Clinical trial NCT02931565
Conditions: Achalasia
Sponsor: Cyclerion Therapeutics
Start date: 2017-04-06; Completion date: 2018-05-01
Enrollment: 9 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
The objectives of this study are as follows:

In participants with primary Type I or II achalasia, following a single 5-mg dose of olinciguat (IW-1701),

* To assess the safety and tolerability
* To determine the effects on measures of esophageal function by high-resolution impedance manometry (HRIM)
* To determine the pharmacokinetic (PK) parameters

Detailed description:

Arms: arm 1: Single 5-mg dose of IW-1701 administered orally arm 2: Matching placebo administered orally
Interventions: intervention 1: Olinciguat intervention 2: Matching Placebo
Intervention descriptions: intervention 1: oral tablet intervention 2: oral tablet

Protocol documents:
Prot_000.pdf: 


IW-1701 Tablets, 1 mg 
 
Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
03 October 2017 
 
Confidential 
Page 2 of 83 
STUDY IDENTIFICATION 
A summary of key study participants is provided in Table 1. All study contact details will be 
provided prior to the Site Initiation Visit. 
Table 1. 
Key Study Participants 
Role 
Contact Information 
Medical Monitor: 
 
 
 
Clinical Laboratory: 
 
 
 
 
 
Contract Research Organization 
(CRO) Contact: 
 
 
 
 
   
 
 
CRO Medical Monitor 
 
 
 
 
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IW-1701 Tablets, 1 mg 
 
Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
03 October 2017 
 
Confidential 
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SYNOPSIS 
Sponsor 
Ironwood Pharmaceuticals, Inc. 
Name of Finished Product 
IW-1701 Tablets, 1 mg 
Name of Active Ingredient 
IW-1701 
Study Title 
A Multicenter, Randomized, Double-blind, Placebo-controlled, Parallel-group, Single-dose, 
Phase 2a Study to Evaluate the Safety, Tolerability, Pharmacokinetics, and Pharmacodynamics 
of IW-1701 in Patients with Achalasia  
Study Number 
C1701-201 
Study Phase: 2a 
Objectives  
In patients with primary Type I or II achalasia, following a single 5-mg oral dose of IW-1701, 
• To assess safety and tolerability  
• To determine the effects on measures of esophageal function by high-resolution impedance 
manometry (HRIM) 
• To determine the pharmacokinetic (PK) parameters, Cmax, Tmax, AUClast. 
Study Design 
This multicenter, randomized, double-blind, placebo-controlled, parallel-group, single-dose 
study will randomize up to 20 patients to receive study drug (15 patients to IW-1701 and 5 to 
matching placebo). 
Study Population  
The study will randomize patients diagnosed with primary Type I or II achalasia with an 
integrated relaxation pressure (IRP) > 15 mm Hg by baseline HRIM. Up to 20 patients are 
planned. 
See Eligibility Criteria for full inclusion and exclusion criteria. 
Test Product, Dose, and Mode of Administration  
Test product (IW-1701 Tablets, 1 mg) will be 1 mg oral tablets; the dose will be 5 mg 
IW-1701 (5 tablets) administered orally. 
Reference Therapy, Dosage, and Mode of Administration 
Placebo will match the IW-1701 oral tablets. The number of placebo tablets administered will 
match the number of tablets used for the IW-1701 treatment arm. 
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IW-1701 Tablets, 1 mg 
 
Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
03 October 2017 
 
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Study Drug Administration 
On the morning of Day 1, after an overnight fast of ≥ 8 hours and after the baseline HRIM 
procedure, randomized patients will receive 5 mg IW-1701 or placebo in a tablet formulation. 
Study drug will be administered with ~ 240 mL (8 fluid ounces) of water; patients should be 
encouraged to consume all ~ 240 mL of water. (Note: Patients are allowed to take multiple 
tablets together, and additional water is allowed to complete dosing.) Fluids will not be 
allowed for at least 1 hour before study drug administration, and except for the water given 
during HRIM study procedures, fluids will not be allowed from at least 1 hour before HRIM 
catheter insertion until after catheter removal. Solid food will not be allowed from Day -1 until 
at least 4 hours postdose. See Exclusion Criteria for prohibited concomitant medications. Oral 
medications not prohibited by the protocol may be taken on Day 1; administration should be 
timed to avoid interfering with study procedures. 
Stopping Criteria 
Dosing of additional patients will be suspended if any of the following occur:  
• Drug-related SAEs in 2 or more patients (per causality and SAE definitions in this 
protocol)  
• Drug-related AE in 1 or more patients that is life threatening or requires urgent 
intervention || 
• An overall pattern of clinically significant treatment-emergent AEs that may appear minor 
in terms of an individual event but collectively represents a safety concern 
• Treatment-emergent AEs judged to be drug-related and reported per the description and in 
the number of patients indicated in the table below. These events are based on clinical 
experience with IW-1701 and the prescribing information for riociguat.  
 
Treatment-emergent,  
Drug-related Event 
Event 
Description 
# of patients that 
trigger stopping 
Symptomatic hypotension-related 
events (eg, syncope, presyncope) 
Medical intervention or hospitalization indicated* 
2 
Vomiting 
≥ 6 episodes (separated by at least 5 minutes) in 
24 hours† 
2 
Spontaneous bleeding events ‡ 
(including hemoptysis, vaginal 
hemorrhage, ovarian hemorrhage, 
subdural hemorrhage, hematemesis, 
hematochezia)  
Moderate symptoms, medical intervention 
indicated § 
2 
Life-threatening consequences; urgent 
intervention indicated || 
1 
* Consistent with National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE) 
Version 4.0, Grade 3 
† CTCAEv4 Grade 3 
‡ Based on riociguat prescribing information  
§ Consistent with CTCAEv4 Grade 2 
|| Consistent with CTCAEv4, Grade 4  
 
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Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
03 October 2017 
 
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Page 7 of 83 
If stopping criteria are met, the study may continue only after the study protocol has been 
amended, if appropriate, and approved by the IRB. 
(Note: This is a single-dose study; therefore, there are no individual subject stopping rules.) 
Discharge Criteria 
Patients may be discharged on Day 2 at the Investigator’s discretion and at least 30 hours after 
study drug administration if the following criteria are met: 
• Systolic BP ≥ 90 mm Hg and diastolic BP ≥ 60 mm Hg 
• No unresolved AEs that the Investigator judges to be drug related 
 
Duration of Treatment  
Patients will receive a single oral dose of IW-1701 and will be followed in the clinic for at 
least 30 hours after dosing. Total patient participation will be 15 to 56 days, including the 
Screening, Clinic, and Follow-up Periods. 
Pharmacodynamic Assessments 
HRIM 
HRIM Methods: After calibration, the HRIM catheter will be inserted transnasally to record 
manometric data from the hypopharynx to the stomach. After insertion and ~ 2-minute 
baseline (relaxed) recordings to assess basal esophagogastric junction (EGJ) pressure, the 
following set of swallowing protocols will be recorded: 
• 10 supine (at ~ 30-degree incline) ~ 5-mL saline swallows 
• 5 upright ~ 5-mL saline swallows (after upright and relaxed for ~ 1 minute) 
• (Optional) Standing ~ 200-mL saline bolus challenge (at both baseline and postdose) 
This procedure will be performed 2 times, once at baseline and once after dosing. The baseline 
and postdose procedures should be identical. For the postdose recording, the swallowing 
protocol should begin at 3 hours (+15 minutes) postdose.  
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IW-1701 Tablets, 1 mg 
 
Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
03 October 2017 
 
Confidential 
Page 8 of 83 
HRIM Measurements   
HRIM measurements for PD analyses will be made by central, blinded reader(s). 
• Bolus Flow Time (BFT) – time in seconds of bolus (~ 5 mL swallow) transit through the 
EGJ from the 10-swallow supine and 5-swallow upright protocols; median measurement 
from the 10-swallow supine and 5-swallow upright protocols 
• Integrated Relaxation Pressure (IRP) – mean of the 4s of maximal deglutitive relaxation 
in the 10-s window beginning at upper esophageal sphincter relaxation. Contributing times 
can be contiguous or non-contiguous (eg, interrupted by diaphragmatic contraction). 
Referenced to gastric pressure; median measurement from the 10-swallow supine and 
5-swallow upright protocols 
• Impedance bolus height (IBH) – height in esophagus of ~ 200 mL saline bolus at ~ 1, 
~ 2, and ~ 5 minutes postbolus from the standing ~ 200-mL bolus challenge (if performed) 
 
Patient-reported Symptom Questions 
The patient will complete a symptom assessment just after removal of the catheter at both the 
baseline and the postdose HRIM procedures. 
Pharmacokinetic Assessments  
If systemic levels of IW-1701 are detectable, the following PK parameters will be calculated, 
when appropriate: 
• AUClast: Area under the plasma concentration time curve from time zero to Tlast, the time at 
which the last measurable plasma concentration (Clast) is observed 
• Cmax: Maximum observed plasma concentration, occurring at Tmax 
• Tmax: Time of maximum observed plasma concentration 
Additional Questionnaires 
• Health status as assessed employing the EQ-5D-3L 
• Health-related quality of life as assessed employing the SF-12v2 
• Screening Patient-reported Symptom Questions  
Safety Assessments 
Adverse event recording, physical examination, clinical laboratory tests, vital sign parameters, 
and ECGs 
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IW-1701 Tablets, 1 mg 
 
Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
03 October 2017 
 
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Statistical Methods:  
Sample Size Determination 
For this Phase 2a study, a sample size of up to 20 patients is planned. The sample size of 
20 was determined outside of statistical considerations and is considered reasonable based on 
precedent set by prior studies of similar nature and design.  
Analysis Populations 
All patients who receive study drug will be included in the Safety Population. 
All patients who receive study drug and have at least 1 postdose PK parameter assessment will 
be included in the PK Population. 
All patients who receive study drug and have at least 1 postdose PD assessment will be 
included in the PD Population. 
Statistical Analysis 
Descriptive statistics (n, mean, standard deviation, minimum, median, and maximum) will be 
calculated to summarize continuous variables. Frequency and percent of patients in each 
category will be calculated to summarize categorical variables. 
Pharmacodynamic Analyses 
Change from baseline will be calculated for continuous variables and summarized by 
treatment group. 
Safety Analyses 
Adverse events will be summarized by system organ class (SOC), preferred term (PT), and 
treatment group. Listings will be provided for pretreatment AEs, treatment-related AEs, severe 
AEs, drug-related AEs, serious AEs (SAEs), and AEs leading to study discontinuation. 
Descriptive statistics will be provided for the safety parameters. 
ECGs, vital signs, and clinical laboratory tests will be summarized at each time point and 
listings will be provided for patients with abnormal values. 
Pharmacokinetic Analyses 
If plasma concentrations of IW-1701 are detected, the PK parameters will be determined and 
tabulated, and summary statistics will be reported. 
 
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Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
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ELIGIBILITY CRITERIA 
INCLUSION CRITERIA 
Patients must meet all of the following criteria to be eligible for enrollment in this study: 
1. Patient has signed an ICF before any study-specific procedures are performed.  
2. Patient is a male or female between 18 and 75 years old at the Screening Visit. 
3. Female patient is not pregnant or breastfeeding at the time of the Screening Visit and must 
have a negative serum pregnancy test at the Screening Visit and a negative urine pregnancy 
test on Day 1 confirmed negative before Randomization.  
4. Patient has a diagnosis of primary Type I or II achalasia per Investigator judgement, and 
median IRP > 15 mm Hg from supine 10-swallow protocol on baseline HRIM procedure as 
read by the Investigator. 
5. Patient must have an endoscopy documented in their medical history that was performed 
within the 90 days before Day 1. 
6. Patient is in generally good health, is in medically stable condition, and has no clinically 
significant findings on a physical examination, 12-lead ECG, and clinical laboratory tests 
(serum chemistry, hematology, coagulation, urine drug screen, and urinalysis) after signing 
the ICF but before receiving the dose of study drug. (Note: The Investigator will determine if 
a particular finding is clinically significant. In making this determination, the Investigator 
will consider whether the particular finding could prevent the patient from performing any of 
the protocol-specified assessments, could represent a condition that would exclude the patient 
from the study, could represent a safety concern if the patient participates in the study, or 
could confound the protocol-specified assessments). 
7. Patient has seated systolic BP from 100 to 160 mm Hg and seated diastolic BP from 60 to 
100 mm Hg at the Screening Visit. (Note: BPs are the averages of 3 measurements obtained 
with an appropriately sized cuff at 2-minute intervals after the patient has been sitting quietly 
for ≥ 5 minutes.) 
8. Patient has platelet count, prothrombin time, International Normalized Ratio (INR), and 
activated partial thromboplastin time (aPTT) within laboratory normal ranges at the 
Screening Visit; if values are out of normal ranges, test may be repeated to determine 
eligibility. 
9. Patient has no contraindications to the performance of the baseline and postdose HRIM 
procedures per Investigator discretion. 
10. Female patient must be postmenopausal or surgically sterile (ie, bilateral oophorectomy, 
hysterectomy, or tubal ligation); must agree to completely abstain from heterosexual 
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Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
03 October 2017 
 
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intercourse; or, if heterosexually active, must agree to use 1 of the following methods of birth 
control from the date she signs the ICF until 90 days after the dose of study drug: 
a. Combination of 2 highly effective birth control methods (eg, condom with spermicide 
plus intrauterine device, condom with spermicide plus a diaphragm or cervical cap, 
hormonal contraceptive [including progesterone implant] combined with a barrier 
method) 
b. Maintenance of a monogamous relationship with a male partner who has been surgically 
sterilized by vasectomy (vasectomy procedure must have been conducted ≥ 60 days 
before the Screening Visit or confirmed via sperm analysis) plus a hormone or barrier 
method 
11. Male patient must be surgically sterile by vasectomy (vasectomy procedure conducted ≥ 60 
days before the Screening Visit or confirmed via sperm analysis), must agree to completely 
abstain from heterosexual intercourse, or, if heterosexually active, must agree to use a 
combination of 2 highly effective birth control methods (eg, condom with spermicide plus 
intrauterine device, condom with spermicide plus a diaphragm or cervical cap, hormonal 
contraceptive [including progesterone implant] combined with a barrier method) from date 
they sign the ICF until 90 days after the dose of study drug. 
12. Patient is fluent and literate in English. 
 
EXCLUSION CRITERIA 
Patients who meet any of the following criteria will not be eligible to participate in the study: 
1. Patient has any autoimmune disease, peptic esophagitis, or evidence of esophageal rings, per 
Investigator judgement. 
2. Patient has malignant or premalignant esophageal lesions. 
3. Patient has history of migraine requiring treatment in the last 6 months. 
4. Patient has had any of the following: 
• Any prior esophageal, periesophageal, or gastric surgery, or treatment with sclerosing 
agent  
• More than 1 pneumatic dilation procedure to a diameter of > 2 cm 
• Pneumatic dilation procedure to a diameter of > 2 cm within 1 year prior to 
randomization  
• Prior esophageal injection of botulinum toxin (Botox) within 6 months prior to 
randomization  
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Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
03 October 2017 
 
Confidential 
Page 12 of 83 
• More than 2 esophageal Botox injection procedures in his/her lifetime  
Note: Prior bougie dilation(s) or pneumatic dilation(s) of ≤ 2 cm are allowed. 
5. Patient has known megaesophagus (esophageal diameter > 5 cm), sigmoid esophagus, or 
esophageal diverticulum, per Investigator judgement.  
6. Patient has history of platelet dysfunction, hemophilia, von Willebrand disease, coagulation 
disorder, or other bleeding diathesis condition(s), even if patient has normal complete blood 
count, prothrombin time, and aPTT. 
7. Patient has used PDE5 inhibitors (including sildenafil and tadalafil), mono- or 
di-nitroglycerine products, riociguat, calcium-channel blockers, opiates, or aspirin in the 
7 days before Check-in on Day 1. Patient has used bisphosphonates in the 3 months before 
Check-in on Day 1. These medications are not allowed until after the Follow-up Call. 
Patient has used CYP3A4 inhibitors or inducers in the 7 days before Check-in on Day 1. 
CYP3A4 inhibitors or inducers are not allowed until after the End of Treatment Visit.  
8. Patient has taken any supplements for the treatment of erectile dysfunction during the 7 days 
before Check-in/Day 1. Supplements for the treatment of erectile dysfunction are not allowed 
until after the Follow-up Call. 
9. Patient has a history of active alcoholism or drug addiction during the year before the 
Screening Visit or has a positive result for illicit drugs at the Screening Visit. Alcoholic 
beverages are not permitted from the Screening Visit until after the Follow-up Call (Day 4 
± 1 day).  
10. Patient has taken any drug that can affect GI motility in the 72 hours before Check-in through 
Discharge from the clinic. 
11. Patient has a 12-lead ECG demonstrating severe bradycardia (heart rate < 40 beats per 
minute) or average QTcF ≥ 450 msec for male patients or ≥ 470 msec for female patients at 
the Screening Visit. (Note: If on initial ECG, QTcF exceeds criteria, the ECG will be 
repeated 2 more times, and the average of the 3 QTcF values will be used to determine the 
patient's eligibility). 
12. Patient has elevated (> 1.5 times the upper limit of normal as defined by the laboratory) 
levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), or creatinine at 
the Screening Visit. 
13. Patient has history of cancer within the past 5 years, other than basal cell carcinoma. 
14. Patient has active hepatitis B, hepatitis C, or history of human immunodeficiency virus 
infection. 
15. Patient has hepatic impairment defined as Child-Pugh A, B, C. 
16. Patient has known eating disorder (eg, anorexia nervosa, binge eating, bulimia). 
17. Patient is chronic, heavy smoker (> 25 cigarettes/day). 
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Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
03 October 2017 
 
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18. Patient has a history of clinically significant hypersensitivity or allergies to any of the 
inactive ingredients contained in the active or placebo drug products. 
19. Patient has previously received IW-1701 in a study, or received an investigational drug 
during the 30 days or 5 half-lives of that investigational drug (whichever is longer) before the 
Screening Visit, or is planning to receive another investigational drug at any time during the 
study. 
20. Patient has undergone a surgery during the 30 days before Check-in except minor dental and 
dermatological procedures. 
21. Female patient who may wish to become pregnant and/or plan to undergo egg donation or 
egg harvesting for current or future in vitro fertilization during the course of the study and for 
at least 30 days after the dose of study drug. 
22. Male patient unwilling to refrain from sperm donation during the study and for at least 90 
days after the dose of study drug. 
23. Patient has an acute or chronic condition that, in the Investigator’s opinion, would limit the 
patient’s ability to complete or participate in this clinical study. 
24. Patient is involved in the conduct and administration of this study as an Investigator, 
sub-Investigator, study coordinator, other study staff, or Sponsor member. 
 
