQ), Parkinson Fatigue Scale 
(PFS-16), University Miami Parkinson Disease Hallucination Questionnaire (UM-PDHQ), NMS 
questionnaire (NMS), MMSE and Trail-making A and B), respectively.    
Primary Efficacy End-point:  


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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The primary efficacy end-point will be regularization of bowel function as determined by 
stool frequency. This will be defined as an increase in frequency by one or more spontaneous, 
complete bowel movement per week compared to baseline or three or more spontaneous, complete 
bowel movements per week. Assessment will occur at the end of the fixed-dose period (Period 
3.2) and baseline will be obtained at the end of the run-in period (Period 3.1) and all patients (n=40) 
will be compared with their baseline bowel function. During the randomized period (Period 4) the 
treatment group will be compared to the placebo group using the same criteria (n=20 in each 
group). 
Summary of success criteria: (at least one of the following is met) 
• 
Complete spontaneous stool frequency increase by 1 or more over baseline or 
• 
Spontaneous, complete stool frequency of three or more per week 
Exploratory End-points:  
The exploratory end-points include increase in colonic motility during the randomized 
period over baseline values and comparison of motility between treatment and control groups 
during the randomized period. Colonic motility will be measured by the mean colonic transit 
(MCT) time based on a single film estimate (Metcalf et al., 1987). 
Other Exploratory End-points:  
Sleep diaries will provide subjective assessment of sleep parameters. BDI will provide 
subjective assessment of mood. PFS-16, NMSQ and UM-PDHQ will provide subjective 
assessments of fatigue and hallucinations. Non-motor symptoms (NMS) questionnaire will 
provide subjective assessment of overall NMS burden and will be supplemented with the RBD 
and PDSS questionnaires. PAC-SYM and PAC-QOL will provide subjective assessment of bowel 
function and QOL issues. MMSE and Trail-making A and B will provide objective assessment of 
memory and executive function. The I-Button will provide objective assessments of skin 
temperature diurnal variations and will be a surrogate assessment of delays in sleep onset, awake 
periods and Circadian rhythm.  
• 
Sleep diary 
• 
I-Button 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
22 
 
• 
BDI 
• 
MMSE 
• 
Trail-making A and B 
• 
UM-PDHQ 
• 
PFS-16 
• 
NMSQ 
• 
RBDQ 
• 
PDSS 
• 
PAC-QOL 
• 
PAC-SYM 
3 
STUDY DESCRIPTION 
The study will consist of 7 periods. Total duration will be 21-29 weeks (maximum 
treatment period for each individual is 4-8 weeks). 
 
 
 
 
Period 1
Period 2.1 Period 2.2 Period 3.1
Period 3.2
Period 4
Period 5
Run-in 
sentinel 
group 
(Cohort 1)
Single 
dose 
escalation 
sentinel 
group  
Washout 
Run-in 
(Cohort 2)
Identification 
of prokinetic 
dose  using a 
10 subject 
sentinel group 
followed by 
the remaining 
subjects
1:1 
Randomization 
onto placebo 
or prokinetic 
dose
Washout 
2 weeks
3 weeks
2 weeks
2 weeks
3-5 weeks
2 weeks
2 weeks
N=10
N=10
N=10
N=40
N=40
N=40
N=40
STAGE 1
STAGE 2A
≤17days 
6 days 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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3.1 
Periods 
PERIOD 1 (Stage 1):  
Run-in period for Sentinel Group (n=10) 
Duration: 2 weeks 
Conducted at home with 1visit to the clinical trial facility and a telephone call at day 14 
 
Patients who meet the inclusion/exclusion criteria and who sign an informed consent will 
be enrolled in the study. The sequence of patients entering the study will be used to generate the 
patient identifying number for the conduct of the study (e.g., all case report forms, laboratory 
assessments). At the beginning of the run-in period, patients will be asked to stop benzodiazepines 
and other sedatives, hypnotics, opiates, proton pump inhibitors. Patients will remain off these 
medications throughout the study.  They will be allowed to continue antihistamines, anti-
cholinergics, anti-psychotics, dopamine, and anti-depressant medication at stable doses. Use of 
laxatives and suppositories will be documented in the stool diary. Laxatives, softeners, Dulcolax 
suppositories and enemas will be disallowed during the 24-hour period before the start of the 
treatment period.  They will keep daily diaries and record stool frequency, consistency, ease of 
evacuation and use of suppositories, laxatives, softeners, fiber, chia or flax seeds, as well as a sleep 
diary and record various sleep parameters. At the end of the Period, they will be re-evaluated using 
the daily stool chart to assess the severity of constipation. A stool occurring within 24 hours of 
laxative, stool softener, bulking agent or suppository use will not be counted. Only spontaneous 
stools will be counted. A spontaneous stool is a stool occurring in the absence of a laxative, stool 
softener, bulking agent or suppository. 
Exploratory Sleep Assessment:  
Between the 9th and 14th days (range 8-15 days), patients will wear an I-Button 
continuously except during periods of bathing (See Appendix). Measurement of extremity 
temperature has been validated against polysomnography as a surrogate for measuring sleep onset 
and fragmentation. Other baseline testing during this period will include NMSQ, PFS-16, BDI, 
UM-PDHQ, RBDQ, PDSS, MMSE, and Trail-making A and B, PAQ-SYM and PAC-QOL. 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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Symptoms of Parkinson’s disease will be assessed using the Unified Parkinson's Disease Rating 
Scale (UPDRS) during ON states. ON-state will be reached one hour after dopamine intake. 
Visit Day 1: Informed Consent 
The eligible subjects will be asked to sign the informed consent after the study has been 
explained to them and after all their questions have been answered. 
At the beginning of the run-in period, the sentinel group of 10 patients will have screening 
visits. In the screening visit inclusion and exclusion criteria will be accurately reviewed and 
constipation history assessed by detailed history about stool frequency, consistency, ease of 
evacuation, duration of constipation, use of laxatives, stool softeners, bulking agents and also  
using the Rome IV criteria; Patients with a history of constipation of longer than 6 months, with 
two or fewer spontaneous, complete bowel movements a week, unresponsive to milk of magnesia 
and requiring Miralax or another laxative or softener at least once a week will qualify. Patients 
will have a physical examination, their vitals will be checked and their past and present medical 
history as well as their medication intake will be reviewed.  PAC-SYM and PAQ-QOL will be 
administered. Detailed information re smell, depression, anxiety, hallucinations, memory, 
swallowing, dyspepsia and sleep patterns will be collected. BDI, RBDQ, PDSS, MMSE, Trail-
making A and B, NMSQ, UM-PDHQ and PFS-16 will be administered. Baseline blood chemistry 
will be checked. Patients with liver enzymes over 1.5x will be excluded from the study. Women 
with childbearing potential will have a negative pregnancy test to enter in the study and within 48 
hours of each of the studies. They will also be instructed as to the use of daily diaries and I-Buttons 
and told to use them during a five-day period during the second week as described above and 
continue for the remainder of the study. The stool chart will contain daily information relating to 
bowel movement, stool consistency, ease of passage and use of laxatives, softeners, bulking agents 
and suppositories.  
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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Phone call Day 14:  
Assessment of constipation severity off medication  
Participants will receive a phone call and be asked about adverse events. They will also 
provide their daily stool chart to assess the severity of the constipation and use of laxatives, 
softeners, bulking agents and suppositories. Patients with two or fewer complete, spontaneous 
bowel movements will qualify for the treatment period. A bowel movement occurring within 24 
hours of suppository, softener, bulking agent or enema use will NOT be counted. A spontaneous 
stool is a stool occurring in the absence of a laxative, softener, bulking agent or suppository. 
PAC-SYM and PAC-QOL to be administrated on day 14, or at the clinic at Dose 1 visit the 
following day if they fail to bring the completed survey. 
PERIOD 2.1 (Stage 1):  Single dose 
Safety, tolerability and pharmacokinetics of orally administered single-dose ENT-01 in 
Sentinel Group 
Duration: 22-57 days  
The 10 patients in the sentinel group will be assigned to Cohort 1 and will participate in 8 
single dosing periods.  
Subjects 
D1 
D4-8 
D7-15 
D10-22 
D13-29 
D16-36 
D19-43 
D22-50 
D25-57 
1-2 
25mg 
50mg 
75mg 
100mg 
125mg 
150mg 
175mg 
200mg 
 
3-4 
 
25mg 
50mg 
75mg 
100mg 
125mg 
150mg 
175mg 
200mg 
5-6 
 
25mg 
50mg 
75mg 
100mg 
125mg 
150mg 
175mg 
200mg 
7-8 
 
25mg 
50mg 
75mg 
100mg 
125mg 
150mg 
175mg 
200mg 
9-10 
 
25mg 
50mg 
75mg 
100mg 
125mg 
150mg 
175mg 
200mg 
All 10 patients will be pre-selected in advance of check-in for their first scheduled dosing 
period. All laxatives, suppositories, softeners, bulking agents and enemas will be disallowed 
during the 24-hour period before or after the administration of study medication. Patients will wear 
the I-Button continuously for the entire dosing period. The first two patients (subjects 1-2) will 
first be admitted and given a single dose. The study will be conducted at a hospital-based clinical 
unit and patients will be monitored for 24 hours after the dose. Unless there are any safety 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
26 
 
concerns, patients will be released at this time and brought back for the next dosing on Day 4-8 
(D4-8), after a minimum of a 72 hour observation period. Dopamine will be taken prior to the 
clinic visit on an empty stomach. Patients will self-administer the study medication during their 
clinic visit. The dose will be taken on an empty stomach along with 8 oz. of water after baseline 
observations in the clinic. The patient will not be allowed to ingest any food for at least 60 minutes 
after study medication. 
Each dose period will be staggered, so that patients 1-2 will be administered a single dose 
of the drug at the lowest dose of 25mg. Once 24 hours have elapsed, and provided there are no 
safety concerns, they will be sent home and brought back on day 4-8 for the next dose.  During the 
days the patients are home, they will complete the daily diaries and e-mail them to the study 
coordinators. The study coordinators will call the patients every other day starting on the day 
following discharge for each dose and determine with the patient if a suppository or laxative is 
needed, as well as inquiring about any adverse events.  Diary information will also be discussed 
with the patients during the telephone calls. Patients 3-10 will be dosed after the first two patients 
have been observed for 72 hours, i.e. on Day 4. Patients 1-2 will also be brought back on Day 4-8 
and will be given a single dose of 50mg. Once another 24 hours have elapsed and provided there 
are no safety concerns, they will all be sent home and instructed to return on Day 7 for the next 
dosing level. This single dosing regimen will be continued until each patient has been given a 
single dose of 200mg or has reached a dose limiting toxicity (DLT). DLT will be the dose which 
induces repeated vomiting, diarrhea, abdominal pain or symptomatic postural hypotension within 
24 hours of dosing. Maximum tolerated dose (MTD) will be the highest dose for which there 
was no more than 1 DLT endpoint (refer to Section 9.8, DLT endpoint criteria). A review of 
safety, tolerability and adverse events will occur prior to administration of each dose level and at 
the end of Period 2 with DSMB. After the last dose of medication, BDI, MMSE, Trail-making A 
and B, NMSQ, UM-PDHQ, PFS-16, RBDQ, PDSS, PAC-SYM and PAC-QOL will be 
administered. 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
27 
 
Schematic for Stage 1, Period 2.1: Hypothetical dosing regimens 
 
 
 
Pharmacokinetics: Timed blood samples (5.0 mL) will be collected into Li-Heparin 
treated tubes for analysis of ENT-01 plasma concentrations. Plasma samples will be obtained for 
pharmacokinetics before dose administration and at 1, 2, 4, 8, and 24 hours. 
 
Once collected, PK samples will be stored in a -20ºC or -70ºC freezer until time of 
shipment.  Labels and appropriate shipping materials will be provided to sites. The concentration 
of study drug will be determined from the plasma samples using a validated analytical method.  
Details of the method validation and sample analysis will be included with the final clinical study 
report. 
Individual stopping criteria: 
The complete panel of Blood Chemistries described in Appendix 1 are to be drawn 24 
hours after each dose.  Results will be reviewed the evening before the following dose. In addition, 
any patient with at least 1 DLT endpoint (see Section 9.8), or with another non-DLT 
gastrointestinal AE > grade 3 that is at least possibly attributable to ENT-01, or elevated AST, 
ALT, or total bilirubin > 3x ULN or a reduction of body weight of 10% or more from baseline will 
be terminated.  ENT-01 has very little systemic absorption, and hence adverse events are expected 
to be within the gastrointestinal system.    
MTD:  Maximum Tolerated Dose
ID: Intolerant Dose
prokinetic response
MTD
ID
Subject Day 1
Day4
Day7
Day10
Day13
Day16
Day19
Day22
Day 25
1
25mg
50mg
75mg
100mg
125mg
150mg
175mg
2
25mg
50mg
75mg
100mg
125mg
150mg
175mg
200mg
3
25mg
50mg
75mg
100mg
125mg
150mg
175mg
4
25mg
50mg
75mg
100mg
125mg
150mg
5
25mg
50mg
75mg
100mg
125mg
150mg
175mg
200mg
6
25mg
50mg
75mg
100mg
125mg
7
25mg
50mg
75mg
100mg
125mg
150mg
175mg
8
25mg
50mg
75mg
100mg
125mg
150mg
9
25mg
50mg
75mg
100mg
125mg
150mg
175mg
200mg
10
25mg
50mg
75mg
100mg
125mg
150mg
175mg
200mg
STDE 
STDE DOSE in this example is 75 mg, that being the 
lowest dose that is safe and tolerable in all subjects and 
has a prokinetic effect in at least some
No 
response
 Stage 1     
Period 2.1  
(COHORT 1)     
Single dose 
safety, 
tolerability and 
identification of 
prokinetic dose 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
28 
 
Refer to Section 9.12 for Adverse Event Grading and to Section 9.8 for DLT Endpoint 
Criteria. 
Go/No-go decision:  
Safety: We are assuming that no more than 1 patient out of ten (10%) will have an adverse event 
of grade 4 or 5 that is at least possibly related to ENT-01 in dose levels thought to be below 
therapeutic effect. Should there be more than 1 patient with AE grade 4 or 5 that is at least possibly 
related to ENT-01 for doses less than 200 mg, the study will be put on an immediate safety hold 
until an emergency DSMB meeting can be held to evaluate the study for possible discontinuation. 
Dose-finding: Beyond the safety and tolerability, this Period will allow us to determine the dose 
which is safe and tolerable in all 10 patients (STD) and which has a pro-kinetic effect in at least 
some patients (STDE: Safe and Tolerable dose in all and pro-kinetic Effect in at least some). We 
will start Period 3 with the STDE. 
PERIOD 2.2 (Stage 1):  Washout 
Duration: 2 weeks 
Patients will be observed off study drug and adverse events will be recorded. They will 
continue to keep a daily diary throughout the period. They will continue to wear the I-Button for 
five 24 hour periods between Days 1 and 5. During the second week, they will be restarted on all 
original medications but will continue their diaries until the termination of the study. They will be 
seen prior to study termination and BDI, MMSE, Trail making A and B, NMSQ, UM-PDHQ, PFS-
16, RBDQ, PDSS, PAQ-SYM and PAC-QOL will be administered. 
Visit 2: Day 14 
Patients will be reviewed prior to discharge and any adverse events noted. 
DSMB review of Stage 1: The DSMB will determine whether to proceed to Stage 2. 
PERIOD 3.1 (Stage 2a)  
 Run-in period for Cohort 2 (n=40) 
Duration: 2 weeks 
Conducted at home. 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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Forty patients will be screened and included in the study on the assumption that roughly 
15% will drop-out, leaving 34 patients who complete the study. Ten sentinel patients will have 
been pre-selected and ready to go before the study is initiated. Patients who fit the inclusion criteria 
and who agree to sign an informed consent will be included in the study. At the beginning of the 
run-in period, they will be asked to stop benzodiazepines and other sedatives, hypnotics, opiates, 
protein pump inhibitors and all laxatives, bulking agents, softeners and suppositories. They will 
be allowed to continue antihistamines, anti-cholinergics, anti-psychotics, dopamine and anti-
depressant medication at stable dose and to use rescue medication for constipation as needed as 
long as they note it in the diary. Patients will remain off these medications throughout the study. 
Patients who fulfill Rome IV criteria will be included in the run-in period. Patients will be required 
to be constipated for over 6 months, have two or fewer complete, spontaneous bowel movements 
per week, unresponsive to milk of magnesia and requiring Miralax or another laxative, bulking 
agent or softener at least once per week. Rescue medication for constipation will consist of 10mg 
suppository of Dulcolax, taken if no bowel movement has occurred for three consecutive days. 
Rescue medication will be taken on the fourth day in the morning. Dulcolax suppository can be 
repeated up to three times and will be followed by the use of an enema if the Dulcolax was 
ineffective. BDI, PFS-16, NMSQ, UM-PDHQ, RBDQ, PDSS, MMSE and Trail-making A and B, 
PAC-SYM and PAC-QOL will be administered. They will keep daily diaries and record stool 
frequency, consistency, ease of evacuation, completeness and use of suppositories (if it occurs) 
and various sleep parameters. The diaries will be reviewed daily by the study coordinator to ensure 
compliance. Patients with two or fewer complete, SPONTANEOUS bowel movements will have 
the Sitzmarks motility assessment, complete the run-in period and proceed to the treatment 
period. Bowel movements occurring within 24 hours of a suppository, (or accidentally used 
laxative, bulking agent, softener) or enema will not be counted. At the end of the first week, 
assuming they are allowed to proceed to the treatment period, they will also have Sitzmarks 
motility assessment to assess baseline colonic motility. Symptoms of Parkinson’s disease will be 
assessed using the Unified Parkinson's Disease Rating Scale (UPDRS) during ON states. ON-state 
will be reached one hour after dopamine intake.  
 Sleep Assessment: Patients will wear an I-Button continuously from Day 8 of the Run-In 
Period to Day 5 of the Wash-Out period except during periods of bathing.  


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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Visit Day 1: Informed Consent 
The eligible volunteers will be asked to sign the informed consent after explanations of the 
study and after all their questions have been answered. Patients will be registered to the study after 
they have provided informed consent. 
At the beginning of the run-in period, the new cohort of 40 patients will have a screening 
visit. In the screening visit inclusion and exclusion criteria will be accurately reviewed and 
constipation history assessed using the Rome IV criteria. Patients will be included who have been 
constipated for over 6 months, who have two or fewer complete spontaneous bowel movements 
per week unresponsive to milk of magnesia and requiring Miralax or another laxative, bulking 
agent, softener or suppository at least once weekly. Details relating to depression, anxiety, 
memory problems, hallucinations, loss of smell, voice difficulties and sleep patterns, thrashing 
and speaking out loud during sleep, swallowing difficulties and dyspepsia will be recorded. 
Patients will have a physical examination, their vitals will be checked and their past and present 
medical history as well as their medication intake will be reviewed. Baseline blood chemistry will 
be checked. Women with childbearing potential will have a negative pregnancy test to enter in the 
study and within 48 hours of each of the studies. After they have signed the consent form, 
participants will be scheduled for the study-days and entry into the randomization procedure. They 
will also be instructed as to the use of daily diaries and I-Buttons. BDI, PFS-16, NMSQ, PDHQ, 
RBDQ, PDSS, MMSE, UPDRS and Trail-making A and B, PAC-SYM and PAC-QOL will be 
administered. They will be given Sitzmarks capsules to take on Days 8, 9, and 10.  
Day 8, Phone call  Assessment of constipation severity off medication and of colonic transit  
Daily stool diaries which they will have been filling in for a week will be reviewed and those 
patients with two or fewer complete, spontaneous bowel movements per week will proceed to 
Sitzmarks assessment. This will trigger shipment of study drug. 
Sitzmarks procedure Days 8, 9, and 10: 
• The patient will self-administer the Sitzmarks capsule (contains 24 radiopaque markers; 
Konsyl Pharmaceuticals, Texas, USA) by mouth with water on Days 8,9, and 10 and will 
consume their usual diet during this period. The patient will abstain from using suppositories 
or enemas until they return on Day 11. An abdominal XR will be scheduled on Day 11 


 
 
 
 
 
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Day 11 – Site Visit 2 
An abdominal XR will be obtained to determine the location and the extent of elimination 
of the radiopaque markers. Patients who expel at least 80% (19 or more of the markers) have 
grossly normal colonic transit. If remaining markers are scattered about the colon, the condition is 
most likely hypomotility or colonic inertia. If remaining markers are accumulated in the rectum or 
rectosigmoid, the condition is most likely functional outlet delay. Radiographs will be collected in 
electronic format and subsequently analyzed in blinded manner by a centralized reader (Michael 
Camilleri MD, and Duane Burton, MHA, Division of Gastroenterology and Hepatology, Mayo 
Clinic). The patient will be given study medication (B) and a PAC-SYM and PAC-QOL to take 
home. This will trigger randomization to treatment or placebo and to PK or no PK. 
 
Day 14- Phone call  
 
Patients will be asked to complete the PAC+SYM and PAC-QOL questionnaires. They 
will be instructed to take the medication the following morning on an empty stomach and not to 
eat anything for at least 60 minutes thereafter. Baseline complete, spontaneous bowel movements 
(CSBM) will be calculated from the diaries at the 14 day phone call as CSBM/entries x 
7.Baseline PAC-SYM and PAC-QOL and baseline ease of passage and consistency will also be 
obtained at this time-point. 
PERIOD 3.2 (Stage 2): Multiple doses 
Safety, tolerability and pro-kinetic effect of orally administered multiple-dose ENT-01 in 
Sentinel Group followed by Remaining Cohort 
Cohort 2 (40 patients, of which 10 comprise the sentinel group):  
Duration: 17-19 days 
The purpose of this period is to demonstrate the continued pro-kinetic effect beyond that 
of a single-dose, to confirm the effective pro-kinetic dose for each individual and to establish the 
safety, tolerability and PK of multiple doses of ENT-01. A pro-kinetic dose is defined as the dose 
that results in a spontaneous, complete stool on at least two of the three dosing days. Once the 
pro-kinetic dose is established, and assuming there are no safety concerns, each patient will be 


 
 
 
 
 
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kept on that fixed dose for a further 2-4 dosing days (3 days if it falls on a Saturday, 4 days if it 
falls on a Sunday). 
The sentinel group will be a group of 10 patients who did not participate in Stage 1. Patients 
will be given three consecutive doses of the dose which was found to be safe and tolerable in 
everybody (STDE) and effective in at least some patients in Stage 1 (75 mg in schematic) at home. 
Patients will self-administer the study medication daily for 3 doses (3 days at each dosing level). 
The dose will be taken upon awakening on an empty stomach along with 8 oz. of water 
simultaneously to dopamine). The patient will not be allowed to ingest any food for at least 60 
minutes after study medication. The daily diary will be completed and results verbally transmitted 
and emailed to the study coordinator every evening including on week-end days and entered into 
the electronic data capture system the same night or first thing in the morning. The coordinator 
will check in with the patients daily, at the end of the day and determine whether there have been 
any adverse events or CSBM. If the patient has not produced a CSBM within 24 hours of a dose, 
on at least two of the three days at a given dose, then the patient will be told to proceed to the 
higher dose of STDE+25mg (eg.100mg qd) and also to use a suppository followed by an enema if 
necessary on the evening of the third day (See rescue diagram below). At the end of the second 
dosing period (100mg for 3 days), the same criteria will be applied to make a determination as to 
whether to proceed to STDE+50mg (e.g. 125 mg qd) or not. The incremental dosing will continue 
up to the consistently clinically effective (pro-kinetic) dose, or to the DLT, whichever comes first. 
Patients who do not have pro-kinetic effect at DLT will be treated with the highest dose at which 
they did not have a DLT, if it is at least the STDE dose; if the DLT occurred at the STDE dose, 
then the patient will be terminated. Should a pro-kinetic response not occur at 175 mg, patients 
will take 175 mg for the 2-4 additional fixed-dose dosing days and 6 randomized dosing days 
(assuming randomization is to treatment). DLT will be the dose inducing repeated vomiting or 
diarrhea within 24 hours of dosing. The lowest dose producing a CSBM on 2/3 consecutive doses 
will be noted as the pro-kinetic dose (LSTDEi i.e. consistently clinically effective safe and 
tolerable dose for that particular individual). Patients will be kept on the lowest dose producing 
a pro-kinetic effect for another 2-4 days prior to randomization in Period 4. The Enterin CMO will 
review the stool diaries prior to randomization to ensure dosing adequacy. PAC-QOL and PAC-
SYM will be repeated prior to randomization at Visit 3. CSBM during the fixed dose period will 


 
 
 
 
 
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be calculated as CSBM/#fixed dose days x 7. The primary end-point is obtained by comparing 
this number to the baseline CSBM obtained at the end of the run-in period. 
2: SUPPOSITORY/ENEMA RESCUE USE DURING DOSE ESCALATION IN STAGE 
 
Figure 1 
 
 
Patients will wear I-Buttons starting with the lowest dose (e.g. 75 mg) and will remove 
them on day 5 of the wash-out period.  The remaining 30 patients will start on STDE (e.g. 75mg) 
as soon as they have completed the Run-in Period and will follow the same incremental dosing 
regimen until they reach their LSTDEi or maximum tolerated dose. These regimens and 
adjustments are shown in the schematic below.  
Once the LSTDEi has been established, each patient will remain on that dose for two to 
four additional days prior to randomization in Period 4 (5 to 7 consecutive days on prokinetic 
dose) 
N.B. Primary end-point is obtained by comparing stool diary at this point with baseline 
diary on rescue medication at the end of the run-in period. 
Visit 3 for non-PK patients (Day before Randomization).  
Non- PK patients will come to the site for a visit to return any unused study drug (B), receive 6 
daily doses of randomized drug (A or C), Sitzmarks capsules and will complete all the 
questionnaires listed below. They will be sent home with 6 daily doses of A or C and Sitzmarks 
capsules and will return for Visit 4 on Day 6 of randomization to complete all the questionnaires. 
Go/No-go decisions:  
Safety: We are assuming that no more than 1 patients out of forty (2.5%) will have an adverse 
event of grade 4 or 5 that is at least possibly related to ENT-01 in this Stage. Should there be more 
than 1 patient with an AE grade 4 or 5 that is at least possibly related to Enterin-0 in this cohort 
75 
75 
75 
100 
100 
100 
125 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
34 
 
of 40 patients, the study will be put on an immediate safety hold until an emergency DSMB meeting 
can be held to evaluate the study for possible discontinuation. 
Tolerability: We are assuming that all patients will exhibit a pro-kinetic effect at a dose lower 
than DLT and less than or equal to 175mg. Patients reaching DLT before a pro-kinetic effect will 
be terminated if the highest dose for the patient without a DLT was the STDE dose. Otherwise the 
patient will be treated at the highest dose at which they did not experience a DLT if they are 
randomized to the treatment arm (or will receive the equivalent dose of placebo). 
Efficacy: We are assuming that a consistent pro-kinetic effect will be observed in all patients. 
Patients in whom a pro-kinetic effect is not observed at the maximum dose of 175mg will continue 
to take 175mg for another 2 days and enter the randomized period. If they are randomized to 
treatment, they will be given 175mg/day for the 6 randomized dosing days.  
Individual stopping criteria: 
Patients who reach DLT before a pro-kinetic effect, exhibit LFT elevation above 3xULN, 
have a 10% or greater reduction in body weight or those with a non-DLT gastrointestinal AE > 
grade 3 within 24 hours of taking ENT-01 that is at least possibly attributable to ENT-01 will be 
terminated.  
Pharmacodynamics: Patient will record bowel movements, ease of passage, consistency and 
suppository usage in the daily diary throughout the Period and transmit the information to the 
coordinator every evening. 
Sleep assessment: Patients will wear the I-Button throughout the dosing period and remove it  
only during bathing periods. 


 
 
 
 
 
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Schematic of Stage 2, Period 3.2 with hypothetical dosing regimen 
 
DSMB: The DSMB will meet at the end of the fixed dose period for the ten sentinel patients.  
PERIOD 4  
(Stage 2): Randomized, double-blinded treatment period to determine the effect of orally 
administered ENT-01 on pharmacodynamics and motility. 
Duration:  6 days 
Visit 3 for PK patients (Day 1 of randomization): 
PK subjects will come to the site on Day 1 of randomization ON AN EMPTY STOMACH 
and return their unused study drug at that time point. They will receive the first dose of medication 
at the site following the first PK blood draw, and complete the questionnaires listed below. PKs 
will also be performed at 1, 2, 4, and 8 hours post-dose.  A sample for liver function tests will be 
drawn with the 8-hour post-dose PK draw. They will receive 4 daily doses to take home, then 
return on Day 6 to receive their 6th dose at the site after their first PK draw of the day. PKs will 
again be obtained post-dose at 1, 2, 4 and 8 hours, a sample for liver function tests will be drawn 
with the 8-hour post-dose PK draw, and all the questionnaires completed.  
Following the 5-7 day course on the pro-kinetic dose, ALL patients will be randomized to 
drug or placebo and enter Period 4. Randomization will be 1:1 to treatment vs. placebo. 
Randomization will occur in accordance with the randomization schedule generated by Dr. Karla 
Ballman at Weill-Cornell Medical College. Patients and study staff will be blinded to actual study 
Subject Day1
Day2
Day3
Day4
Day5
Day6
Day7
Day8
Day9
Day10 Day11 Day 12 Day13 Day14 Day15 Day16 Day17 Day18 Day19 Day20 Day21 Day22
1
75 mg 75 mg 75 mg 100mg 100mg 100mg 100mg 100mg 100mg 100mg
2
75mg
75mg
75mg
100mg 100mg 100mg 100mg 100mg 100mg 100mg
3
75mg
75mg
75mg
100mg 100mg 100mg 125mg 125mg 125mg 125mg 125mg 125mg 125mg
4
75mg
75mg
75mg
100mg 100mg 100mg 125mg 125mg 125mg 150mg 150mg 150mg 150mg 150mg 150mg 150mg
5
75mg
75mg
75mg
100mg 100mg 100mg 100mg 100mg 100mg 100mg
6
75mg
75mg
75mg
100mg 100mg 100mg 125mg 125mg 125mg 150 mg 150mg 150mg 150mg 150mg 150mg 150mg
7
75mg
75mg
75mg
75mg
75mg
75mg
75mg
8
75 mg 75 mg 75 mg 100mg 100mg 100mg 100mg 100mg 100mg 100mg
9
75mg
75mg
75mg
100mg 100mg 100mg 125mg 125mg 125mg 150mg 150mg 150mg 150mg 150mg 150mg 150mg
10
75mg
75mg
75mg
100mg 100mg 100mg 125mg 125mg 125mg 150mg 150mg 150mg 175mg 175mg 175mg 175mg 175mg 175mg 175mg
COHORT 2 
GROUP, N=32
RUN-IN PERIOD FOR SUBJECTS 11-40
Day7
Day21
Day 23 Day24 Day25 Day26 Day27 Day28 Day29 Day30 Day31 Day32 Day 33 Day34 Day35 Day36 Day37 Day38 Day39 Day40 Day41 Day42 Day43 Day44
COHORT 2 
GROUP, N=32 11-40
75mg
75mg
75 mg 100mg 100mg 100mg
LSTDEi LSTDEi LSTDEi LSTDEi
STDE DOSE in this example is 75 mg, that being the 
lowest dose that is safe and tolerable in all subjects and 
has a prokinetic effect in at least some
LSTDEi is the lowest safe and tolerable dose that produces a 
prokinetic response reached by an individual  in at least 2 of 3 
days at that dose
LSTDEi
Stage 2,  
PERIOD 3.2, 
(COHORT 1)  
Safety and 
tolerability of 
3 day dosing, 
and extension 
to a total of 5 
doses 
 STDE 
DOSE
COHORT 1, 
SENTINEL 
GROUP  N=10


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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treatment and will remain blinded throughout the remainder of the study. BDI, MMSE, Trail 
making A and B, NMSQ, UM-PDHQ, PFS-16, RBDQ, PDSS, PAC-SYM and PAC-QOL will be 
administered prior to randomization.  The UPDRS will be administered and weight and vital signs 
will be taken). 
Assuming randomization is to treatment group, the patient will remain on that dose daily 
for a 6 day period (6 dosing days). Patients randomized to placebo will be switched to the same 
number of placebo tablets as they were taking prior to switching. All patients will continue to keep 
daily diaries during this period. The use of rescue medication (suppositories and enemas) for 
constipation will be permitted as needed if a bowel movement has not occurred for 3 days and 
recorded in the diary. Patients will be contacted daily. The suppository will be taken on the evening 
of the third day as shown in Figure 1 above. Any bowel movement produced as a result of rescue 
medication will not be counted.  
Patients reaching their tolerability limit at a dose lower than a clinically effective dose 
(STDE) will be terminated from the study. We have included 40 patients in this Stage to allow for 
15% drop-outs for reasons including tolerability. 
Patients will wear the I-Button continuously throughout the randomized period except 
during bathing periods. Stool and sleep diaries will be completed daily throughout the period. BDI, 
MMSE, Trail making A and B, NMSQ, UM-PDHQ, PFS-16, RBDQ, PDSS, PAC-SYM and PAC-
QOL will be administered on the first (Visit 3) and last day (Visit 4). 
 
Pharmacodynamics WSW:  
On Day 6 of this randomized period, patients will have a second set of motility tests 
(Sitzmarks capsules) to determine whether there has been any improvement in colonic motility. 
Patients will be compared against their own baseline motility and the placebo group will also be 
compared to the treatment group. The protocol will be identical to the baseline motility assessment. 
Patients will self-administer one capsule per day for three days beginning on Day 3 of the 
randomization period with subsequent capsules taken on Day 4 and Day 5. An abdominal XR will 
be scheduled on Day 6. Motility at the end of the randomized period will be compared with 


 
 
 
 
 
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baseline for all 40 patients and motility will also be compared between treatment and placebo 
groups (20 patients per group). 
Safety and tolerability will be reassessed throughout the randomized treatment period. The 
study coordinator will call each patient at the end of each day and patients will be told to call the 
coordinator whenever they need to.  
 
Summary of Stage 2, Period 4 
 
Pharmacokinetics and LFTs: On the randomization Day 1, 7 patients from the treatment group 
and 3 patients from the control group will undergo blood draws for PK analysis at pre-dose, 1, 2, 
4 and 8 hours after a dose and LFTs will be obtained on the last blood draw.  These same 10 
patients will undergo repeat blood draws for PK analysis and LFTs during their visit to the clinical 
trial unit on randomization Day 6 for the abdominal x-ray and repeated patient 
questionnaires/surveys. Study medication will be administered by study staff on randomization 
Day 1 and again on Day 6 after the first blood draw. 
PERIOD 5 (Stage 2): Wash-out period 
Duration: 2 weeks 
Patients will be taken off study drug. They will continue to keep a daily diary throughout the 
period. They will be contacted by phone daily during Week One to check diaries and laxative use. 
During the first week of the wash-out period they will only use rescue medication as needed. 
During the second week, they will be restarted on all original medications including laxatives but 
will continue their diaries until the termination of the study. They will continue to wear the I-
Button for the first 5 days of this period. During the last visit, the UPDRS, BDI, Trail-Making A 
and B, MMSE, PFS-16, NMSQ, UM-PDHQ, RBDQ, PDSS, PAC-SYM and PAC-QOL will be 
administered 
6 daily doses
LSTDEi
Randomized 
1:1
Stage 2  
PERIOD 4 
Double 
blinded, 
randomized
All cohorts, 
N=40
After 4th  
dose at the 
LSTDEi
PLACEBO


 
 
 
 
 
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4 
STUDY POPULATION 
Patients will be selected by neurologists specializing in movement disorders from among 
PD patients in tertiary referral centers. Patients will have PD and constipation as defined by the 
inclusion criteria. PD will be diagnosed according to the UK Parkinson’s Disease Society Brain 
Bank criteria. 
4.1 
Number of Patients  
Roughly 50 patients will be included. Ten patients will be included in Stage 1 and 40 
patients in Stage 2. Assuming a 15% drop-out rate, 8-9/10 patients will complete Stage 1 and 34/40 
patients will complete Stage 2. 
4.2 
Selection Criteria 
Patients will be selected by individual site PIs from among PD patients seen at the tertiary 
referral center. Patients will be screened for the presence of non-motor symptoms of PD, 
specifically constipation and REM behavior disorder (RBD). The population will be enriched 
with respect to NMS, specifically sleep delay and sleep fragmentation problems, and RBD. The 
presence of constipation at baseline will be established according to the Rome IV criteria and other 
inclusion criteria below. Onset of constipation before or after a diagnosis of PD will specifically 
be noted and its duration and severity established. The daily stool diary will be analyzed at the end 
of the two week run-in period to establish the presence of severe constipation on rescue medication 
alone (does not apply to Stage 1). Patients with two or fewer complete, spontaneous bowel 
movements per week will be entered into the treatment phase. RBD will be determined by history 
and questionnaire. The use of anti-depressants, anti-cholinergics and anti-psychotics will be 
permitted as long as patients are on a stable dose of medication. 
4.3 
Inclusion Criteria  
• Patients aged 30-86 year old, both genders 
• Subjects must be diagnosed with Parkinson's disease (PD), according to the UK 
Parkinson’s Disease Society Brain Bank criteria, by a neurologist specializing in 
movement disorders. 


 
 
 
 
 
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• There are insufficient criteria for IBS 
• Constipation which has been present for over 6 months and is unresponsive to first line, 
typically over the counter treatments such as milk of magnesia (1 gram), Miralax (17g 
in 8 ounces of water) or the equivalent at least once weekly with an inconsistent 
response over a 6 week period. 
• Body mass index of 18-40 kg/m2 
• Subjects must fulfill Rome IV criteria for functional constipation which includes 2 or 
more of the following:  
o Straining during at least 25% of defecations 
o Lumpy or hard stools in at least 25% of defecations 
o Sensation of incomplete evacuation for at least 25% of defecations 
o Sensation of anorectal obstruction/blockage for at least 25% of defecations 
o Manual maneuvers to facilitate at least 25% of defecations (e.g., digital evacuation, 
support of the pelvic floor) 
• Fewer than 3 spontaneous, complete defecations a week. 
• Loose stools are rarely present without the use of laxatives 
• Patients must provide written informed consent and be willing and able to comply with 
study procedures. 
• Patients must be able to read, understand, and accurately record data into the diary to 
guarantee full participation in the study. 
• Female patients must have negative serum or urine pregnancy tests and must not be 
lactating. For females able to bear children, a hormonal (i.e., oral, implantable, or 
injectable) and single-barrier method, or a double-barrier method of birth control must 
be used throughout the study. A vasectomized partner will be allowed as one in 
conjunction with another single-barrier method. 
• Female patients unable to bear children must have this documented in the case report 
form (i.e., tubal ligation, hysterectomy, or postmenopausal [defined as a minimum of 


 
 
 
 
 
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one year since the last menstrual period]). Post-menopausal status will be confirmed 
by follicle stimulating hormone (FSH) in women less than 60 years of age 
4.4 
Exclusion Criteria 
• Unable or unwilling to provide informed consent or to comply with study procedures. 
• Diagnosis of secondary constipation beyond that of PD 
• Structural or metabolic diseases that affect the GI system 
• Functional GI disorder 
• Unable to withdraw the following medications 2 weeks prior to the dose-escalation 
period and throughout the study: Laxatives, opiates, sedatives, hypnotics, protein pump 
inhibitors or any medications which may cause constipation 
• History of recent major surgery (within 60 days of screening) 
• Any clinically significant abnormalities on screening laboratories or physical 
examination requiring further evaluation or treatment. 
• Neurological disorder other than PD 
• On treatment with intra-jejunal dopamine or carbidopa/levodopa (i.e. Duopa) 
• Treatment with COMT inhibitors (entacapone, tolcapone, Stalevo) 
• Patients starting a new PD medication or modifying an existing medication within 4 
weeks prior to enrollment 
• Unable to maintain a stable diet regimen 
• Patients with a cognitive impairment that preclude them from understanding the 
informed consent 
• Patients placed under legal guardianship 
• Acute GI illness within 2 weeks of the baseline period 
• History of major GI surgery (e.g. previous abdominal surgery, including 
Cholecystectomy), except that patients with uncomplicated appendectomy are allowed. 
• ALT or AST > 1.5 X upper limit of normal (ULN) during screening 


 
 
 
 
 
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• Females who are pregnant or breastfeeding 
• History of excessive alcohol use or substance abuse 
• Patient or caregiver unable to administer daily oral dosing. 
• Participation in an investigational clinical study within the 6 months prior to dosing in 
the present study. 
• Any other reason, which in the opinion of the Investigator would confound proper 
interpretation of the study. 
4.5 
Discontinuation Criteria 
Patients may withdraw voluntarily from participation in the study at any time and for any 
reason. They may want to withdraw because of increased constipation off laxatives, nausea, 
vomiting or diarrhea.  
Patients may be withdrawn on the basis of the medical monitor safety and tolerability 
review or the Investigator’s clinical judgment, protocol deviation, change in dose of medications 
unrelated to study medication or loss to follow-up. 
This study may be terminated at the discretion of the Enterin, Inc. or of any regulatory 
agency for reasons including safety and/or efficacy.  
An Investigator may elect to discontinue or stop the study at his or her site for any reason 
including safety. 
When a patient withdraws or is withdrawn before completing the study, the date and reason 
for withdrawal are to be documented in the eCRF.  The study site must immediately notify the 
medical monitor.  Patients who withdraw prematurely are to attend an early discontinuation visit 
if possible, at which time they will complete all assessments.  
In the event that a patient is withdrawn prematurely due to an AE or serious AE, the AE or 
serious AE will be followed until it resolves or stabilizes, or until it is judged by the Investigator 
to be no longer clinically significant. 


 
 
 
 
 
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4.6 
Concomitant Medication 
• Preventive laxatives will be discontinued during the run-in period. Throughout the study, 
patients will be allowed to take constipation rescue medication on demand, and only if 
they have not had a bowel movement in three consecutive days.  
• In stage 1, rescue medication (laxative, bulking agent, softener, suppository) can be taken 
as needed and as stated above, and noted in the stool chart. Rescue medication cannot be 
taken during the 24 hrs before or after study medication. Any bowel movement produced 
within 24 hours of rescue medication will not be counted.  
In Stage 2, rescue medication (Dulcolax suppository and/or enema) can be taken on demand 
during the run-in period as stated above and noted in the stool diary. During the dosing period, 
rescue medication is taken on the evening of the third dose at any given level if a stool has not 
been produced in response to three days of study medication. Enterin CMO should be consulted 
for any questions related to rescue medication. Rescue medications will be recorded in the daily 
diary. Bowel movement produced within 24 hours of rescue medication will not be counted as 
either spontaneous bowel movement or complete spontaneous bowel movement. 
 
• In both stages, protein pump inhibitors will be discontinued during the run-in period and 
reinstated during the wash-out period.  
•  Benzodiazepines, hypnotics, other sedatives, barbiturates, and opiates will not be allowed. 
All medications shall be reviewed and dis/approved by the investigator on a case-by-case 
basis. 
• Selective serotonin reuptake inhibitor (SSRI), SNRI, tricyclic antidepressants anti-
cholinergics, anti-histamines and anti-psychotics are permissible at stable doses.  
• Dopamine agonists and amantadine will be allowed if on a stable dose for at least 4 weeks 
preceding the study. 
• Deep brain stimulation is allowed at the PI’s discretion. 