Note: Screening Visit assessments may take place over more than 1 day. Patients may be 
rescreened.  
At the Investigator’s discretion and after consultation with the Medical Monitor, during the 
Screening Period, laboratory values that are outside specified ranges may each be repeated once 
to confirm eligibility 
 
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Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
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SCHEDULE OF EVENTS 
Study Period 
Screening Period 
Clinic Period 
Follow-up Period 
Visit Days → 
Study Procedure ↓ 
Screening Visit  
Day -28 to -1 
Dosing 
Day 1  
Discharge  
Day 2 
Follow-up Call 
Day 4 (±1) 
End of Trial Visit 
Day 21 (±7) 
Informed consent signed 
X 
 
 
 
 
Inclusion/exclusion evaluation 
X 
X 
 
 
 
Demographics 
X 
 
 
 
 
Medical history 
X 
 
 
 
 
Drug screen (a) 
X 
 
 
 
 
HBsAg, HCV, & HIV Screen 
X 
 
 
 
 
Pregnancy test (b) 
X 
X 
 
 
X 
Weight (W) & height (H) 
W, H 
W 
 
 
W 
Physical exam 
X 
 
 
 
X 
12-lead ECG (c) 
X 
predose 
pd:  4h (±30m) 
pd:  24h (±30m) 
 
X 
Vital signs (oral temperature, 
respiratory rate, seated BP & pulse) (d) 
X 
pre: 0 (≤ 15m) 
BP & pulse only: 
pd:  0.5h (±10m), 1, 2, 3, 4, 
6, 8, 12h (±15m) 
pd:  24h (±30m) 
At discharge (30 h 
or later): BP only 
 
 
X 
Clinical chemistry, coagulation, 
hematology, urinalysis (e) 
X 
 
pd:  24h (±30m) 
 
X 
Adverse event evaluations 
X 
X 
X 
X 
X 
Prior & concomitant medications 
X 
X 
X 
X 
X 
EQ-5D-3L 
X 
 
 
 
X 
SF-12v2 
X 
 
 
 
X 
Screening patient-reported symptom 
questions 
X 
 
 
 
 
Optional genotyping sample (f) 
 
 
pd:  24h (±30m) 
 
 
PK/PD blood samples (g) 
 
pre: 0 (≤ 15m) 
pd:  0.5h (±2m),1,2,3,4,5 
6,8h (±5m),12h (±15m) 
pd: 17h (±15m), 
24h (±30m) 
 
X 
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Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
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Study Period 
Screening Period 
Clinic Period 
Follow-up Period 
Visit Days → 
Study Procedure ↓ 
Screening Visit  
Day -28 to -1 
Dosing 
Day 1  
Discharge  
Day 2 
Follow-up Call 
Day 4 (±1) 
End of Trial Visit 
Day 21 (±7) 
Randomization 
 
X 
 
 
 
Study drug administration (h) 
 
X 
 
 
 
High resolution impedance manometry 
(HRIM) procedure (i) 
 
predose 
pd: 3h (+15m) 
 
 
 
Patient-reported symptom questions  
 
after pre HRIM (+15m) 
after pd HRIM (+15m) 
 
 
 
Discharge (j) 
 
 
X 
 
 
Study completion 
 
 
 
 
X 
0=time of dose (ie, swallowing of initial tablet[s]); BP = blood pressure; ECG = electrocardiogram; h = hour; H = height; HBsAg = hepatitis B surface antigen; 
HCV = hepatitis C virus; HIV = human immunodeficiency virus; m = minute; msec = millisecond; pd = postdose; PK = pharmacokinetic; 
PD = pharmacodynamic; pre = predose; W = weight 
a. 
Urine drug screen for selected illicit drugs  
b. For female patients, a negative serum pregnancy test must be documented at the Screening and Follow-up Visits; a negative urine pregnancy test must be 
documented on Day 1 with results available before Randomization. 
c. 
Patients must be supine for ≥ 5 m before the ECG recording (Note:  If on initial ECG, QTcF is ≥ 450 msec for male patients or is ≥ 470 msec for female 
patients, the ECG will be repeated 2 more times, and the average of the 3 QTcF values will be used to determine the patient's eligibility). Before blood draws 
and vital signs where applicable. The predose ECG may be completed any time on Day 1 prior to dose administration. 
d. Oral temperature (°C), respiratory rate, systolic and diastolic BP, and pulse after the patient has been sitting for ≥ 5 m. At the Screening Visit, BPs will be 
the average of 3 measurements obtained at 2-m intervals after the patient has been sitting quietly for ≥ 5 m; vital sign measurements will be taken before 
HRIM procedures and before blood draws where applicable. 
e. 
Patients must fast for ≥ 8 h before sample collection in the morning. If the Screening Visit is Day -2 or -1, local labs results may be used to determine 
eligibility.  
f. 
Optional:  one 4-mL blood sample for genotyping 
g. One ~ 6 mL blood sample at each timepoint for PK and exploratory PD (plasma renin activity) assessment, each sample divided into 2 equal plasma 
aliquots; samples taken before HRIM procedures when applicable; record exact time of blood draw 
h. In the morning, after overnight fast of ≥ 8 h, after baseline HRIM procedure and randomization, study drug will be administered with ~ 240 mL (~ 8 fluid 
ounces) of water; patients should be encouraged to consume all ~ 240 mL of water. (Note:  Patients are allowed to take multiple tablets together, and 
additional water is allowed to complete dosing.) Fluids will not be allowed for at least 1 h before dosing. Solid food will not be allowed from the morning of 
Day -1 until at least 4 hours postdose. 
i. 
All recordings will include supine 10-swallow and upright 5-swallow protocols, and may include an optional 200 mL bolus challenge. Baseline and postdose 
procedures should be identical. The postdose swallowing protocol should begin at 3 h (+15 m) postdose. Fluids are not allowed for at least 1 h before 
catheter insertion until after catheter removal. 
j. 
Patients may be discharged at the Investigator’s discretion a minimum of 30 h postdose if discharge criteria are met. 
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Ironwood Pharmaceuticals, Inc. 
Clinical Trial Protocol: C1701-201-P-06 
 
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TABLE OF CONTENTS 
STUDY IDENTIFICATION ...........................................................................................................2 
SYNOPSIS .......................................................................................................................................4 
ELIGIBILITY CRITERIA.............................................................................................................10 
INCLUSION CRITERIA.........................................................................................................10 
EXCLUSION CRITERIA .......................................................................................................11 
SCHEDULE OF EVENTS ............................................................................................................14 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS ................................................22 
1. 
INTRODUCTION .................................................................................................................25 
1.1 
ACHALASIA ..............................................................................................................25 
1.2 
THE NO-SGC-CGMP PATHWAY ............................................................................25 
1.3 
RATIONALE FOR USE OF AN SGC STIMULATOR IN ACHALASIA................25 
1.4 
IW-1701 .......................................................................................................................26 
1.4.1 
Nonclinical .........................................................................................................26 
1.4.2 
Clinical Studies ..................................................................................................28 
1.4.3 
IW-1701 Tablet Formulation .............................................................................28 
2. 
STUDY OBJECTIVES ..........................................................................................................30 
3. 
INVESTIGATIONAL PLAN ................................................................................................31 
3.1 
OVERALL STUDY DESIGN AND PLAN ................................................................31 
3.2 
DISCUSSION OF STUDY DESIGN, INCLUDING THE CHOICE OF 
CONTROL GROUPS ..................................................................................................32 
3.3 
STUDY DURATION ..................................................................................................33 
3.4 
STUDY POPULATION SELECTION .......................................................................33 
3.4.1 
Study Population ................................................................................................33 
3.4.2 
Removal of Patients from Therapy or Assessment ............................................33 
3.5 
STUDY TREATMENT(S) ..........................................................................................34 
3.5.1 
Description of Treatment ...................................................................................34 
3.5.1.1 
Investigational Product ............................................................................34 
3.5.1.2 
Placebo .....................................................................................................34 
3.5.1.3 
Packaging and Labeling ...........................................................................34 
3.5.1.4 
Storage and Accountability ......................................................................35 
3.5.2 
Method of Assigning Patients to Treatment Groups..........................................35 
3.5.3 
Selection of Dosage in the Study .......................................................................35 
3.5.4 
Selection and Timing of Dose for Each Patient .................................................36 
3.5.5 
Blinding..............................................................................................................37 
3.5.6 
Concomitant Therapy.........................................................................................38 
3.5.7 
Restrictions ........................................................................................................38 
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3.5.7.1 
Prohibited Medicines ...............................................................................38 
3.5.7.2 
Prohibited Supplements ...........................................................................38 
3.5.7.3 
Fluid and Food Restrictions .....................................................................39 
3.5.7.4 
Patient Activity Restrictions ....................................................................39 
3.6 
STOPPING CRITERIA ...............................................................................................40 
3.7 
DISCHARGE CRITERIA ...........................................................................................41 
3.8 
STUDY PROCEDURES .............................................................................................41 
3.8.1 
Informed Consent...............................................................................................41 
3.8.2 
Medical History .................................................................................................41 
3.8.3 
Questionnaires....................................................................................................42 
3.8.3.1 
EQ-5D-3L ................................................................................................42 
3.8.3.2 
SF-12v2 Health Survey ............................................................................42 
3.8.3.3 
Screening Patient-reported Symptom Questions .....................................42 
3.8.4 
Pharmacodynamic Assessments ........................................................................44 
3.8.4.1 
High Resolution Impedance Manometry .................................................44 
3.8.4.2 
Predose (Baseline) and Postdose Patient-reported Symptom 
Questions..................................................................................................45 
3.8.4.3 
Exploratory Biomarker ............................................................................47 
3.8.5 
Safety Assessments ............................................................................................47 
3.8.5.1 
Physical Examination...............................................................................47 
3.8.5.2 
Vital Signs ................................................................................................47 
3.8.5.3 
Electrocardiograms ..................................................................................48 
3.8.5.4 
Clinical Laboratory Tests .........................................................................48 
3.8.5.5 
Adverse Events ........................................................................................50 
3.8.5.6 
Serious Adverse Events ...........................................................................51 
3.8.5.7 
Recording Adverse Events .......................................................................52 
3.8.5.8 
Reporting Serious Adverse Events ..........................................................53 
3.8.6 
Pharmacokinetic Assessments ...........................................................................54 
3.8.7 
Genotyping .........................................................................................................54 
3.9 
STUDY ACTIVITIES .................................................................................................55 
3.9.1 
Screening Period (Day -28 to Day -1) ...............................................................55 
3.9.1.1 
Screening Visit (Days -28 to -1) ..............................................................55 
3.9.2 
Clinic Period (Day 1 to Day 2) ..........................................................................56 
3.9.2.1 
Dosing (Day 1) .........................................................................................56 
3.9.2.2 
Discharge (Day 2) ....................................................................................56 
3.9.3 
Follow-up Period (Day 2 at Discharge to Day 21 ± 7) ......................................57 
3.9.3.1 
Follow-up Phone Call (Day 4 ± 1) ...........................................................57 
3.9.3.2 
End of Trial Visit (Day 21 ± 7) ................................................................57 
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3.10 
STATISTICAL METHODS ........................................................................................58 
3.10.1 
Determination of Sample Size ...........................................................................58 
3.10.2 
Analysis Populations ..........................................................................................58 
3.10.3 
Statistical Methods .............................................................................................59 
3.10.3.1 
General Considerations ............................................................................59 
3.10.3.2 
Patient Disposition, Demographics, and Baseline Characteristics ..........59 
3.10.3.3 
Pharmacodynamic Analyses ....................................................................59 
3.10.3.4 
Safety Analyses ........................................................................................61 
3.10.3.5 
Pharmacokinetic Analyses .......................................................................62 
3.10.3.6 
Interim Analysis .......................................................................................62 
3.10.3.7 
Computer Methods...................................................................................62 
3.11 
CHANGES IN THE CONDUCT OF THE STUDY OR PLANNED ANALYSES ...62 
4. 
ETHICAL CONSIDERATIONS ...........................................................................................63 
4.1 
INSTITUTIONAL REVIEW BOARD........................................................................63 
4.2 
PATIENT INFORMATION AND INFORMED CONSENT .....................................63 
5. 
INVESTIGATORS AND STUDY ADMINISTRATIVE STRUCTURE ............................65 
5.1 
GENERATION OF STUDY RECORDS ....................................................................65 
5.2 
DATA QUALITY ASSURANCE ...............................................................................66 
5.3 
ELECTRONIC CASE REPORT FORMS AND DATA MANAGEMENT ...............66 
5.4 
STUDY MONITORING .............................................................................................66 
6. 
STUDY SPONSORSHIP ......................................................................................................68 
6.1 
INVESTIGATOR AND STUDY TERMINATION ...................................................68 
6.2 
REPORTING AND PUBLICATION ..........................................................................68 
7. 
INVESTIGATOR OBLIGATIONS ......................................................................................69 
7.1 
DOCUMENTATION ..................................................................................................69 
7.2 
PERFORMANCE ........................................................................................................71 
7.3 
USE OF INVESTIGATIONAL MATERIALS ...........................................................71 
7.4 
RETENTION AND REVIEW OF RECORDS ...........................................................71 
7.5 
PATIENT CONFIDENTIALITY ................................................................................72 
8. 
REFERENCE LIST ...............................................................................................................73 
9. 
SPONSOR SIGNATURE ......................................................................................................75 
10. INVESTIGATOR’S STATEMENT ......................................................................................76 
11. APPENDICES .......................................................................................................................77 
 
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LIST OF IN-TEXT TABLES 
Table 1. 
Key Study Participants .........................................................................................2 
Table 2. 
Adverse Event Stopping Criteria .......................................................................40 
Table 3. 
Clinical Laboratory Tests ...................................................................................49 
 
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LIST OF IN-TEXT FIGURES 
Figure 1. 
Study Schematic ................................................................................................31 
 
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LIST OF APPENDICES 
Appendix 1 
EQ-5D-3L ..........................................................................................................78 
Appendix 2 
SF-12v2 ..............................................................................................................81 
 
 
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LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS 
Abbreviation 
Term 
ADP 
adenosine diphosphate 
AE 
adverse event 
ALT 
alanine aminotransferase 
AST 
aspartate aminotransferase 
AUClast 
area under the plasma concentration time curve from time zero to the last 
observation 
BFT 
bolus flow time 
BP 
blood pressure 
BUN 
blood urea nitrogen 
CBC 
complete blood count 
cGMP 
cyclic guanosine 3’, 5’-monophosphate 
Cmax 
maximum observed plasma concentration 
ECG 
electrocardiogram 
eCRF 
electronic case report form 
EDC 
electronic data capture 
EGJ 
esophagogastric junction 
FDA 
Food and Drug Administration 
GCP 
good clinical practice 
GGT 
gamma glutamyl transferase   
GLP 
good laboratory practice 
HDPE 
high-density polyethylene 
HPF 
high power field 
HRIM 
high-resolution impedance manometry 
IBH 
impedance bolus height 
IC50 
half maximal inhibitory concentration 
ICF  
informed consent form 
ICH 
International Conference on Harmonization 
IRB 
Institutional Review Board 
IRP 
integrated relaxation pressure 
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Abbreviation 
Term 
IWRS 
interactive web response system 
kg 
kilogram 
kg/m2 
kilograms/meters squared (body mass index) 
LDH 
lactate dehydrogenase  
LES 
lower esophageal sphincter 
MAD 
multiple ascending dose 
MCH 
mean corpuscular hemoglobin 
MCHC 
mean corpuscular hemoglobin concentration 
MCV 
mean corpuscular volume 
MedDRA 
Medical Dictionary for Regulatory Activities 
mg 
milligram 
mL 
milliliter 
mm Hg 
millimeters of mercury 
MPV 
mean platelet volume 
msec 
millisecond 
nNOS 
neuronal NO synthase 
NO 
nitric oxide 
NOAEL 
no-observed-adverse-effect level 
NOEL 
no-observed-effect level 
pd 
postdose  
PD 
pharmacodynamic(s) 
PDE5 
phosphodiesterase 5 
PEG 
polyethylene glycol 
PK 
pharmacokinetic(s) 
pre 
predose 
PT 
preferred term 
QTcF 
QT interval corrected using Fridericia’s formula 
RBC 
red blood cell 
RDW 
red blood cell distribution width 
SAD 
single ascending dose 
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Abbreviation 
Term 
SAE 
serious adverse event 
SAS® 
Statistical Analysis System 
sGC 
soluble guanylate cyclase 
SNP 
single nucleotide polymorphisms  
SOC 
system organ class 
Tmax 
time of maximum observed plasma concentration 
TRAP 
thrombin receptor activating peptide 
UGT 
uridine diphosphate–glucuronosyl transferase  
WBC 
white blood cell 
 
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1. 
INTRODUCTION 
1.1 
ACHALASIA 
Achalasia is a primary motility disorder characterized by impaired esophageal peristalsis and 
insufficient relaxation of the lower esophageal sphincter (LES) during swallowing. Symptoms of 
the disorder include dysphagia, heartburn, chest pain, and regurgitation.(1) Patients may also 
aspirate undigested food leading to respiratory complications and symptoms including 
pneumonia, cough, hoarseness, and wheezing.(2) Patients with more severe symptoms may also 
have weight loss, esophageal dilation, or retention of food in the esophagus. The disease is 
chronic, and symptoms worsen over time. The condition substantially affects patients’ 
quality-of-life, interfering with their attendance and productivity at work and limiting their 
lifestyle.(3) 
1.2 
THE NO-SGC-CGMP PATHWAY 
Soluble guanylate cyclase (sGC) is an enzyme that catalyzes the formation of cyclic guanosine 
3’, 5’-monophosphate (cGMP) from guanosine 5’-triphosphate (GTP) in response to binding and 
activation by nitric oxide (NO).(4) The NO-sGC-cGMP signaling pathway plays an important 
role in several physiological processes including smooth muscle relaxation.(4) Reduced NO 
bioavailability and reduced responsiveness to endogenous NO have been implicated in the 
pathogenesis of many disease processes.(5,6) sGC stimulators are compounds that directly 
stimulate sGC and synergize with endogenous NO to increase cGMP production.(7) 
1.3 
RATIONALE FOR USE OF AN SGC STIMULATOR IN ACHALASIA 
Esophageal peristalsis and LES tone are regulated by excitatory and inhibitory neurons, which 
release smooth muscle constrictors (such as acetylcholine) resulting in contraction and release 
neurotransmitters (such as NO) resulting in relaxation. In achalasia, reductions in NO signaling 
are believed to result in an imbalance between excitatory and inhibitory neurons during 
swallowing, leading to impaired LES relaxation.  
Evidence from animal models as well as from achalasia patients supports the role of NO in this 
disorder. Mice lacking neuronal NO synthase (nNOS) show achalasia-like symptoms including 
LES hypertension with impaired relaxation.(8) Consistent with this animal model, some 
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achalasia patients have polymorphisms of genes encoding NO synthase (NOS).(9) Low nNOS 
activity has also been observed in biopsies of the muscularis externa of the esophagus from 
achalasia patients.(10) The reported benefit of treatment (off-label) with nitrate donors and 
phosophodiesterase 5 (PDE5) inhibitors provides further evidence supporting the potential of the 
NO-sGC-cGMP pathway in achalasia. Both nitrates, which increase NO concentration, and the 
PDE5 inhibitor sildenafil, which blocks the degradation of cGMP, have been shown to reduce 
LES pressure in achalasia patients.(11-13) Recent genetic evidence also supports the role of sGC 
in achalasia: 9 individuals with early onset achalasia from 3 unrelated consanguineous families 
were found to have loss-of-function mutations in a gene that encodes sGC.(14) 
In total, these data strongly suggest that achalasia can be caused by a failure of the NO-sGC-
cGMP signaling pathway to relax the LES. Thus, in achalasia patients, the augmentation of 
cGMP production by sGC stimulators in response to impaired NO signaling may ameliorate 
excessive pressure in the LES and potentially elsewhere in the esophageal body, and 
consequently may improve the symptoms of achalasia. 
1.4 
IW-1701 
Ironwood Pharmaceuticals is developing IW-1701, an orally administered sGC stimulator, for 
the treatment of conditions associated with deficient NO-sGC-cGMP signaling, including 
achalasia.  
1.4.1 
Nonclinical 
 
 
 
 
 
 
 
 
 
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2. 
STUDY OBJECTIVES 
In patients with primary Type I or II achalasia, following a single 5-mg oral dose of IW-1701, 
• To assess safety and tolerability  
• To determine the effects on measures of esophageal function by high-resolution impedance 
manometry (HRIM) 
• To determine the PK parameters, Cmax, Tmax, AUClast 
 