 
 
 
 
 
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5 
STRATIFICATION AND RANDOMIZATION (STAGE 2) 
 
Participants will be randomly assigned to study drug or placebo in block sizes of 4 
according to a schedule provided by the study statistician. Patients will be stratified according to 
constipation severity. We will compare results in patients with average complete spontaneous 
stool frequency of 0-1 per week with those of patients with an average frequency of 1-2 per week. 
Patients will be blinded throughout the study until all data are analyzed and locked. 
6 
UNBLINDING 
 
Patients will be unblinded under the following circumstances: 
• Patient is terminated from the study 
• Patient drops-out of the study 
• A SAE occurs 
7 
MATERIALS 
7.1 
Study Drug 
 
All study medications will be supplied by Enterin Inc. Ent-01 will be supplied as 25mg tablets 
packaged in white opaque high density polyethylene (HDPE) bottles with child-resistant closures. 
The placebo tablets will be identical in configuration and packaged similarly. 
7.2 
Packaging and Labeling 
 
 
 
The study medication will come packaged in a white bottle containing 10 tablets. Tablets can 
be divided between patients as appropriate. To ensure blinding the packages will be identified as A, 
B, or C, and each will contain 10 tablets, all equal in appearance.  
7.3 
Storage, Dispensing and Reconciliation of Study Drug 
 
All study medication must be stored in a locked, limited access area.  Both the used and unused 
study medication should be stored at 2 - 8°C (35.6 - 46.4°F). 
Identity of Investigational Products 
Product Name 
ENT-01 
Placebo 


 
 
 
 
 
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Dosage form 
Tablets (mg) 
Tablet 
Dosage levels(mg) 
25 
Identical, but absent active 
Route/dosage 
Oral. Doses above 25 mg will 
require multiple tablets:    
25 mg=1 tablet 
50 mg=2 tablets 
75 mg=3 tablets 
100 mg=4 tablets 
125 mg=5 tablets 
150mg=6 tablets 
175mg=7 tablets 
Oral 
Dosing Instructions 
Take 60 min before breakfast 
with 8 oz. water 
Take 60 min before breakfast 
with 8 oz. water 
8 
WARNINGS AND PRECAUTIONS 
Patients will be advised to stay at home for 4-6 hours after taking the first dose of 
medication in case they need to evacuate urgently. Beyond the first dose, patients themselves will 
be in a position to know if and when to leave the house.  
9 
STUDY PROCEDURES 
9.1 
Sitzmarks assessment of colonic motility in Stage 2 
 
On Day 8 of the run-in period and again on Day 3 of the randomized period, patients will 
be reminded to self-administer a Sitzmarks capsule (contains 24 radiopaque markers; Konsyl 
Pharmaceuticals, Texas, USA). The patient will be instructed to abstain from using suppositories 
or enemas if possible until they return on Day 11 (or day 6 of randomized period). If they have to 
use suppositories on day 11 (or Day 6 of randomized period), make sure it’s recorded in the daily 
diaries. An abdominal XR will be scheduled for Day 11 of run-in and Day 6 of randomized period. 
On Day 11 of the run-in Period and again on Day 6 of the randomized period, an abdominal 
XR will be performed to determine the location and the extent of elimination of the radioisotope 
markers. Patients who expel at least 80% (19 or more of the markers) have grossly normal colonic 
transit. If remaining markers are scattered about the colon, the condition is most likely 
hypomotility or colonic inertia. If remaining markers are accumulated in the rectum or 


 
 
 
 
 
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rectosigmoid, the condition is most likely functional outlet delay. For patients whose markers 
accumulate in the rectosigmoid or when markers are retained diffusely, instruct patient to 
maximize liquid intake for the next two weeks.  
9.2 
Observations and Measurements 
All the questionnaires will be filled in according to the schedule. In Trail-Making B, the 
Trail will be completed twice, and both results will be reported. If the patient fails to complete the 
Trail, it will be reported as failed to complete. 
Stool diaries and sleep diaries: Patients will be instructed how to fill the diaries daily 
throughout the study. They will be told to mark use of suppositories or enemas. 
           Vital signs: Per study procedures listings and for PK subjects Randomization Day 1 and 
Day 6 before dose and 2 hours after dose. 
An I-Button which measures skin temperature and attached to a wrist band will be worn 
for 5 consecutive 24 hr periods from day 8-13 during Period 1, throughout the dosing period and 
randomized period and for the first five days of the wash-out period, and will not be removed 
except for brief periods during a bath/shower. Patients will be shown how to wear the I-Buttons 
with the button facing the skin and overlying the radial artery.  
9.3 
Informed Consent  
The Principal Investigator(s) at each center will ensure that the patient is given full and 
adequate oral and written information about the nature, purpose, possible risk and benefit of the 
study.  Patients must also be notified that they are free to discontinue from the study at any time.  
The patient should be given the opportunity to ask questions and allowed time to consider the 
information provided. 
9.4 
Instructions to Patients 
Prior to entry into the study, patients and their spouse/companion will be taught how to fill 
in the daily diaries for stool and sleep. They will be instructed about the use of rescue medication 
and told to record it in the diary. They will also be shown how to wear the   I-Button and how to 
use the actigraphy device (See Appendix). 


 
 
 
 
 
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9.5 
Clinical Laboratory Procedures 
The following tests will be obtained at baseline and at the end of each Period. 
• 
CBC 
• 
Chemistry 
• 
PT/PTT 
• 
Pregnancy test 
• 
Urinalysis 
PK Samples for 10 selected subjects: 
Timed blood samples (5.0 mL) will be collected into potassium-EDTA treated tubes for 
analysis of ENT-01 and its metabolite plasma concentrations.  
Blood samples for PK analysis will be collected at the following times before and after 
dosing on the day before randomization and on Day 6 of the randomized period:  0 (pre-dose), 60 
min, 2, 4, 8 hrs. 
Once collected, PK samples will be stored in a -20ºC or -70ºC freezer until time of 
shipment.  Labels and appropriate shipping materials will be provided to sites.   
The concentration of study drug will be determined from the plasma samples using a 
validated analytical method.  Details of the method validation and sample analysis will be included 
with the final clinical study report. 
9.6 
Other Procedures 
 
Patients will fill in the following daily diary for stools (example inserted): 
Date Dose mg 
BM 
Yes/No 
 
Consistency 
Modified Bristol 
Scale 1-7 
1.Hard lumps 
2.Lumpy sausage 
3.Cracked 
sausage 
Ease of passage 
scale 1-7 
1.Manual 
disimpaction 
2.Enema needed 
3.Straining 
needed 
4.Normal 
Complete 
evacuation 
Yes/No 
 
Suppository 
or laxative  
Yes/No/list 
med 


 
 
 
 
 
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4.Smooth 
sausage 
5.Soft lumps 
6.Mushy 
7.Watery 
5.Urgent without 
pain 
6.Urgent with pain 
7.Incontinent 
3/20 
- 
Yes 
1 
2 
No 
Yes/suppos 
3/21 
100 
Yes 
3 
4 
No 
No 
3/22 
100 
Yes 
3 
4 
Yes 
No 
3/23 
100 
Yes 
4 
4 
Yes 
No 
3/24 
100 
Yes 
4 
4 
Yes 
No 
3/25 
100 
Yes 
3 
4 
Yes 
No 
9.7 
CTCAE Definitions of Dose Limiting Adverse Events  
Adverse 
event 
Grade 1 
Grade 2 
Grade 3 
Grade 4 
Grade 5 
Diarrhea  
(watery 
stool) 
Increase of <4 
stools a day 
over baseline 
Increase of 4-6 
stools a day 
over baseline 
Increase of >7 
stools a day; 
incontinence; 
hospitalization 
indicated 
Life 
threatening 
complications; 
urgent 
intervention 
indicated 
Death 
Vomiting 
1-2 episodes 
per 24 hrs 
3-5 episodes per 
24 hrs 
6 or more 
episodes in 24 
hrs; tube 
feeding, TPN or 
hospitalization 
indicated 
Life-
threatening 
consequences; 
urgent 
intervention 
indicated 
Death 
Abdominal 
pain 
Mild pain 
 
Moderate pain; 
limiting 
instrumental1 
ADL 
 
Severe pain; 
limiting self-
care ADL2 
 
- 
- 
Postural 
hypotension3 
Mild, transient 
lightheadedness 
or dizziness 
Moderate 
lightheadedness, 
dizziness or 
Severe 
lightheadedness, 
dizziness or 
Life-
threatening 
and urgent 
Death 


 
 
 
 
 
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upon rising 
from lying or 
sitting position 
that does not 
interfere with 
ADL 
fainting upon 
rising from 
lying or sitting 
position that 
limits 
instrumental 
ADL; Non-
urgent medical 
intervention 
indicated 
fainting upon 
rising from 
lying or sitting 
position that 
limits self-care 
ADL; Medical 
intervention or 
hospitalization 
indicated 
intervention 
indicated 
1Preparing meals, shopping for groceries or clothes, using the telephone, managing money, etc. 
2Bathing, dressing and undressing, feeding self, using the toilet, taking medications, and not bedridden 
3Definition adapted from combining CTCAE definitions for Hypotension and Dizziness 
9.8 
Criteria for DLT Endpoints: 
Recurrent vomiting: 3-5 episodes of vomiting within 24 hours of taking ENT-01 (Grade 2 AE) 
Recurrent diarrhea: 4-6 episodes of diarrhea within 24 hours of taking ENT-01 (Grade 2 AE) 
Abdominal pain: Moderate pain that limits instrumental activities of daily living within 24 hours 
of taking ENT-01 (Grade 2 AE) 
Postural hypotension: Moderate dizziness, lightheadedness or fainting upon rising from lying to 
sitting or standing and severe enough to require medical intervention within 24 hours of taking 
ENT-01 (Grade 2 AE) or either a systolic blood pressure less than 80mmHg or diastolic blood 
pressure less than 40mm Hg.  
9.9 
Pre-Existing Medical Conditions 
All subjects enrolled in the study will have Parkinson’s disease. Specific pre-existing 
medical conditions that will be excluded are listed in patient exclusion criteria. 
9.10 
Treatment Emergent Adverse events  
Treatment-emergent adverse event (TEAE) is defined as any event not present prior to the 
initiation of the treatments or any event already present that worsens in either intensity or 
frequency following exposure to the treatments. 
AE is typically collected after signing the informed consent form and could be related or 
unrelated to the study drug. TEAE is for after the subject actually taking the study drug. 


 
 
 
 
 
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AEs may be called “baseline-emergent adverse event” which is defined as any event which occurs 
or worsens during the staged screening process (after informed consent) including the 
randomization visit. We will include separate summaries for AEs that occur prior to the initiation 
of the treatment and AEs that occur after the initiation of the treatment (i.e., summary of treatment 
emergent adverse events). 
9.11 
Laboratory Abnormalities 
 
Clinical labs will be performed by a local laboratory.  Labs to be drawn during the study 
include: serum chemistries, a hematology panel and urinalysis.  A serum pregnancy test must be 
performed and the result must be negative prior to the entry of women of child bearing potential.  The 
investigator must obtain verification that the local laboratory meets the standards for quality and 
consistency set by the College of American Pathologists. 
 
Clinical laboratory reports must be reviewed by a physician for out-of-range values within 24 
hours of receipt.   Out-of-range values will be evaluated using the following notations: 
 
• NS 
Not clinically significant 
 
• LE 
Laboratory Error 
 
• PT 
Patient abnormal; relates to the patient's usual state of health 
 
• SG 
Significant, other.  This value cannot be explained by any of the other flags. 
 
By definition a lab value flagged as "SG" must be entered on the adverse clinical laboratory 
event page in the case report form.  A laboratory test flagged "SG" must be repeated as soon as 
possible.  The investigator may use his/her own judgment as to whether the abnormal finding is 
sufficient reason to immediately withdraw the patient from the study. 
 
If a laboratory value is considered to be serious and life-threatening the patient should be 
immediately discontinued from the study and appropriate therapy started.  Refer to sections 9.7-9.12 
for definition of a serious adverse event and related terms, and to sections 9.13-9.16 for details on 
reporting a serious adverse event. 
9.12 
AE Assessment and Recording 
All adverse events, exacerbations of concomitant illnesses, or events known to be related 
to underlying disease processes or concomitant medications are to be recorded on the case report 


 
 
 
 
 
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form throughout the study.  If a pre-existing condition worsens on study, the date on which the 
exacerbation began should be recorded.  Onset dates for study treatment-related adverse events 
must be on or after the date of initial study treatment use.  The need to record an adverse event on 
the case report form is not dependent on whether the adverse event is associated with the use of 
the study medication.  In order to avoid vague, ambiguous or colloquial expressions, the adverse 
event should be recorded in standard medical terminology. 
Adverse event recording will include the date of onset, severity, duration, whether or not 
the study medication was discontinued or its dosage changed because of the event, the treatment 
given and the outcome.  The investigator must also assess whether the event was related to the 
study medication, concurrent drug therapy, underlying disease, a combination of these factors, or 
if it is unknown.  Patients with an adverse event should be carefully followed to determine 
outcome.   
The investigator will use the NCI definitions to grade the severity of the event. 
Grade 1: Mild; asymptomatic or mild symptoms; clinical or diagnostic observations 
only; intervention not indicated 
Grade 2: Moderate; minimal, local, or non-invasive intervention indicated; limiting age-
appropriate ADL 
Grade 3: Severe or medically significant but not immediately life-threatening; 
hospitalization or prolongation of hospitalization indicated; disabling; limiting 
self-care ADL. 
Grade 4: Life-threatening consequences; urgent intervention indicated 
Grade 5: Death 
The relationship or association of the study medication in causing or contributing to the 
adverse event will be characterized as remote, possible or probable as defined below: 
Not related: Evidence indicates no plausible direct relationship to the study medication 


 
 
 
 
 
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Remote: Suggests other conditions are reasonably likely to account for the event including 
concurrent illness, progression or expression of the disease state, or reaction to concurrent 
medication 
Possible: Suggests that the association of the event with the study medication is unknown; 
however, the adverse event is not reasonably supported by other conditions 
Probable: Suggests that a reasonable temporal sequence of the event with medication 
administration exists and, based upon the investigator's clinical experience, the association 
of the event with study medication seems likely 
Definite: Suggests that based upon the investigator’s experience, the association of the 
event with the study medication seems very certain.  
Procedures such as surgery should not be recorded as adverse events.  However, the 
medical condition for which the procedure was performed should be reported if it meets the 
definition of adverse event. 
9.13 
Reporting Requirements 
 
Any adverse event, including both observed or volunteered problems, complaints, or 
symptoms that begins any time between the start of the first dose and within 30 days after the end of 
the last dose are to be recorded briefly on the appropriate case report form and in detail in the source 
documents.   A check list of adverse events may not be used during this study. 
 
The following are specific definitions that are relevant to meeting your reporting obligations 
and which are included in the FDA Regulations, 21CFR Part 312.32, and International Conference 
on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use 
(ICH) Guidelines: 
Adverse Event:  
Any untoward medical occurrence in a patient administered a 
pharmaceutical product and which does not necessarily have to have a causal relationship with 
this treatment.  An adverse event can be any unfavorable sign (including an abnormal 
laboratory finding), symptom, or disease temporally associated with the use of the 
investigational drug, whether or not considered related to the investigational drug. 
Serious Adverse Event: 
An untoward event or reaction that at any dose: 


 
 
 
 
 
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• 
results in death 
• 
is life-threatening 
• 
requires inpatient hospitalization 
• 
prolongs existing hospitalization 
• 
results in permanent or significant disability or incapacity 
• 
is a congenital anomaly/birth defect 
• 
requires intervention to prevent permanent impairment/damage 
Life-threatening: 
An event which a patient was at risk of death at the time of event. 
 
There is a distinction between the severity and the seriousness of an adverse event.  Severe is 
a measurement of intensity, thus a severe reaction is not necessarily a serious adverse event.  For 
example, a headache may be severe in intensity, but would not be serious unless it met one of the 
criteria for serious adverse events listed previously. 
9.14 
Serious Adverse Events 
Adverse events (AEs) and serious adverse events (SAEs) will be collected from the start 
of study treatment until the follow-up contact. Medical occurrences that began prior to the start of 
study treatment, but after obtaining informed consent were recorded on the Medical 
History/Current Medical Conditions CRF. The investigator or site staff will be responsible for the 
detection and documentation of events meeting the criteria and definition of an AE or SAE, as 
provided in the study protocol, 
However, any SAEs assessed as related to study participation (e.g., dosing, protocol 
mandated procedures, invasive tests, or change in existing therapy) or related to a concomitant 
medication will be recorded from the time a subject consents to participate in the study up to and 
including any follow-up contact. 
In the event of an AE or SAE, it will be the responsibility of the Investigator to review all 
documentation (e.g., hospital progress notes, laboratory, and diagnostics reports) relative to the 
event and attempt to establish a diagnosis of the event based on signs, symptoms, and other clinical 
information. The diagnosis was to be documented as the AE/SAE and not the individual’s 
signs/symptoms. Once an Investigator becomes aware that an SAE has occurred in a study subject, 
they are' to report the information to Enterin within 24hrs and provide an assessment of causality. 


 
 
 
 
 
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9.15 
Notification of Serious Adverse Events 
Under IND regulations, 21 CFR Part 312.64, investigators are required to notify the 
Sponsor promptly, within 24 hours of the sites’ notification of any serious adverse events, deaths, 
or life-threatening problems with the investigational drug.  This regulation also requires that if the 
adverse event is alarming, the investigator must notify the Sponsor immediately. 
The Sponsor must be notified as detailed in section 9.16, "Reporting a Serious Adverse 
Event."  The Sponsor, in turn, will report all serious adverse events to regulatory agencies as 
required.  In addition to the serious adverse events described in section 9.7, other events that in the 
investigator's opinion suggest a significant hazard, contraindication, or precaution should be 
considered serious.  This includes, but is not limited to, blood dyscrasias, endocrine disturbances, 
hemorrhage from any site, or severe skin disorder. Additional examples are intensive treatment in 
an emergency room or at home for allergic bronchospasm, blood dyscrasias or convulsions that do 
not result in hospitalization, and development of drug dependency or drug abuse.  Hospitalization 
for elective surgery is not considered a serious adverse event. In addition, pregnancy is not a 
Serious Adverse Event, but is reportable and must be reported per IND regulations and timelines, 
21 CFR Part 312.64, on the SAE reporting form and submitted to the sponsor. 
Patients who experience a serious adverse event must be given appropriate examinations 
and treatment.  The investigator must provide written information to the sponsor as soon as 
possible. 
When an investigator is in doubt when to report an event, the investigator should err on the 
side of caution and contact the Sponsor. 
9.16 
Reporting a Serious Adverse Event 
Any serious adverse event, including death due to any cause that occurs during this study, 
whether or not believed to be related to the study medication, must be reported immediately (within 
one business day) via telephone/email to: 
Safety@EnterinInc.com 
and 
 
 
 
Denise Barbut MD FRCP 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
54 
 
 
 
denise@enterininc.com 
                       d.barbut@enterininc.com 
 
 
Cell 9179751377 
 
And 
 
 
Michael Zasloff MD PhD 
 
 
M.zasloff@enterininc.com  
 
 
Cell 484 433 7807 
 
And  
 
 
Heidi Marcus Moran 
 
 
Hm.moran@enterininc.com 
 
 
Cell 609.412.8411 
Enterin Inc: Phone Number:  1-215-966-6083 
Specific medical questions can be addressed to the Chief Medical Officer: 
 
 
Denise Barbut, M.D, FRCP. 
 
 
President and Chief Medical Officer 
 
 
Phone (9179751377) 
 
The back-up medical monitor, to be contacted in instances when the medical monitor is not 
available, is: 
 
 
Michael Zasloff, MD, Ph.D 
 
 
Chairman and CEO 
 
 
Phone (484 433 7807) 
 
The initial report should contain as much information as is available concerning the event 
in order to permit the Sponsor to file a report that satisfies regulatory requirements.  Initial 
telephone reports of serious adverse events must be followed-up by a fax of a completed SAE 


 
 
 
 
 
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report form or an appropriate event narrative.  The event should also be entered into the source 
documents and case report form, as appropriate. When additional information is available, the 
investigator should fax a follow-up SAE form or an appropriate supplementary event narrative to 
the Drug Safety Associate. 
 
All appropriate serious adverse events will be reported immediately to appropriate 
regulatory authorities by the Sponsor.  A copy of all FDA reportable serious adverse events will 
be mailed to all investigators participating in ongoing clinical studies with the study medication in 
order to permit prompt notification of the appropriate institutional review board (IRB). 
9.17 
Serious Adverse Experiences: After Study Participation 
Preclinical data to date do not point to specific classes of adverse events for which patients may 
be at risk after completion of study participation.  In order to monitor for unanticipated adverse 
events occurring after study participation, the following must be complied with: 
• 
Any occurrence of a patient death is to be reported any time that the investigator 
becomes aware of the event 
• 
A congenital anomaly in an offspring of a patient where that offspring has been 
born after the patient used study medications should be reported 
• 
Any serious adverse event, for which, in the investigator's opinion, there is a 
reasonable possibility that the event could have been caused by the study drug, should be 
reported 
• 
Any other serious adverse event should be reported if it occurred within thirty days 
following the last dose of study medication 
9.18 
Departure from Protocol for Emergency or Adverse Event 
In medical emergencies, the investigator should use medical judgment and remove the 
patient from immediate hazard.  As soon as possible after removing the patient from hazard, the 
investigator must contact Enterin Inc. by telephone to permit a decision as to whether the patient 
may continue in the study.  The IRB should also be notified as to the type of emergency and the 
course of action.  The case report form for the patient must describe the departure from the protocol 
and state the reason. 


 
 
 
 
 
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9.19 
Data Safety and Monitoring Board 
A Data Safety and Monitoring Committee (DSMB) has been established to monitor the 
safety of the subjects during the course of the study.  The DSMB includes members with relevant 
clinical expertise, including a good understanding of the safety of medications for Parkinson’s 
disease. These members include a statistician, a gastroenterologist and a neurologist. The 
methodology and the operating procedures for the safety reviews will be developed by the 
Chairperson in collaboration with the sponsor and will be documented in the DSMB Charter. 
9.20 
Review of Stage 1 Data by Data Safety Monitoring Board 
When the last subject in Stage 1 has completed the course of dose escalation safety, PD 
and PK data will be reviewed by a DSMB, which will then provide a recommendation, based on 
guidelines established in the DSMB Charter, whether to advance to Stage 2.  The starting dose 
for Stage 2 will be that dose which is safe and tolerable in all subjects and has a prokinetic effect 
in at least some (“STDE dose”). When the first ten subjects in Stage 2 have completed the course 
of dose escalation, data will be reviewed by the DSMB, which will then provide a 
recommendation, based on guidelines established by the DSMB Charter, whether to advance to 
the randomized controlled Period. 
9.21 
Stopping Rules 
The DSMB will monitor the safety of the subjects during the course of the study.  The 
DSMC can recommend changes to the protocol or temporary stopping of the trial at any time if 
there are significant safety concerns.  The DSMC Charter will include recommended safety 
stopping rules. 
9.22 
Follow-Up and Final Reports 
The investigator shall provide Enterin, Inc. with an accurate final report within 1 month 
after completion, termination or discontinuation of the study.  The final report may not precede 
retrieval of case report forms which have not been monitored. 
9.23 
Regulatory Aspects 
Neither the investigator nor Enterin Inc. shall modify this protocol without first obtaining 
concurrence of the other in writing.  All changes must be submitted to the IRB.  Protocol 


 
 
 
 
 
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modifications which impact patient safety or the validity of the study must be approved by the IRB 
and submitted to the FDA before implementation.  In the case of a medical emergency to increase 
safety of patients, a change may be made after discussion with the sponsor.  In these instances the 
IRB and FDA will be notified as soon as possible. 
10 DATA MANAGEMENT AND STATISTICS  
This is a trial with two stages. The Stage 1 portion will consist of 10 patients who will be 
dose escalated in single doses. Each patient will proceed through all the dose escalations starting 
at 25 mg and ending at 200 mg or at the dose at which s/he experiences a dose limiting toxicity 
(DLT). The doses to be tested are 25mg, 50mg, 75mg, 100mg, 125mg, 150mg, 175mg and 200mg. 
In the Stage 2 portion, all patients will go through a multiple dose escalation (3 dosing days at each 
dose) and the dose at which the patient experiences a consistent pro-kinetic effect (spontaneous, 
complete stool in at least 2/3 consecutive doses) without a DLT will be the dose for that patient. 
Patients who experience a DLT prior to reaching their pro-kinetic dose and at a dose equal to or 
less than the STDE dose will be terminated. Otherwise the patient will be assigned the highest 
dose at which they did not experience a DLT. If the patient reached 175 mg without a DLT and 
without a pro-kinetic dose, her/his dose will be 175 mg. The primary endpoint for the Stage 1 study 
is to determine whether the dose levels explored are safe and tolerable. The primary endpoint for 
the Stage 2 study is the regularization of bowel function as determined by stool frequency at the 
end of the fixed dose period compared to baseline. The Stage 2 portion will enroll 40 patients, with 
a target sample size of 34 evaluable patients. Six extra patients are enrolled to allow for patient 
withdrawals or terminations prior to the completion of the trial.  
10.1 
Sample Size/Power Considerations 
10.1.1  Stage 1 
There will be 10 patients enrolled in Stage 1. Two patients will be started on the 25mg 
dose. If no DLTs are observed, they will be escalated to the next dose and the remaining 8 patients 
will be started at 25mg. Each dose level will be a single dose and patients will be observed for 
three days before being given the next higher single dose. It is intended that each patient receive a 
single dose of each of the levels: 25mg, 50mg, 75mg, 100mg, 125mg, 150mg, 175mg and 200mg. 
However, if a patient experiences a DLT, they will not be further escalated.  


 
 
 
 
 
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A DLT is defined in Section 9.8 using the Common Terminology Criteria for Adverse 
Events (CTCAE), as a grade 2 vomiting within 24 hours of taking ENT-01 grade 2 diarrhea within 
24 hours of taking ENT-01, grade 2 abdominal pain within 24 hours of taking ENT-01, grade 2 
postural hypotension within 24 hours of taking ENT-01. Additionally, an LFT elevation > 3xULN 
within 24 hours of taking ENT or any gastrointestinal AE > grade 3 within 24 hours of taking 
ENT-01 that is at least possibly attributable to ENT-01 will be considered dose-limiting toxicity 
for the purpose of analyses. 
There is no power calculation for the Stage 1 portion because it is merely trying to 
determine whether ENT-01 is safe and tolerable for these levels. The PK analyses will be 
descriptive and are meant to understand how ENT-01 is metabolized. 
10.1.2 Stage 2 
The target sample size for Stage 2 is 34 evaluable patients and an additional 6 patients will 
be accrued to allow for patient drop-outs and terminations; these 40 patients will be different from 
the 10 patients accrued to Stage 1. A patient is evaluable if s/he has baseline measurements 
(obtained at the end of the run-in Period, 3.1) and measurements at the end of the fixed dose period 
(end of Period 3.2). The primary endpoint for Stage 2 is average complete spontaneous stool 
frequency increase by 1 point over baseline, OR 3 or more complete spontaneous stools per week. 
A patient will be labeled as a “success” if s/he meets one or more of the criteria above, otherwise 
the patient will be labeled as a “failure”. The primary analysis compares the proportion of patients 
deemed a success at the end of the fixed-dose period (end of period 3.2) to 0.10, which is the 
highest proportion of spontaneous resolution of constipation assuming no treatment. A patient is 
deemed a success or failure by comparing the assessment at the end of the fixed-dose period 
(Period 3.2) to baseline (obtained at the end of the run-in period, Period 3.1). There could be 
between 34 and 40 patients with these measurements and each patient will be labeled as a “success” 
or “failure”. It is assumed that without treatment, the success rate would be 10% (null hypothesis). 
An exact binomial test with a 0.05 two-sided significance level will have 80% power to detect the 
difference between 0.10 (proportion expected if patients are not treated), p0, and an ENT-01 
proportion, pE, of 0.29 (0.27) when the sample size is 34 (40). It is assumed that most patients will 
be deemed a success on the ENT-01 arm and so the study is adequately powered. 


 
 
 
 
 
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10.2 
Data Collection and Case Report Form Monitoring 
Reports will be generated on a weekly basis to monitor data completeness. Sites and 
patients who are missing data will be contacted and measures will be taken to resolve any obstacles 
regarding the data submission. 
10.3 
Endpoint definitions and measurements 
10.3.1 Assessment of Safety 
Safety assessments (vital signs, physical examinations, interim medical history, vital signs, 
EKG (when clinically warranted for any unscheduled EKG), AEs, clinical laboratory results 
(routine hematology and biochemistry) are to be performed at protocol-specified visits, as 
specified in the Schedule of Procedures and Assessments.  
 
Vital Signs 
Vital signs (body temperature, respiration rate, heart rate, systolic and diastolic blood 
pressure measurements) will be evaluated at the visits indicated in the Schedule of Procedures and 
Assessments.  Blood pressure will be taken twice.  The first time, the patient shall be lying down.  
The second blood pressure will be taken within 5 minutes of sitting or standing from the recumbent 
position.  All other vital signs will be measured after the patient has been resting in a sitting 
position for at least 5 minutes.  Blood pressure measurements are to be taken in the same arm for 
the duration of the study.  Body weight (without shoes) will be recorded whenever vital signs are 
recorded, and height (without shoes) will be recorded at screening only. 
Vital sign measurements will be repeated if clinically significant or machine/equipment 
errors occur.  Any confirmed, clinically significant vital sign measurements must be recorded as 
AEs. 
 
Physical Examination 
A complete physical examination (head, eyes, ears, nose and throat, heart, lungs, abdomen, 
skin, cervical and axillary lymph nodes, neurological, and musculoskeletal systems) will be 
performed at Visit 1.  Physical examinations will be performed by a physician.  In addition, 
medical history will be recorded at screening, including smoking history, if applicable.  


 
 
 
 
 
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A limited (brief) physical examination to verify continued patient eligibility and to follow 
up any change in medical history will be performed at the visits indicated in the Schedule of 
Procedures and Assessments.  Symptom-driven limited physical examinations will be performed 
as clinically indicated at any study visit.  All changes not present at baseline or described in the 
past medical history and identified as clinically noteworthy must be recorded as AEs. 
 
Electrocardiogram (EKG) 
A 12-lead resting EKG will be obtained at the visits indicated in the Schedule of Procedures 
and Assessments. 
  An assessment of normal or abnormal will be recorded and if the EKG is considered 
abnormal, the abnormality will be documented on the CRF. EKGs will be repeated if clinically 
significant abnormalities are observed or artifacts are present. 
 
Laboratory Assessments 
Laboratory assessment samples are to be obtained at designated visits as detailed in the 
Schedule of Procedures and Assessments.   
All clinically significant laboratory values should be captured on the CRF as AEs.  
Blood and urine samples will be analyzed at the laboratory facility at the Investigational 
site. Urine samples will be analyzed by dipstick, and a microscopic analysis will be performed if 
the results of dipstick indicate abnormalities to be further investigated.  All laboratory reports must 
be reviewed, signed, and dated by the Investigator. A legible copy of all reports must be filed with 
both the patient's CRF and medical record (source document) for that visit. Any laboratory test 
result considered by the Investigator to be clinically significant should be considered an AE 
(clinically significant AEs include those that require an intervention). Clinically significant 
abnormal values occurring during the study will be followed until repeat test results return to 
normal, stabilize, or are no longer clinically significant. 
Adverse events 
All reported AEs will be coded using the CTCAE version 4.03 terminology.  The incidence 
of treatment-emergent AEs (TEAEs; events with onset dates on or after the start of the study drug) 
and treatment-related AEs will be summarized by system organ class (SOC) and preferred term 


 
 
 
 
 
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(PT).  Events with missing onset dates will be included as treatment-emergent.  If a patient 
experiences more than 1 occurrence of the same AE, the occurrence with the greatest severity and 
the closest association with the study drug will be used in the summary tables.  SAEs and AEs that 
result in treatment discontinuation will be summarized.  In addition, AEs of special interest will 
be summarized by SOC and PT; categories of AEs of special interest include: nausea, vomiting, 
abdominal pain, diarrhea, and rectal bleeding. 
All AEs will be listed by patient, along with information regarding onset, duration, 
relationship and severity to study drug, action taken with study drug, treatment of event, and 
outcome. 
10.3.2 Efficacy Endpoints 
 
Frequency of bowel movements: 
The frequency of bowel movements will be determined via a daily diary where the passage 
of a bowel movement or absence thereof will be recorded daily throughout the study. Bowel 
movements that are the consequence of a rescue medication or suppository will not be counted 
if they occur within 24 hours of the rescue medication or suppository. Episodes of incontinence 
will also be recorded.  
Success will be defined as an average complete spontaneous stool frequency increase by 
1/week or more compared to baseline or 3 or more complete spontaneous stools/week. Baseline 
will be the run-in period weekly average and treatment period will be determined as the weekly 
average during the fixed dose (40 patients) treatment period. 
Colonic Motility:  
Colonic motility will be determined using the Sitzmarks technique at the beginning of the 
run-in period and again at the end of the randomized treatment period during Stage 2. The primary 
measure of interest is the mean colonic transit (MCT) time estimated with a single film (Metcalf 
et al., 1987). 


 
 
 
 
 
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10.4 
Analysis Plan 
10.5 
Stage 2 
Primary analysis: The AE information for the Stage 2 portion of the trial will be 
summarized as frequency and relative frequency by each dose level and for the LSTDEi for each 
person.  
The primary efficacy outcome variable is whether or not a patient is a “success” or 
“failure”. This is an endpoint based on patient diary entries for the week prior to the endpoint 
assessment defined as average complete stool frequency increase by 1 or more over baseline, OR 
3 or more complete spontaneous stools/week. The patient is deemed a “success” if s/he meets one 
or more of the criteria listed above, otherwise the patient will be deemed a “failure”. The primary 
analysis will be based on all patients with a baseline assessment (the end of the run-in period, 
Period 3.1) and an assessment at the end of the fixed-dose period (Period 3.2) and will be a 
comparison of the proportion of successes with 0.10 (the null hypothesis corresponding to no 
treatment effect). The proportion of patients for whom the drug was a success at the end of period 
3.2 will be estimated with a binomial point estimate and corresponding 95% confidence interval. 
The observed proportion of successes will be compared to 0.10 using an exact binomial test. ENT-
01 will be deemed as demonstrating sufficient efficacy if the two-sided p-value is less than 0.05 
for this comparison.  
Secondary analyses:  
A secondary analysis will compare the proportions of patients who are deemed a success 
at the end of the randomized period (Period 4) as compared to baseline (end of the run-in period, 
Period 3.1) between those randomized to the ENT-01 arm and those randomized to the placebo 
arm. Patients to be included in this analysis will be those who have a baseline assessment (end of 
the run-in period) and an assessment at the end of the randomization period of the trial, which 
could be between 34 and 40 patients (17 to 20 in each arm). A Fisher’s exact test will be used to 
compare the proportions of patients who are deemed a success at the end of randomization period 
(period 4) between the two randomized arms. Another secondary analysis will compare the 
proportions of patients who are a success (compared to baseline) between the end of the first week 
of the washout period (period 5) to the end of the randomization period (period 4), for those 


 
 
 
 
 
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patients who were randomized to receive ENT-01 (between 17 and 20 patients). This analysis will 
consist only of those patients with assessments at baseline (end of run-in period, Period 3.1), the 
end of Period 4, and the end of Period 5. This analysis will have between 17 and 20 patients and 
will be done using an exact sign test of equality of paired proportions.  
Exploratory analyses:  
The first exploratory analysis will of the colonic motility values as measured by an estimate 
of the mean MCT times. Comparisons will be made within each patient at different time points 
(i.e. baseline and the end of fixed period) as well as between patient groups (i.e. values observed 
at the end of randomization period, period 4 for the placebo and the ENT-01 treated groups). The 
primary endpoint for this analysis is the MCT based on a single film estimate. From a previous 
study (Ashraf et al., 1997), and estimate for the average MCT for constipated patients with 
Parkinson’s disease is 53.4 hours and an estimate of the standard deviation is 15.9 hours. A paired 
t-test will be used to compare the mean change in MCT at the end of the fixed period from baseline. 
A sample size of 34 (40) patients will have 80% power to detect a mean change of 11.1 hours (10.2 
hours) with a 0.05 two-sided level of significance and using a paired t-test. This calculation 
assumed a conservative estimate of 22.4 hours for the standard deviation of the change in MCT, 
which is the square root of 2 times the variance of the baseline standard deviation. The comparison 
of the average MCT between the control group and the group randomized to ENT-01 at the end of 
the randomization period will be made with a two-group t-test. 
Exploratory analyses will also be done with respect to the sleep data, the body temperature 
data, mood, fatigue, hallucinations, overall NMS scale and all other questionnaires and neuro-
cognitive tests. These parameters will be summarized numerically and graphically. There will also 
be comparisons done of changes over time within a patient (i.e. between baseline, end run-in 
period, and at the end of the fixed dose period, Period 3.2) and differences between groups (i.e. 
between patients in the Enterin arm and in the placebo arm at the end of period 4). As before, 
continuous measurements within a patient will be compared with a paired t-test (transforming the 
data if necessary) and continuous measurements between patient groups will be compared with a 
two-group t-test (transforming the data if necessary). In addition, these data will be analyzed using 
a mixed linear model since there are multiple measurements over time. Categorical data will be 


 
 
 
 
 
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compared with a chi-squared test or a Fisher’s exact test if the expected cell counts are too small 
for a chi-squared test.  
11 ESTIMATED DURATION OF THE STUDY 
This study has an estimated duration of 8 weeks for each patient during each stage. 
Maximum dosing days for each patient will be no more than 8 days during Stage 1 and 25 days 
during Stage 2. The study duration from first patient in to last patient out is expected to be 9-12 
months. 
12 ADMINISTRATIVE PROCEDURES AND ETHICAL CONSIDERATIONS 
12.1 
Medical Care 
This study has an estimated duration of 21 weeks. During this period, patients will be on 
medication for a maximum of eight weeks. None of the patients will be given medication during 
the run-in period or the wash-out period. 
12.2 
Patient Information and Informed Consent 
The Principal Investigator(s) at each center will ensure that the patient is given full and 
adequate oral and written information about the nature, purpose, possible risk and benefit of the 
study.  Patients must also be notified that they are free to discontinue from the study at any time.  
The patient should be given the opportunity to ask questions and allowed time to consider the 
information provided. 
The patient’s signed and dated informed consent must be obtained before conducting any 
study procedures. 
The Principal Investigator(s) must maintain the original, signed Informed Consent Form.  
A copy of the signed Informed Consent Form must be given to the patient. 
12.3 
Study Monitoring 
Before an investigational site can enter a patient into the study, a representative of Enterin, Inc. 
will visit the investigational study site to: 
• Determine the adequacy of the facilities 


 
 
 
 
 
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• Discuss with the investigator(s) and other personnel their responsibilities with regard to 
protocol adherence, and the responsibilities of Enterin, Inc. or its representatives.  This will 
be documented in a Clinical Study Agreement between Enterin, Inc. and the investigator. 
During the study, a monitor from Enterin, Inc. or representative will have regular contacts with 
the investigational site, for the following: 
• Provide information and support to the investigator(s) 
• Confirm that facilities remain acceptable 
• Confirm that the investigational team is adhering to the protocol, that data are being 
accurately recorded in the case report forms, and that investigational product accountability 
checks are being performed 
• Perform source data verification.  This includes a comparison of the data in the case report 
forms with the patient’s medical records at the hospital or practice, and other records 
relevant to the study.  This will require direct access to all original records for each patient 
(e.g. clinic charts). 
• Record and report any protocol deviations not previously sent to Enterin, Inc. 
• Confirm AEs and SAEs have been properly documented on CRFs and confirm any SAEs 
have been forwarded to Enterin, Inc. and those SAEs that met criteria for reporting have 
been forwarded to the IRB. 
The monitor will be available between visits if the investigator(s) or other staff needs 
information or advice. 
12.4 
Audits and Inspections 
Authorized representatives of Enterin, Inc., a regulatory authority, an Independent Ethics 
Committee or an Institutional Review Board may visit the site to perform audits or inspections, 
including source data verification.  The purpose of an Enterin, Inc. audit or inspection is to 
systematically and independently examine all study-related activities and documents to determine 
whether these activities were conducted, and data were recorded, analyzed, and accurately reported 
according to the protocol, Good Clinical Practice guidelines of the International Conference on 


 
 
 
 
 
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Harmonization, and any applicable regulatory requirements. The investigator should contact 
Enterin, Inc. immediately if contacted by a regulatory agency about an inspection. 
12.5 
Ethics Committee Review 
The final study protocol, including the final version of the Informed Consent Form, must 
be approved or given a favorable opinion in writing by an IRB or IEC as appropriate.  The 
investigator must submit written approval to Enterin Inc. before he or she can enroll any 
patient/subject into the study. 
The Principal Investigator is responsible for informing the IRB or IEC of any amendment 
to the protocol in accordance with local requirements.  In addition, the IRB or IEC must approve 
all advertising used to recruit patients for the study.  The protocol must be re-approved by the IRB 
or IEC upon receipt of amendments and annually, as local regulations require. 
The Principal Investigator is also responsible for providing the IRB with reports of any 
reportable serious adverse drug reactions from any other study conducted with the investigational 
product. Enterin, Inc. will provide this information to the Principal Investigator. 
Progress reports and notifications of serious adverse drug reactions will be provided to the 
IRB or IEC according to local regulations and guidelines. 
12.6 
Standards 
The study will be performed in accordance with ethical principles that have their origin in 
the Declaration of Helsinki and are consistent with ICH/Good Clinical Practice, applicable 
regulatory requirements and Enterin, Inc.’s policy on Bioethics. 
12.7 
Confidentiality 
Any research information obtained about the subject in this study will be kept confidential.  
A subject will not be identified by name, only by his/her initials.  The subject’s name or any 
identifying information will not appear in any reports published as a result of this study. 
However, information obtained from individual subject’s participation in the study may be 
disclosed with his/her consent to the health care providers for the purpose of obtaining appropriate 
medical care.  The subject’s medical records/charts, tests with his/her name on them may be made 


 
 
 
 
 
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available to the appropriate contract research organization (CRO), the sponsor, its potential 
eventual partners, and any other regulatory authorities.  This is for the purpose of verifying 
information obtained for this study.  Confidentiality will be maintained throughout the study within 
the limits of the law. 
A subject’s name will not be given to anyone except the researchers conducting the study, 
who have pledged an oath of confidentiality.  All identifying information will be kept behind 
locked doors, under the supervision of the study Investigator and will not be transferred outside of 
the investigator site. 
A subject may take away his/her permission to collect, use and share information about 
him/her at any time.  If this situation occurs, the subject will not be able to remain in the study.  
No new information that identifies the subject will be gathered after that date.  However, the 
information about the subject that has already been gathered and transferred may still be used and 
given to others as described above in order to preserve the scientific integrity and quality of the 
study. 
12.8 
Protocol Adherence 
The pharmacist or other designated individual will maintain records of study treatment 
delivered to the study site; the inventory at the site; the distribution to and use by each patient; and 
the return of materials to Enterin, Inc. for storage or disposal. These records should include dates, 
quantities, batch/serial numbers, expiration dates, in-clinic temperature log, and unique code 
numbers assigned to the product and study patients.   
At each visit after initiation of treatment, site staff will record compliance of patients with 
their assigned regimen.  Patients will be instructed to bring their diaries and cartons containing 
unused/partially used/empty vials back for inspection at each study visit. Patients are to be 
reminded of the importance of compliance with their assigned regimen, with an emphasis on taking 
their study drug on schedule, and maintaining the prescribed interval between doses. 
Investigator will maintain records that document adequately that the patients were provided 
with the correct study drug and will reconcile the products received from the drug dispensing 
center. Investigational product will not be returned to Enterin, Inc. until accountability has been 
fully monitored. 


 
 
 
 
 
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Medication containers must be returned at each visit, as compliance will be assessed by 
tablet counts. Noncompliance is defined as taking less than 80% or more than 120% of study drug 
during any outpatient evaluation period (visit to visit). Discontinuation for noncompliance is at the 
Investigator’s discretion and is to be noted on the case report form (CRF). 
12.9 
Amendments to the Protocol 
Modifications to the protocol are only possible by approved protocol amendments 
authorized by the study sponsor.  All protocol amendments will be approved by the appropriate 
regulatory authorities as well as each institutional review board prior to implementation.  The 
Investigator must not implement any deviations from, or changes to the protocol, except where it 
is necessary to eliminate an immediate hazard to the study subject. 
12.10 Protocol Deviations 
The protocol must be read thoroughly and the instructions followed exactly.  The sponsor 
and/or designee will not grant waivers for protocol deviations.  Any deviation to the protocol has 
to be reported as soon as possible to the sponsor.  The governing reporting guidelines for protocol 
deviations must be adhered to by the Investigator. 
12.11 Study Termination 
This study may be prematurely terminated, if in the opinion of the Investigator or Enterin, there 
is sufficient reasonable cause.  Written notification documenting the reason for study termination 
will be provided to the Investigator or Enterin by the terminating party.  Circumstances that may 
warrant termination include, but are not limited to: 
• 
Determination of unexpected, significant, or unacceptable risk to subjects. 
• 
Failure to enroll subjects at an acceptable rate. 
• 
Insufficient adherence to protocol requirements. 
• 
Insufficient complete and/or evaluable data. 
• 
Plans to modify, suspend, or discontinue the development of ENT-01. 
Should the study be closed prematurely, all study materials must be returned to Enterin. 


 
 
 
 
 
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12.12 Data handling and record keeping 
12.13 Inspection of Records 
 
Enterin, Inc. will be allowed to conduct site visits to the investigation facilities for the 
purpose of monitoring any aspect of the study.  The Investigator agrees to allow the monitor to 
inspect the drug storage area, study drug stocks, drug accountability records, subject charts and 
study source documents, and other records relative to study conduct. 
12.14 Data Management 
 
All data relating to study procedures will be entered on to electronic case report forms 
provided by the sponsor.  All forms must be completed electronically. All requested information 
must be entered on the electronic case report form.  If an item is not available or not applicable this 
fact should be indicated by the use of "NA".  Spaces should not be left blank.  Electronic case report 
forms will be reviewed during the monitoring visits.  Data will be entered into a database.  Data entry 
and management and the production of the clinical study report will be the responsibility of the 
sponsor or a designated agent. 
 
The data cut-off for the study will be 60 days after entry of the last patient.  If patients remain 
on study thereafter, a revised final study report will be issued after the last patient has completed the 
final study evaluation. 
12.15 Data Capture and Management 
The sponsor will provide the study sites with an electronic case report system. 
Electronic CRFs will be completed for each subject.  It is the Investigator’s responsibility 
to ensure the accuracy, completeness, and timeliness of the data entered in each subject’s eCRF.  
Source documentation supporting the eCRF data should indicate the subject’s participation in the 
study and document the dates and details of study procedures, AEs, and subject status. 
The Investigator, or designated representative, should complete the eCRF as soon as 
possible after information is collected, preferably on the same day that a subject is seen for an 
examination, treatment, or any other study procedure. The information collected from the daily 
sleep and stool diaries will be entered by the sponsor or a designated representative on an on-going 


 
 
 
 
 
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basis. Any outstanding entries must be entered immediately after the final examination.  An 
explanation should be given for all missing data. 
12.16 Liability and Insurance 
The sponsor has subscribed to an insurance policy covering, in its terms and conditions, its 
legal liability for certain injuries to participating persons arising out of this research performed 
strictly in accordance with the scientific protocol as well as with applicable law and professional 
standards. 
12.17 Retention of Records 
Study records and source documents must be preserved for at least 15 years after the 
completion or discontinuation of/withdrawal from the study or 2 years after the last approval of a 
marketing application in an ICH region, whichever is the longer time period. 
The Investigator agrees to comply with all applicable federal, state, and local laws and 
regulations relating to the privacy of patient health information, including, but not limited to, the 
Standards for Individually Identifiable Health Information, 45 CFR, Parts 160 and 164 (the Health 
Insurance Portability Accountability Act of 1996 [HIPAA] Privacy Regulation). The Investigator 
shall ensure that study patients authorize the use and disclosure of protected health information in 
accordance with HIPAA Privacy Regulation and in a form satisfactory to the Enterin, Inc. 
12.18 Data Quality Assurance 
As per GCP guidelines, the sponsor or designee will be responsible for implementing and 
maintaining quality assurance and quality control systems for this study. 
Participating sites, the study database, and study documentation including subject medical 
records may be subject to a quality assurance audit during the course of the study.  In addition, 
inspections may be conducted by regulatory bodies at their discretion. 
If sites receive a request for an inspection or written or oral inquiries regarding any aspect 
of institution’s or Investigator’s activities related to this study from a regulatory authority, the 
Investigator must immediately notify the sponsor and the appropriate CRO of the request.  
Following this inspection and/or audit, the Investigator must notify the sponsor of any violation or 
deficiency noted by the regulatory authority. 