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catheter will be removed, and patients will complete a baseline symptom assessment. Patients 
with confirmed Type I or II achalasia including an IRP > 15 mm Hg will be randomized to 
receive a single 5-mg dose of IW-1701 or matching placebo (see Section 3.5.1.1). Dosing of 
study drug should occur such that the patient may be observed in the clinic for at least 30 hours 
after dosing (see Section 3.5.4, Selection and Timing of Dose for Each Patient). Following study 
drug administration, the HRIM catheter will be reinserted for the postdose HRIM procedure 
following the protocol-specific swallowing protocol. The HRIM catheter will be removed after 
the postdose recording, and patients will complete a postdose symptom assessment. Safety, PK, 
and PD assessments, including blood collections, will be performed at prespecified times (see 
Schedule of Events). At the Investigator’s discretion, patients may be discharged 30 hours after 
dosing if they meet discharge criteria (Section 3.7).  
Follow-up Period:  The Follow-up Period will begin immediately after the patient is discharged 
from the clinic. Patients will have a Follow-up Call on Day 4 (±1 day) to review AEs. The 
Follow-up Period will end after the End of Trial Visit on Day 21 (± 7 days). 
3.2 
DISCUSSION OF STUDY DESIGN, INCLUDING THE CHOICE OF 
CONTROL GROUPS 
A double-blind, randomized, placebo-controlled, parallel-group study design was chosen in order 
to provide comparable treatment groups. Patients, Investigators, Sponsor personnel involved in 
study conduct, and central reader(s) of the HRIM recordings are blinded to individual treatment 
assignment to minimize the chance of bias during selection and treatment of patients as well as 
during data collection and assessment. Randomization, use of placebo control, and blinding in 
this study are consistent with the concepts in International Conference on Harmonisation (ICH) 
E10, Choice of Control Groups and Related Issues in Clinical Trials (International Conference 
on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human 
Use, 2001). (See Section 3.5.5, Blinding.) 
A 3:1 drug:placebo randomization ratio will be used in order to maximize exposure to study 
medication while allowing for a sufficient placebo control.  
The parameters used to assess the efficacy of study medication will be the HRIM measurements. 
HRIM is the gold standard for the diagnosis and assessment of achalasia. Centralized analysis of 
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HRIM recordings will be performed in a blinded manner to maintain objectivity and provide a 
uniform standard for interpretation of results. 
3.3 
STUDY DURATION  
Patients will receive a single oral dose of IW-1701 and will be followed in the clinic for at least 
30 hours after dosing. Total patient participation will be 15 to 56 days, including the Screening, 
Clinic, and Follow-up Periods. 
3.4 
STUDY POPULATION SELECTION 
3.4.1 
Study Population 
This study will be conducted in patients diagnosed with primary Type I or II achalasia with an 
IRP >15 mm Hg by baseline HRIM procedure. Up to 20 patients will be randomized in a 3:1 
ratio, IW-1701 to placebo. The study will be conducted at up to 10 study centers in the United 
States. 
Refer to Eligibility Criteria for full inclusion and exclusion criteria. 
3.4.2 
Removal of Patients from Therapy or Assessment 
A premature discontinuation will occur when a patient who has signed the ICF and has been 
randomized ceases participation in the study, regardless of circumstances, before completion of 
the End of Trial Visit. A patient will be considered to have completed the study after completing 
the End of Trial Visit. 
Patients will be informed that they are free to withdraw from the study at any time and for any 
reason. The Investigator may remove a patient from the study if, in the Investigator’s opinion, it 
is not in the best interest of the patient to continue the study. Patients may also be discontinued 
from the study by the Investigator or the Sponsor at any time for any reason, including the 
following: 
• Adverse event(s) 
• Protocol violation, including lack of compliance 
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• Lost to follow-up (every effort must be made to contact the patient; a certified letter must be 
sent) 
• Withdrawal of consent (attempts should be made to determine the reason for the patient 
withdrawing consent if possible) 
• Study termination by the Sponsor 
• Other reasons (eg, administrative reasons or pregnancy) 
 
The Sponsor will be notified of any patient discontinuation after randomization. The date the 
patient is withdrawn from the study and the reason for discontinuation will be recorded on the 
study termination form of the electronic case report form (eCRF). Patients who discontinue from 
the study will be followed until resolution of all of their AEs or until the unresolved AEs are 
judged by the Investigator to have stabilized. 
If a patient does not return for a scheduled visit, the study center should contact the patient. An 
effort must be made to contact the patient, including sending a certified letter. In every case, the 
patient outcome, including lost to follow-up information, will be documented. 
3.5 
STUDY TREATMENT(S) 
3.5.1 
Description of Treatment 
3.5.1.1 
Investigational Product 
The investigational product, IW-1701 Tablets, consists of 1 mg of IW-1701 in a 200-mg oral 
tablet. The dose will be 5 mg IW-1701 (5 tablets) administered orally. 
3.5.1.2 
Placebo 
Placebo will match IW-1701 Tablets in appearance. The number of placebo tablets administered 
will match the number of tablets used for the IW-1701 treatment arm. 
3.5.1.3 
Packaging and Labeling 
IW-1701 Tablets and placebo to match will be supplied to the site in 60cc high-density 
polyethylene (HDPE) induction-sealed bottles, 35 tablets per bottle. Bottles will be supplied 
open label to the clinical site.   
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3.5.1.4 
Storage and Accountability 
IW-1701 Tablets and placebo to match will be shipped at ambient temperature. 
IW-1701 Tablets and placebo to match must be stored under controlled room temperature 
conditions, 15 °C to 30 °C (59 °F to 86 °F) per the instructions in the study Pharmacy Manual. 
Any deviation from these storage conditions must be reported to Ironwood and use of the study 
drug suspended until authorization for its continued use has been provided by Ironwood. 
The Investigator’s duly trained designee must ensure that the receipt and use of all study drug 
supplied is recorded and must supervise the storage and allocation of these supplies. All study 
drug supplies must be retained in a locked room that may only be accessed by the pharmacist or 
other duly designated persons. Study drug must not be used outside the context of this protocol, 
and under no circumstances should the Investigator or study center personnel allow the supplies 
to be used other than as directed by this protocol without prior authorization from Ironwood. 
3.5.2 
Method of Assigning Patients to Treatment Groups 
Patients who meet all of the inclusion criteria and none of the exclusion criteria will be 
randomized to treatment on Clinic Day (Day 1). Patients will be randomized through central 
randomization in a 3:1 ratio to receive either 5 mg IW-1701 or placebo.  
The computer-generated randomization schedule will be prepared by an independent statistician 
not otherwise associated with the study. 
Patients, Investigators, and central reader(s) of the HRIM recordings will be blinded to 
individual treatment assignments in this study. 
3.5.3 
Selection of Dosage in the Study 
In this Phase 2a study, C1701-201, a single, oral dose of 5 mg IW-1701 Tablet will be studied. 
This dose was chosen based on final PD and safety data from ICP-1701-101, the Phase 1 SAD 
study in healthy subjects, and based on preliminary, blinded AE and Day 1 PK data from 
ICP-1701-102, the Phase 1 MAD study in healthy subjects.  
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water given during HRIM study procedures, fluids will not be allowed from at least 1 hour 
before HRIM catheter insertion until after catheter removal. Solid food will not be allowed from 
Day -1 until at least 4 hours postdose. Oral medications not prohibited by the protocol may be 
taken on Day 1; administration should be timed to avoid interfering with study procedures. 
3.5.5 
Blinding 
This study is double-blind and placebo-controlled. The patients, Investigators, and site study 
staff, with the exception of specifically designated unblinded Study Center pharmacy staff, will 
be blinded to individual treatment assignments. Sponsor personnel involved in the conduct of the 
study (eg, study medical monitor, study statistician, study data manager, and clinical operations 
study personnel) will be blinded to individual patient treatment assignments; however, Sponsor 
personnel may be unblinded to data at the treatment-group level. In addition, the HRIM 
recordings will be interpreted by central, blinded reader(s) for PD analyses. Further, the 
investigational product and placebo will consist of identical oral tablets. 
Unblinding of an individual patient’s treatment assignment to an Investigator is restricted to 
emergency situations where knowledge of the treatment is necessary for the safety of the patient. 
Except in a medical emergency, the Investigator and blinded study center staff will remain 
blinded to patient treatment assignments during the conduct of the study and until, at a minimum, 
all discrepancies in the clinical database are resolved (ie, at the time of the database lock). 
Individual patient treatment assignment unblinding is available to the Investigator through the 
interactive web response system (IWRS) in the event of an emergency. The Investigator should 
make all reasonable efforts to notify and discuss the circumstances requiring unblinding with the 
Medical Monitor or designee in advance of breaking the blind. If the treatment blind is broken, 
the reason and the date should be recorded and signed by the Investigator and information 
regarding the unblinding should be submitted as soon as possible to the Sponsor. If the 
Investigator is unblinded to the treatment assignment of a patient, the patient will be immediately 
withdrawn from study drug dosing.  
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3.5.6 
Concomitant Therapy 
At the Screening Visit, the following information will be recorded for each patient: 
• All medications the patient is taking (ongoing) 
• All prior medications taken during the 30 days before the Screening Visit 
 
Beginning at the Screening Visit, any medication taken by a patient during the course of the 
study (including medication for sedation/anesthesia, new medications added, or changes in 
medication previously reported) and the reason for use will be documented in the source 
documents and the eCRF. 
3.5.7 
Restrictions 
3.5.7.1 
Prohibited Medicines 
• CYP3A4 inhibitors (including but not limited to itraconazole, ketoconazole, fluconazole, 
clarithromycin, telithromycin, erythromycin, nefazodone, suboxone, and protease inhibitors) 
are prohibited from 7 days before Check-in on Day 1 until after the End of Treatment Visit 
on Day 21 (± 7 days). 
• CYP3A4 inducers (including but not limited to carbamazepine, enzalutamide, mitotane, 
phenytoin, phenobarbital, rifampin/rifampicin, rifabutin, rifapentine, and St. John’s wort) are 
prohibited from 7 days before Check-in on Day 1 until after the End of Treatment Visit on 
Day 21 (± 7 days). 
• PDE5 inhibitors (including sildenafil and tadalafil), mono- or di-nitroglycerine products, 
riociguat, calcium-channel blockers, opiates, and aspirin are prohibited from 7 days before 
Check-in on Day 1 through the Follow-up Call on Day 4 (± 1 day). Bisphosphonates are 
prohibited from 3 months before Check-in on Day 1 through the Follow-up Call on 
Day 4 (± 1 day). 
 
3.5.7.2 
Prohibited Supplements 
Any supplements for the treatment of erectile dysfunction are prohibited from 7 days before 
Check-in on Day 1 through the Follow-up Call on Day 4 (± 1 day). 
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3.5.7.3 
Fluid and Food Restrictions  
Patients must fast overnight before sample collections for clinical chemistry, coagulation, 
hematology, and urinalysis assessments at the Screening and End of Trial Visits. 
Patients must adhere to a liquid diet on Day -1 and then fast overnight until at least 4 hours after 
study drug administration. Fluids will not be allowed for at least 1 hour before study drug 
administration, and except for the water given during HRIM procedures, fluids will not be 
allowed from at least 1 hour before HRIM catheter insertion until after catheter removal. 
Alcoholic beverages are prohibited from the Screening Visit until after the Follow-up Call 
(± 1 day). 
3.5.7.4 
Patient Activity Restrictions 
Female patients of childbearing potential (ie, women who are not postmenopausal or who have 
not had a bilateral oophorectomy, hysterectomy, or tubal ligation) must agree to completely 
abstain from heterosexual intercourse or, if heterosexually active, must agree to use 1 of the 
following methods of birth control from the date they sign the ICF until 90 days after the dose of 
study drug: 
a. Combination of 2 highly effective birth control methods (eg, condom with spermicide plus 
intrauterine device, condom with spermicide plus a diaphragm or cervical cap, hormonal 
contraceptive [including progesterone implant] combined with a barrier method) 
b. Maintenance of a monogamous relationship with a male partner who has been surgically 
sterilized by vasectomy (vasectomy procedure must have been conducted ≥ 60 days before 
the Screening Visit or confirmed via sperm analysis) plus a hormone or barrier method 
 
In addition, female patients may not donate or harvest eggs during the course of the study 
through at least 30 days after the dose of study drug. 
Male patients who are not surgically sterile by vasectomy (vasectomy procedure conducted ≥ 60 
days before the Screening Visit or confirmed via sperm analysis) must agree to completely 
abstain from heterosexual intercourse, or, if heterosexually active, must agree to use a 
combination of 2 highly effective birth control methods (eg, condom with spermicide plus 
intrauterine device, condom with spermicide plus a diaphragm or cervical cap, hormonal 
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contraceptive [including progesterone implant] combined with a barrier method) from date they 
sign the ICF until 90 days after the dose of study drug. 
In addition, male patients may not donate sperm during the study through at least 90 days after 
the dose of study drug. 
3.6 
STOPPING CRITERIA 
Dosing of additional patients will be suspended if any of the following occur:  
• Drug-related SAEs in 2 or more patients (per causality and SAE definitions in this protocol)  
• Drug-related AEs in 1 or more patients that are life threatening or require urgent intervention|| 
• An overall pattern of clinically significant treatment-emergent AEs that may appear minor in 
terms of an individual event but collectively represents a safety concern 
• Treatment-emergent AEs judged to be drug-related and reported per the description and in 
the number of patients indicated in Table 2. These events are based on clinical experience 
with IW-1701 and the prescribing information for riociguat.  
 
Table 2. 
Adverse Event Stopping Criteria 
Treatment-emergent,  
Drug-related Event 
Event 
Description 
# of patients that 
trigger stopping 
Symptomatic hypotension-related 
events (eg, syncope, presyncope) 
Medical intervention or hospitalization indicated* 
2 
Vomiting 
≥ 6 episodes (separated by at least 5 minutes) in 
24 hours† 
2 
Spontaneous bleeding events ‡ 
(including hemoptysis, vaginal 
hemorrhage, ovarian hemorrhage, 
subdural hemorrhage, hematemesis, 
hematochezia)  
Moderate symptoms, medical intervention 
indicated § 
2 
Life-threatening consequences; urgent 
intervention indicated || 
1 
* Consistent with National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE) 
Version 4.0, Grade 2 (15) 
† CTCAEv4 Grade 3 
‡ Based on riociguat prescribing information  
§ Consistent with CTCAEv4 Grade 2 
|| Consistent with CTCAEv4, Grade 4  
 
If stopping criteria are met, the study may continue only after the study protocol has been 
amended, if appropriate, and approved by the IRB. 
(Note: This is a single-dose study; therefore, there are no individual subject stopping rules.) 
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3.7 
DISCHARGE CRITERIA 
Patients may be discharged at the Investigator’s discretion a minimum of 30 hours after study 
drug administration if the following criteria are met: 
• Systolic BP ≥ 90 mm Hg and diastolic BP ≥ 60 mm Hg 
• No unresolved AEs that the Investigator judges to be drug related 
 
3.8 
STUDY PROCEDURES 
3.8.1 
Informed Consent 
Informed consent procedures will comply with the Code of Federal Regulations (CFR) 21 CFR, 
Parts 50 and 312. 
The written ICF must be approved by the Institutional Review Board (IRB) for the purposes of 
obtaining and documenting consent. 
Before entry into the study, each patient will be provided with a written explanation of the study. 
It is the responsibility of the Investigator or appropriately trained health professional to give each 
patient full and adequate information regarding the objectives and procedures of the study and 
the possible risks involved. Patients will then be given the opportunity to ask questions and the 
Investigator will be available to answer questions as needed. Patients will be informed of their 
right to withdraw from the study at any time without prejudice. After this explanation and before 
entering the study, the patient will voluntarily sign an ICF. The patient should receive a copy of 
the signed and dated ICF. The Investigator must retain each patient’s original signed ICF. 
If new information becomes available that may be relevant to the patient’s consent and 
willingness to participate in the study, the ICF will be revised. The revised ICF must be 
submitted to the IRB for review and approval prior to its use. 
3.8.2 
Medical History 
A complete medical history will be recorded at the Screening Visit. 
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3.8.3 
Questionnaires 
3.8.3.1 
EQ-5D-3L 
The EuroQol (EQ)-5D-3L (Appendix 1) is a generic measure of health status widely used in 
Europe.(16) The first component consists of 5 questions assessing the following dimensions: 
mobility, self-care, usual activities, pain/discomfort, and anxiety/depression. Responses to the 
5 questions define a health state for which a utility index can be derived from published 
algorithms.(17) The second component of the EQ-5D is a visual analogue scale asking patients 
to rate their health from 0 to 100 (0 represents worst imaginable health state and 100 represents 
best imaginable health state). Patients will complete the EQ-5D-3L at the Screening and End of 
Trial Visits. 
3.8.3.2 
SF-12v2 Health Survey 
The SF-12v2 (Appendix 2) is a widely used generic measure of health-related quality of life and 
measures 8 concepts of health: physical functioning, role limitations due to physical health 
problems, bodily pain, general health, vitality (energy/fatigue), social functioning, role 
limitations due to emotional problems, and mental health (psychological distress and 
psychological well-being). These 8 scales are aggregated into 2 summary measures: the physical 
component and mental component summary scores.(18) Patients will complete the SF-12v2 at 
the Screening and End of Trial Visits. 
3.8.3.3 
Screening Patient-reported Symptom Questions  
Patients will answer the following Screening Patient-reported Symptom Questions at the 
Screening Visit; responses will be entered into the respective patient’s eCRF. 
 
 
 
 
 
 
ere 
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3.8.4 
Pharmacodynamic Assessments 
The assessments used to determine the pharmacodynamic parameters will include baseline and 
postdose HRIM recordings and patient-reported symptom questions. 
3.8.4.1 
High Resolution Impedance Manometry 
On Clinic Day (Day 1), the HRIM procedure will be performed 2 times, once at baseline and 
once after study drug administration. Catheter insertion should be timed to minimize the total 
duration of catheter placement during the procedure; baseline and postdose procedures, including 
the timing of prerecording placement of the catheter and use of sedation/anesthesia, should be 
identical (± 10 minutes). For the postdose recording, the swallowing protocol should begin at 
3 hours (+ 15 minutes) postdose. HRIM measurements for PD analyses will be made by central, 
blinded reader(s).  
During HRIM procedures, patients may experience gagging, watery eyes, slight nosebleed, 
discomfort in nose and throat. Afterwards, patients may have mild side effects including sore 
throat, stuffy nose, and minor nosebleed. Rare severe complications may include irregular 
heartbeat, aspiration, and perforation of the esophagus. 
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HRIM procedure  
• Calibrate the HRIM catheter per manufacturer’s instructions. 
• Administer sedation (optional per Investigator standard protocol). 
• Anesthetize transnasally with lidocaine (2% jelly) and/or oropharyngeally with Ketocaine® 
per Investigator standard protocol. 
• Insert catheter transnasally to record manometric data from the hypopharynx to the stomach.  
• Recline patient into a supine position (at ~ 30-degree incline). 
• Begin recording with ~ 2-minute resting measurement to assess basal esophagogastric 
junction (EGJ) pressure without swallowing.  
• With patient supine, have patient swallow ~ 5 mL saline in a single swallow; repeat 9 times 
allowing ~ 30 seconds for each swallow. Mark each swallow and encourage patient not to 
swallow between sips; repeat any technically inadequate swallows. 
• With patient upright, repeat the ~ 2-minute resting measurement without swallowing. 
• With patient upright, have patient swallow ~5 mL saline in a single swallow; repeat 4 times 
allowing ~ 30 seconds for each swallow. Mark each swallow and encourage patient not to 
swallow between sips. 
• (Optional) With patient standing, have patient drink a ~ 200-mL saline bolus as quickly as 
possible. Patient should be encouraged to drink the full bolus. 
• Remove catheter and save the data. 
 