 
 
 
 
 
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13 USE OF INFORMATION     
Both the use of data and the publication policy are detailed within the clinical study 
agreement. Intellectual property rights (and related matters) generated by the Investigator and 
others performing the clinical study will be patient to the terms of a clinical study agreement that 
will be agreed between the Institution and Enterin, Inc. or their designee. With respect to such 
rights, Enterin, Inc. or its designee will solely own all rights and interests in any materials, data, 
and intellectual property rights developed by Investigators and others performing the clinical study 
described in this protocol, patient to the terms of any such agreement. In order to facilitate such 
ownership, Investigators will be required to assign all such inventions directly to Enterin, Inc. or 
its designee, as will be set forth in the clinical study agreement. 
14 INVESTIGATOR AGREEMENT 
I have read the foregoing protocol “A Multicenter, Single-dose, Multiple-dose, Randomized, 
Double-blind, Placebo-controlled study to evaluate Safety, Tolerability, Pharmacokinetics and 
Pharmacodynamics of orally administered ENT-01 for Constipation in Parkinson’s disease” and 
agree to conduct the study as described therein. 
 
[name] 
 
 
 
Investigator’s Name  
 
 
 
 
 
 
 
 
 
[dd/mm/yyyy] 
 
Investigator’s Signature 
 
Date 
 
 
 
 


 
 
 
 
 
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APPENDIX 1:  CLINICAL LABORATORY DETERMINATION 
Hematology 
Serum chemistry 
Urine analysis 
(dipstick) 
Full and differential blood 
count 
Hematocrit (Hct) 
Hemoglobin (Hb) 
Mean corpuscular hemoglobin 
(MCH) 
Mean corpuscular hemoglobin 
concentration (MCHC) 
Mean corpuscular volume 
(MCV) 
Platelet count 
Red blood cell (RBC) count  
White blood cell (WBC) count 
with differential  
 
Albumin 
Alanine aminotransferase (ALT) 
Alkaline phosphatase (ALP) 
Aspartate aminotransferase (AST) 
Blood Urea Nitrogen (BUN) or 
Urea 
Carbon dioxide (CO2) 
Creatinine 
Creatine kinase and subtypes 
Electrolytes (Na, K, Cl, Ca, P) 
Gamma--glutamyl transpeptidase 
(GGT) 
Glucose  
Lactate dehydrogenase (LDH)? 
Total bilirubin 
Direct bilirubin 
Total cholesterol 
Triglycerides 
Appearance 
pH 
Protein 
Glucose 
Ketone bodies 
Indicators of blood 
and WBCs 
Specific gravity 
Urine human 
chorionic 
gonadotropin (HCG) 
(pre-menopausal 
females only)  
Urobilinogen 
Coagulation 
 
 
Prothrombin time (PT) 
Activated partial 
thromboplastin time (PTT) 
 
 
Pregnancy test: A pregnancy test will be performed on all female patients of child-bearing 
potential at the screening visit. 
 
 


 
 
 
 
 
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APPENDIX 2: SCHEDULES OF EVENTS 
 
STAGE 1 
 
Period 
1 
Run-In 
Sentinel 
2 wks 
2.1 
Single 
Dose 
3-8 wks 
2.2 
Wash-Out 
2 wks 
 
D1 
D14 
D1 
 
Consent 
x 
 
 
 
Demographics 
x 
 
 
 
Inclusion/exclusion 
x 
x 
 
 
Medical history 
x 
 
 
 
Physical exam 
x 
 
x 
x 
Vital signs1 
x 
 
x 
x 
Rome-IV 
x 
 
 
 
Blood chemistry 
x 
 
x 
x 
Hematology 
x 
 
 
 
Urinalysis 
x 
 
 
x 
Fecal occult blood 
x 
 
x 
x 
EKG 
x 
 
x 
x 
Pregnancy test 
x 
 
 
 
Monitoring 
 
 
x 
x 
Discontinue medications 
x 
 
 
 
Re-start medications 
 
 
 
x 
Diary entries 
x 
x 
x 
x 
UPDRS 
x 
 
X4 
x2 
I-Button3 
x 
x 
X 
 
PAC-QOL 
x 
x 
X4 
x2 
PAC_SYM 
x 
x 
X4 
x2 
RBDQ 
x 
 
X4 
x2 
NMSQ 
x 
 
X4 
x2 
PFS-16 
x 
 
X4 
x2 
UM-PDHQ 
x 
 
X4 
x2 


 
 
 
 
 
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BDI 
x 
 
X4 
x2 
Trail-making 
A and B 
x 
 
X4 
x2 
MMSE 
x 
 
X4 
x2 
Adverse events 
 
x 
X 
x 
PK samples 
 
 
X5 
 
Schedule visit days 
x 
 
 
 
Administer ENT Dose 
 
 
x 
 
Record Medications 
x 
x 
x 
x 
Daily phone calls to on non-visit days 
 
x 
x 
x 
1Vital signs taken before dose and 2 hours after dose  
2End of period 2 
3I-Button placed days 9 of run-in periods and worn continuously through day 5 of wash-out periods.   
4Surveys taken at last dosing visit  
5PK samples taken pre-dose and 1, 2, 4, 8 and 24 hours post-dose  
 
 
 


 
 
 
 
 
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STAGE 2 


 
 
 
 
 
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1For PK patients, vital signs taken pre-dose and 2 hours after dose.  Blood pressure will be taken for all patients twice, first lying down and then  
after 5 minutes of sitting or standing from the recumbent position.  All other vital signs will be measured after the patient has been resting  
in a sitting position for at least 5 minutes. 
2Sitzmark capsules are dispensed to patients at Visit 1 with instructions to take one a day on Days 8, 9, and 10 of the Run-In period for the  
abdominal X-ray on Day 11.  Patients will receive more Sitzmark capsules at Visit 3 with instructions to take one a day on Days 3, 4,  
and 5 of the Randomized Treatment period for the abdominal X-ray on Day 6.   
3Subjects are reminded during the Day 7 call of the Run-In period and Day 2 call of the Randomized Treatment period to begin taking the  
Sitzmark capsules the following day to prepare for the abdominal X-ray on Day 11 of the Run-In period and Day 6 of the Randomized  
Treatment period. 
4Patients will wear an I-Button continuously from Day 8 of the Run-In period to Day 5 of the Wash-Out period except during periods of bathing. 
5PK samples are taken pre-dose and 1, 2, 4, and 8 hours post-dose.  A sample for liver function tests will be taken with the 8-hour post-dose PK draw. 
6Visit 3 occurs on the day before randomization in non-PK subjects.


 
 
 
 
 
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APPENDIX 3: NUMERIC RATING SCALES FOR PAIN AND SWELLING 
 
 
 


 
 
 
 
 
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APPENDIX 4: ROME-IV CRITERIA FOR CONSTIPATION 
 
Must include two or more of the following: 
Straining during at least 25% of defecations 
Lumpy or hard stools in at least 25% of defecations 
Sensation of incomplete evacuation for at least 25% of defecations 
Sensation of incomplete evacuation for at least 25% of defecations 
Sensation of anorectal obstruction/block age for at least 25% of defecations 
Manual maneuvers to facilitate at least 25%of defecations (e.g., digital evacuation, support of the 
pelvic floor) 
Fewer than three defecations per week 
Loose stools are rarely present without the use of laxatives 
 
 
 


 
 
 
 
 
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APPENDIX 5: EASE OF EVACUATION SCALE 
 
 
1.Manual disimpaction 
2.Enema needed 
3.Straining needed 
4.Normal 
5.Urgent without pain 
6.Urgent with pain 
7.Incontinent 
 
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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APPENDIX 6: BRISTOL STOOL CHART 
 
The assessment of stool consistency is a validated surrogate of intestinal motility (Lewis and 
Heaton, 1997). We have chosen the Bristol Stool Chart as a patient-friendly means of categorizing 
stool characteristics.  
 
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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APPENDIX 7: SLEEP DIARY 
Participants will complete a sleep diary on a daily basis throughout the study.  
The diaries will include time into bed and estimated time to sleep as well as wake time and duration 
during the night.  
An example of the sleep diary is shown below: 
Date 
Dose 
(mg) 
Time 
to bed 
(hr) 
Sleep 
onset 
(hr) 
Morning 
awakeni
ng (hr) 
Wakes 
at 
night/ 
duration 
(min) 
Total 
Hours 
sleep 
Leg 
thrashi
ng/screa
ming/vi
vid 
dreams 
I-Button 
on 
3/20 
- 
12 
1.30 
7.0 
2 
(30min 
each) 
4.5 
Yes 
No 
3/21 
100 
12 
1.0 
7.0 
1 (60 min) 
5.0 
Some 
Yes,  
3/22 
100 
11 
12.30 
7.30 
2 (60 min, 
60min) 
5.0 
Yes 
Yes,  
3/23 
100 
12 
12.30 
6.30 
1 (30min) 
5.5 
Some 
Yes,  
3/24 
150 
- 
11.0 
7.30 
1 (30min) 
8.0 
No 
Yes,  
3/25 
150 
- 
11.0 
7.0 
- 
8.0 
No 
Yes,  
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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APPENDIX 8: I-BUTTON 
The I-Button is a small rugged self-sufficient system that measures temperature and records 
the results in a protected memory section. The Thermochron I-Button DS1921H (Maxim 
Integrated, Dallas TX) used for skin temperature measurement.  The I-Button DS1921H is 
regulated by Federal Communications Commission (FCC) as a class B digital radiofrequency 
device that is an unintentional radiator (i.e., CFR 47, Part 15, Subpart B), and conforms to the 
national standards tested under ANSI C63.4-2001. The  I-Button is composed of a semiconductor 
temperature sensor, a real timer, a memory and a lithium battery, all enclosed in a 16-mm diameter 
by 6-mm thick cylinder. Manufacturing specifications include: temperature range from +15 to +46 
°C, and 1 °C of accuracy with a precision of 0.125 °C. The memory (NV RAM) allows it to record 
up to 2048 values taken at regular intervals of 1 to 255 min. I-Buttons will be programmed to 
sample every 10 min, and attached to a double-sided cotton sport wrist band using Velcro®, with 
the sensor face of the I-Button being placed over the inside of the wrist, on the radial artery of the 
dominant hand.   Subjects will remove and replace the data logger when necessary (i.e., to have a 
bath or shower). After the period of monitoring, the information stored in the I-Button will be 
transferred through an adapter (DS1402D-DR8, IDC, Spain) to a personal computer using I-Button 
Viewer v. 3.22© 1992–2005 Dallas Semiconductor MAXIM software provided by the 
manufacturer. The value of skin temperature assessment in sleep research is that the endogenous 
skin warming resulting from increased skin blood flow is functionally linked to sleep propensity 
(Sarabia et al., 2008; van Marken Lichtenbelt et al., 2006). 
      From the collected data we will calculate the mesor, amplitude,acrophase (time of peak 
temperature), Rayleigh test (an index of interdaily stability), mean waveforms (averaged per 
subject, and then for each experimental group) using an analysis package for temporal series. 
Briefly, interdaily phase stability will be calculated as follows: firstly, daily acrophases for subjects 
at baseline and following treatment will be obtained using least-squares data fitting to a cosine 
function with a period of 24 h. The acrophase distribution within a 24-h period will be assessed 
using a Rayleigh test (CircStat: Matlab toolbox for circular statistical package). This test provides 
an r vector with its origin at the center of a circumference of radius one. The r vector length 
(between 0 and 1) is proportional to the degree of phase homogeneity during the period analyzed, 
and can be considered to be a measure of the rhythm's phase stability during successive days. 


 
 
 
 
 
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Statistical differences between baseline and treatment values will be analyzed by two-way repeated 
measures ANOVA(Sarabia et al., 2008). 


 
 
 
 
 
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APPENDIX 9: NON-MOTOR SYMPTOMS QUESTIONNAIRE (NMSQ) 
 
 


 
 
 
 
 
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APPENDIX 10: BECK DEPRESSION INVENTORY (BDI) 
 
1.  
0 I do not feel sad.  
1 I feel sad  
2 I am sad all the time and I can't snap out of it.  
3 I am so sad and unhappy that I can't stand it.  
2.  
0 I am not particularly discouraged about the future.  
1 I feel discouraged about the future.  
2 I feel I have nothing to look forward to.  
3 I feel the future is hopeless and that things cannot improve.  
3. 
0 I do not feel like a failure.  
1 I feel I have failed more than the average person.  
2 As I look back on my life, all I can see is a lot of failures.  
3 I feel I am a complete failure as a person.  
4.  
0 I get as much satisfaction out of things as I used to.  
1 I don't enjoy things the way I used to.  
2 I don't get real satisfaction out of anything anymore.  
3 I am dissatisfied or bored with everything.  
5. 
0 I don't feel particularly guilty  
1 I feel guilty a good part of the time.  
2 I feel quite guilty most of the time.  
3 I feel guilty all of the time.  
6.  
0 I don't feel I am being punished.  
1 I feel I may be punished.  


 
 
 
 
 
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2 I expect to be punished.  
3 I feel I am being punished.  
7.  
0 I don't feel disappointed in myself.  
1 I am disappointed in myself.  
2 I am disgusted with myself. 
3 I hate myself.  
8.  
0 I don't feel I am any worse than anybody else.  
1 I am critical of myself for my weaknesses or mistakes.  
2 I blame myself all the time for my faults.  
3 I blame myself for everything bad that happens.  
9.  
0 I don't have any thoughts of killing myself.  
1 I have thoughts of killing myself, but I would not carry them out.  
2 I would like to kill myself.  
3 I would kill myself if I had the chance.  
10.  
0 I don't cry any more than usual.  
1 I cry more now than I used to.  
2 I cry all the time now.  
3 I used to be able to cry, but now I can't cry even though I want to.  
11.  
0 I am no more irritated by things than I ever was.  
1 I am slightly more irritated now than usual.  
2 I am quite annoyed or irritated a good deal of the time.  
3 I feel irritated all the time.  
12. 
 0 I have not lost interest in other people.  
1 I am less interested in other people than I used to be.  


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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2 I have lost most of my interest in other people.  
3 I have lost all of my interest in other people.  
13.  
0 I make decisions about as well as I ever could.  
1 I put off making decisions more than I used to.  
2 I have greater difficulty in making decisions more than I used to.  
3 I can't make decisions at all anymore.  
14.  
0 I don't feel that I look any worse than I used to.  
1 I am worried that I am looking old or unattractive.  
2 I feel there are permanent changes in my appearance that make me look unattractive  
3 I believe that I look ugly.  
15.  
0 I can work about as well as before.  
1 It takes an extra effort to get started at doing something.  
2 I have to push myself very hard to do anything.  
3 I can't do any work at all.  
16.  
0 I can sleep as well as usual.  
1 I don't sleep as well as I used to.  
2 I wake up 1-2 hours earlier than usual and find it hard to get back to sleep.  
3 I wake up several hours earlier than I used to and cannot get back to sleep.  
17.  
0 I don't get more tired than usual.  
1 I get tired more easily than I used to.  
2 I get tired from doing almost anything.  
3 I am too tired to do anything.  
 
18.  
0 My appetite is no worse than usual.  


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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1 My appetite is not as good as it used to be.  
2 My appetite is much worse now.  
3 I have no appetite at all anymore.  
19.  
0 I haven't lost much weight, if any, lately.  
1 I have lost more than five pounds.  
2 I have lost more than ten pounds.  
3 I have lost more than fifteen pounds.  
20.  
0 I am no more worried about my health than usual.  
1 I am worried about physical problems like aches, pains, upset stomach, or constipation.  
2 I am very worried about physical problems and it's hard to think of much else.  
3 I am so worried about my physical problems that I cannot think of anything else.  
21.  
0 I have not noticed any recent change in my interest in sex.  
1 I am less interested in sex than I used to be.  
2 I have almost no interest in sex.  
3 I have lost interest in sex completely.  
 
INTERPRETING THE BECK DEPRESSION INVENTORY Now that you have completed the 
questionnaire, add up the score for each of the twenty-one questions by counting the number to 
the right of each question you marked. The highest possible total for the whole test would be sixty-
three. This would mean you circled number three on all twenty-one questions. Since the lowest 
possible score for each question is zero, the lowest possible score for the test would be zero. This 
would mean you circles zero on each question. You can evaluate your depression according to the 
Table below. Total Score____________________ 
Levels of Depression 1-10____________________These ups and downs are considered normal  
11-16___________________ Mild mood disturbance  
17-20___________________Borderline clinical depression  


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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21-30___________________Moderate depression  
31-40___________________Severe depression  
over 40__________________Extreme depression 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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APPENDIX 11: UNIFIED PARKINSON’S DISEASE RATING SCALE 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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CP1001 v0.9, Version Date: 27JUL2017 
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APPENDIX 12: MINI MENTAL STATE EXAMINATION 
 
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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APPENDIX 13: TRAIL MAKING A & B 
 
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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CP1001 v0.9, Version Date: 27JUL2017 
46 
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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APPENDIX 14: UM-PDHQ 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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CP1001 v0.9, Version Date: 27JUL2017 
49 
 
APPENDIX 15: PARKINSON’S DISEASE FATIGUE SCALE (PFS-16) 
 
 
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
50 
 
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
51 
 
APPENDIX 16: PATIENT ASSESSMENT OF CONSTIPATION SYMPTOMS 
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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APPENDIX 17: PATIENT ASSESSMENT OF CONSTIPATION QUALITY OF LIFE  
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
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CP1001 v0.9, Version Date: 27JUL2017 
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CP1001 v0.9, Version Date: 27JUL2017 
55 
 
APPENDIX 18: REM SLEEP BEHAVIOR DISORDER SCREENING 
QUESTIONNAIRE 
 
1. I sometimes have very vivid dreams. yes/no  
2. My dreams frequently have an aggressive or action-packed content. yes/no  
3. The dream contents mostly match my nocturnal behavior. yes/no  
4. I know that my arms or legs move when I sleep. yes/no  
5. It thereby happened that I (almost) hurt my bed partner or myself. yes/no  
6. I have or had the following phenomena during my dreams:  
6.1. Speaking, shouting, swearing, laughing loudly yes/no  
6.2. Sudden limb movements, “fights” yes/no  
6.3. Gestures, complex movements, that are useless during sleep, e.g., to wave, to salute, 
to frighten mosquitoes, fall off the bed yes/no  
6.4. Things that fell down around the bed, e.g., bedside lamp, book, glasses yes/no  
7. It happens that my movements awake me. Yes/no  
8. After awakening I mostly remember the content of my dreams well. Yes/no  
9. My sleep is frequently disturbed. Yes/no  
10. I have/had a disease of the nervous system (e.g., stroke, head trauma, Parkinsonism, RLS, 
narcolepsy, depression, epilepsy, inflammatory disease of the brain), which? yes/no 
 
 
 
 


 
 
 
 
 
CP1001 v0.9, Version Date: 27JUL2017 
56 
 
APPENDIX 19: PARKINSON’S DISEASE SLEEP SCALE 
 
 

SAP_001.pdf: 
Enterin Inc. 
 
 
Protocol No: ENT-01 
 
       Statistical Analysis Plan 
 
Confidential 
 
 
 
     Page 1 | 47 
 
 
STATISTICAL ANALYSIS PLAN: ENT-01 
 
Study Title: 
A Multicenter, Single-Dose, Multiple-Dose, Randomized, Double-
Blind, Placebo-Controlled Study to Evaluate Safety, Tolerability, 
Pharmacokinetics and Pharmacodynamics of Orally Administered 
ENT-01 for the Treatment of Parkinson’s Disease Related 
Constipation 
Protocol Number: 
ENT-01 
Protocol Version/ Date: 
27JUL2017 and 02FEB2018 
Study Phase: 
1/2a 
Product Name: 
ENT-01  
 
 
Sponsor: 
Enterin Inc. 
Sponsor Contact: 
2005 Market Street, Suite 3125 
Philadelphia, PA 19103 
 
 
Date 
Statistical Analysis Plan Date: 
04May2018 
Statistical Analysis Plan Version: 
V1.0 
SAP Amendment 1 Date: 
03Jul2018 
SAP Amendment 1 Version: 
V2.0. 
 
 
 
 
 
 
Confidentiality Statement 
The information in this document contains trade secrets and commercial information that are 
privileged or confidential and may not be 
disclosed unless such disclosure is required by law or regulations. In any event, persons to whom the 
information is disclosed must be 
informed that the information is privileged or confidential and may not be further disclosed by them. 


Enterin Inc. 
 
 
Protocol No: ENT-01 
 
       Statistical Analysis Plan 
 
Confidential 
 
 
 
     Page 2 | 47 
SPONSOR SIGNATURE PAGE 
 
Protocol Title: 
A Multicenter, Single-Dose, Multiple-Dose, Randomized, Double-
Blind, Placebo-Controlled Study to Evaluate Safety, Tolerability, 
Pharmacokinetics and Pharmacodynamics of Orally Administered 
ENT-01 for the Treatment of Parkinson’s Disease Related 
Constipation 
Protocol Number: 
ENT-01 
Sponsor: 
Enterin Inc. 
By signing this document, I acknowledge that I have read the document and approve of the 
planned statistical analyses described herein. I agree that the planned statistical analyses are 
appropriate for this study, are in accordance with the study objectives, and are consistent with 
the statistical methodology described in the protocol, clinical development plan, and all 
applicable regulatory guidance and guidelines. 
Author 
 
Weining Volinn, PhD  
Director of Biostatistics 
LLX Solutions, LLC 
Signature: 
Date: 
Approver 
 
Denise Barbut, MD 
FRCP 
President and CMO 
Enterin Inc. 
Signature: 
Date: 


Enterin Inc. 
 