3.8.4.2 
Predose (Baseline) and Postdose Patient-reported Symptom Questions 
Following removal of the HRIM catheter after both the baseline and postdose procedures, the 
patient will answer a set of questions related to their most recent swallowing protocols; the 
answers will be entered into the patient’s eCRF.  
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3.8.4.3 
Exploratory Biomarker  
Blood samples for determination of plasma renin activity will be collected according to the 
Schedule of Events. 
3.8.5 
Safety Assessments 
3.8.5.1 
Physical Examination 
A complete physical examination will be performed according to the Schedule of Events. The 
physical examination of each patient should include examination and assessment of the 
following: 
General appearance 
Head, eyes, ears, nose, and throat 
Cardiovascular system 
Neck 
Respiratory system 
Musculoskeletal system 
Abdomen/liver/spleen 
Nervous system 
Lymph nodes 
Skin 
Neurologic status 
Mental status 
 
Breast, genitourinary, and rectal examinations are optional and may be performed at the 
discretion of the Investigator. Any new, clinically significant abnormal findings from the 
physical examination will be reported as an AE. 
Each patient’s weight will be recorded at each study visit; height will only be recorded at the 
Screening Visit. 
3.8.5.2 
Vital Signs 
Vital signs will be measured according to the Schedule of Events and documented on the eCRF. 
Vital sign measurements include oral temperature (°C), respiratory rate, BP, and pulse. 
Respiratory rate, BP, and pulse measurements will be taken after the patient has been seated for 
at least 5 minutes. Where applicable, vital signs should be measured before blood draws. 
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3.8.5.3 
Electrocardiograms 
A 12-lead ECG will be performed according to the Schedule of Events and documented on the 
eCRF. Electrocardiograms should be obtained after the patient has been supine for at least 
5 minutes. (Note:  If on initial ECG, QTcF is ≥ 450 msec for male patients or is ≥ 470 msec for 
female patients, the ECG will be repeated 2 more times, and the average of the 3 QTcF values 
will be used to determine the patient's eligibility.) 
3.8.5.4 
Clinical Laboratory Tests 
3.8.5.4.1 
Laboratory Parameters 
Blood and urine samples for clinical laboratory tests will be collected according to the laboratory 
study procedures at the days and times defined in the Schedule of Events. Patients must have 
fasted for at least 8 hours before sample collections. 
The clinical laboratory evaluations will include the serum chemistry, hematology, coagulation, 
urinalysis and urine drug tests presented in Table 3. 
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Table 3. 
Clinical Laboratory Tests 
Serum Chemistry 
Hematology (CBC) 
Complete Urinalysis 
Albumin 
Hematocrit 
Color and appearance 
Alkaline Phosphatase 
Hemoglobin 
pH and Specific Gravity 
ALT 
Platelet count 
Bilirubin 
AST 
    MPV 
Glucose 
Bicarbonate 
RBC count 
Ketones 
BUN 
WBC count 
Leukocytes 
Calcium 
WBC differential 
Nitrites 
Chloride 
  (% & absolute): 
Occult blood 
Cholesterol 
    Basophils 
Protein 
Creatinine 
    Eosinophils 
Urobilinogen 
GGT 
    Lymphocytes 
Microscopic  
Glucose 
    Monocytes 
Including bacteria, RBCs, WBCs 
HDL-c 
    Neutrophils 
per HPF if dipstick is abnormal 
LDH 
RBC indices 
 
LDL-c (calculated) 
    MCH 
 
Magnesium 
    MCHC 
Additional Urine Tests 
Phosphorus 
    MCV 
Urine Cotinine 
Potassium 
RDW 
 
Sodium 
 
 
Total Bilirubin 
Coagulation 
 
Total Protein 
aPTT 
 
Triglycerides 
Prothrombin time 
 
 Uric acid 
INR 
 
ALT = alanine aminotransferase; aPTT = activated partial thromboplastin time; AST = aspartate 
aminotransferase; BUN = blood urea nitrogen; CBC = complete blood count; GGT = gamma glutamyl 
transferase; HPF = high power field; INR = International Normalized Ratio; LDH = lactate dehydrogenase; 
MCH = mean corpuscular hemoglobin; MCHC = mean corpuscular hemoglobin concentration; MCV = mean 
corpuscular volume; MPV = mean platelet volume; RBC = red blood cell; RDW = red blood cell distribution 
width; WBC = white blood cell.  
 
For female patients, a negative serum pregnancy test must be documented at the Screening and 
Follow-up Visits; a negative urine pregnancy test must be documented on Day 1 with results 
available before Randomization. In the event of a positive pregnancy test, the test will be 
repeated. If pregnancy is confirmed, see Section 3.8.5.7. 
Screens for a hepatitis panel (including HBsAg and anti-HCV) and HIV antibody will be 
performed at the Screening Visit. 
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A urine drug screen for the following illicit drugs will be performed at the Screening Visit: 
Amphetamines 
Cocaine 
Opiates 
Barbiturates 
Phencyclidine (PCP) 
Benzodiazepines 
Marijuana 
Propoxyphene 
 
3.8.5.5 
Adverse Events  
All patients will be monitored for AEs throughout the study. All AEs will be recorded in 
accordance with the procedures outlined in this section. 
An AE is any untoward medical occurrence in a patient or clinical investigation subject 
administered a pharmaceutical product and which does not necessarily have to have a causal 
relationship with this treatment.  
An AE, therefore, can be any unfavorable and unintended sign (including an abnormal laboratory 
finding, for example), symptom, or disease temporally associated with the use of a medicinal 
product, whether or not considered related to the medicinal product. 
An AE includes, but is not limited to, the following: 
• Any unfavorable changes in general condition 
• Any clinically significant worsening of a preexisting condition 
• Any intercurrent diseases and accidents 
 
Note: A procedure is not an AE, but the reason for a procedure may be an AE. 
3.8.5.5.1 
Causality Assessment 
For all AEs, the Investigator must provide an assessment of causal relationship to study drug. 
The causality assessment must be recorded in the patient’s source documentation and on the AE 
page of the subject’s eCRF. Causal relationship must be assessed according to the following: 
Related:   
An event where there is a reasonable possibility of a causal relationship between 
the event and the study drug 
Unrelated:   Any other event 
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3.8.5.5.2 
Severity Assessment 
The Investigator will provide an assessment of the severity of each AE by recording a severity 
rating in the patient’s source documentation and on the AE page of the patient’s eCRF. Severity, 
which is a description of the intensity of manifestation of the AE, is distinct from seriousness, 
which implies a patient outcome or AE-required treatment measure associated with a threat to 
life or functionality. Severity will be assessed according to the following scale: 
Mild: 
A type of AE that is usually transient and may require only minimal treatment or 
therapeutic intervention. The event does not generally interfere with usual 
activities of daily living. 
Moderate: 
A type of AE that is usually alleviated with additional specific therapeutic 
intervention. The event interferes with usual activities of daily living, causing 
discomfort but poses no significant or permanent risk of harm to the research 
participant. 
Severe: 
A type of AE that interrupts usual activities of daily living, or significantly affects 
clinical status, or may require intensive therapeutic intervention. 
 
3.8.5.6 
Serious Adverse Events 
An SAE is defined as any AE occurring at any dose that results in any of the following 
outcomes: 
• Death 
• Life-threatening:  the patient was at immediate risk of death from the reaction as it occurred 
(ie, it does not include a reaction that hypothetically might have caused death if it had 
occurred in a more severe form) 
• Hospitalization or prolongation of existing hospitalization 
• Persistent or significant disability/incapacity:  a substantial disruption of a person’s ability to 
conduct normal daily functions 
• Congenital anomaly/birth defect 
• Important medical events:  events that may not result in death, be life threatening, or require 
hospitalization. Such an event may be considered serious when, based on appropriate medical 
judgment, it may jeopardize the subject and may require medical or surgical intervention to 
prevent 1 of the outcomes listed in this definition. Examples of such medical events include 
allergic bronchospasm requiring intensive treatment in an emergency department or at home, 
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blood dyscrasias or convulsions that do not result in inpatient hospitalization, or the 
development of drug dependency or drug abuse. 
Emergency room visits that do not result in admission to the hospital should be evaluated for 1 of 
the other serious outcomes (eg, life-threatening, other serious [medically important] event). 
3.8.5.7 
Recording Adverse Events 
Adverse events will be collected and recorded from the time the patient signs the ICF at the 
Screening Visit through the End of Trial Visit. All AEs, regardless of the assumption of a causal 
relationship with study procedures or study medication, must be recorded in the patient’s source 
documentation and subsequently on the appropriate AE page of the patient’s eCRF. This record 
includes AEs the patient reports spontaneously, those observed by the Investigator, and those 
elicited by the Investigator in response to open-ended questions during the study, such as “Have 
you had any health problems since your last visit?” 
For every AE, the Investigator must: 
• Provide an assessment of the severity, causal relationship to the study medication, and 
seriousness of the event 
• Document all actions taken with regard to the study medication (ie, no action taken, 
treatment temporarily interrupted, or treatment discontinued)  
• Detail any other treatment measures taken for the AE, including concomitant medications 
and/or procedures 
 
Pretreatment AEs will be collected from the time the patient signs the ICF until the patient 
receives study drug. Pretreatment AEs will be captured in the patient’s source documentation but 
will only be entered for randomized patients on the AE page of the patient’s eCRF. 
Laboratory abnormalities and changes in vital signs, physical examination findings, and 12-lead 
ECG parameters should be considered AEs and reported on the AE page of the patient’s eCRF if 
the Investigator considers them clinically significant and/or they necessitate intervention. 
Any medical condition that is present when a patient is screened and does not worsen in severity 
and/or frequency should be reported as Medical History and not as an AE. However, if the 
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condition does deteriorate in severity and/or frequency at any time during the study, it should be 
reported as an AE. 
3.8.5.8 
Reporting Serious Adverse Events 
An AE that meets any of the serious criteria must be reported to Ironwood within 24 hours from 
the time that site personnel first learn of the event, using the SAE Report form provided for the 
study. Regardless of causality, all SAEs must be reported and will be collected and recorded 
from the time the subject signs ICF at the Screening Visit until the End of Trial Visit. All SAEs 
must also be recorded in the subject’s source documentation and on the AE page of the subject’s 
eCRF. 
The initial report should include at least the following information: 
• Patient identification number 
• Description and onset of the event 
• Serious criteria 
• Causality assessment to study drug 
 
Special Situation:  Exposure to Study Drug during Pregnancy 
In the event that a pregnancy occurs in a patient, study personnel must report the pregnancy as 
soon as possible (within 24 hours after notification) on the pregnancy notification form provided 
for this study. The study personnel must follow the pregnancy until the end and report the 
pregnancy outcome on the pregnancy outcome form provided for this study. If the pregnancy is 
associated with an SAE (eg, if the mother is hospitalized), a separate SAE form must be 
completed. 
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All relevant SAE or pregnancy information should be emailed to Ironwood Drug Safety and 
Pharmacovigilance. 
All SAE Report Forms should be emailed to: 
 
Clinical Drug Safety & Pharmacovigilance 
Ironwood Pharmaceuticals, Inc. 
 
 
 
If follow-up is obtained, or requested by Ironwood, the additional information should be emailed 
on an SAE Report Form to Ironwood, in a timely manner according to the procedures outlined 
above. Copies of discharge summaries, consultant reports, autopsy reports, and any other 
relevant documents may also be requested. 
Appropriate remedial measures should be taken by the Investigator using his/her best medical 
judgment to treat the SAE. These measures and the subject’s response to these measures should 
be recorded. All SAEs regardless of relationship to study drug will be followed by the 
Investigator until satisfactory resolution, until the Investigator deems the SAE to be chronic or 
stable, or until the subject is lost to follow-up. Clinical, laboratory, and diagnostic measures 
should be employed by the Investigator as needed to adequately determine the etiology of the 
event. 
The Investigator will be responsible for reporting all SAEs to the IRB. Ironwood will be 
responsible for reporting to the regulatory authorities. 
3.8.6 
Pharmacokinetic Assessments 
Blood samples for PK and exploratory PD assessments (Section 3.8.4.3) will be collected 
according to the Schedule of Events.   
3.8.7 
Genotyping 
An optional single blood sample for genotyping of single nucleotide polymorphisms (SNPs) that 
may be related to interindividual differences in drug disposition or PD response (eg, uridine 
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diphosphate–glucuronosyl transferase [UGT] drug metabolizing enzymes) may be collected 
according to the Schedule of Events. 
3.9 
STUDY ACTIVITIES 
3.9.1 
Screening Period (Day -28 to Day -1) 
3.9.1.1 
Screening Visit (Days -28 to -1) 
• Signing of ICF 
• Review of inclusion and exclusion criteria 
• Demographics 
• Medical history 
• Weight and height 
• Physical examination 
• 12-lead ECG 
• Seated vital signs (oral temperature, respiratory rate, blood pressure and pulse) 
• Prior medication recording (all medicines taken during the 30 days before Screening Visit) 
• Collection of blood and urine samples for: 
- 
Clinical chemistry 
- 
Hematology 
- 
Coagulation 
- 
Urinalysis 
- 
Urine cotinine test 
- 
Hepatitis and HIV screen 
- 
Drug screen 
- 
Serum pregnancy test for all females (must be confirmed negative) 
• AE evaluation 
• EQ-5D-3L 
• SF-12v2 
• Screening patient-reported symptom questions 
 
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3.9.2 
Clinic Period (Day 1 to Day 2) 
3.9.2.1 
Dosing (Day 1) 
• Review of inclusion and exclusion criteria 
• Prior and concomitant medication recording 
• Weight  
• Urine pregnancy test for all females (must be confirmed negative before Randomization) 
• 12-lead ECG prior to dosing and at 4 hours (± 30 minutes) postdose 
• Oral temperature and seated respiratory rate predose (≤ 15 minutes) 
• Seated blood pressure and pulse predose (≤ 15 minutes) and at 0.5 hours (± 10 minutes), 1, 2, 
3, 4, 6, 8, and 12 hours (± 15 minutes) postdose 
• Collection of blood samples for: 
- 
PK and exploratory PD assessments predose (≤ 15 minutes) and at 0.5 hours (± 2 
minutes), 1, 2, 3, 4, 5, 6, 8 hours (± 5 minutes) and 12 hours (± 15 minutes) postdose 
• AE evaluations throughout day 
• Baseline HRIM procedure  
• Baseline patient-reported symptom questions 
• Review of IRP eligibility criterion 
• Randomization 
• Study drug administration 
• Postdose HRIM procedure (the swallowing protocol should begin at 3 hours [+ 15 minutes] 
postdose) 
• Postdose patient-reported symptom questions 
 
3.9.2.2 
Discharge (Day 2) 
• 12-lead ECG at 24 hours (± 30 minutes) postdose 
• Seated vital signs (oral temperature, respiratory rate, blood pressure, and pulse) at 24 hours 
(± 15 minutes) postdose 
• Collection of blood samples for: 
- 
Clinical chemistry at 24 hours (± 30 minutes) postdose 
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- 
Hematology at 24 hours (± 30 minutes) postdose 
- 
Coagulation at 24 hours (± 30 minutes) postdose 
- 
Urinalysis at 24 hours (± 30 minutes) postdose 
- 
Genotyping at 24 hours (± 30 minutes) postdose (optional) 
- 
PK and exploratory PD assessments at 17 (± 15 minutes) and 24 hours (± 30 minutes) 
postdose 
• AE evaluations throughout day 
• Concomitant medication recording 
• Discharge at least 30 hours postdose, per Investigator discretion and discharge criteria 
(Section 3.7) 
• BP prior to discharge (Section 3.7) 
 
3.9.3 
Follow-up Period (Day 2 at Discharge to Day 21 ± 7) 
3.9.3.1 
Follow-up Phone Call (Day 4 ± 1) 
• Concomitant medication recording 
• AE evaluation 
 
3.9.3.2 
End of Trial Visit (Day 21 ± 7) 
• Weight  
• EQ-5D-3L 
• SF-12v2 
• Physical examination 
• 12-lead ECG 
• Seated vital signs (oral temperature, respiratory rate, blood pressure, and pulse) 
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• Collection of blood and urine samples for: 
- 
Clinical chemistry 
- 
Hematology 
- 
Coagulation 
- 
Urinalysis 
- 
PK and exploratory PD assessments  
- 
Serum pregnancy test for all females 
• Concomitant medication recording 
• AE evaluation 
 
3.10 
STATISTICAL METHODS 
Detailed statistical methods will be provided in the Statistical Analysis Plan (SAP), to be 
finalized prior to the clinical database lock. 
3.10.1 
Determination of Sample Size 
For this Phase 2a study, a total sample size of up to 20 patients is planned (subjects will be 
randomized to either IW-1701 or placebo in a 3:1 ratio, resulting in approximately 15 patients 
receiving IW-1701 and approximately 5 patients receiving placebo). For this exploratory Phase 
2a study, the sample size of up to 20 patients was determined outside of statistical considerations 
and is based on precedent set by prior studies of similar nature and design.(11,20-24) 
3.10.2 
Analysis Populations  
Safety Population 
All patients who receive study drug will be included in the Safety Population. Patients in the 
Safety Population will be evaluated for safety according to the treatment they actually received. 
Pharmacokinetic Population 
All patients who receive study drug and have at least 1 postdose PK parameter assessment will 
be included in the PK Population. 
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Pharmacodynamic Population 
All patients who receive study drug and have at least 1 postdose PD assessment will be included 
in the PD Population. 
3.10.3 
Statistical Methods 
3.10.3.1 
General Considerations 
Descriptive statistics (n, mean, standard deviation, minimum, maximum, median, and 
interquartile range [IQR]) will be calculated to summarize continuous variables. Frequency and 
percentage of patients in each category will be calculated to summarize categorical variables. 
Due to the exploratory nature of the trial, no adjustments will be made for multiplicity. Details of 
the data handling methods will be specified in the SAP. Inferential statistics will be used for 
descriptive purposes only. If not otherwise specified, the baseline value is defined as the last 
nonmissing value measured before administration of study drug on Day 1. All statistical analyses 
will be performed using SAS® Version 9.3 (or later) for Windows. 
3.10.3.2 
Patient Disposition, Demographics, and Baseline Characteristics 
The overall number of patients screened and the number of patients randomized to each 
treatment group will be presented. The number and percentage of randomized patients included 
in the Safety Population as well as the PK and PD Populations will be presented by treatment 
group and overall. The distribution of randomized patients by completion status, as well as 
reason for discontinuation, will also be presented by treatment group and overall. 
Demographic parameters (eg, age, race, ethnicity, sex, weight, and height) and other baseline 
characteristics will be summarized by treatment group for the PD Population. 
This table will be repeated for the Safety Population if it is different from the PD Population. 
3.10.3.3 
Pharmacodynamic Analyses 
All PD analyses will be conducted using the PD Population. 
3.10.3.3.1 
HRIM Measurements 
HRIM measurements for PD analyses will be made by central, blinded reader(s). 
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• Bolus Flow Time (BFT) – time in seconds of bolus (5 mL swallow) transit through the EGJ 
from the 10-swallow supine and 5-swallow upright protocols; median measurement from the 
10-swallow supine and 5-swallow upright protocols 
• Integrated Relaxation Pressure (IRP) – mean of the 4s of maximal deglutitive relaxation in 
the 10-s window beginning at upper esophageal sphincter relaxation. Contributing times can 
be contiguous or non-contiguous (eg, interrupted by diaphragmatic contraction). Referenced 
to gastric pressure; median measurement from the 10-swallow supine and 5-swallow upright 
protocols 
• Impedance bolus height (IBH) – height in esophagus of 200 mL saline bolus at 1, 2, and 
5 minutes postbolus from the standing 200-mL bolus challenge 
 
A single HRIM procedure consists of a set of swallowing protocols. For postdose recordings, the 
swallowing protocols begin at 3 hours (+ 15 minutes) postdose. The procedures at baseline and 
postdose should be identical.  
 The following PD parameters will be derived for each subject from the HRIM measurements: 
• Supine BFT3h: median of the measurements from the 10 supine swallows at baseline and at 
3 hours postdose 
• Upright BFT3h: median of the measurements from the 5 upright swallows at baseline and at 
3 hours postdose 
• Supine IRP3h: median of the measurements from the 10 supine swallows at baseline and at 
3 hours postdose 
• Upright IRP3h: median of the measurements from the 5 upright swallows at baseline and at 
3 hours postdose 
• IBH1m: height in esophagus of standing 200 mL saline bolus at 1-minute postbolus 
(if performed for the subject) 
• IBH2m: height in esophagus of standing 200 mL saline bolus at 2 minutes postbolus 
(if performed for the subject) 
• IBH5m: height in esophagus of standing 200 mL saline bolus at 5 minutes postbolus 
(if performed for the subject) 
 