 
Protocol No: ENT-01 
 
       Statistical Analysis Plan 
 
Confidential 
 
 
 
     Page 3 | 47 
TABLE OF CONTENTS 
STATISTICAL ANALYSIS PLAN: ENT-01 ................................................................................ 1 
SPONSOR SIGNATURE PAGE ................................................................................................... 2 
TABLE OF CONTENTS ................................................................................................................ 3 
TABLES INCLUDED IN THE TEXT ........................................................................................... 7 
ABBREVIATIONS ........................................................................................................................ 8 
1 
INTRODUCTION ................................................................................................................. 11 
2 
STUDY DESIGN OVERVIEW ............................................................................................ 12 
2.1 
Overall Study Design ..................................................................................................... 12 
2.2 
Sample Size/ Power Calculation .................................................................................... 20 
2.3 
Randomization and Blinding .......................................................................................... 20 
3 
STUDY OBJECTIVES ......................................................................................................... 21 
3.1 
Objectives for Stage 1 .................................................................................................... 21 
3.1.1 
Primary Objective ............................................................................................................... 21 
3.1.2 
Secondary Objective ........................................................................................................... 21 
3.1.3 
Exploratory Objective ......................................................................................................... 21 
3.2 
Objectives for Stage 2 .................................................................................................... 21 
3.2.1 
Primary Objectives .............................................................................................................. 21 
3.2.2 
Exploratory Objective ......................................................................................................... 21 
4 
STUDY ENDPOINTS AND EVALUATIONS .................................................................... 22 
4.1 
Pharmacokinetic Parameters .......................................................................................... 22 
4.1.1 
Pharmacokinetic Parameters for Stage 1............................................................................. 22 
4.1.2 
Pharmacokinetic Parameters for Stage 2............................................................................. 22 
4.2 
Safety Evaluations .......................................................................................................... 22 
4.3 
Tolerability Evaluations ................................................................................................. 22 
4.4 
Pharmacodynamic Evaluations ...................................................................................... 23 
4.4.1 
Stool Diary Data.................................................................................................................. 24 
Stage 1 Pharmacodynamics Parameters......................................................................................... 24 
Stage 2 Pharmacodynamics Parameters: ....................................................................................... 25 
4.4.2 
Sleep Diary Data ................................................................................................................. 27 
4.4.3 
Questionnaire Data .............................................................................................................. 27 


Enterin Inc. 
 
 
Protocol No: ENT-01 
 
       Statistical Analysis Plan 
 
Confidential 
 
 
 
     Page 4 | 47 
4.4.3.1 
Non-Motor Symptoms Questionnaire ................................................................................. 28 
4.4.3.2 
Beck Depression Inventory II ............................................................................................. 28 
4.4.3.3 
Mini Mental State Examination .......................................................................................... 28 
4.4.3.4 
The University of Miami Parkinson’s Disease Hallucinations Questionnaire .................... 28 
4.4.3.5 
Parkinson’s disease Fatigue Scale ....................................................................................... 29 
4.4.3.6 
Patient Assessment of Constipation Symptoms .................................................................. 29 
4.4.3.7 
Patient Assessment of Constipation Quality of Life ........................................................... 30 
4.4.3.8 
REM Sleep Behavior Disorder Screening Questionnaire ................................................... 30 
4.4.3.9 
Unified Parkinson’s disease Rating Scale ........................................................................... 30 
4.4.3.10 Other Questionnaires........................................................................................................... 31 
5 
PLANNED ANALYSES ....................................................................................................... 32 
5.1 
Changes from planned analyses in the Protocol............................................................. 32 
5.2 
Interim Analyses ............................................................................................................ 32 
5.3 
Final Analyses and Reporting ........................................................................................ 32 
6 
ANALYSIS POPULATIONS ............................................................................................... 33 
6.1 
Pharmacokinetic Population ........................................................................................... 33 
6.1.1 
Pharmacokinetic Population for Stage 1 ............................................................................. 33 
6.1.2 
Pharmacokinetic Population for Stage 2 ............................................................................. 33 
6.2 
Safety Population ........................................................................................................... 33 
6.3 
Full Analysis Set ............................................................................................................ 33 
6.4 
Randomized population.................................................................................................. 33 
7 
STATISTICAL CONSIDERATIONS .................................................................................. 34 
7.1 
General Statistical Procedures ........................................................................................ 34 
7.2 
Enrollment and Disposition ............................................................................................ 34 
7.2.1 
Subjects Enrollment ............................................................................................................ 34 
7.2.2 
Subject Disposition ............................................................................................................. 34 
7.2.3 
Subject Disposition for Stage 1 ........................................................................................... 34 
7.2.4 
Subject Disposition for Stage 2 ........................................................................................... 35 
7.3 
Protocol Deviations ........................................................................................................ 35 
7.4 
Demographics and Baseline Characteristics .................................................................. 35 
7.4.1 
Demographics and Baseline Characteristics ....................................................................... 35 
7.4.2 
Baseline Disease Characteristics ......................................................................................... 36 


Enterin Inc. 
 
 
Protocol No: ENT-01 
 
       Statistical Analysis Plan 
 
Confidential 
 
 
 
     Page 5 | 47 
7.4.3 
Medical History................................................................................................................... 36 
7.5 
Prior and Concomitant Medications ............................................................................... 36 
7.6 
Analysis of Pharmacokinetic Data ................................................................................. 37 
7.6.1 
Summary of Pharmacokinetic Concentration Data for Stage 1 .......................................... 37 
Summary of Pharmacokinetic Concentration Data for Stage 2 ...................................................... 37 
7.6.2 
Analyses of Pharmacokinetic Parameters ........................................................................... 37 
7.6.3 
Analyses of Pharmacokinetic Parameters for Stage 1 ......................................................... 37 
7.6.4 
Analyses of Pharmacokinetic Parameters for Stage 2 ......................................................... 37 
7.7 
Analysis of Pharmacodynamics Data ............................................................................. 37 
7.7.1 
Analysis of Continuous Pharmacodynamics Data .............................................................. 38 
7.7.2 
Analysis of Categorical Pharmacodynamics Data .............................................................. 38 
7.8 
Analysis of Safety Data .................................................................................................. 39 
7.8.1 
Adverse Events ................................................................................................................... 39 
7.8.2 
Clinical Laboratory Tests .................................................................................................... 40 
7.8.3 
Vital Signs ........................................................................................................................... 41 
7.8.4 
Electrocardiogram (ECG) ................................................................................................... 41 
7.8.5 
Physical Examination .......................................................................................................... 42 
7.8.6 
Analysis of Tolerability Data .............................................................................................. 42 
7.8.7 
Extent of Exposure .............................................................................................................. 42 
7.8.8 
I-BUTTON .......................................................................................................................... 43 
7.9 
Additional Data Presentation as Listing ......................................................................... 43 
8 
DEFINITIONS AND CONVENTIONS FOR DATA HANDLING .................................... 44 
8.1 
Definition of Baseline .................................................................................................... 44 
8.2 
Analysis Visit Window .................................................................................................. 44 
8.3 
Pharmacodynamic Data Handling .................................................................................. 44 
8.4 
Safety Data Handling ..................................................................................................... 44 
8.5 
Handling of Repeated Clinical Laboratory Tests ........................................................... 44 
8.6 
Pharmacokinetic Data Handling..................................................................................... 44 
8.6.1 
Pharmacokinetic Plasma Drug Concentration Data ............................................................ 44 
8.7 
Handling of Partial Dates for AEs.................................................................................. 45 
8.8 
Handling of Partial Dates for Medications ..................................................................... 45 
9 
Changes to the Planned Analyses in the Protocol(s) ............................................................. 46 


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10 
REFERENCES .................................................................................................................. 47 
 


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TABLES INCLUDED IN THE TEXT 
 
Table 1: Overall Study Design 
Table 2: Schedule of Assessments of Stage 1 
Table 3: Schedule of Assessments of Stage 2 under Protocol Dated 27 July 2017 
Table 4: Schedule of Events of Stage 2 under Protocol Dated 02 February 2018 
 
 
 
 
 
 
 
 


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ABBREVIATIONS  
Abbreviation 
Definition of Terms 
AE 
Adverse Events  
AUC0-24 
Area under the concentration versus time curve from time 0 to the 
concentration at 24 hours; calculated using linear trapezoid rule  
BDI 
Beck Depression Inventory 
BLQ  
Below the Quantification Limit  
BMI 
Body mass index 
CL/F  
Clearance defined as: Dose/AUC0-24   
Cmax  
Maximum or peak measured plasma concentration  
Cmin 
Minimum or trough measured plasma concentration  
CSR 
Clinical Study Report 
CV 
Coefficient of Variation 
DLT 
Dose limiting Toxicity 
ECG 
Electrocardiogram 
eCRF 
Electronic Case Report Form 
FAS 
Full analysis set 
LFT 
Liver Function Test 
LSTDEi 
Consistently lowest clinically effective safe and tolerable dose for that 
particular individual 
MCT 
Mean Colonic Transit 
MDRP 
Medical Data Review Plan 
MedDRA 
Medical Dictionary for Regulatory Activities 
MMSE 
Mini Mental State Examination  
NC  
Not Calculated 


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Abbreviation 
Definition of Terms 
NMSQ 
Non-Motor Symptoms Questionnaire 
PAC-QoL 
Patient Assessment of Constipation Quality of Life 
PAC-SYM 
Patient Assessment of Constipation Symptoms 
PD 
Pharmacodynamics 
PDSS 
Parkinson’s Disease Sleep Scale 
PFS-16 
Parkinson’s Disease Fatigue Scale  
PK 
Pharmacokinetics 
PT 
Preferred Term 
Q1 
First Quartile 
Q3 
Third Quartile 
RBD 
REM behavior disorder 
RBDQ 
Rem Sleep Behavior Disorder Screening Questionnaire 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SD 
Standard Deviation 
STDE 
Safe and Tolerable dose in all and pro-kinetic Effect in at least some subjects 
SOC 
System Organ Class 
t1/2 
 
Apparent terminal elimination half-life; calculated as ln(2)/ λz. The number 
of data points included in the regression will be determined by visual 
inspection, but a minimum of three data points in the terminal phase, 
excluding Cmax, will be required to estimate λz.  
TEAEs 
Treatment Emergent Adverse Events 
Tmax  
Time of maximum or peak measured plasma concentration  
ULN 
Upper limit of normal 


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Abbreviation 
Definition of Terms 
UM-PDHQ 
The University of Miami Parkinson’s Disease Hallucinations Questionnaire 
UPDRS 
Unified Parkinson’s Disease Rating Scale 
WHO 
World Health Organization  
WHO-DD 
World Health Organization Drug Dictionary  
λz 
Apparent terminal rate constant 
 


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1 INTRODUCTION  
This statistical analysis plan (SAP) is designed to outline the statistical methods to be used for 
analyses and data presentation for reporting pharmacokinetics (PK), pharmacodynamics (PD), 
safety and tolerability, and any exploratory objectives for study protocol ENT-01. This document 
has been prepared based on Study Protocol dated 27 July 2017 and Protocol amendment 02 
February 2018.  
This SAP supersedes the statistical considerations identified in the protocol. If additional 
analyses are required to supplement the planned analyses described in this SAP, they may be 
performed and will be identified in the CSR. Any substantial deviations from this SAP will be 
agreed upon between Enterin Inc. and LLX Solutions, LLC and identified in the CSR. 
This SAP amendment was done for 
1. Adding details on the scoring methods for each questionnaire to be analyzed in the study; 
2. Deleting two secondary analyses for “Success” endpoint:  
• Due to the protocol amendment, the sample size for randomization period became too 
small to do statistical analysis. Therefore, it was deleted from this SAP amendment. 
• During the trial, it became apparent that the period for “Success” at the end of the first 
week in washout period is not consistent with the washout period used for other 
pharmacodynamics parameters. Therefore, this analysis is also deleted in the SAP 
amendment. 
3. Clarifying the summaries and analyses should be provided for fixed dosing or fixed doing 
period as applicable.  
 
 


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2 STUDY DESIGN OVERVIEW 
2.1 Overall Study Design 
This is a single-dose, multiple-dose, randomized, double-blinded, placebo-controlled, two-staged 
study in Parkinson’s disease subjects with constipation. The study will consist of two stages, 
each with run-in period and dosing periods and followed by a wash-out period of 2 weeks. Stage 
1 will consist of 10 subjects while Stage 2 will consist of 40 subjects to ensure at least 34 
evaluable subjects are obtained. During Stage 1, after providing informed consent, subjects will 
be screened for eligibility. Qualified subjects will proceed through all the dose escalations of 
ENT-01 starting at 25 mg and ending at 200 mg or at the dose at which s/he experiences a dose 
limiting toxicity (DLT). Stage 1 is staggered, where the first two subjects will be administered a 
single dose of 25 mg first. The rest of the subjects will be dosed after the first two subjects have 
been observed for 72 hours with no safety concerns. The doses to be tested are 25mg, 50mg, 
75mg, 100mg, 125mg, 150mg, 175mg and 200mg. Subjects will then be observed off study drug 
for two weeks during the wash-out period. STDE (Safe and Tolerable dose in all and pro-kinetic 
Effect in at least some) dose will be identified and carried over to Stage 2. 
In Stage 2 under protocol dated 27 July 2017, subjects who signed the informed consent and are 
eligible will go through a multiple dose escalation (3 dosing days at each dose) period. The dose 
range is 75 mg up to 175 mg with an increment of 25 mg for next higher dose. Ten subjects will 
be in the sentinel group and will be dosed first. The dose at which the subject experiences a 
consistent pro-kinetic effect (spontaneous, complete stool in at least 2/3 consecutive doses) 
without a DLT will be the dose selected going forward for that subject. This will be the LSTDEi 
(i.e. consistently clinically effective safe and tolerable dose for that particular individual). After 
the LSTDEi is established, subjects will continue to take this dose for 2-4 days, followed by 
randomization on the LSTDEi dose with a 1:1 ratio to ENT-01 or placebo in block sizes of 4 for 
a 6-day period (period 4). Seven subjects from the ENT-01 group and three subjects from the 
control group will be in the PK group and have the PK measurement taken. The rest will be in 
the non-PK group. All the subjects will then enter the wash-out period for two weeks. 
The protocol dated 27 July 2017 was amended during the trial. And the protocol amendment was 
finalized on 02 February 2018. In this amendment, Stage 2 dose range is from 100 mg to 250 mg 
with an increment 25 mg for next higher dose. It also removed randomization period. 
Except for the above changes, there were no other changes from protocol dated 27 July 2017 


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Table 1: Overall Study Design 
 


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Table 2: Schedule of Assessments of Stage 1 
Period 
1 
Run-In 
Sentinel 
2 wks 
2.1 
Single 
Dose 
3-8 wks 
2.2 
Wash-Out 
2 wks 
D1 
D14 
D1 
Consent 
X 
Demographics 
X 
Inclusion/exclusion 
X 
X 
Medical history 
X 
Physical exam 
X 
X 
X 
Vital signs1 
X 
X 
X 
Rome-IV 
X 
Blood chemistry 
X 
X 
X 
Hematology 
X 
Urinalysis 
X 
X 
Fecal occult blood 
X 
X 
X 
EKG 
X 
X 
X 
Pregnancy test 
X 
Monitoring 
X 
X 
Discontinue medications 
X 
Re-start medications 
X 
Diary entries 
X 
X 
X 
X 
UPDRS 
X 
X4 
X2 
I-Button3 
X 
X 
X 
PAC-QOL 
X 
X 
X4 
X2 
PAC_SYM 
X 
X 
X4 
X2 
RBDQ 
X 
X4 
X2 
NMSQ 
X 
X4 
X2 
PFS-16 
X 
X4 
X2 
UM-PDHQ 
X 
X4 
X2 
BDI 
X 
X4 
X2 
Trail-making A and B 
X 
X4 
X2 


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Table 2: Schedule of Assessment of Stage 1 
 
Period 
1 
Run-In 
Sentinel 
2wks 
2.1 
Single 
Dose 
3-8 wks 
2.2 
Wash-Out 
2 wks 
          D1 
D14 
D1 
MMSE 
X 
X4 
X2 
Adverse events 
X 
X 
X 
PK samples 
X5 
Schedule visit days 
X 
Administer ENT Dose 
X 
Record Medications 
X 
X 
X 
X 
Daily phone calls to on non-visit days 
X 
X 
X 
1Vital signs taken before dose and 2 hours after dose 
2End of period 2 
3I-Button placed days 9 of run-in periods and worn continuously through day 5 of wash-out periods. 
4Surveys taken at last dosing visit 
5PK samples taken pre-dose and 1, 2, 4, 8 and 24 hours post-dose 
 
 


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Table 3: Schedule of Assessments of Stage 2 under Protocol Dated 27 July 2017 
 Scheduled Assessments 
Period 3.1 
Run-In  
Period 3.2 
Multiple Dose 
Period 4  
Randomized Treatment 
  Period 5 
Wash-Out 
Day 1 
Visit 1 
Days 2-7, 9- 
10, 12-13 
Calls 
Day8  
Call 
Day 11 
Visit 2 
Day l4 
Call 
Days. 1-13, 
15-16 
Calls 
Day l4 
Call 
 
Day 1  
Visit 36 
Days 2-5 
Calls 
Day6 
Visit 4 
Days 1-7 
Calls 
Day l4 
Visit 5 
Informed Consent 
X 
Inclusion/Exclusion Criteria 
X 
Demographics/Medical History 
X 
Prior and Con Meds 
X 
Complete Physical Exam 
X 
X 
Height 
X 
Weight 
X 
X 
X 
X 
X 
Vital Signs1 
X 
X 
X 
X
X 
ECG 
X 
X 
X 
Local Labs -Chemistry, Hematology 
X 
X 
X 
X 
Local Labs - Urinalysis, Serum Pregnancy 
X 
Fecal Occult Blood 
X 
X 
X 
Randomization 
X 
Study Drug Dispense, Accountability, or 
Collection 
X 
X 
       X 
Reminder to Begin Taking Study Drug the Next 
Morning 
X 
Instructions to Discontinue Laxatives and other 
BM Medication 
X 
Instructions to Discontinue Laxatives and other 
BM medications 
X 
Stool and Sleep eDiary Instruction/Review 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
AE Review 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Rome IV questionnaire 
X 
X 
UPDRS 
X 
X 
X 
PAC-QOL and PAC-SYM 
X 
X 
X 
X 
X 
X 
Questionnaires- BDI, NMSQ, PFS-16, UM- 
PDHQ, MMSE, Trail-Making A and B, RBDQ 
and PDSS 
X 
X 
X 
X 
Sitzmarks Capsule Distributions2 
X 
X 
Reminder to Take Sitzmarks Capsules3 
 
X 
 
 
 
X 
 
 
 
Abdominal X-Ray  
X 
 
 
X 
 
 
Dispense I-Button4 
X 
 
 
 
 
 
 
 


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           Page 17 | 47 
 Scheduled Assessments 
Period 3.1 
Run-In  
Period 3.2 
Multiple Dose 
Period 4  
Randomized Treatment 
  Period 5 
Wash-Out 
Day 1 
Visit 1 
Days 2-7, 9- 
10, 12-13 
Calls 
Day8  
Call 
Day 11 
Visit 2 
Day l4 
Call 
Days. 1-13, 
15-16 
Calls 
Day l4 
Call 
 
Day 1  
Visit 36 
Days 2-5 
Calls 
Day6 
Visit 4 
Days 1-7 
Calls 
Day l4 
Visit 5 
PK Draws & LFT5 
 
 
 
 
 
 
 
X 
 
X 
 
Phone Call 
 
X 
X 
 
X 
X 
X 
 
X 
 
X 
1
For PK patients, vital signs taken pre-dose and 2 hours after dose.  Blood pressure will be taken for all patients twice, first lying down and then after 5 minutes 
of sitting or standing from the recumbent position.  All other vital signs will be measured after the patient has been resting in a sitting position for at least 5 
minutes. 
2Sitzmark capsules are dispensed to patients at Visit 1 with instructions to take one a day on Days 8, 9, and 10 of the Run-In period for the abdominal X-ray on 
Day 11.  Patients will receive more Sitzmark capsules at Visit 3 with instructions to take one a day on Days 3, 4, and 5 of the Randomized Treatment period for 
the abdominal X-ray on Day 6. 
3Subjects are reminded during the Day 7 call of the Run-In period and Day 2 call of the Randomized Treatment period to begin taking the Sitzmark capsules the 
following day to prepare for the abdominal X-ray on Day 11 of the Run-In period and Day 6 of the Randomized Treatment period. 
4Patients will wear an I-Button continuously from Day 8 of the Run-In period to Day 5 of the Wash-Out period except during periods of bathing. 
5PK samples are taken pre-dose and 1, 2, 4, and 8 hours post-dose.  A sample for liver function tests will be taken with the 8-hour post-dose PK draw. 
6Visit 3 occurs on the day before randomization in non-PK subject 


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Table 4: Schedule of Events of Stage 2 under Protocol Dated 02 February 2018   
Scheduled Assessments 
Period 3.1 Run In 
Period 3.2 Dose 
Escalation 
Period 4 Fixed Dose
Period 5 Wash Out 
Day 1 
Visit 1 
Screening 
Days 2-7, 
9,10,12,13 
Phone Calls 
Day 8 
Phone 
Call 
Day 11 
Visit 2 
Day 14 
Phone 
Call 
Days 1-13 
Phone 
Calls 
Day 14 
Phone 
Call 
Days 1-3 
Phone 
Calls 
Day 4 
Visit 3 
Days 1-7 
Phone Calls 
Day 14 
Visit 4 
Day 28 
FU Call 
Informed Consent 
X 
Inclusion/Exclusion Criteria 
X 
Demographics/Medical History 
X 
Concomitant Medications 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Complete Physical Exam 
X 
X 
Height 
X 
Weight 
X 
X 
X 
X 
Vital Signs (1) 
X 
X 
X 
X 
EKG 
X 
X 
X 
Local Labs: Chemistry*, Hematology* 
X 
X 
X 
Local Labs: Coagulation Panel* 
X 
Local Labs: Serum Pregnancy (2a) or FSH (2b) 
X 
Urinalysis* (dipstick) 
X 
X 
X 
Urine Pregnancy Test (3) 
X 
X 
X 
Fecal Occult Blood (4) 
X 
X 
X 
Study Drug Dispense, Accountability, Collection 
X 
X 
Reminder to Begin Taking Study Drug the Next 
X 
Instruction to Discontinue Laxatives and 
Other Restricted Medications (5) 
X 
Instruction to Resume Laxatives and 
Other Restricted Medications 
X (6) 
Stool and Sleep Diary Instruction/Review 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
AE Assessment and Review 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Rome IV Questionnaire 
X 
X 
UPDRS 
X 
X 
X 
PAC-QOL and PAC-SYM 
X 
X 
X 
X 
X 
BDI-II, NMSQ, PFS-16, UM-PDHQ, MMSE, 
Trail-Making A and B, RBDQ, PDSS 
X 
X 
X 
I-Button 
X (7) 
X 
X (9) 
X (10) 
 


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* Refer to Appendix 1 for full list of clinical laboratory assessments 
(1) 
 The first blood pressure and heart rate will be taken after the subject has been in the supine position for at least 5 minutes. The second 
blood pressure and heart rate reading will be taken while the subject is standing, within 5 minutes of standing from the supine position. All 
other vital signs (respiration rate and body temperature) will be taken after the subject has been sitting or standing for 5 minutes. 
(2) 
(2a) For women of child-bearing potential and (2b) For post-menopausal women less than 60 years of age. 
(3) 
For women of child-bearing potential, done on site with urine pregnancy kit provided 
(4) 
Subjects will be provided a home testing kit with instructions; to be used at home on the first bowel movement following the visit, and 
results to be given to the study coordinator at subsequent phone call visit. 
(5) 
Subjects will be instructed to discontinue all laxatives, bulking agents, softeners and suppositories as well as clonazepam, opiates, etc. as 
specified in the protocol. Proton pump inhibitors and antacids may be continued until the end of the Run-In Period. 
(6) 
Subjects will be instructed to resume use of their regular regimen laxatives, bulking agents, softeners, proton pump inhibitors, and antacids 
after week 1 of wash out. 
(7) 
Dispense I-Button 
(8) 
Instruct subjects to begin wearing the I-Button 
(9) 
Instruct subjects to stop wearing the I-Button at the end of Day 5 of Wash Out 
(10) Collect I Button 


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2.2 Sample Size/ Power Calculation  
No formal sample size calculation was performed for Stage 1. The number of subjects is based 
on feasibility and is considered sufficient to meet the objectives of the study.  
For Stage 2, assuming the highest proportion of spontaneous resolution of constipation with no 
treatment to be 0.10, 34 evaluable subjects who have measurements at both baseline and at the 
end of the fixed dose period will provide 80% power to detect the difference between 0.10 
(proportion expected if patients are not treated), p0, and an ENT-01 proportion, pE, of 0.29. 
Assuming a drop-out rate of 15%, a total of 40 subjects will ensure at least 34 evaluable subjects 
will be obtained.   
2.3 Randomization and Blinding 
No randomization is performed for Stage 1. 
During randomization period of Stage 2 under the protocol dated 27 July 2017, subjects will be 
randomly allocated in equal proportion (1:1) to 1 of 2 double-blind treatment groups: 
• ENT-01 – Patients will remain at dose level  taken in period 3.2 of Stage 2 
• Placebo – Patients will be switched to the same number of placebo tablets as the dose 
level  taken in period 3.2 of Stage 2 
 
 
 


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3 STUDY OBJECTIVES 
3.1 Objectives for Stage 1 
3.1.1 Primary Objective 
To investigate the safety and tolerability of single oral doses of ENT-01 in patients with 
Parkinson’s disease related constipation. 
3.1.2 Secondary Objective 
To evaluate the pharmacokinetic (PK) characteristics of single oral doses of ENT-01 in patients 
with Parkinson’s disease and constipation. 
3.1.3 Exploratory Objective 
To determine the pharmacodynamic (PD) characteristics of ENT-01 in patients with Parkinson’s 
disease related constipation. 
3.2 Objectives for Stage 2 
3.2.1 Primary Objectives 
• To determine the Safety, Tolerability of repeated oral doses of ENT-01 in patients with 
Parkinson’s disease related constipation.  
• To determine the PK and PD characteristics of repeated oral doses of ENT-01 in patients 
with Parkinson’s disease related constipation. 
 