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For both baseline and postdose HRIM procedures, the central reader will assess the HRIM 
recording and determine the presence (or absence) of achalasia and, if present, will define the 
achalasia type (I, II, III).(25)  
Descriptive statistics will be presented by treatment group for each parameter at baseline and 
postdose. Mean and median change from baseline for each parameter will be presented by 
treatment group. Percent reduction (from baseline to postdose) in IRP parameters will also be 
calculated and summarized by treatment group. 
3.10.3.3.2 
Patient-reported Symptom Questions 
Descriptive statistics will be presented by treatment group for each question at baseline and 
postdose.  
3.10.3.4 
Safety Analyses 
All safety analyses will be conducted using the Safety Population. 
3.10.3.4.1 
Adverse Events 
AEs will be coded using the most current version of Medical Dictionary for Regulatory 
Activities (MedDRA) available at the start of the study. Treatment-emergent AEs (TEAEs) are 
AEs that first occurred or worsened in severity after study drug administration. AEs occurring 
more than 72 hours after administration of study drug will not be considered treatment emergent. 
Incidence (number and percentage) of TEAEs will be presented by system organ class (SOC), 
preferred term (PT), and treatment group. Listings, including verbatim terms, will be provided 
for TEAEs, drug-related AEs, SAEs, and AEs leading to study discontinuation. 
3.10.3.4.2 
Vital signs, ECGs, and Clinical Laboratory Tests 
ECGs, vital signs, and clinical laboratory tests at each assessment time point and the change 
from baseline at each postbaseline time point will be summarized by treatment group. Listings 
will be provided for patients with abnormal values. 
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3.10.3.5 
Pharmacokinetic Analyses 
If systemic levels of IW-1701 are detectable, the following PK parameters will be calculated, 
when appropriate: 
• AUClast: Area under the plasma concentration time curve from time zero to Tlast, the time at 
which the last measurable plasma concentration (Clast) is observed 
• Cmax: Maximum observed plasma concentration, occurring at Tmax 
• Tmax: Time of maximum observed plasma concentration 
 
3.10.3.6 
Interim Analysis 
No interim analysis is planned for the study. 
3.10.3.7 
Computer Methods 
Statistical analyses will be performed using version 9.3 (or newer) of SAS® for Windows. 
3.11 
CHANGES IN THE CONDUCT OF THE STUDY OR PLANNED 
ANALYSES 
Any amendment to this protocol will be provided to the Investigator in writing by Ironwood or 
its designee. Prior to implementation, any protocol amendment regarding reportable deviations 
(as defined by the IRB) must be approved by the IRB and the signature page must be signed by 
the Investigator and received by Ironwood or its designee, with the following exception: If the 
protocol is amended to eliminate or reduce the risk to patients, the amendment may be 
implemented before IRB review and approval. However, the IRB must be informed in writing of 
such an amendment, and approval must be obtained within reasonable time limits. 
Deviating from the protocol is permitted only if absolutely necessary for the safety of the 
patients and must immediately be reported to Ironwood or its designee. 
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4. 
ETHICAL CONSIDERATIONS 
4.1 
INSTITUTIONAL REVIEW BOARD 
Prior to the study onset, the protocol, any protocol amendments, ICFs, advertisements to be used 
for patient recruitment, and any other written information regarding this study to be provided to a 
patient or patient’s legal guardian must be approved by the IRB. 
All IRB approvals must be dated and signed by the IRB Chairman or his or her designee and 
must identify the IRB by name and address, the clinical protocol by title and/or protocol number, 
and the date upon which approval or favorable opinion was granted for the clinical research. 
Copies of IRB approvals should be forwarded to Ironwood. All correspondence with the IRB 
should be maintained in the Investigator File. 
No drug will be released to the site to dose a patient until written IRB approval has been received 
by Ironwood. 
The Investigator is responsible for obtaining continuing review of the clinical research at least 
annually or more often if specified by the IRB. The Investigator must supply Ironwood with 
written documentation of the approval of the continued clinical research. 
The IRB must be constituted in accordance with Federal and ICH Good Clinical Practice (GCP) 
guidelines and any relevant and applicable local regulations. 
Major changes in this research activity, except those to remove an apparent immediate hazard to 
the patient, must be reviewed and approved by Ironwood and by the IRB that approved the study. 
Amendments to the protocol must be submitted in writing to the Investigator’s IRB for approval 
prior to patients being enrolled into the amended protocol. 
4.2 
PATIENT INFORMATION AND INFORMED CONSENT 
Informed consent procedures will comply with the Code of Federal Regulations (CFR) 21 CFR, 
Parts 50 and 312. 
The written ICF must be approved by the IRB for the purposes of obtaining and documenting 
consent. 
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Before entry into the study, each patient will be provided with a written explanation of the study. 
It is the responsibility of the Investigator or appropriately trained health professional to give each 
patient full and adequate information regarding the objectives and procedures of the study and 
the possible risks involved. Patients will then be given the opportunity to ask questions and the 
Investigator will be available to answer questions as needed. Patients will be informed of their 
right to withdraw from the study at any time without prejudice. After this explanation and before 
entering the study, the patient will voluntarily sign an ICF. The patient should receive a copy of 
the signed and dated ICF. The Investigator must retain each patient’s original signed ICF. 
If new information becomes available that may be relevant to the patient’s consent and 
willingness to participate in the study, the ICF will be revised. The revised ICF must be 
submitted to the IRB for review and approval prior to its use. 
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5. 
INVESTIGATORS AND STUDY ADMINISTRATIVE 
STRUCTURE 
This study will be performed at approximately 10 study centers in the US. The Investigator at 
each study center will be responsible for ensuring that the study is conducted according to the 
signed Clinical Trial Agreement, the protocol, IRB requirements, and GCP guidelines. 
The Investigator will be responsible for the oversight of the site’s conduct of the study, which 
will consist of completing all protocol assessments, maintaining the study file and the patient 
records, drug accountability, corresponding with the IRB, and completing the eCRF. 
5.1 
GENERATION OF STUDY RECORDS 
Ironwood or its designated representative will conduct a study center visit to verify the 
qualifications of each Investigator, inspect study center facilities, and inform the Investigator of 
responsibilities and procedures for ensuring adequate and correct study documentation. 
The Investigator is required to prepare and maintain adequate and accurate case histories 
designed to record all observations and other data pertinent to the study for each study patient. 
All information recorded in the eCRFs for this study must be consistent with the patient’s source 
documentation. 
During the course of the study, the Clinical Site Monitor will make study center visits to review 
protocol compliance, compare eCRFs and individual patient’s medical records, assess drug 
accountability (in a blinded manner), and ensure that the study is being conducted according to 
pertinent regulatory requirements. All eCRFs will be verified with source documentation. The 
review of medical records will be performed in a manner that ensures patient confidentiality is 
maintained. 
The Clinical Site Monitor will discuss instances of missing or uninterpretable data with the 
Investigator for resolution. Any changes to the study data will be made to the eCRF and 
documented via an electronic audit trail associated with the affected eCRF. 
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5.2 
DATA QUALITY ASSURANCE 
Ironwood performs quality control and assurance checks on all of its clinical studies. Section 5.4 
provides details regarding study monitoring procedures. 
The study may be subject to audit by Ironwood, its representatives, or regulatory authorities. In 
the event of an audit, the Investigator must agree to allow Ironwood, representatives of 
Ironwood, or the FDA or other regulatory agencies access to all study records. 
5.3 
ELECTRONIC CASE REPORT FORMS AND DATA MANAGEMENT 
All data relating to the study will be recorded in the patient’s source documentation and eCRF to 
be provided by Ironwood or designee via the electronic data capture (EDC) system. Source 
documentation supporting the eCRF data should indicate the patient’s participation in the study 
and should document the dates and details of study procedures, AEs, all observations, and patient 
status. The Investigator is responsible for verifying that all data entries on the eCRFs are accurate 
and correct and ensuring that all data are entered in a timely manner, as soon as possible after the 
information is collected. An explanation should be provided for any missing data. The 
Investigator as well as the central reader(s) must provide his or her formal approval through the 
EDC system of all the information in the eCRFs and changes to the eCRFs to endorse the final 
submitted data for each patient. 
Ironwood will retain the final eCRF data and corresponding audit trails. A copy of the final 
archival eCRF in the form of a compact disc or other electronic media will be placed in the 
Investigator’s study file. 
A record of screen failures and pretreatment failures will be maintained for patients who do not 
qualify for randomization, including the reason for the failure. 
5.4 
STUDY MONITORING 
Ironwood performs quality control and assurance checks on all of its clinical studies. Before any 
patients are enrolled in the study, a representative of Ironwood or its authorized designee will 
meet with the Investigator and his/her staff to review relevant and important study-related 
information including, but not limited to, the protocol, the Investigator’s Brochure, the eCRFs 
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and instructions for their completion using the EDC system, the procedure for obtaining 
informed consent, and the procedure for reporting AEs and SAEs. 
An Ironwood representative, the Clinical Site Monitor, will monitor the progress and conduct of 
the study by periodically conducting monitoring visits and by frequent communications 
(telephone, e-mail, letter, and fax) with the study centers. The site monitor will ensure that the 
study is conducted according to the protocol and regulatory requirements. During monitoring 
visits, the information recorded on the eCRFs will be verified against source documents. Upon 
request of the monitor, auditor, IRB, or regulatory authority, the Investigator should make all 
requested study-related records available for direct access. 
All aspects of the study will be carefully monitored by Ironwood or its designee for compliance 
with applicable government regulations with respect to GCP and current standard operating 
procedures. 
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6. 
STUDY SPONSORSHIP 
6.1 
INVESTIGATOR AND STUDY TERMINATION 
Ironwood may terminate Investigator participation at any institution for any reason. If 
participation is ended at the site by either Ironwood or the Investigator, the Investigator must 
• Return all study medications and any study materials to Ironwood; 
• In cases where the Investigator opts to self-terminate, provide a written statement describing 
why the study was terminated prematurely. 
 
Ironwood may terminate the study in its entirety or at a specific center at any time for any 
reason, including but not limited to the following: 
• Failure to enroll patients 
• Protocol violations 
• Inaccurate or incomplete data 
• Unsafe or unethical practice 
• Questionable safety of the study medication 
• Administrative decision 
 
6.2 
REPORTING AND PUBLICATION 
All data generated in this study will be the property of Ironwood. An integrated clinical and 
statistical report will be prepared at the completion of the study. 
Publication of the results by the Investigator will be subject to mutual agreement between the 
Investigator and Ironwood. 
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7. 
INVESTIGATOR OBLIGATIONS 
7.1 
DOCUMENTATION 
The Investigator must provide the Sponsor with the following documents BEFORE the 
enrollment of any subjects, in accordance with ICH E6 (Note: Ironwood must be notified if there 
are any changes to these documents): 
1. Completed and signed Form FDA 1572 (Statement of Investigator) including all sub-
investigators involved in the study 
2. Financial disclosure form(s) for the Investigator and all sub-investigators listed on Form 
FDA 1572 
3. Current, signed curricula vitae of the Investigator and all sub-investigators 
4. Copy of current medical license of the Investigator and all sub-investigators (as applicable) 
5. Copy of the IRB approval letter for the protocol and ICF 
6. Copy of the IRB-approved ICF to be used 
7. Copy of the IRB approval of recruitment advertising (if applicable) 
8. A list of IRB members and their qualifications, and a description of the committee’s working 
procedures 
9. Investigator’s Statement page, signed by the Investigator 
10. Fully executed Clinical Trial Agreement 
11. Written document containing the name, location, certification number, and date of 
certification of the local laboratory to be used for laboratory assays and those of other 
facilities conducting tests 
12. List of normal laboratory values and units of measurements for all laboratory tests required 
by the protocol. This list is required for each local laboratory to be used during the study. 
 
During the study, the Investigator must maintain the following essential/administrative 
documents related to the study: 
1. Copy of the signed Investigator’s Statement Page 
2. Copy of financial disclosure form(s) for the Investigator and all sub-investigators (as 
applicable) if updated 
3. Curricula vitae of any new Investigator(s) and/or sub-investigators involved in the study 
4. Copy of current medical license of the Investigator and all sub-investigators (as applicable) if 
updated 
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5. Copy of the signed Form FDA 1572 
6. IRB Approval Notification for the following: 
a. Protocol 
b. Informed consent document 
c. Recruitment advertising (if applicable) 
d. Amendment(s) (if applicable) 
e. Annual review of the protocol and the informed consent document 
f. SAEs 
g. Study closure 
7. SAE Reports 
8. Drug Inventory Forms (drug receipts, drug dispensing, and inventory forms) 
9. Name and address of local or central laboratory, list of normal laboratory values and units of 
measurement, as well as laboratory certification or hospital accreditation 
10. Updates of medical/laboratory/technical procedures/tests: 
a. Normal value(s)/ranges(s) 
b. Certification 
c. Accreditation 
d. Established quality control and/or external quality assessment 
e. Other validation (where required) 
11. Record of retained body fluids/tissue samples (if any) 
12. Correspondence with Sponsor 
13. Written assurance of continuing approval (at least annually) as well as a copy of the annual 
progress report submitted to the IRB must also be provided to the Sponsor. Any changes in 
this study or unanticipated problems involving risks to the patients must be reported 
promptly to the IRB. An Investigator must not make any changes in a study without IRB and 
Sponsor approval, except when necessary to eliminate apparent immediate hazards to the 
subjects. All protocol amendments must be submitted to the IRB and approved. 
14. Responsibility Log 
15. Other logs (eg, prescreening/screening, enrollment) 
16. Signed ICFs 
17. Patient source documentation 
18. eCRFs 
19. Audit certificate(s), if applicable 
 
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7.2 
PERFORMANCE 
The Investigator must demonstrate reasonable efforts to obtain qualified patients for the study. 
The Sponsor may terminate the study with any Investigator for any reason, including, but not 
limited to, Investigator nonperformance or Investigator noncompliance. 
7.3 
USE OF INVESTIGATIONAL MATERIALS 
The Investigator will acknowledge that the drug supplies are investigational and as such must be 
used strictly in accordance with the protocol and only under the supervision of the Investigator or 
sub-investigators. Study medication must be stored in a safe and secure temperature-monitored 
location. The Investigator’s duly trained designee must maintain adequate records documenting 
the receipt and disposition of all study supplies. The study center must record the date the study 
medication was received and maintain a dispensing record in which to record each patient’s use. 
A complete reconciliation of study medication will be performed at the site close-out visit, with a 
final accountability report provided to Ironwood as part of the site close-out report. Written 
instructions for return of all unused and reconciled study medication to an appropriate waste 
handler will be provided prior to the end of the study. No study medication may be destroyed by 
study centers without prior written permission of Ironwood. 
7.4 
RETENTION AND REVIEW OF RECORDS 
Records and documents pertaining to the conduct of this study, including eCRFs, source 
documents, ICFs, laboratory test results, and medication inventory records, must be retained by 
the Investigator in accordance with locally applicable regulatory requirements; and, in any event, 
for a minimum period of 5 years.  
No study records shall be destroyed without notifying the Sponsor and giving the Sponsor the 
opportunity to take such study records or authorizing in writing the destruction of records after 
the required retention period. 
If the Investigator retires, relocates, or otherwise withdraws from the responsibility of keeping 
the study records, custody must be transferred to another person (Ironwood, IRB, or other 
Investigator) who will accept the responsibility. Ironwood must be notified of and agree to the 
change. 
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7.5 
PATIENT CONFIDENTIALITY 
All data collected in the context of this study will be stored and evaluated in such a way as to 
guarantee patient confidentiality in accordance with the legal stipulations applying to 
confidentiality of data. All patient records will be identified only by initials and patient 
identification (PID) number. Patient names are not to be transmitted to Ironwood or its 
authorized designee. The Investigator will keep a Master Patient List on which the PID number 
and the full name, address, and telephone number of each patient is listed. 
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8. 
REFERENCE LIST 
1. Patel DA, Kim HP, Zifodya JS, Vaezi MF. Idiopathic (primary) achalasia: a review. 
Orphanet J Rare Dis 2015;10:89. 
2. Allaix ME, Patti MG. What is the best primary therapy for achalasia: medical or surgical 
treatment? Who owns achalasia? J Gastrointest Surg 2013 Sep;17(9):1547-9. 
3. Nenshi R, Takata J, Stegienko S, Jacob B, Kortan P, Deitel W, et al. The cost of achalasia: 
quantifying the effect of symptomatic disease on patient cost burden, treatment time, and 
work productivity. Surg Innov 2010 Dec;17(4):291-4. 
4. Derbyshire ER, Marletta MA. Structure and Regulation of Soluble Guanylate Cyclase. Annu 
Rev Biochem 2012;81:533-59. 
5. Nossaman BD, Kadowitz PJ. Stimulators of soluble guanylyl cyclase: future clinical 
indications. The Ochsner journal 2013;13(1):147-56. 
6. Stasch JP, Pacher P, Evgenov OV. Soluble guanylate cyclase as an emerging therapeutic 
target in cardiopulmonary disease. Circulation 2011;123(20):2263-73. 
7. Follmann M, Griebenow N, Hahn MG, Hartung I, Mais FJ, Mittendorf J, et al. The 
Chemistry and Biology of Soluble Guanylate Cyclase Stimulators and Activators. Angew 
Chem Int Ed Engl 2013;52(36):9442-62. 
8. Sivarao DV, Mashimo HL, Thatte HS, Goyal RK. Lower esophageal sphincter is achalasic in 
nNOS(-/-) and hypotensive in W/W(v) mutant mice. Gastroenterology 2001 Jul;121(1):34-
42. 
9. Gyawali CP. Achalasia: new perspectives on an old disease. Neurogastroenterol Motil 2016 
Jan;28(1):4-11. 
10. Hoshino M, Omura N, Yano F, Tsuboi K, Kashiwagi H, Yanaga K. Immunohistochemical 
study of the muscularis externa of the esophagus in achalasia patients. Dis Esophagus 2013 
Jan;26(1):14-21. 
11. Bortolotti M, Mari C, Lopilato C, Porrazzo G, Miglioli M. Effects of sildenafil on esophageal 
motility of patients with idiopathic achalasia. Gastroenterology 2000 Feb;118(2):253-7. 
12. Nassri A, Ramzan Z. Pharmacotherapy for the management of achalasia: Current status, 
challenges and future directions. World J Gastrointest Pharmacol Ther 2015 Nov 6;6(4):145-
55. 
13. Eherer AJ, Schwetz I, Hammer HF, Petnehazy T, Scheidl SJ, Weber K, et al. Effect of 
sildenafil on oesophageal motor function in healthy subjects and patients with oesophageal 
motor disorders. Gut 2002 Jun;50(6):758-64. 
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14. Herve D, Philippi A, Belbouab R, Zerah M, Chabrier S, Collardeau-Frachon S, et al. Loss of 
alpha1beta1 soluble guanylate cyclase, the major nitric oxide receptor, leads to moyamoya 
and achalasia. Am J Hum Genet 2014 Mar 6;94(3):385-94. 
15. National Cancer Institute. Common Terminology Criteria for Adverse Events v4.0.. May 29, 
2009. NCI, NIH, DHHS; NIH Publications #09-7473.  
16. EuroQol Group. EuroQol--a new facility for the measurement of health-related quality of 
life. The EuroQol Group. Health Policy 1990;16(3):199-208. 
17. Shaw JW, Johnson JA, Coons SJ. US valuation of the EQ-5D health states: development and 
testing of the D1 valuation model. Med Care 2005;43(3):203-20. 
18. Ware J, Kosinski M, Keller SD. A 12-Item Short-Form Health Survey: construction of scales 
and preliminary tests of reliability and validity. Med Care 1996;34(3):220-33. 
19. Urbach DR, Tomlinson GA, Harnish JL, Martino R, Diamant NE. A measure of disease-
specific health-related quality of life for achalasia. Am J Gastroenterol 2005 
Aug;100(8):1668-76. 
20. Robertson CS, Hardy JG, Atkinson M. Quantitative assessment of the response to therapy in 
achalasia of the cardia. Gut 1989;30(6):768-73. 
21. Gelfond M, Rozen P, Gilat T. Isosorbide dinitrate and nifedipine treatment of achalasia: a 
clinical, manometric and radionuclide evaluation. Gastroenterology 1982;83(5):963-9. 
22. Traube M, Dubovik S, Lange RC, McCallum RW. The role of nifedipine therapy in 
achalasia: results of a randomized, double-blind, placebo-controlled study. The American 
journal of gastroenterology 1989;84(10):1259-62. 
23. Traube M, Hongo M, Magyar L, McCallum RW. Effects of nifedipine in achalasia and in 
patients with high-amplitude peristaltic esophageal contractions. JAMA 1984;252(13):1733-
6. 
24. Bortolotti M, Labò G. Clinical and manometric effects of nifedipine in patients with 
esophageal achalasia. Gastroenterology 1981;80(1):39-44. 
25. Kahrilas PJ, Bredenoord AJ, Fox M, Gyawali CP, Roman S, Smout AJ, et al. The Chicago 
Classification of esophageal motility disorders, v3.0. Neurogastroenterol Motil 2015 
Feb;27(2):160-74. 
 
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10. 
INVESTIGATOR’S STATEMENT 
Study Title: 
A Multicenter, Randomized, Double-blind, Placebo-controlled, Parallel-
group, Single-dose, Phase 2a Study to Evaluate the Safety, Tolerability, 
Pharmacokinetics, and Pharmacodynamics of IW-1701 in Patients with 
Achalasia 
Study Number: 
C1701-201 
Final Date: 
03 October 2017  
I have read the protocol described above. I agree to comply with all applicable regulations and to 
conduct the study as described in the protocol.  
 
 
Signed: 
 
 
 
Date: 
 
 
 
 
Investigator Name: 
 
 
 
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11. 
APPENDICES 
 
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APPENDIX 2 
SF-12V2 
Protocol Number: C1701-201             Study Period:                                            Subject ID:                  
                                                             Visit Date: 
S
S
(
 
 
 
 
 
 
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Protocol Number: C1701-201             Study Period:                                            Subject ID:                  
                                                             Visit Date: 
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 Protocol Number: C1701-201             Study Period:                                            Subject ID:                  
                                                             Visit Date: 
 
 
 
 
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1. 
TABLE OF CONTENTS 
1. 
TABLE OF CONTENTS .........................................................................................................2 
2. 
LIST OF ABBREVIATIONS ..................................................................................................5 
3. 
INTRODUCTION ...................................................................................................................7 
3.1 
STUDY DESIGN SUMMARY .....................................................................................7 
3.2 
STUDY OBJECTIVES ..................................................................................................8 
3.3 
SCHEDULE OF EVENTS ............................................................................................9 
4. 
STATISTICAL METHODS, STUDY ASSESSMENTS AND ENDPOINTS, AND 
DETERMINATION OF SAMPLE SIZE ..............................................................................12 
4.1 
STATISTICAL AND ANALYTICAL PLAN ............................................................12 
4.1.1 
General Methods ................................................................................................12 
4.1.1.1 
Analysis Populations ................................................................................12 
4.1.1.2 
Study Medication .....................................................................................13 
4.1.1.3 
Statistical Methods ...................................................................................13 
4.1.1.4 
Missing Data ............................................................................................13 
4.1.1.5 
Study Periods ...........................................................................................13 
4.1.2 
Interim Analyses ................................................................................................14 
4.1.3 
Demographics ....................................................................................................14 
4.1.3.1 
Disposition of Subjects ............................................................................14 
4.1.3.2 
Protocol Deviations ..................................................................................14 
4.1.3.3 
Demographics and Baseline Characteristics ............................................15 
4.1.3.4 
Medical History .......................................................................................16 
4.1.3.5 
Prior and Concomitant Medication ..........................................................16 
4.1.3.6 
Achalasia Disease History .......................................................................16 
4.1.3.7 
Study Drug Compliance ...........................................................................17 
4.1.3.8 
Achalasia Diagnosis .................................................................................17 
4.1.4 
Pharmacodynamic Assessments and Analyses ..................................................17 
4.1.4.1 
High Resolution Impedance Manometry (HRIM) ...................................17 
4.1.4.2 
Esophagogastric Junction (EGJ) Pressure ...............................................19 
4.1.4.3 
Exploratory Biomarker Analyses .............................................................20 
4.1.4.4 
Multiple Comparisons/Multiplicity .........................................................20 
4.1.5 
Pharmacokinetic Assessments and Analyses .....................................................20 
4.1.5.1 
PK Parameters ..........................................................................................20 
4.1.5.2 
PK Analysis .............................................................................................21 
4.1.6 
Graphical Presentation of PK Data ....................................................................22 
4.1.7 
Patient-reported Symptom Assessments and Analyses .....................................22 
4.1.8 
Health Outcomes Assessments and Analyses ....................................................24 
4.1.8.1 
EuroQol-5 Dimension (EQ-5D-3L) .........................................................24 


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4.1.8.2 
Short Form 12 Health Survey Version 2 (SF-12v2) ................................25 
4.2 
SAMPLE SIZE JUSTIFICATION ..............................................................................26 
4.3 
SAFETY ANALYSIS ..................................................................................................26 
4.3.1 
Adverse Events ..................................................................................................26 
4.3.2 
Clinical Laboratory Data ....................................................................................28 
4.3.3 
Vital Signs Parameters .......................................................................................29 
4.3.4 
ECG Parameters .................................................................................................30 
4.3.5 
Physical Examination.........................................................................................30 
5. 
CHANGES FROM ANALYSES PLANNED IN THE PROTOCOL ...................................31 
6. 
DATA HANDLING CONVENTIONS .................................................................................32 
6.1 
REPEATED OR UNSCHEDULED ASSESSMENTS OF SAFETY 
PARAMETERS ...........................................................................................................32 
6.2 
CONVENTIONS FOR SUMMARIZING ADVERSE EVENTS ...............................32 
6.3 
MISSING DATE INFORMATION FOR ADVERSE EVENTS ................................32 
6.4 
MISSING SEVERITY ASSESSMENTS FOR ADVERSE EVENTS .......................32 
6.5 
MISSING RELATIONSHIP TO STUDY DRUG FOR ADVERSE EVENTS ..........33 
6.6 
MISSING DATE INFORMATION FOR PRIOR OR CONCOMITANT 
MEDICATIONS ..........................................................................................................33 
6.7 
HANDLING OF MISSING PK DATA .......................................................................33 
7. 
REFERENCES ......................................................................................................................35 
8. 
APPENDICES .......................................................................................................................36 
8.1 
ADMINISTRATIVE INTERIM STATISTICAL ANALYSIS PLAN .......................36 
8.2 
HEALTH OUTCOMES SURVEYS ...........................................................................43 
8.2.1 
Short Form 12 Health Survey Version 2............................................................43 
8.2.2 
EQ-5D-3L ..........................................................................................................46 
8.3 
SITE DOCUMENTATION .........................................................................................48 
 
 
 


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LIST OF TABLES 
 
 
Table 1. 
Schedule of Events .............................................................................................10 
Table 2. 
Statistical Methods .............................................................................................13 
Table 3. 
Study Periods .....................................................................................................14 
Table 4. 
Disposition Analyses .........................................................................................14 
Table 5. 
Important Protocol Deviations ...........................................................................15 
Table 6. 
Demographics ....................................................................................................15 
Table 7.  
Baseline Characteristics (PD Population) ..........................................................16 
Table 8. 
Prior and Concomitant Medication ....................................................................16 
Table 9. 
Pharmacodynamic Endpoints and Analyses ......................................................19 
Table 10. 
PK Parameters ....................................................................................................21 
Table 11. 
Patient Symptom Endpoints and Analyses ........................................................23 
Table 12. 
EQ-5D-3L Endpoints and Analyses ..................................................................24 
Table 13. 
EuroQol Quality-of-Life Questionnaire Utility Index .......................................25 
Table 14. 
SF-12v2 Endpoints and Analyses ......................................................................25 
Table 15. 
AE Classifications and Definitions ....................................................................27 
Table 16. 
AE Summary Tables Specifications ..................................................................28 
Table 17. 
Clinical Laboratory Summary Specifications ....................................................29 
Table 18. 
Vital Signs Summaries .......................................................................................29 
Table 19. 
Potentially Clinically Significant Post-Baseline Vital Signs .............................30 
 


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2. 
LIST OF ABBREVIATIONS 
Abbreviation 
Full Term 
ADO 
adverse event leading to drop out 
AE 
adverse event 
ATC 
Anatomical Therapeutic Chemical 
AUClast  
area under the plasma concentration time curve from time zero to the last 
observation 
BFT 
bolus flow time 
BLQ 
below the limit of quantitation 
BP  
blood pressure 
bpm 
beats per minute 
Clast 
last quantifiable plasma concentration 
Cmax 
maximum observed plasma concentration 
CS 
clinically significant 
CSR 
clinical study report 
ECG  
electrocardiogram 
eCRF 
electronic case report form 
EDC 
electronic data capture 
EGJ 
esophagogastric junction 
EOT 
end of treatment 
HDL-c 
high density lipoprotein cholesterol 
HRIM 
high-resolution impedance manometry 
IBH 
impedance bolus height 
ICH 
International Conference on Harmonisation 
IPD 
important protocol deviation 
IQR 
interquartile range 
IRP 
integrated relaxation pressure 
IWRS 
interactive web response system 
kg 
kilogram 
kg/m2  
kilograms per meters squared (body mass index) 
LDH  
lactate dehydrogenase 


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Abbreviation 
Full Term 
LDL-c 
low density lipoprotein cholesterol 
LES  
lower esophageal sphincter 
LLN 
lower limit of normal 
LLQ 
lower level of quantification 
MCV  
mean corpuscular volume 
MedDRA  
Medical Dictionary for Regulatory Activities 
mg  
milligram 
mL  
milliliter 
mmHg  
millimeters of mercury 
MPV 
mean platelet volume 
msec  
millisecond 
NCS 
not clinically significant 
PCS 
potentially clinically significant 
pd  
postdose 
PD  
pharmacodynamic(s) 
PK  
pharmacokinetic(s) 
pre  
predose 
PT  
preferred term 
QTcF 
QT interval corrected using Fridericia’s formula 
RBC 
red blood cell 
RDW 
red blood cell distribution width 
SAE 
serious adverse event 
SAS® 
Statistical Analysis System 
SE 
standard error 
SOC 
system organ class 
Tmax 
time of maximum observed plasma concentration 
TEAE 
treatment-emergent adverse event 
WBC 
white blood cell 
WHO 
World Health Organization 


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3. 
INTRODUCTION 
This statistical analysis plan (SAP) provides a more technical and detailed elaboration of the 
statistical analyses of the pharmacokinetics (PK), pharmacodynamics (PD), and safety data as 
outlined and/or specified in the final protocol of Study C1701-201 (Amendment 5, dated October 
3, 2017). Specifications for the tables, figures, and data listings are contained in a separate 
document. 
3.1 
STUDY DESIGN SUMMARY 
Study C1701-201 is a Phase 2a, multicenter, randomized, double-blind, placebo-controlled, 
parallel-group, single-dose study consisting of 3 distinct periods (Screening, Clinic, and Follow-
up); see Figure 1. The study will enroll subjects aged 18-75 who have been diagnosed with Type 
I or Type II achalasia, with an integrated relaxation pressure (IRP) >15 mmHg as assessed by 
baseline high-resolution impedance manometry (HRIM). As originally designed, approximately 
20 patients will be randomized to either IW-1701 or placebo in a 3:1 ratio.  
 
On the morning of Day 1, after an overnight fast of ≥8 hours and after a baseline HRIM 
procedure, randomized patients will receive 5 mg IW-1701 or placebo in a tablet formulation. 
Safety, PK, and PD assessments, including blood collections, will be performed at prespecified 
times (see Table 1). Following all study assessments and at the Investigator’s discretion, patients 
will be discharged on Day 2, at least 30 hours after study drug administration. Patients will have 


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a Follow-up Call on Day 4 (±1 day) to review adverse events (AEs). The Follow-up Period will 
end after the End of Trial Visit on Day 21 (±7 days). 
3.2 
STUDY OBJECTIVES  
In patients with primary Type I or II achalasia, following a single 5-mg oral dose of IW-1701, 
• To assess safety and tolerability 
• To determine the effects on measures of esophageal function by high-resolution impedance 
manometry (HRIM) 
• To determine the PK parameters, Cmax, Tmax, AUClast 
 
 


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3.3 
SCHEDULE OF EVENTS 
The schedule of events for Study C1701-201 is presented in Table 1. 


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Table 1. 
Schedule of Events 
 


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4. 
STATISTICAL METHODS, STUDY ASSESSMENTS AND 
ENDPOINTS, AND DETERMINATION OF SAMPLE SIZE 
4.1 
STATISTICAL AND ANALYTICAL PLAN 
Statistical analyses defined in this SAP will be performed using SAS Version 9.4 or newer. 
4.1.1 
General Methods 
Descriptive statistics (n, mean, standard deviation, minimum, maximum, median, and 
interquartile range [IQR]) will be calculated to summarize continuous variables. Frequency and 
percentage of patients in each category will be calculated to summarize categorical variables. 
Additionally, the geometric mean and geometric CV% will be calculated for PK parameters 
when data allows (i.e., concentration >0). Due to the exploratory nature of the trial, no 
adjustments will be made for multiplicity. Inferential statistics will be used for descriptive 
purposes only. If not otherwise specified, the baseline value is defined as the last nonmissing 
value measured before administration of study drug on Day 1. 
4.1.1.1 
Analysis Populations 
Screened Population 
All screened patients who have signed the informed consent form for the study and received a 
patient identification number will be included in the Screened Population. 
Safety Population 
All patients who receive study drug will be included in the Safety Population. Patients in the 
Safety Population will be evaluated for safety according to the treatment they actually received. 
Pharmacokinetic Population 
All patients who receive IW-1701 and have at least 1 evaluable postdose PK parameter 
assessment will be included in the PK Population. 
Protocol deviations or events that may affect the evaluability of the PK results are described 
under Section 4.1.5.2 . 


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Pharmacodynamic Population 
All patients who receive study drug and have at least 1 postdose PD assessment will be included 
in the PD Population. Protocol deviations or events that may affect IW-1701 systemic exposure 
(described in Section 4.1.5.2) will be evaluated prior to database lock to determine if PD results 
should be excluded. 
4.1.1.2 
Study Medication 
During the clinic period, patients will receive a single dose of either 5-mg of IW-1701 or 
matching placebo.  
4.1.1.3 
Statistical Methods 
For descriptive summaries, N will represent the total number of patients within the summarized 
population and n will represent the number of non-missing patients for the summarized endpoint. 
All summary statistics will be presented by treatment and based on non-missing endpoint values, 
unless otherwise specified.  
Table 2. 
Statistical Methods 
Method 
Definition 
Categorical Summaries 
• Number and percent of patients in each category (Percent based on number of 
non-missing values) 
• Number of missing patients in each category 
Continuous Summaries 
• Number of non-missing patients, mean, median, standard deviation, 
minimum, maximum, interquartile range 
• Geometric mean and geometric CV% (PK parameters only) 
 
4.1.1.4 
Missing Data 
No imputation will be performed for missing PD observations. All safety and tolerability data 
will be summarized and analyzed when data are available for a patient. Data handling for 
missing dates and other key safety data are described in Section 5. Refer to Section 5.7 for 
details regarding handling of missing PK data. 
4.1.1.5 
Study Periods 
The study periods are outlined in Table 3. 


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Table 3. 
Study Periods 
Period 
Start Date 
End Date 
Screening 
Date of Signed ICF 
Day -1 
Clinic 
Check-in (morning of Day 1) 
Discharge (Day 2) 
Follow-up Period* 
Immediately after Discharge 
End of Treatment Visit Date (Day 21±7) 
*Includes Follow-up Call on Day 4 ±1. 
 
4.1.2 
Interim Analyses 
At least one administrative interim analysis will be conducted. For details, please refer to the full 
administrative interim analysis plan in the appendices of this document. 
4.1.3 
Demographics  
4.1.3.1 
Disposition of Subjects  
Subject disposition will be presented as outlined in Table 4. 
Table 4. 
Disposition Analyses 
Analysis Population 
Description 
Summary Method 
Screening, PK, PD, Safety 
Disposition by investigational site, overall 
Counts 
Screening 
Reason for screening and pretreatment failure 
Categorical summary  
PD, Safety 
Premature discontinuation 
• Completed study 
• Prematurely discontinued 
- 
Reason for discontinuation 
Categorical summary  
 
4.1.3.2 
Protocol Deviations 
Protocol deviations and important protocol deviations (IPD) will be identified and documented 
for all randomized patients prior to unblinding through select individual data reviews and 
possible programmatic checks of the study data. IPDs will be determined based on review of all 
protocol deviations and protocol deviation categories (i.e., Use of Prohibited Medication) by 
members of the Ironwood Clinical Trials team, including but not limited to the Sponsor Medical 
Monitor and study Biostatistician.  


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Table 5. 
Important Protocol Deviations 
Analysis Population 
Description 
Summary Method 
Safety 
Important Protocol Deviations 
• those who entered the study even though they did not 
satisfy the entry criteria  
• those who developed discontinuation criteria during the 
study but were not withdrawn  
• those who received the wrong treatment or incorrect dose  
• those who received a prohibited concomitant treatment 
Listing 
 
4.1.3.3 
Demographics and Baseline Characteristics 
Patient demographics will be summarized by treatment arm and overall for the PD and, if 
different, the Safety Population as shown in Table 6. Baseline characteristics will be summarized 
for the PD population as shown in Table 7. 
Table 6. 
Demographics 
Endpoint 
Description 
Summary Method 
Age 
Age at informed consent date 
Continuous summary 
Weight 
Weight (kg) 
Height 
Height (m) 
BMI 
Weight (kg)/ [Height (m)]2 
Age category 
<40 years 
40 to <65 years 
≥65 years 
Categorical summary 
 
Sex 
Male 
Female 
Race 
• 
White 
• 
Non-white 
- Black or African American 
- Asian 
- American Indian or Alaska Native 
- Native Hawaiian or Other Pacific Islander 
- Other 
Ethnicity 
• 
Hispanic or Latino 
• 
Not Hispanic or Latino  
 


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Table 7.  
Baseline Characteristics (PD Population) 
Endpoint 
Description 
Summary Method 
Baseline Pharmacodynamic 
Parameters 
• 
Bolus Flow Time (BFT) 
o 
Upright 
o 
Supine 
• 
Integrated Relaxation Pressure 
(IRP) 
o 
Upright 
o 
Supine 
• 
Impedance Bolus Height (IBH) 
o 
~1 minute 
o 
~2 minutes 
o 
~5 minutes 
Continuous summary 
 
4.1.3.4 
Medical History 
Medical History (abnormalities and surgeries) reported at screening will be coded using 
MedDRA version 20.0. A complete listing by subject of medical and surgical history will be 
presented.  
4.1.3.5 
Prior and Concomitant Medication 
Reported medication will be coded using the World Health Organization (WHO) Drug 
Dictionary (version December 2016 LT) to the respective Anatomical Therapeutical Chemical 
(ATC) class and PT (Table 8). A complete listing by subject of prior and concomitant 
medications will be presented.  
Table 8. 
Prior and Concomitant Medication 
Parameter 
Description 
Summary 
Prior medications 
Medication taken prior to first dose 
of study medication date 
Listing 
Concomitant medication 
Medication taken on or after first 
dose of study medication date 
 
4.1.3.6 
Achalasia Disease History 
A complete listing by subject of Achalasia Disease History reported at Screening will be 
presented. 