3.2.2 Exploratory Objective 
• To determine the effect of study drug on Quality of Life Measures including sleep, 
temperature, memory, energy, mood, fatigue, hallucinations, memory and executive 
function in patients with Parkinson’s disease related constipation. 
 
 


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4 STUDY ENDPOINTS AND EVALUATIONS  
4.1 Pharmacokinetic Parameters 
4.1.1 Pharmacokinetic Parameters for Stage 1 
The following PK parameters will be determined where possible: 
Parameter 
Definition 
Cmax  
Maximum or peak measured plasma concentration  
Cmin 
Minimum or trough measured plasma concentration  
Tmax  
Time of maximum or peak measured plasma concentration  
t1/2   
Apparent terminal elimination half-life; calculated as ln(2)/ λz. 
The number of data points included in the regression will be 
determined by visual inspection, but a minimum of three data 
points in the terminal phase, excluding Cmax, will be required to 
estimate λz.  
AUC0-24 
Area under the concentration versus time curve from time 0 to 
the concentration at 24 hours; calculated using linear trapezoid 
rule  
CL/F  
Clearance defined as: Dose/AUC0-24   
λz 
Apparent terminal rate constant 
Additional pharmacokinetic parameters may be determined where appropriate. 
4.1.2 Pharmacokinetic Parameters for Stage 2 
Pharmacokinetic parameters will not be calculated. 
4.2 Safety Evaluations 
The safety evaluations for both stages include:  
• Treatment emergent adverse events (TEAEs) 
• Clinical laboratory assessments   
• Vital signs  
• Physical examination findings 
• Electrocardiogram 
4.3 Tolerability Evaluations 
Tolerability will be measured by the following DLT endpoints: Dose Limiting Toxicity (DLT) 
will be the dose which induces repeated vomiting, diarrhea, abdominal pain or symptomatic 
postural hypotension within 24 hours of dosing. It is defined using the Common Terminology 
Criteria for Adverse Events (CTCAE) as: 
• Recurrent vomiting: 3-5 episodes of vomiting within 24 hours of taking ENT-01 (i.e., 3-5 
episodes of Grade 2 vomiting) 


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• Recurrent diarrhea: 4-6 episodes of diarrhea within 24 hours of taking ENT-01 (will be 
recoded as a Grade 2 diarrhea)  
• Moderate Abdominal pain that limits instrumental activities of daily living within 24 
hours of taking ENT-01 (Grade 2 AE) 
• postural hypotension including Moderate dizziness, lightheadedness or fainting upon 
rising from lying to sitting or standing and severe enough to require medical intervention 
within 24 hours of taking ENT-01 (Grade 2 AE) or either a systolic blood pressure less 
than 80mmHg or diastolic blood pressure less than 40mm Hg 
In addition, the following will also be considered dose limiting toxicity for the purpose of 
analyses: 
• 
Elevation of LFTs > 3 times the upper limit of normal (ULN)  
• 
A reduction of body weight of 10% or more 
4.4   Pharmacodynamic Evaluations  
Unless otherwise stated, the following pharmacodynamic (PD) variables are evaluated for both 
Stage 1 and Stage 2. All PD parameters will be defined for each period. The periods under 
consideration for summary and analyses are run-in (i.e., baseline), each dosing period or 
treatment period (as applicable), and washout period (i.e., follow-up period).  
For Stage 1 stool diaries, the baseline PD parameters will be calculated using all diaries from 
run-in period; dosing period PD parameters will be calculated on dosing day immediately 
following each dose (for CSBM and SBM after dose, need to consider the day before dosing for 
the rescue medications use); and washout PD parameters will be calculated using diaries 
excluding the first day diaries (i.e., this one day diaries will be attributed to last dosing period).    
For Stage 2 stool diaries, all the diaries for each dosing period will be used for the calculation of 
PD parameters. The washout period started 24 hours of last dose. 
For Stage 1 sleep diaries, PD parameters will be calculated using all diaries from run-in period; 
up to last dosing period. The first 7-day diaries after last dosing period will be attributed to last 
dosing period and the second 7-day diaries after last dosing period will be attributed to washout 
period.   
For Stage 2 sleep diaries, PD parameters will be calculated using all diaries from run-in period; 
all diaries in each dosing period up to last dosing period. The washout period started 24 hours of 
last dose. 


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4.4.1 Stool Diary Data 
 Stage 1 Pharmacodynamics Parameters 
• Stage 1 complete spontaneous bowel movement (CSBM) is defined as CSBM (Yes) 
without rescue meds within 24 hours based on dosing day diary.  
• Stage 1 spontaneous bowel movements (SBM) is defined as SBM (Yes) without rescue 
meds within 24 hours based on dosing day diary 
• CSBM days per week will be calculated as follows: 
a. Run-in and washout period: calculation will be based on all diaries in Run-in and 
Washout period (excluding the first day diary in the period). 
 
CSBM Days per Week = Sum of days with CSBM  without rescue meds within 24 hours
Number of Diary Days
 × 7 days 
b. Treatment period: calculation will be based on Stage 1 CSBM as: 
 
CSBM Days per Week = Sum of days with Stage 1 CSBM 
Number of Dosing Days
× 7 days 
• SBM days per week will be calculated as follows: 
a. Run-in and washout period: calculation will be based on all diaries in Run-in and 
Washout period (excluding the first day diary in the period). 
SBM Days per Week = Sum of days with SBM  without rescue meds within 24 hours
Number of Diary Days
 × 7 days 
b. Treatment period: calculation will be based on Stage 1 SBM as: 
SBM Days per Week = Sum of days with Stage 1 SBM 
Number of Dosing Days
× 7 days 
• Average ease of passage for each period is defined as 
a. Run-in and washout period: calculation will be based on all diaries in Run-in and 
Washout period (excluding the first day diary in the period). 
b. Treatment period: calculation will be based on diaries on dosing days. 
Aberage ease of passage = 𝑆𝑆𝑆𝑆𝑆𝑆 𝑜𝑜𝑜𝑜 𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒 𝑠𝑠𝑠𝑠𝑠𝑠𝑠𝑠𝑠𝑠  𝑤𝑤/𝑜𝑜 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑤𝑤𝑤𝑤𝑤𝑤ℎ𝑖𝑖𝑖𝑖 24 ℎ𝑟𝑟𝑟𝑟
𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁 𝑜𝑜𝑜𝑜 𝐵𝐵𝐵𝐵𝐵𝐵 𝑤𝑤/𝑜𝑜 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑤𝑤𝑤𝑤𝑤𝑤ℎ𝑜𝑜𝑜𝑜𝑜𝑜 24 ℎ𝑟𝑟𝑟𝑟  


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• Average stool consistency for each period is defined as 
a. Run-in and washout period: calculation will be based on all diaries in Run-in and 
Washout period (excluding the first day diary in the period). 
b. Treatment period: calculation will be based on diaries on dosing days. 
Average stool consistency = Sum of stool consistency  w/o rescue meds within 24 hrs
Number of BMs w/o rescue meds without 24 hrs 
 
• Days using rescue med per week for each period is defined as 
a. Run-in and washout period: calculation will be based on all diaries in Run-in and 
Washout periods (excluding the first day diary in the period). 
Days using rescue meds per Week = Sum of days using rescue meds
Number of Diary Days
 × 7 days 
b. Treatment period: calculation will be based on diaries on dosing days. 
Days using rescue meds per Week = Sum of days using rescue meds 
Number of Dosing Days
× 7 days 
These first two parameters will be treated as categorical variables while other parameters will be 
treated as continuous variables. 
Stage 2 Pharmacodynamics Parameters: 
• CSBM days per week for each dosing period is defined as: 
CSBM Days per Week = Sum of days with CSBM  w/o rescue meds within 24 hours
Number of Diary Days
× 7 days 
• CSBM episodes  per week for each period is defined as: 
CSBM epidoses per Week = Sum of  daily CSBM episodes w/o rescue meds within 24 hours
Number of Diary Days
× 7 days 
where daily CSBM episodes = ൜#of CSBM ocuurences w/o rescue meds within 24 hrs for each diary day
0 if none CSBM w/o rescue meds within 24 hrs for each diary day                
• SBM days per week for each dosing period is defined as: 
SBM Days per Week = Sum of  Days with SBM w/o rescue meds within 24 hrs 
Number of Diary Days
× 7 days 


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• SBM episodes per week for each period is defined as: 
SBM episodes per Week = Sum of  Daily SBM Episodes w/o rescue meds within 24 hrs 
Number of Diary Days
× 7 days 
𝑤𝑤ℎ𝑒𝑒𝑒𝑒𝑒𝑒 𝐷𝐷𝐷𝐷𝐷𝐷𝐷𝐷𝐷𝐷 𝑆𝑆𝑆𝑆𝑆𝑆 𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸
= ൜#𝑜𝑜𝑜𝑜 𝑆𝑆𝑆𝑆𝑆𝑆 𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜 𝑤𝑤𝑤𝑤𝑤𝑤ℎ𝑜𝑜𝑜𝑜𝑜𝑜 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑤𝑤𝑤𝑤𝑤𝑤ℎ 24 ℎ𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜 𝑓𝑓𝑓𝑓𝑓𝑓 𝑒𝑒𝑒𝑒𝑒𝑒ℎ 𝑑𝑑𝑑𝑑𝑑𝑑 𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑
0 𝑖𝑖𝑖𝑖 𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑆𝑆𝑆𝑆𝑆𝑆 𝑤𝑤𝑤𝑤𝑤𝑤ℎ𝑜𝑜𝑜𝑜𝑜𝑜 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑓𝑓𝑓𝑓𝑓𝑓 𝑒𝑒𝑒𝑒𝑒𝑒ℎ 𝑑𝑑𝑑𝑑𝑑𝑑 𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑                                          
• Average ease of passage for each period is defined as 
Average ease of passage = Sum of ease scale  w/o rescue meds within 24 hrs 
Number of BMs w/o rescue meds without 24 hrs  
• Average stool consistency for each period is defined as 
Average stool consistency = Sum of  stool consistency wi/o rescue meds within 24 hrs 
Number of BMs w/o rescue meds within 24 hrs
 
• Days using rescue meds (i.e., laxative/suppository/enema) per week for each period is 
defined as 
Days using rescue meds = 𝑆𝑆𝑆𝑆𝑆𝑆 𝑜𝑜𝑜𝑜 𝐷𝐷𝐷𝐷𝐷𝐷𝐷𝐷 𝑤𝑤𝑤𝑤𝑤𝑤ℎ 𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑢𝑢𝑢𝑢𝑢𝑢 
𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁𝑁 𝑜𝑜𝑜𝑜 𝑑𝑑𝑑𝑑𝑎𝑎𝑎𝑎𝑎𝑎 𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑
× 7 𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑 
• Rescue med episodes per week for each period is defined as 
Rescue med episodes per week = Sum of daily rescue meds use 
Number of diary days
× 7𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑 
𝑤𝑤ℎ𝑒𝑒𝑒𝑒𝑒𝑒 𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑢𝑢𝑢𝑢𝑢𝑢= ൜#𝑜𝑜𝑜𝑜 𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒𝑒  𝑜𝑜𝑜𝑜 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑢𝑢𝑢𝑢𝑢𝑢 𝑓𝑓𝑓𝑓𝑓𝑓 𝑒𝑒𝑒𝑒𝑒𝑒ℎ 𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑 𝑑𝑑𝑑𝑑𝑑𝑑   
0 𝑖𝑖𝑖𝑖 𝑛𝑛𝑛𝑛 𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟𝑟 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚 𝑢𝑢𝑢𝑢𝑢𝑢 𝑓𝑓𝑓𝑓𝑓𝑓 𝑒𝑒𝑒𝑒𝑒𝑒ℎ 𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑𝑑 𝑑𝑑𝑑𝑑𝑑𝑑                         
 
All of the above parameters are treated as continuous variables. 
• Success (Yes/No) based on CSBM days  will be defined as “Yes” if either one criteria is 
met: 
a. if increase of one or more in CSBM days per week compared to baseline 
b. 3 or more of CSBM with CSBM per week 
Success based on CSBM will be treated as categorical variable and is primary endpoint. 


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4.4.2 Sleep Diary Data 
Sleep diary data are considered as exploratory nature.  For both stages, the PD parameters for 
each period from sleep diary are as below: 
• Total sleep time (hours) per night  
Total sleep time (hrs) per night = Sum of  daily total sleep time  
Number of Diary Days
 
• Delay (minutes)  to  sleep onset per night 
Delay to sleep onset (minutes) per night = Sum of  daily delay to sleep onset  
Number of Diary Days
 
• Total awake time (hours) per night 
Total awake time (hours) per night = Sum of  daily total awake time   
Number of Diary Days
 
• Number of wakes per night 
Average number of wakes per night = Sum of  daily number of wakes   
Number of Diary Days
 
•  Number of arm or leg thrashing days per week will be calculated as 
Number of arm or leg thrashing days per Week =  Sum of  days with arm or leg thrashing  
Number of Diary Days
× 7 days 
Arm or leg thrashing is defined as either arm or leg fidget or leg twitching from sleep 
behavior question. 
Based on number of arm or leg thrashing days per week,  
a. Disappearance of  arm or leg thrashing from sleep diary – defined as zero days of arm 
or leg thrashing per week 
Except for disappearance of arm or leg thrashing, all the above parameters will be treated as 
continuous outcomes.  
4.4.3 Questionnaire Data 
Questionnaire data will be considered as exploratory nature.  


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4.4.3.1 Non-Motor Symptoms Questionnaire 
Non-Motor Symptoms Questionnaire (NMSQ) comprises 30-item questionnaire. The answer for 
each question is Yes/No (coded as 1/0). The total score will be calculated as a summed score 
over 30-items [1].  
4.4.3.2 Beck Depression Inventory II 
Beck depression inventory II (BDI-II) consists of 21 items. Each item is rated on a 4 point scale 
ranging from 0-3.  For Question 16 and 18, the response categories are 0, 1a, 1b, 2a, 2b, 3a, or 
3b. For the calculation of total score, the letters a and b will be dropped for these questions. 
BDI-II total score will be calculated by summing the ratings for the 21 items based on the 
manual for Beck Depression Inventory-Second Edition.  The maximum total score is 63. 
Furthermore, total score will be classified into following categories. 
BDI-II Total Scores 
BDI-II Depression categories 
0-13 
Minimal 
14-19 
Mild 
20-28 
Moderate 
29-63 
Severe 
 
4.4.3.3 Mini Mental State Examination 
Mini mental state examination (MMSE) consists of 11 types of questions/activity. In each type, 
the number of questions can range from 1 to 5.  Score one point for each correct response for 
each question or activity within each type.  
According to MMSE manual, the total score will be calculated by summing the item scores 
across all 11 types of questions/activities. The possible maximum total score is 30.    
4.4.3.4 The University of Miami Parkinson’s Disease Hallucinations Questionnaire 
The University of Miami Parkinson’s Disease Hallucinations Questionnaire (UM-PDHQ) 
consists of 6 quantitative questions (Q1 to Q6) regarding severity of hallucinations and 14 
qualitative questions regarding quality of hallucination (Q7 to Q20). Question 1 is a gating 
question to assess the presence or absence of hallucination.  
The scoring ranges are not uniform in each item (Q3 and Q4: 0-2; Q2:0-4; Q5: 1-3; and Q6:0-3). 
According to the information on the questionnaire (total score [min=0, max=14]), the total score 
from 6 quantitative questions  will be calculated by summing the response scores over Q2 to Q6 


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if the answer of Question 1 is 1 or 2 while total score is equal to zero if the answer for Question 1 
is zero (i.e. no hallucinations). 
4.4.3.5 Parkinson’s disease Fatigue Scale 
Parkinson’s disease fatigue scale (PFS-16) consists of 16 questions. According to Appendix 15 
of protocol, the responses to each question are “Strongly disagree”, “Disagree”, “Neither agree 
nor disagree”, “Agree”, and “Strongly agree”. The scoring method used for the study is  
PFS-16 Response 
Score 
Strongly disagree 
1 
Disagree 
2 
Neither agree nor disagree 
3 
Agree 
4 
Strongly agree 
5 
 
The total score will be calculated by summing scores over 16 questions. [2, 3] The possible total 
scores range from 16 to 80. 
4.4.3.6 Patient Assessment of Constipation Symptoms 
Patient Assessment of Constipation Symptoms (PAC-SYM) is about constipation in the past two 
weeks and consists of 12 items. The scores for each item is  
PAC-SYM Response 
Score 
Absent 
0 
Mild 
1 
Moderate 
2 
Severe 
3 
Very Severe 
4 
There are total score and 3 subdomain scores: abdominal symptoms (Item 1-4), rectal symptoms 
(Item 5-7), and Stool symptoms (Item 8-12). Following the information booklet (2nd Edition, 
May 2015) from Mapi Research Trust, a subscale score is calculated as the average item score of 
non-missing items within that subscale. Total score is the average score of non-missing items 
within the instrument. 


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4.4.3.7 Patient Assessment of Constipation Quality of Life 
Patient Assessment of Constipation Quality of Life (PAC-QoL) is to measure the impact 
constipation has had on subject’s daily life over the past 2 weeks.  
PAC-QoL: 2 Sets of Responses separated 
by “/” Depending on questions 
Score 
Not at all / Never 
0 
A little bit / Rarely 
1 
Moderately / Sometimes 
2 
A lot / Often 
3 
Extremely / Always 
4 
PCAC-QoL contains 28 items grouped into 4 subscales: Physical discomfort (Item 1-4, 
Psychosocial discomfort (Item 5-12), Worries and concerns (Item 13-23), and satisfaction (Item 
24-28).  
Following the information booklet (4th Edition, May 2015) from Mapi Research Trust, scores of 
items 18 and 25 to 28 are reversed meaning that the score corresponding to the response:  
• “Not at all”/”None of the time” will be 4  
• “A little bit”/”A little of the time” will be 3    
• “Moderately”/”Some of the time” will be 2  
• “Quite a bit”/”Most of the time” will be 1  
• “Extremely”/”All of the time” will be 0  
Then, subscale score are calculated as the average item scores of non-missing items within that 
subscale. Total score is calculated as the mean of 28 item score within the instrument. When 
more than 50% of items are missing for a subscale score, then that subscale score is set to 
missing. If a subscale is missing, the PAC-QoL total score is set to missing. 
4.4.3.8 REM Sleep Behavior Disorder Screening Questionnaire 
REM sleep behavior disorder screening questionnaire (RBDQ) comprises 10 items. Each item is 
answered as yes/no (yes=1 or no =0). According to Scaling and Scoring (Version 1.0: Sept 2016) 
from Mapi Research Trust, total score will be summed over all items/questions. The maximum 
score s 13 points. A score of 0 is given to item/question is not answered. The multiple answers 
for an item will be considered as missing data.  
4.4.3.9 Unified Parkinson’s disease Rating Scale 
Unified Parkinson’s disease rating scale (UPDRS) consists of  


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• Part I: Non-Motor Aspects of Experiences of Daily Living (nM-EDL) - Q1.1 to Q1.6 
completed by rater; Q1.7 to Q1.13 completed by subject 
• Part II: Motor Aspects of Experiences of Daily Living (M-EDL) - Q2.1 to Q2.13 
completed by subject 
• Part III: Motor Examination - Q3.1 to Q3.18 completed by examiner 
• Part IV: Motor Complications - Q4.1 to Q4.6 by rater 
 
Rating for items is from 0 (normal) to 4 (severe). Score for each part is obtained from the sum of 
corresponding item scores.  Total score is the summed score from the scores of 4 parts, 
4.4.3.10 Other Questionnaires 
Trail make A& B and Parkinson’s disease Sleep Scale (PDSS) will not be used for analyses but 
will be listed only. 
ROME-IV Criteria for Constipation (ROME-IV) is only collected at screening. It will be listed. 
 


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5 PLANNED ANALYSES 
5.1 Changes from planned analyses in the Protocol 
There are no changes from planned Analyses in the Protocol. 
5.2 Interim Analyses 
After Stage 1 data completed, Stage 1 analyses will be performed.  
5.3 Final Analyses and Reporting 
All final planned analyses per protocol and this SAP will be performed only after database lock. 
 