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4.1.3.7 
Study Drug Compliance 
This is an in-clinic dosing study and, therefore, an assessment of compliance is not necessary. A 
data listing containing the study drug dosing information will be provided and will include the 
number of tablets each patient swallowed. 
4.1.3.8 
Achalasia Diagnosis 
Per Protocol, a diagnosis of achalasia is determined from supine 10-swallow protocol on baseline 
HRIM procedure as read by the Investigator at the site. Subjects enrolled under C1701-201-P-05 
or earlier will have two sets of predose swallows; diagnosis will be determined from the first set 
of predose swallows. In the event that the first set of predose swallows is missing, the diagnosis 
will be determined from the second set of predose swallows. The EDC allows the Investigator at 
the site to enter a diagnosis only after the first predose set of swallows. A patient with a missing 
first set of predose swallows, but an applicable diagnosis on the second set of predose swallows 
will be randomized and included in analysis. Because a diagnosis from a second set of swallows 
cannot be entered into the EDC, correspondence with the site and documentation of the Type I or 
Type II achalasia diagnosis will be included as an Appendix in this SAP (see Appendix 7.3). 
A complete by subject listing of Achalasia diagnosis will be presented. 
4.1.4 
Pharmacodynamic Assessments and Analyses 
4.1.4.1 
High Resolution Impedance Manometry (HRIM) 
HRIM Methods: After calibration, the HRIM catheter will be inserted transnasally to record 
manometric data from the hypopharynx to the stomach. After insertion and ~2-minute baseline 
(relaxed) recordings to assess basal esophagogastric junction (EGJ) pressure, the following set of 
swallowing protocols will be recorded. 
Timing 
• 10 supine (at ~30-degree incline) ~5-mL saline swallows 
• 5 upright ~5-mL saline swallows (after upright and relaxed for ~1 minute) 
• (Optional) Standing ~200-mL saline bolus challenge (at both baseline and postdose) 
 


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Measurement 
• Bolus Flow Time (BFT) – time in seconds of bolus (~5 mL swallow) transit through the 
EGJ from the 10-swallow supine and 5-swallow upright protocols 
• Integrated Relaxation Pressure (IRP) – mean of the 4s of maximal deglutitive relaxation in 
the 10-s window beginning at upper esophageal sphincter relaxation. Contributing times can 
be contiguous or non-contiguous (e.g., interrupted by diaphragmatic contraction).  
• Impedance Bolus Height (IBH) – height in esophagus of ~200 mL saline bolus at ~1, ~2, 
and ~5 minutes postbolus from the standing ~200-mL bolus challenge (if performed) 
 
HRIM measurements will be recorded at each site and then sent to a central reader for analysis. 
For all PD analyses, the values determined by the central reader will be used. 
Subjects randomized under protocol version C1701-201-P-05 or earlier will have 2 sets of 
swallowing protocols both pre- and postdose. For subjects with 2 sets of predose swallowing 
protocols, the first predose recording will be used to obtain the baseline BFT and IRP values. For 
subjects with 2 sets of postdose swallowing protocols, the second postdose recording will be 
used to define the postdose BFT and IRP values. If a subject randomized under C1701-201-P-05 
or earlier is missing the first predose recording, the second predose recording will be used to 
obtain the baseline BFT and IRP values. Similarly, if a subject randomized under C1701-201-P-
05 or earlier is missing the second postdose recording, the first postdose recording will be used 
to define the postdose BFT and IRP values. 
Descriptive statistics will be calculated at baseline and each postdose assessment. Mean and 
median change from baseline for each parameter will be presented by treatment group. Percent 
change (from baseline to postdose) in IRP parameters will also be calculated and summarized by 
treatment group. A shift table from baseline to postdose will also be created for BFT to 
summarize improvement in flow (BFT=0 and BFT>0). Refer to Table 9 for a full description of 
endpoints. All pre- and postdose values (not just the median) will be plotted for each HRIM 
measure (BFT and IRP) by patient and timepoint. A complete listing of all HRIM measurements 
provided from both the site (only available for predose assessments) and central read will be 
provided for each patient. 


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Table 9. 
Pharmacodynamic Endpoints and Analyses 
Endpoint 
Description 
Timing 
Summary 
Method 
Change from 
Baseline in Supine 
Bolus Flow Time 
(BFT) 
Supine BFT defined as the median measurement 
from the 10 available swallows in supine position as 
determined by the central read (seconds; longer 
times=more severe achalasia). Change=(postdose 
supine BFT – predose supine BFT). 
Predose, 
postdose (3 
hours +15 
minutes after 
dosing), and 
change. 
 
Continuous 
summary 
 
 
Change from 
Baseline in 
Upright BFT) 
Upright BFT defined as the median measurement 
from the 5 available swallows in the upright position 
as determined by the central read (seconds; longer 
times=more severe achalasia). Change = (postdose 
upright BFT – predose upright BFT). 
Change from 
Baseline in Supine 
Integrated 
Relaxation 
Pressure (IRP) 
Supine IRP defined as the median measurement from 
the 10 available swallows in supine position as 
determined by the central read (mmHg; higher 
pressure=more severe achalasia). Change = (postdose 
supine IRP – predose supine IRP). Percent change = 
((postdose supine IRP – predose supine IRP) ÷ 
(predose supine IRP)) × 100  
Change from 
Baseline in 
Upright IRP 
Upright IRP defined as the median measurement 
from the 5 available swallows in the supine position 
as determined by the central read (mmHg; higher 
pressure=more severe achalasia). Change = (postdose 
upright IRP – predose upright IRP). Percent change = 
((postdose upright IRP – predose upright IRP) ÷ 
(predose upright  IRP)) × 100 
Change from 
Baseline in 
1-minute 
Impedance Bolus 
Height (IBH) (if 
performed) 
1-minute IBH defined by the height in esophagus of 
200 mL saline bolus 1-minute post-bolus as 
determined by the central read (cm; greater 
height=more severe achalasia). Change = (postdose 
height 1 min IBH – predose height 1 min IBH) 
Change from 
Baseline in 
2-minute IBH (if 
performed) 
2-minute IBH defined by the height in esophagus of 
200 mL saline bolus 2 minutes post-bolus as 
determined by the central read (cm; greater 
height=more severe achalasia). Change = (postdose 
height 2 min IBH – predose height 2 min IBH). 
Change from 
Baseline in 
5-minute IBH (if 
performed) 
5-minute IBH defined by the height in esophagus of 
200 mL saline bolus 5 minutes post-bolus as 
determined by the central read (cm; greater 
height=more severe achalasia). Change = (postdose 
height 5-min IBH – predose height 5-min IBH) 
 
4.1.4.2 
Esophagogastric Junction (EGJ) Pressure 
Basal EGJ pressure will be recorded to assess the resting pressure without swallowing. For 
subjects randomized under C1701-201-P-05 and earlier resting EGJ pressure will be recorded 


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after insertion of HRIM catheter and every 15 minutes for 1 hour (baseline, 15 min, 30 min, 45 
min, and 60 min EGJ pressure) both predose and postdose. Subjects randomized under C1701-
201-P-06 only have a single measure of EGJ pressure assessed following insertion of the 
catheter. These values will be presented by subject and timepoint in a data listing. 
4.1.4.3 
Exploratory Biomarker Analyses 
Blood samples for determination of plasma renin activity will be collected according to the 
schedule of events. As data permit, summaries for each postdose timepoint and change from 
baseline may be calculated. Results may be presented outside of the clinical study report (CSR). 
4.1.4.4 
Multiple Comparisons/Multiplicity 
Due to the exploratory nature of the trial, no adjustments will be made for multiplicity. 
4.1.5 
Pharmacokinetic Assessments and Analyses 
4.1.5.1 
PK Parameters 
The PK parameters listed in Table 10 will be calculated for each patient, whenever possible, if 
systemic levels of IW-1701 are quantifiable. PK parameter calculations will be performed using 
noncompartmental methods with Phoenix® WinNonlin® Version 6.4, or higher, (Certara LP, 
Princeton, NJ).  
Plasma concentrations that are below the lower level of quantification (LLQ) or missing will be 
treated as described in Section 4.1.5.2 for the calculation of PK parameters. Actual sampling 
times relative to IW-1701 dosing times will be used in calculations of the PK parameters. Area 
under the concentration-time curve will be calculated by linear up/log down trapezoidal 
summation. 


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Table 10. 
PK Parameters 
 
Parameter* 
Description 
AUClast 
Area under the plasma concentration time curve from time zero to Tlast, the time at 
which the last quantifiable plasma concentration (Clast) is observed. 
Tmax 
Time of maximum observed plasma concentration, obtained directly from the 
concentration-time profile.   
Cmax 
Maximum observed plasma concentration, occurring at Tmax    
*To be calculated relative to the morning dose, unless otherwise noted. 
 
4.1.5.2 
PK Analysis 
Plasma concentration of IW-1701 will be summarized using the PK Population for each 
assessment timepoint for the active treatment group. Concentrations that are below LLQ (BLQ) 
will be treated as zero for the computation of descriptive statistics.  A by-subject listing of all 
concentration-time data and scheduled sample collection time will be presented.  A listing of PK 
blood sample collection times, and elapsed times relative to dose, and time deviations will be 
provided. 
Derived PK parameters for IW-1701 will be summarized using the PK Population. A by-subject 
listing of all calculated PK parameters will be presented. 
Changes to protocol procedures or events which many impact the quality of the PK data or alter 
the evaluation of the PK will be reviewed on a case-by-case basis to determine if PK data may be 
excluded from relevant PK summaries.  Not all the events may be considered a protocol 
deviation; as an event may be secondary to an AE or to a circumstance outside of the control of 
the study site or the patient.  Examples include, but may not be limited to, vomiting following 
oral dosing occurring within the time frame of 2 times the median time to Tmax, sample 
processing errors that lead to inaccurate bioanalytical results, inaccurate dosing or administration 
of a prohibited medication expected to impact IW-1701 exposure.  Deviations to procedures or 
events which do not impact the quality of the PK data such as a missed blood sample or 
deviations from blood collection times will not be considered for exclusion from PK summaries. 


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4.1.6 
Graphical Presentation of PK Data 
Geometric mean plasma concentrations will be plotted over time on both a linear and a semi-
logarithmic scale. Individual drug plasma concentrations will also be plotted over time.  
4.1.7 
Patient-reported Symptom Assessments and Analyses 
Patients will answer a number of Screening Patient-reported Symptom Questions at the 
Screening Visit. Refer to Table 11 for full description. 
Following removal of the HRIM catheter after both the baseline and postdose procedures, the 
patients will answer a set of questions related to their most recent swallowing protocols; the 
answers will be entered into each patient’s eCRF.  
Descriptive statistics will be calculated at baseline and each postdose assessment for the patient-
reported symptoms data. Change from baseline for each symptom will be calculated and 
presented by treatment group. Refer to Table 11 for full endpoint descriptions. 


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Table 11. 
Patient Symptom Endpoints and Analyses 
Endpoint 
Description 
Timing 
Summary 
Method 
Analysis 
Population 
Severity of difficulty 
swallowing food 
during/between 
mealsa 
During the last 7 days, how would you rate the 
severity of your difficulty swallowing food 
during/between meals? 0=Did not have, 1=Very 
mild, 2=Mild, 3=Moderate, 4=Moderatively 
severe, 5=Severe 
Screening 
Continuous 
summary 
Screened 
population; 
PD 
population 
Severity of pain 
while swallowing 
food during/between 
mealsa 
During the last 7 days, how would you rate the 
severity of your pain while swallowing food 
during/between meals? 0=Did not have, 1=Very 
mild, 2=Mild, 3=Moderate, 4=Moderatively 
severe, 5=Severe 
Frequency of cough 
while swallowing 
food during/between 
mealsa 
During the last 7 days, how often did you 
experience cough while swallowing food 
during/between meals? 0=Never, 1=Rarely, 
2=Sometimes, 3=Often, 4=Very often 
Impact of achalasia/ 
swallowing disorder 
symptoms on 
what/when you ateb 
During the last 7 days, how much did your 
achalasia/swallowing disorder symptoms impact 
what/when you ate? 0=Not at all, 1=A little bit, 
2=Somewhat, 3=Quite a bit, 4=A lot 
Overall severity of 
achalasia/ 
swallowing disorder 
symptoms 
How would you rate the overall severity of 
achalasia/swallowing disorder symptoms over 
the past 7 days? 0=Did not have, 1=Very mild, 
2=Mild, 3=Moderate, 4=Moderately severe, 
5=severe 
Ability to swallow 
raw hard fruits and 
vegetables/rice/ 
clear fluids (water, 
juice, coffee, tea)c 
Please indicate which of the following types of 
foods you are able to swallow without 
experiencing any problems such as pain or food 
“sticking” as it goes down. Check one box for 
each type of food. Options: Can swallow without 
problem; can swallow, but with some difficulty; 
Can swallow with great difficulty or not at all 
Categorical 
summary 
Change from 
baseline in severity 
of difficulty 
swallowing during 
swallowing tests 
Severity of difficulty swallowing during 
swallowing tests (0=Did not have, 1=Very mild, 
2=Mild, 3=Moderate, 4=Moderately severe, 
5=Severe); Change=(postdose severity – predose 
severity). 
Within 15 
minutes of 
predose 
HRIM; 
within 15 
minutes of 
postdose 
HRIM; 
Change 
Continuous 
summary 
PD 
population 
Change from 
baseline in severity 
of pain while 
swallowing during 
swallowing tests 
Severity of pain while swallowing during 
swallowing tests (0=Did not have, 1=Very mild, 
2=Mild, 3=Moderate, 4=Moderately severe, 
5=Severe); Change=(postdose severity – predose 
severity). 
Change from 
baseline in 
frequency of 
coughing while 
swallowing during 
swallowing tests 
Frequency of cough while swallowing during 
swallowing tests (0=Never, 1=Rarely, 
2=Sometimes, 3=Often, 4=Very often); 
Change=(postdose frequency – predose 
frequency). 
Relief of achalasia 
symptoms 
Overall achalasia symptoms since treatment with 
study medication (1=Significantly relieved, 
2=Moderately relieved, 3=Somewhat relieved, 
Within 15 
minutes of 
Continuous 
summary 


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Endpoint 
Description 
Timing 
Summary 
Method 
Analysis 
Population 
4=Unchanged, 5=Somewhat worse, 
6=Moderately worse, 7=Significantly worse) 
postdose 
HRIM 
aTwo separate endpoints; one for during meals and one for between meals. 
bTwo separate endpoints; one for what you ate and one for when you ate. 
cThree separate endpoints; one for fruits and vegetables, one for rice, and a third for clear fluids. 
4.1.8 
Health Outcomes Assessments and Analyses 
4.1.8.1 
EuroQol-5 Dimension (EQ-5D-3L) 
Patients will complete the EQ-5D-3L at Baseline (Screening Visit) and the EOT visit. Patient 
responses to the descriptive system (e.g., health state) will be converted to the corresponding 
utility score and the descriptive statistics (n, mean, SD, median, minimum and maximum) will be 
presented for utility score by treatment group at each timepoint for the PD Population. The same 
statistics will be calculated for the analog scale reflecting the patient’s preference for their health 
state. 
Table 12. 
EQ-5D-3L Endpoints and Analyses 
 
Endpoint 
Description 
Timing  
Summary 
Method 
Analysis 
Population 
Utility Score 
Multi-component, single measure derived 
from 5 general health items: mobility, self-
care, usual activities, pain/discomfort, and 
anxiety/depression. Each item measured on 0 
to1 scale.  
Screening, 
EOT Visit 
Continuous 
descriptive 
PD Population 
Visual Analog 
Scale 
Rating of Current Health as of today (0-100): 
Higher score indicates better health 
 
The 5-item responses of the EQ-5D-3L define a health state that can be converted into a single 
summary utility index that assigns weights (i.e., value) to each level. Different weights are 
available. For this study, the US TTO (time trade-off) weights will be used.(1) 
The EQ-5D-3L index is computed according to the following equation as:  
 
 
 
 


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Table 13. 
EuroQol Quality-of-Life Questionnaire Utility Index 
If any dimension is missing in the EQ-5D-3L, the EQ-5D-3L index will not be calculated and 
will be set to missing. 
4.1.8.2 
Short Form 12 Health Survey Version 2 (SF-12v2)  
Patients will complete the SF-12v2 at Baseline (Screening Visit) and the EOT visit. SF-12v2 
scores will be calculated using QualityMetric Health Outcomes(™) Scoring Software 5.0. 
Summaries will be provided by treatment for the PD population at each timepoint as well as the 
change from baseline as described in Table 14. 
Table 14. 
SF-12v2 Endpoints and Analyses 
Endpoint 
Description 
Timing  
Summary 
Method 
Analysis 
Population 
SF-12v2 Physical 
Component 
Summary Score 
8 concepts of health: physical functioning, 
role limitations due to physical health 
problems, bodily pain, general health, vitality 
(energy/fatigue), social functioning, role 
limitations due to emotional problems, and 
mental health (psychological distress and 
psychological well-being); aggregated into 2 
summary measures: the physical component 
and the mental component summary scores. 
Screening, 
EOT Visit, 
change 
from 
baseline 
Continuous 
descriptive 
PD 
population 
SF-12v2 Mental 
Component 
Summary Score 


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The SF-12 is a multipurpose, short-form health survey consisting of 12 questions designed for 
use in clinical practice and research, health policy evaluations, and general population surveys. 
The SF-12, Version 2.0, contains 12 items that, when scored, yield an 8-scale profile of 
functional health and well-being, as well as physical and mental summary measures.  
Two distinct, higher-order summary norm-based scores: Physical Component Summary 
(PCS-12) and Mental Component Summary (MCS-12) scores are standardized to have a mean of 
50 and an SD of 10 in the general US Population. These two scores will be summarized as 
described in Table 14. 
The steps involved in scoring SF-12 can be found at Ware et al. (2,3) 
If any item is missing in SF-12, then the PCS-12 and MCS-12 scores will not be calculated and 
will be set to missing. 
4.2 
SAMPLE SIZE JUSTIFICATION 
For this exploratory Phase 2a study, a total sample size of up to 20 patients is planned (subjects 
will be randomized to either IW-1701 or placebo in a 3:1 ratio, resulting in approximately 15 
patients receiving IW-1701 and approximately 5 patients receiving placebo). The sample size 
was determined outside of statistical considerations and is based on precedent set by prior studies 
of similar nature and design.(3, 4) 
4.3 
SAFETY ANALYSIS 
4.3.1 
Adverse Events 
All adverse events reported during the study will be coded using MedDRA version 20.0 and will 
be classified by MedDRA system organ class (SOC) and preferred term (PT) as outlined in Table 
15. 
 
 


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Table 15. 
AE Classifications and Definitions 
AE classification 
Definition 
Treatment-
emergent Adverse 
Event (TEAE) 
TEAE if: 
• Treatment Start date/time ≤ AE Start Date/Time ≤ (Date of Last Treatment +72 hours) 
And 
• AEs that first occurred or worsened in severity after study drug administration 
Serious Adverse 
Events 
SAEs will be recorded from the time the subject signs the ICF at the Screening Visit through 
the End of Trial Visit (Day 21 ±7). 
 
An SAE is defined as any AE occurring at any dose that results in any of the following 
outcomes: 
• Death 
• Life-threatening: the patient was at immediate risk of death from the reaction as it 
occurred 
• Hospitalization or prolongation of existing hospitalization 
• Persistent or significant disability/incapacity: a substantial disruption of a person’s 
ability to conduct normal daily functions 
• Congenital anomaly/birth defect 
• Important medical events: events that may not result in death, be life threatening, or 
require hospitalization. Such an event may be considered serious when, based on 
appropriate medical judgment, it may jeopardize the subject and may require medical or 
surgical intervention to prevent 1 of the outcomes listed in this definition. Examples of 
such medical events include allergic bronchospasm requiring intensive treatment in an 
emergency department or at home, blood dyscrasias or convulsions that do not result in 
inpatient hospitalization, or the development of drug dependency or drug abuse. 
 
These criteria will be identified by AE Serious criteria = Yes 
Patients reporting multiple incidence of an adverse event mapping to the same PT will be 
counted only once overall, at the highest severity level, and at the closest relationship to 
treatment. AEs will be summarized as described in Table 16. 


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Table 16. 
AE Summary Tables Specifications 
 
Summaries 
Description 
Summary 
Method 
Analysis 
Population 
Overall Summary  
• TEAEs 
• Deaths 
• SAEs 
• Withdrawals Due to TEAEs (ADOs) 
• Premature Discontinuations 
•  
Categorical 
summary 
 
Safety 
population 
TEAEs 
• At least 1 TEAE  
• By SOC 
• By PT within SOC 
 
TEAEs by Severity 
TEAEs by Relationship to Study 
Drug 
Severity 
Overall Summary by Severity Level 
• 
Mild 
• 
Moderate 
• 
Severe 
Relationship to Study Drug 
Overall Summary by Relationship to Study 
Drug 
• 
Related 
• 
Unrelated 
Listings of All Adverse Events 
(including pretreatment) for 
Patients with: 
1. SAEs 
2. ADOs 
3. Fatal 
For Patients meeting the defined criteria, list 
all of the patient’s Adverse Events  
Data listing 
– See table 
shells for 
variables to 
include 
Listing of Screening Period 
Adverse Events 
AEs occurring prior to first dose of study drug 
Data listing 
4.3.2 
Clinical Laboratory Data 
Clinical laboratory results recorded over the course of the study will be summarized as defined in 
Table 17. Baseline is defined as the last non-missing laboratory assessment reported prior to 
dosing with study medication. Complete clinical laboratory listings will be provided for each 
patient. Central laboratory results are preferred. If patients have both central and local labs for a 
given assessment, the central laboratory results will be used. 


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Table 17. 
Clinical Laboratory Summary Specifications 
 
 
Endpoint 
Description 
Timing 
Summary Method 
Analysis 
Population 
Shift from 
baseline 
Summary by laboratory category 
and parameter 
• 
Normal 
• 
Abnormal NCS 
• 
Abnormal CS 
Postdose Day 1, Day 
2 (Discharge), Day21 
(End of Trial Visit) 
Categorical 
summary 
(denominator based 
on patients with 
both baseline and 
follow-up measure) 
Safety 
population 
 
4.3.3 
Vital Signs Parameters 
Vital signs endpoints will be defined and summarized as described in Table 18. Criteria for 
potentially clinically significant post-baseline vital signs is summarized in Table 19. Baseline is 
defined as the last nonmissing vital sign value prior to dosing with study medication. Complete 
vital sign listings will be provided for each patient.  
Table 18. 
Vital Signs Summaries 
Endpoint 
Description 
Timing 
Summary Method 
PCS Vital 
Signs 
Summarized by vital 
signs category (Table 19) 
and parameter 
Baseline (predose values Day 1), Day 1 
(postdose: 0.5h ±10m, 1h, 2h, 3h, 4h, 6h, 
8h, 12h±15m), Day 2 (postdose: 24h 
±30m; at discharge (BP only): 30h or 
later), EOT visit (Day 21±7) 
Categorical summary 
(denominator based on 
patients with both 
baseline and follow-up 
measure) 
Descriptive 
• 
Summary BP and 
Pulse at each 
timepoint measured 
• 
Change from baseline 
Baseline (predose values Day 1), Day 1 
(postdose: 0.5h ±10m, 1h, 2h, 3h, 4h, 6h, 
8h, 12h±15m), Day 2 (postdose: 24h 
±30m; at discharge (BP only): 30h or 
later), EOT visit (Day 21±7)  
Continuous summary 
• 
Summary oral 
temperature and 
respiratory rate 
Baseline (predose values Day 1), Day 2 
(24h ±30m), EOT visit (Day 21 ±7) 
Continuous summary 
 


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Table 19. 
Potentially Clinically Significant Post-Baseline Vital Signs 
Vital Sign Parameter 
Flag 
Criteria* 
Observed Value 
Change from Study Baseline 
Seated systolic blood pressure 
(mmHg) 
High 
≥ 180 
Increase of ≥ 20 
Low 
≤ 90 
Decrease of ≥ 20 
Seated diastolic blood 
pressure (mmHg) 
High 
≥ 105 
Increase of ≥ 15 
Low 
≤ 50 
Decrease of ≥ 15 
Seated pulse rate 
(bpm) 
High 
≥ 110 
Increase of ≥ 15 
Low 
≤ 50 
Decrease of ≥ 15 
*A postbaseline value is considered as a notable value if it meets both criteria for observed value and change from 
baseline. 
 
4.3.4 
ECG Parameters 
Baseline ECG is defined as the last nonmissing value prior to dosing with study medication. 
Shift tables will be provided to show the categorical shift from baseline at each postdose 
assessment (Normal, Abnormal NCS, Abnormal CS). Complete ECG listings will be provided 
for each patient. 
4.3.5 
Physical Examination 
Physical examination results for all subjects will be presented in a data listing for the Safety 
Population. 


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5. 
CHANGES FROM ANALYSES PLANNED IN THE 
PROTOCOL 
The following analyses are added to those specified in the study protocol: 
1. The addition of a Screened Population to the Analysis Populations. 
2. Descriptive statistics for chemistry, vital signs, and ECG parameters have been removed 
and shift tables only will be presented due to the small number of subjects enrolled 
(N=9). 


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6. 
DATA HANDLING CONVENTIONS 
6.1 
REPEATED OR UNSCHEDULED ASSESSMENTS OF SAFETY 
PARAMETERS 
If a patient has repeated safety assessments (e.g., for a lab parameter) prior to dosing of study 
drug, then the results from the final non-missing assessment made prior to the start of study drug 
will be used as baseline. If there is more than one safety measurement (e.g., for a lab parameter) 
at a postbaseline timepoint, only the last measurement will be used. All postbaseline assessments 
including unscheduled assessments, if any, will be used for notable value determination, and all 
assessments will be presented in data listings.  
6.2 
CONVENTIONS FOR SUMMARIZING ADVERSE EVENTS 
The following conventions will be followed in summarizing TEAEs within a treatment group: 
• For patient incidence summaries, each patient will be counted only once within each 
SOC, PT, or the overall AE summary 
• If a patient reported more than one AE within an SOC or PT, then the TEAE with the 
highest severity or strongest study drug relationship within each SOC and each PT will be 
included in the summaries by severity or relationship, respectively.   
6.3 
MISSING DATE INFORMATION FOR ADVERSE EVENTS 
If it is not possible to determine when an AE started due to incomplete start date/times, it will be 
assumed to be treatment emergent. 
6.4 
MISSING SEVERITY ASSESSMENTS FOR ADVERSE EVENTS 
If severity is missing for an AE that started prior to the dosing of study drug, all efforts should be 
made to obtain the severity from the Investigator. If it is still missing after all efforts, then a 
severity of “Mild” will be assigned. If the severity is missing for an AE that started on or after 
the date of dosing of study drug, then a severity of “Severe” will be assigned. The imputed 
values for the missing severity assessment will be used for the incidence summary, while the 
actual missing values will be presented in data listings. 


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6.5 
MISSING RELATIONSHIP TO STUDY DRUG FOR ADVERSE 
EVENTS 
If the relationship to study drug is missing for an AE that started on or after the date of dosing of 
study drug, all efforts should be made to obtain the relationship from the Investigator. If it is still 
missing after all efforts, a causality of “Related” will be assigned in the corresponding analysis-
derived data set. The imputed values for the missing relationship to study drug will be used only 
for incidence summary, while the actual missing values will be presented in data listings. 
6.6 
MISSING DATE INFORMATION FOR PRIOR OR CONCOMITANT 
MEDICATIONS 
If the start date of a medication is missing or incomplete (i.e., partially missing), then the 
medication will be assumed to be concomitant. 
6.7 
HANDLING OF MISSING PK DATA 
• Handling of predose samples prior to the IW-1701 administration: Concentrations that 
are BLQ or missing will be assigned a numerical value of zero for the calculation of 
AUC. Any anomalous concentration values observed at predose will be included in the 
computation of AUC. If the predose concentration is greater than 5% of Cmax, the data 
will be evaluated on a case-by-case basis to determine if exclusion of the affected profile 
is warranted.   
• Handling of all other BLQ concentrations: Any other BLQ concentrations will be 
assigned a value of zero if at predose or if they precede quantifiable samples in the initial 
portion of the profile. A BLQ value that occurs between quantifiable data points, 
especially prior to Cmax, will be evaluated to determine if an assigned concentration of 
zero makes sense, or if exclusion of the data is warranted. Following Cmax, BLQ values 
embedded between 2 quantifiable data points will be treated as missing when calculating 
AUC. If BLQ values occur at the end of the collection interval (after the last quantifiable 
concentration), these will be set to zero. If consecutive BLQ concentrations are followed 
by quantifiable concentrations in the terminal portion of the concentration curve, these 
quantified values will be excluded from the PK analysis by setting them to missing, 
unless otherwise warranted by the concentration-time profile. 


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• Handling of missing data: In general, missing data in a given profile will not be imputed. 
The affected profile(s) will be evaluated whether sufficient IW-1701 concentrations are 
available to calculate any of the planned PK parameters.  


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7. 
REFERENCES 
1. Shaw JW, Johnson JA, Coons SJ. US valuation of the EQ-5D health states: development and 
testing of the D1 valuation model. Med Care. 2005;43(3):203-20. 
2. Ware J, Jr., Kosinski M, Keller SD. A 12-Item Short-Form Health Survey: construction of 
scales and preliminary tests of reliability and validity. Med Care. 1996;34(3):220-33. 
3. Bortolotti M, Mari C, Lopilato C, Porrazzo G, Miglioli M. Effects of sildenafil on esophageal 
motility of patients with idiopathic achalasia. Gastroenterology. 2000;118(2):253-7. 
4. Bortolotti M, Labò G. Clinical and manometric effects of nifedipine in patients with 
esophageal achalasia. Gastroenterology. 1981;80(1):39-44. 
5. Gelfond M, Rozen P, Gilat T. Isosorbide dinitrate and nifedipine treatment of achalasia: a 
clinical, manometric and radionuclide evaluation. Gastroenterology. 1982;83(5):963-9. 
6. Robertson CS, Hardy JG, Atkinson M. Quantitative assessment of the response to therapy in 
achalasia of the cardia. Gut. 1989;30(6):768-73. 
7. Traube M, Hongo M, Magyar L, McCallum RW. Effects of nifedipine in achalasia and in 
patients with high-amplitude peristaltic esophageal contractions. JAMA. 1984;252(13):1733-
6. 
8. Traube M, Dubovik S, Lange RC, McCallum RW. The role of nifedipine therapy in 
achalasia: results of a randomized, double-blind, placebo-controlled study. The American 
journal of gastroenterology. 1989;84(10):1259-62. 
 




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1. INTRODUCTION 
 
This document describes in detail the analysis plan for the unblinded administrative interim 
analyses being performed for corporate planning, for patients randomized in C1701-201, A 
Multicenter, Randomized, Double-blind, Placebo-controlled, Parallel-group, Single-dose, 
Phase 2a Study to Evaluate the Safety, Tolerability, Pharmacokinetics, and Pharmacodynamics 
of IW-1701 in Patients with Achalasia. Study details are presented in the full protocol, C1701-
201-P-06, finalized on October 3, 2017.  
 
The analyses documented in this plan will be executed by an unblinded statistician (employee of 
Ironwood) not otherwise associated with the day-to-day study execution. As defined in the 
protocol blinding section, Sponsor personnel involved in the conduct of the study (e.g., study 
medical monitor, study statistician, study data manager, and clinical operations study personnel) 
will be blinded to individual patient treatment assignments; however, descriptive and inferential 
statistics will be presented with a treatment identifier. In contrast, patient-level data such as 
minimum, maximum, median, and graphical analysis will not be presented with patient identifiers 
so that study conduct decisions regarding individual patients can be addressed without explicit 
knowledge of the patient’s treatment assignment. A list of study personnel and their 
blinded/unblinded status is presented below. 
 
Safety is monitored blinded in an ongoing basis throughout the course of the study. Safety 
assessments and endpoints will not be part of the administrative interim analysis. 
 
2. STUDY DESIGN 
 
This multicenter, randomized, double-blind, placebo-controlled, parallel-group, single-dose study 
will randomize up to 20 patients to receive study drug (15 patients to IW-1701 and 5 to matching 
placebo). 
 
2.1 Study Population 
 
This study will randomize up to 20 patients diagnosed with primary Type I or II achalasia with an 
integrated relaxation pressure (IRP) >15 mmHg by baseline High Resolution Impedance 
Manometry (HRIM). 
 
2.2 Study Periods 
 
Screening Period: The Screening Period will begin with the signature of the informed consent 
form (ICF) at the Screening Visit (which may occur from Day -28 to Day -1) and will last 1 to 
28 days (see Study Schematic). At the Screening Visit, the patient will undergo screening 
procedures (see Schedule of Events) to determine their preliminary eligibility for the study (see 
Inclusion/Exclusion Criteria). Patients will begin a liquid diet on Day -1 and then will fast 
overnight. The Screening Period will end at Check-in on the morning of Day 1.  
Clinic Period: The Clinic Period will begin at Check-in on the morning of Day 1 and will end at 
Discharge from the clinic on Day 2. To confirm eligibility (see Eligibility Criteria), patients will 


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undergo a baseline protocol-specific HRIM procedure After the procedure, the HRIM catheter 
will be removed and patients will complete a baseline symptom assessment. Patients who meet 
all eligibility criteria in addition to having confirmed Type I or II achalasia and IRP > 15 mmHg 
will be randomized to receive a single 5-mg dose of IW-1701 or matching placebo. Following 
study drug administration, the HRIM catheter will be reinserted for the postdose HRIM procedure 
following the protocol-specific swallowing protocol. The HRIM catheter will be removed after 
the postdose recording, and patients will complete a postdose symptom assessment. Safety, PK, 
and pharmacodynamic (PD) assessments, including blood collections, will be performed at 
prespecified times. Following all study assessments, and at the Investigator’s discretion, patients 
may be discharged on Day 2 at least 30 hours after study drug administration if they meet 
discharge criteria. 
Follow-up Period: The Follow-up Period will begin immediately after the patient is discharged 
from the clinic. Patients will have a Follow-up Call on Day 4 (± 1 day) to review adverse events 
(AEs). The Follow-up Period will end after the End of Trial Visit on Day 21 (± 7 days). 
2.3 Method of Assigning Patients to Treatment Groups 
Patients who meet all the inclusion criteria and none of the exclusion criteria will be randomized 
to treatment on Clinic Day (Day 1). Patients will be randomized through central randomization 
in a 3:1 ratio to receive either 5 mg IW-1701 or placebo. 
 
2.4 Blinding  
 
This study is double-blind and placebo-controlled. The patients, Investigators, and site study 
staff, except for specifically designated unblinded Study Center pharmacy staff, will be blinded 
to individual treatment assignments. Sponsor personnel involved in the conduct of the study 
(e.g., study medical monitor, study statistician, study data manager, and clinical operations study 
personnel) will be blinded to individual patient treatment assignments; however, Sponsor 
personnel may be unblinded to data at the treatment-group level. In addition, the HRIM 
recordings will be interpreted by central, blinded reader(s) for PD analyses.  
 
2.5 Administrative Unblinded Interim Analysis Objective 
 
The objective of this unblinded interim analysis is to determine if a single 5 mg dose of IW-1701 
has an effect on a patient’s IRP measures (supine or upright).  
 
3. STATISTICAL METHODS 
 
For any unblinded interim analysis, descriptive statistics and graphical analyses (e.g., number of 
subjects, mean, standard deviation, median, minimum, and maximum) will be calculated for IRP 
in the supine and upright position at both the predose (baseline) and postdose (HRIM) 
procedures, as well as change from baseline in IRP measured in both the supine and upright 
position. However, for the first interim analysis, number of subjects, standard deviation, 
minimum, and maximum will not be calculated in order to maintain blinding to individual 


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treatment assignments. Summaries will be presented by treatment group and overall. Where 
descriptive statistics or graphical analyses might unblind individual patients due to the sparsity of 
data, select descriptive statistics of graphical analyses may be omitted at the discretion of the 
unblinded statistician. Inferential statistics will not be calculated except if specifically requested 
by the Study Medical Monitor. All statistical analyses will be performed using SAS Version 9.4 
(or later). Analysis will be conducted using the Pharmacodynamic Population. 
 
Pharmacodynamic Population: All patients who receive study drug and have at least 1 postdose 
PD assessment will be included in the PD population. 
 
3.1 Pharmacodynamic methods for unblinded interim analysis 
 
Integrated Relaxation Pressure (IRP): mean of the 4s of maximal deglutitive relaxation in the 10-
s window beginning at the upper esophageal sphincter relaxation measured in mmHg. Contributing 
times can be contiguous or non-contiguous (e.g., interrupted by diaphragmatic contraction). 
Referenced to gastric pressure; median measurement from the 10-swallow supine and 5-swallow 
upright protocols. IRP was measured in both the supine position (a subject’s value is the median 
of their 10 swallows) and the upright position (a subject’s value is the median of their 5 swallows). 
Both the median supine and upright IRP values will be used in the primary IRP analysis. 
 
Predose IRP: For subjects with 2 sets of swallowing protocols (subjects randomized under version 
C1701-201-P-05 or earlier), the first recording, or the IRP from the first set of predose swallowing 
protocol, will be used as the predose IRP values.  
 
Postdose IRP: For subjects with 2 sets of swallowing protocols (subjects randomized under C1701-
201-P-05 or earlier), the second recording, or the median supine and upright IRP values from the 
second set of postdose swallowing protocol will be used for the postdose IRP values. 
 
Mean change from predose (postdose supine IRP - predose supine IRP) will be calculated for 
supine IRP (which is recorded as the median of the 10 supine swallows from each swallowing 
protocol). 
 
Mean change from predose (postdose upright IRP – predose upright IRP) will be calculated for 
upright IRP (which is recorded as the median of the 5 upright swallows from each swallowing 
protocol). 
 
See the appendix 4.1 for a more detailed list of variables used for this administrative interim 
analysis. 
 


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3.2 Sensitivity Analysis 
 
We will perform a sensitivity analysis where the average of a subject’s two supine IRP values is 
calculated for subjects with 2 sets of swallowing protocols (subjects randomized under version 
C1701-201-P-05 or earlier). The mean change from predose for supine will be recalculated for all 
patients using the mean values for patients with 2 sets of swallowing protocols. This analysis will 
also be repeated for upright IRP. 
 
3.3 Timing of Administrative Interim Analyses 
 
Administrative unblinded interim analyses will be performed after the first 5 patients have 
completed both pre- and postdose HRIMs, and then subsequently when approximately 10, and 15 
patients have completed both pre- and postdose HRIMs.  
 
3.4 Roles and Responsibilities 
 
The unblinded statistician will execute the statistical analyses defined in this document in a secure 
location, accessible to him/her only, to ensure the limited access to the unblinded results. The 
unblinded statistician will create a summary report of all interim finding and present the report to 
the Study Medical Monitor. 
 
In turn, the Study Medical Monitor and unblinded statistician will present the results to a Senior 
Management Group consisting of: President of R & D, Chief Development Officer, and SVP of 
Clinical Affairs with assistance from the Head of Biostatistics of VP of Data Science, as needed.  
 
The Study Execution Team (SET), with the exception of the Study Medical Monitor, will remain 
blinded at both the group and individual level. 
 
 
 




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Statistical Analysis Plan: C1701-201-SAP 
 
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4. APPENDIX 
4.1 Key variables for administrative interim analysis 
 
Variable 
Definition 
CR1SIRPM 
IRP in supine position from first recording (this value is the median from 
the first set of 10 supine swallows) 
CR2SIRPM 
IRP in supine position from second recording; not all subjects will have 
this (this value is the median from the second set of 10 supine swallows) 
CR1UIRPM 
IRP in upright position from first recording (this value is the median 
from the first set of 5 upright swallows) 
CR2UIRPM 
IRP in upright position from second recording; not all subjects will have 
this (this value is the median from the second set of 5 upright swallows) 
 










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Ironwood Pharmaceuticals, Inc. 
Statistical Analysis Plan: C1701-201-SAP 
 
July 20, 2018 
 
Confidential 
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