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6 ANALYSIS POPULATIONS  
All data will be presented in listings for the Enrolled population. A subject will be considered 
enrolled when the subject has been consented, screened, and all eligibility criteria have been 
confirmed in the electronic case report form (eCRF). 
6.1 Pharmacokinetic Population 
6.1.1 Pharmacokinetic Population for Stage 1 
The Pharmacokinetic (PK) Population will consist of all subjects who received ENT-01 and for 
whom there are adequate samples collected to provide analyzable PK parameters.  
Any subject who violates the protocol in a way which might influence the plasma concentration 
by time profiles will be excluded from the PK population.  
6.1.2 Pharmacokinetic Population for Stage 2 
Seven subjects from the ENT-01 group and three subjects from the placebo group will be 
selected for the Pharmacokinetic (PK) Population.   
Any subject who violates the protocol in a way which might influence the plasma concentration 
by time profiles will be excluded from the PK population. 
6.2 Safety Population  
For each stage, safety Population will consist of all subjects who receive at least one dose of 
ENT-01 during that stage.  
6.3 Full Analysis Set  
For each stage, full Analysis Set (FAS) includes subjects who are treated with ENT-01 in the 
treatment period and have the stool diary assessments both at baseline and at least one post 
baseline. 
6.4 Randomized population 
For Stage 2, randomized population included all subjects who are randomized and receive at 
least one dose of blinded treatment under the protocol dated 27 July, 2017. 


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7 STATISTICAL CONSIDERATIONS 
All analyses described in this plan are considered a priori analyses in that they have been defined 
prior to locking the database. All other analyses, if any, designed subsequently to locking the 
database, will be considered post hoc analyses and will be described as exploratory analyses in 
the Clinical Study Report.  
All summaries and statistical analysis will be performed by SAS® Version 9.3 or later for each 
stage separately.   
7.1 General Statistical Procedures 
Frequency distributions for categorical variables will be provided as counts of subjects and 
percentages. Percentages will be based on the number of subjects with a non-missing parameter 
unless missing category is presented. Percentages will be reported to one decimal place.  
The descriptive statistics for continuous variables will be number of subjects, arithmetic mean, 
standard deviation (SD), median, quartiles (Q1, Q3), minimum and maximum. Arithmetic mean, 
median, Q1, and Q3 will be reported to 1 more decimal place than the raw data, while the SD 
will be reported to two more decimal places than the raw data.  
In addition to the above descriptive statistics, the coefficient of variation (CV%) will also be 
used for the summaries of drug concentration data and pharmacokinetic parameter values. The 
geometric means will be calculated for AUC and Cmax values. 
All data listings that contain an evaluation date will also contain a relative study day. 
Missing values will not be imputed. 
7.2 Enrollment and Disposition  
7.2.1 Subjects Enrollment 
Subject enrollment will be summarized for each analysis population separately for each stage. 
The number of subjects in each analysis population will be presented. 
Enrollment information will be provided in a data listing. 
7.2.2 Subject Disposition 
7.2.3 Subject Disposition for Stage 1 
Subject disposition will be summarized for each dose of ENT-01 and overall. Number and 
percentage of subjects who complete each dose, number and percentage of subjects who 
discontinued from the study early by primary reasons will be presented. The denominators for 
calculating the percentages will be based on number of subjects enrolled. 


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Discontinued subjects will be provided in a data listing. 
7.2.4 Subject Disposition for Stage 2 
Subject disposition will be summarized for each period and for each analysis population. 
Number and percentage of subjects who complete the full protocol, number and percentage of 
subjects who discontinued from the study early by primary reasons will be presented. The 
denominators for calculating the percentages will be based on number of subjects in the analysis 
population. 
Discontinued subjects will be provided in a data listing. 
7.3 Protocol Deviations 
Important or significant protocol deviations (PD) will be assessed by sponsor personnel 
following Protocol Deviation Guideline outlined in Clinical Management Plan.  
• A PD is classified as important if there is the potential to impact the completeness, 
accuracy, and/or reliability of the study data, or affect a subject’s rights, safety, or well-
being.  
• An important PD is classified as significant if it is confirmed to adversely impact the 
completeness, accuracy, and/or reliability of the study data, or affect a subject’s rights, 
safety, or well-being.  
All PDs will be identified and finalized prior to database lock.   
All protocol deviations will be listed. Protocol deviations will be presented in summary tables 
separately for each stage for the Safety population. 
7.4 Demographics and Baseline Characteristics 
7.4.1 Demographics and Baseline Characteristics  
Demographic and baseline characteristics will be summarized overall for each analysis 
population as applicable.  
The demographic data are: 
• Age  
• Sex 
• Race 
• If female, childbearing potential (Yes/No)  
• Ethnicity 
• Height (cm) 


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• Baseline Weight (kg) 
• BMI (Body mass index, kg/m2, calculated as Weight (kg) / Height (m)2) 
      Conversions for height and weight are as follows: 
 
 Height (cm) = Height (inches) x 2.54 
             Weight (kg) = Weight (lb) x 0.4536 
The demographic data will be listed. 
7.4.2 Baseline Disease Characteristics 
Age at diagnosis of Parkinson’s disease, years with Parkinson’s disease, years of constipations, 
Hoehn and Yahr stage, and constipation severity – CSBM will be summarized as appropriate  for 
each analysis population. 
Baseline disease characteristics will be provided in a listing. 
7.4.3 Medical History 
In Stage 2, general medical history will be collected. The medical history will be coded using the 
current version of the Medical Dictionary for Regulatory Activities (MedDRA). The frequency 
count and percentage of patients experiencing any medical conditions will be tabulated by 
MedDRA system organ class (SOC) and preferred term (PT) for the Safety and Randomized 
population.  The denominators for calculating the percentages will be based on number of 
patients in the analysis population.  
A data listing of medical history will be provided. 
 
7.5 Prior and Concomitant Medications  
All medications as documented by the investigator will be coded using the World Health 
Organization (WHO) Drug Dictionary. The count and percentage of subjects who take 
prior/concomitant medications will be provided using therapeutic class and WHO Drug preferred 
name.  The denominators for calculating percentages will be based on the number of subjects 
overall in the Safety population. For the summary tables, if a subjects took a prior or concomitant 
medication more than once, the subject would be counted only once for each therapeutic class 
and preferred name. 
Prior medications are defined as medications taken prior to the first dose of study drug 
(ENT-01). Concomitant medications are defined as medications taken on or after the first dose 
study drug. Prior and concomitant medications will be summarized overall for the safety 
population.  
For Randomized population, concomitant medications will be summarized by treatment. 
All prior and concomitant medications, including ancillary medication will be listed. 


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7.6 Analysis of Pharmacokinetic Data 
7.6.1 Summary of Pharmacokinetic Concentration Data for Stage 1 
All plasma concentration data will be summarized based on the PK population by timepoint (pre-
dose, 1, 2, 4, 8 and 24 hours post-dose) and dose level in terms of n, mean, SD, CV%, median, 
minimum, maximum, geometric mean, and geometric CV%. 
Individual plasma concentration will be listed by timepoint and dosage. The concentration will 
be listed to the same degree of accuracy as supplied by the analytical laboratory and PK group. 
As well, the units of concentration will be presented as they are received from the bioanalytical 
laboratory and PK group. 
Individual concentration and mean plasma concentration will be displayed graphically.   
Summary of Pharmacokinetic Concentration Data for Stage 2 
Individual plasma concentration will be listed by timepoint for the PK population. The 
concentration will be listed to the same degree of accuracy as supplied by the analytical 
laboratory and PK group. As well, the units of concentration will be presented as they are 
received from the bioanalytical laboratory and PK group. 
Individual concentration will be displayed graphically using actual sampling times. 
7.6.2 Analyses of Pharmacokinetic Parameters 
7.6.3 Analyses of Pharmacokinetic Parameters for Stage 1 
Summary of PK parameters for each dose level will be presented in terms of n, mean, SD, CV%, 
median, minimum, maximum, geometric mean, and geometric CV%. Geometric mean and 
geometric CV% will only be calculated for AUC and Cmax. Individual derived PK parameters 
will also be listed by dose level. 
7.6.4 Analyses of Pharmacokinetic Parameters for Stage 2 
No PK parameters will be derived. 
7.7 Analysis of Pharmacodynamics Data 
Generally speaking, for Stage 1 and Stage 2, the summaries will be provided for active treatment 
ENT-01 or by dose level if deemed as necessary. The comparison is made to baseline for each 
PD variable up to the end of fixed dosing period plus washout period.  
For Randomized population of Stage 2, the descriptive summaries will be provided for active 
treatment and placebo. Due to small sample size resulting from amended protocol, there will be 
no formal statistical comparisons between active treatment and placebo for PD variables. 


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7.7.1 Analysis of Continuous Pharmacodynamics Data  
The following continuous PD parameters and their change from baseline will be summarized by 
period (run-in [baseline), dosing periods (diary) or treatment period (questionnaire data), and 
washout) for each appropriate population, using descriptive statistics as specified in Section 7.1 
(General Statistical Procedures). 
Stool Diary PD parameters: 
• CSBM days  per week 
• CSBM episodes per week (Stage 2 only) 
• SBM days per week 
• SBM episodes per week (Stage 2 only) 
• Average ease of passage 
• Average stool consistency 
• Days using  rescue meds per week 
• Number of rescue meds use per week  
Sleep Diary PD parameters: 
• Total sleep time (hours) per day  
•  Delay (minutes)  in sleep onset per night 
• Total awake time (hours) per night 
• Number of wakes per night 
• Number of arm or leg thrashing days per week  
Questionnaire PD parameters:  
• Total score from NMSQ   
• Total score from BDI-II 
• Total score from MMSE  
• Total score from UM-PDHQ 
• Total scores from PFS-16  
• Total score and 3 subdomain scores from PAC-SYM   
• Total Score and 4 subdomain scores from PAC-QoL 
• Total Score from RBDQ  
• Total score and Part I to III scores from UPDRS  
7.7.2 Analysis of Categorical Pharmacodynamics Data  
The following categorical PD parameters will be summarized by period (run-in [baseline], 
dosing periods [diary] or treatment period [questionnaire data], and washout) for each 


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appropriate population, using frequency distribution as specified in Section 7.1 (General 
Statistical Procedures). 
Stool Diary PD parameters: 
• Stage 1 CSBM based on dosing day diary  
• Stage 1 SBM based on dosing day diary 
• Success based on CSBM days per week (Stage 2 only) - primary efficacy outcome variable 
o The proportion of subjects for whom the drug was a success at the end of fixed 
period will be estimated with a binomial point estimate and corresponding 95% 
confidence interval. The observed proportion of successes will be compared to 
0.10 using an exact binomial test. Success will be defined as an increase in 
CSBM/week of 1 or more OR a CSBM/week of 3 or more. 
 
Sleep Diary PD parameters: 
• Disappearance of arm or leg thrashing (Yes/no) 
Questionnaire PD parameters: 
• 
BDI-II Depression categories 
7.8 Analysis of Safety Data  
7.8.1 Adverse Events 
Adverse events will be coded into SOC and PT using the current version of MedDRA. Severity 
of AEs will be assessed by investigators according to CTCAE (v4.03). In database, Grade 1 is 
labeled as Mild, Grade 2 as Moderate, and Grade 3 as Severe.  
AEs that have a possible, probable or definite relationship to study drug will be defined to be 
related to the study drug while others will be defined as “not related”. AEs with the closest 
relationship to the study drug will be used for summary. 
The number and percentage of subjects who experienced an AE during escalation and fixed 
dosing periods will be summarized by dose level (Stage 1: 75 mg up 200 mg; Stage 2: 75 mg up 
to 250 mg, fixed dose, and washout) and overall for each stage and for the Safety population. 
The denominator for calculating the percentages will be based on the number of subjects ever 
exposed to each dose and overall in the Safety Population.   
Additionally, the number and percentage of subjects who experienced an AE during 
randomization period will be summarized by treatment (Placebo and ENT-01). The 
denominators for calculating the percentages will be based on the number of subjects ever 
exposed to each treatment in the Randomized Population. 


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A treatment emergent adverse event (TEAE) is defined as adverse events with a date of onset (or 
worsening) on or after the start date of a treatment up to the start date of the next treatment. The 
following types of summaries will be provided: 
• Overview of TEAEs 
• TEAEs by SOC and PT 
• TEAEs by SOC, PT, and Maximum CTCAE toxicity grade 
• TEAEs by SOC, PT, and Maximum CTCAE toxicity grade >= 3 
• TEAEs by SOC, PT, and closest relationship to study drug 
• TEAEs by PT in decreasing frequency 
• Drug-related TEAEs by SOC and PT 
• Drug-related TEAEs by PT in decreasing frequency 
• Serious TEAEs by SOC and PT 
• TEAEs leading to study discontinuation by SOC and PT 
• TEAEs of special interest including nausea, diarrhea, and dizziness by SOC and PT.  
If a preferred term or system organ class was reported more than once for a subject, the subject 
would only be counted once in the incidence for that preferred term or system organ class.  
In tabulation by severity (i.e., CTCAE toxicity grade),  
• For a given preferred term, only the most severe preferred term for each subject will be 
included.  
• For a given system organ class, only the most severe system organ class for each subject 
will be included.  
Similarly, in tabulation by relationship, 
• For a given preferred term, the most closely related preferred term to the study drug for 
each subject will be included.  
• For a given system organ class, the most closely related system organ class to the study 
drug for each subject will be included.  
Adverse events including baseline AEs, serious adverse events, and adverse events leading to 
discontinuation from the study will be provided in separate data listings.  
7.8.2 Clinical Laboratory Tests 
Stage 1 and up to end of fixed dosing period in Stage 2 
In this study, clinical laboratory tests were done at local laboratories. Therefore, the reference 
ranges may not be the same. All laboratory test results will be summarized by period (run-in 
period, dose period, and washout period) using frequency distribution for low/normal/high or 
normal/abnormal. The denominators for calculating the percentages will be based on the number 
of subjects with non-missing values in the Safety Population. 


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Shift from baseline in laboratory parameters will be summarized by period for the Safety 
Population by period based on Low/Normal/High classification. The denominators for 
calculating the percentages will be based on the number of subjects with non-missing values at 
both baseline and each post-baseline analysis visit in the Safety Population. 
Randomization period in Stage 2 
Additionally, the above summaries will be provided for viral signs during the randomization 
period by treatment for randomized population. 
All clinical laboratory assessments will be listed. 
7.8.3 Vital Signs 
Stage 1 and up to end of fixed dosing period in Stage 2 
Vital signs (Stage 1 and up to end of fixed dosing period in Stage 2) for  body temperature, 
respiration rate, heart rate and weight will be summarized by period, together with the change 
from baseline. Baseline for body temperature, respiration rate, heart rate and weight will be the 
last assessment during the run-in period.    
Systolic and diastolic blood pressure will be summarized by time point (pre-dose and 2 hours 
post-dose) and dose period separately for each position (lying down and sitting). Baseline for 
predose in each dose period and washout will be the last assessment during the run-in period. 
Baseline for 2 hour post dose in each dose period will be the assessment at predose at that dose 
period. 
All summaries will be provided for the safety population. 
Randomization period in Stage 2 
Additionally, the above summaries will be provided for viral signs during the randomization 
period by treatment for randomized population. 
7.8.4 Electrocardiogram (ECG) 
Stage 1 and up to end of fixed dosing period in Stage 2 
ECG findings will be classified as normal vs abnormal. The number and percentage of each 
category will be summarized using frequency table for run-in, dose period, and wash-out. The 
denominators for calculating the percentages will be based on the number of subjects with non-
missing values in each period for the Safety Population. Baseline will be defined as the most 
recent measurement taken in the run-in period. 


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Randomization period in Stage 2 
Additionally, the above summaries will be provided for ECG during the randomization period by 
treatment for randomized population. 
Detailed ECG results will be provided in a data listing. 
7.8.5 Physical Examination 
Stage 1 and up to end of fixed dosing period in Stage 2 
Summary of physical examinations will present frequency distribution by body system and each 
period. The denominators for calculating the percentages will be based on the number of subjects 
evaluated for a particular body system of each dose level in the Safety Population. 
Randomization period in Stage 2 
Additionally, the above summaries will be provided by treatment for randomized population. 
Details on physical examinations will be provided in a data listing. 
7.8.6 Analysis of Tolerability Data 
Stage 1 and up to end of fixed dosing period in Stage 2 
Tolerability will be measured by DLT endpoints defined in Section 4.3. 
The frequency of each tolerability endpoints including recurrent vomiting, recurrent diarrhea, 
abdominal pain and postural hypertension, and etc. will be summarized by dose (Stage 1) or for 
treatment period (Stage 2). Percentages will be based on the number of subjects in each dose 
level (Stage 1) or the number of subjects (Stage 2) in the safety population. 
Randomization period in Stage 2 
Additionally, the above summaries will be provided during the randomization period by 
treatment for randomized population. 
7.8.7 Extent of Exposure 
For Stage 1, dose administration details by subject will be listed.  
For Stage 2, number of subjects and percentage on each dose level will be presented for period 
3.2. The denominators for percentages will be based on the number of subjects in the Safety 
Population. For each dose period, total dose (mg), extent of exposure (days), and applicable 
compliance (%) captured on the eCRF page will be summarized using descriptive statistics. 


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7.8.8 I-BUTTON 
I-Button data will be analyzed by a separate group. The data will not be included in this SAP. 
7.9 Additional Data Presentation as Listing 
Additional data listings will be provided for the following information as appropriate: 
• Subject eligibility 
• Medical history 
• Constipation history 
• Stool diary 
• Sleep diary 
• Individual questions from questionnaires: 
o NMSQ   
o BDI-II 
o MMSE  
o UM-PDHQ 
o PFS-16  
o PAC-SYM   
o PAC-QoL 
o RBDQ  
o UPDRS 
o ROME-IV 
o Trail make A 
o Trail make B 
o PDSS 
 
 


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8 DEFINITIONS AND CONVENTIONS FOR DATA HANDLING 
8.1 Definition of Baseline 
For all evaluations unless otherwise noted, baseline is defined as the most recent measurement 
prior to the first administration of study drug in Stage 1. Baseline can be the same date as first 
dose, given the measurement is expected prior to first dose when only date information is 
available. 
8.2 Analysis Visit Window 
For safety parameters excluding clinical laboratory data, measurements collected from 
unscheduled visits will not be included in the by-visit summary tables but will be included in the 
listings. Early termination visits for safety measurements will not be mapped to any scheduled 
post-baseline visit, but will be used as the last assessment during treatment period. 
8.3 Pharmacodynamic Data Handling 
All PD parameters from questionnaires will follow instructions or manuals coming in with them. 
If any special handling rules were needed, they would be documented in ADaM data 
specifications.  
For PD parameters from diary data, generally, there will be no imputation for missing diaries. 
However, if there were some partial dates or times not in 24-hour clock present in diary data, 
they might be imputed in order to derive PD parameters, using common sense. These details of 
handling rules would be documented in ADaM data specifications.  
8.4 Safety Data Handling 
For all safety data, only observed data will be used for analyses, and missing data will not be 
imputed. 
8.5 Handling of Repeated Clinical Laboratory Tests 
For laboratories results at unscheduled visits, it will be treated as repeated laboratory results for 
the closest previous visit. The last repeat of laboratory results will be used in the summary tables 
for that visit. All the laboratory test results (original test results and repeated results) will be 
included in the data listings. 
8.6 Pharmacokinetic Data Handling 
8.6.1 Pharmacokinetic Plasma Drug Concentration Data 
Pharmacokinetic concentration values below the quantification limit (BLQ) will be treated as 
zero for the calculation of summary statistics.   


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Individual values that are BLQ will be presented as “BLQ” in the concentration data listing.  
No imputation will be performed for missing concentration.   
8.7 Handling of Partial Dates for AEs 
When determining the treatment emergent AE, partial dates will be handled as follows.  
• If the day of the month is missing, the onset day will be set to the first day of the month 
unless it is the same month and year as first dose date. In this case, the onset date will be 
assumed to be the first date of treatment. 
• If the onset day and month are both missing, the day and month will be assumed to be 
January 1, unless the event occurred in the same year as the study treatment. In this case, 
the event onset will be coded to the day of treatment to conservatively report the event as 
treatment-emergent.  
• A missing onset date will be coded as the day of treatment. If the resulting onset date is 
after a reported date of resolution, the onset date will be set equal to the date of 
resolution.   
• Imputation of partial dates is used only to determine whether an event is treatment-
emergent; data listings will present the partial date as recorded in the eCRF. 
8.8 Handling of Partial Dates for Medications 
A medication with a completely missing start date will be considered a prior medication. A 
medication with a completely missing stop date will be considered to be ongoing.   
When determining prior or concomitant medications, partial start dates of prior and concomitant 
medications will be assumed to be the earliest possible date consistent with the partial date. 
Partial stop dates of prior and concomitant medications will be assumed to be the latest possible 
date consistent with the partial date.  


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9 Changes to the Planned Analyses in the Protocol(s) 
Because protocol amendment dated 02 February 2018 removed randomization portion during the 
trial, there will not be enough samples size to do the following analysis specified in the original 
protocol dated dated 27 July 2017.  Therefore, the following analysis will not be performed for 
CSR. 
• A secondary analysis will compare the proportions of subjects who are deemed a success 
at the end of the randomized fixed-dose period between those randomized to the ENT-01 
arm and those randomized to the placebo arm. Subjects to be included in this analysis 
will only be those who have been randomized. If the number of subjects for this analysis 
is between 20 and 30, a Fisher’s exact test will be used to compare the proportions of 
subjects who are deemed a success at the end of randomization period between the two 
randomized arms. 
 
The protocol also proposed the following analysis as below:  
• Another secondary analysis will compare the proportions of subjects who are a success 
(compared to baseline) between the end of the first week of the washout period (period 5) 
and the end of the fixed-dose period (period 4). This analysis will consist only of those 
subjects with assessments at baseline (end of run-in period, Period 3.1), the end of Period 
4, and the end of Period 5. If the number of subjects for this analysis is between 17 and 
20 subjects, this analysis will be done using an exact sign test of equality of paired 
proportions. 
In this analysis, using the end of the first week of washout is not consistent with the time frame 
for other PD parameters (note: other PD parameters’ are all evaluated using two weeks of 
washout data, i.e., the end of 2 weeks). Therefore, it was determined we would not do this 
analysis for CSR.  
 
 
 
 


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10 REFERENCES 
1. 
Chaudhuri, K.R., et al., International multicenter pilot study of the first comprehensive 
self-completed nonmotor symptoms questionnaire for Parkinson's disease: the NMSQuest 
study. Mov Disord, 2006. 21(7): p. 916-23. 
2. 
Friedman, J.H., et al., Fatigue rating scales critique and recommendations by the 
Movement Disorders Society task force on rating scales for Parkinson's disease. Mov 
Disord, 2010. 25(7): p. 805-22. 
3. 
Grace, J., A. Mendelsohn, and J.H. Friedman, A comparison of fatigue measures in 
Parkinson's disease. Parkinsonism & Related Disorders, 2007. 13(7): p. 443-445.