Conditions: Corneal Edema
Keywords: Corneal Edema Fuchs Corneal Dystrophy Netarsudil Ophthalmic Solution
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 40
Healthy volunteers: no

Brief summary:
Two different dosing regimens (QD and BID) of netarsudil will be studied to evaluate their efficacy in reducing or resolving corneal edema in subjects with FCD.

Detailed description:

Arms:
arm 1: One drop of Netarsudil 0.02% ophthalmic solution in the study eye in the evening for an 8 week period in up to 20 subjects. arm 2: One drop of Netarsudil 0.02% ophthalmic solution in the study eye in the morning and in the evening for an 8 week period in up to 20 subjects.

Interventions: intervention 1: Netarsudil Ophthalmic intervention 2: Netarsudil Ophthalmic
Intervention descriptions: intervention 1: Netarsudil Ophthalmic Solution 0.02% Topical Sterile Ophthalmic Solution intervention 2: Netarsudil Ophthalmic Solution 0.02% Topical Sterile Ophthalmic Solution

Protocol document:
Prot_000.pdf: 
Study Protocol 
 
 
A Randomized, Open-Label, Parallel-Group Study to Evaluate 
the Safety and Efficacy of Two Dosing Regimens of Netarsudil 
Ophthalmic Solution in Patients with Corneal Edema Due to 
Fuchs Corneal Dystrophy 
 
 
NCT 04498169 
 
 
Protocol Amendment 1 (Rev 1) 03 February 2021 


Netarsudil Ophthalmic Solution 0.02% 
Clinical Study Protocol: AR-13324-CS210, Amendment 1 
Aerie Pharmaceuticals, Inc.
 
 
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Page 1 of 55 
 
 
Clinical Study Protocol 
Study Title:  
A Randomized, Open-Label, Parallel-Group Study to Evaluate the 
Safety and Efficacy of Two Dosing Regimens of Netarsudil 
Ophthalmic Solution in Patients with Corneal Edema Due to Fuchs 
Corneal Dystrophy 
Study Number: 
AR-13324-CS210 
Study Phase: 
2 
Product Name: 
Netarsudil Ophthalmic Solution 0.02% 
Indication: 
Corneal edema due to Fuchs Corneal Dystrophy (FCD) 
Investigators: 
Multicenter 
Sponsor: 
Aerie Pharmaceuticals, Inc. 
550 Hills Drive, 3rd Floor 
Bedminster, NJ 07921 
(908) 470-4320 
Medical 
Monitor: 
Richard A. Lewis, MD 
Phone: (916) 768-7703 
Email: rlewis@aeriepharma.com 
 
NCT Number 
04498169 
 
 
Date 
Original Protocol (Rev 0): 
Protocol Amendment 1 (Rev 1) 
14 August 2020 
03 February 2021 
 
 
 
Confidentiality Statement 
This document contains Aerie Pharmaceuticals, Inc.’s (Aerie) information that is 
confidential, a trade secret and/or proprietary in nature.  It is loaned to you for your 
confidential use on behalf of Aerie and is not to be photocopied, disclosed or transmitted to 
any other person or party who is not covered by a Confidential Disclosure Agreement with 
Aerie.  As the Principal Investigator you are responsible for the safekeeping and return of this 
document to Aerie upon request.  You will be sent updated information and/or amendments 
as they become available. 
 
 


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CLINICAL PROTOCOL CONTACT INFORMATION 
Clinical Operations 
Nancy Ramirez, MS 
Director  
Aerie Pharmaceuticals Inc. 
550 Hills Drive, Floor 3 
Bedminster, NJ 07921  
Phone: +1 908-947-3543 
Cell: +1-973-809-6965 
Email: nramirez@aeriepharma.com 
Swetha Mandava, MS 
Clinical Trial Manager 
Aerie Pharmaceuticals Inc. 
550 Hills Drive, Floor 3 
Bedminster, NJ 07921  
Cell: +1-973-204-9467 
Email: smandava@aeriepharma.com 
Clinical Development 
Hayley McKee, PhD 
Director, Clinical Development 
Aerie Pharmaceuticals Inc. 
550 Hills Drive, Floor 3 
Bedminster, NJ 07921  
Phone: +1- 908-947-3445 
Cell: +1-949-838-7607 
Email: hmckee@aeriepharma.com 
Amber Lewis, PharmD, PSM 
Clinical Development Manager 
Aerie Pharmaceuticals Inc. 
2030 Main Street, Suite 1400 
Irvine, CA 92614 
Phone: (949) 392-8252 
alewis@aeriepharma.com 


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CLINICAL PROTOCOL APPROVAL FORM 
 
Protocol Title: A Randomized, Open-Label, Parallel-Group Study to Evaluate the Safety and Efficacy 
of Two Dosing Regimens of Netarsudil Ophthalmic Solution in Patients with Corneal Edema Due to 
Fuchs Corneal Dystrophy 
Study No: AR-13324-CS210 
Original Protocol Date: 14 August 2020 
Protocol Version No: Rev. 1 
Protocol Version Date: 03 February 2021 
Role 
Contact Information 
Aerie Management and Sponsor Safety 
Officer 
 
David Hollander, MD, MBA 
Chief Research & Development Officer 
Aerie Pharmaceuticals, Inc. 
2030 Main Street, Suite 1400  
Irvine, CA 92614 
Phone: +1 949-526-8738 
Fax: 
+1 949-526-8787 
Email: dhollander@aeriepharma.com 
 
 
 
 
 
 
 
 
 
 
 
Signature 
 
 
Date 
 
 
Clinical Operations & Development 
 
Michelle Senchyna, PhD 
VP, Clinical Development and Medical Affairs  
Aerie Pharmaceuticals Inc. 
2030 Main Street, Suite 1400  
Irvine, CA 92614 
Phone: +1 908-947-3551 
Cell: +1 949-237-1325 
Email: msenchyna@aeriepharma.com 
 
 
 
 
 
 
 
 
 
 
Signature 
 
 
Date 


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Medical Monitor 
Richard Lewis, MD 
Chief Medical Officer 
1515 River Park Dr. 
Sacramento, CA 95815 
Cell:  +1 916-768-7703 
Email: rlewis@aeriepharma.com 
Signature 
Date 


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SYNOPSIS 
Sponsor: 
Aerie Pharmaceuticals, Inc. 
Name of Finished Product: 
Netarsudil Ophthalmic Solution 0.02% 
Name of Active Ingredients: 
Netarsudil mesylate 
Study Title: 
A Randomized, Open-Label, Parallel-Group Study to Evaluate the Safety and Efficacy of Two Dosing 
Regimens of Netarsudil Ophthalmic Solution in Patients with Corneal Edema Due to Fuchs Corneal 
Dystrophy 
Study Number: 
AR-13324-CS210 
Study Phase: 
2 
Primary Objective: 
To evaluate the reduction in central corneal thickness (CCT) in eyes dosed with either Netarsudil Ophthalmic 
Solution 0.02% (netarsudil) once-daily (QD) or netarsudil twice-daily (BID) for the treatment of corneal 
edema due to Fuchs corneal dystrophy (FCD). 
Secondary Objectives: 
(1) To evaluate the proportion of subjects demonstrating improvement in vision following treatment for FCD
with netarsudil QD or netarsudil BID.
(2) To evaluate the safety of netarsudil QD and netarsudil BID in subjects with FCD.
Study Design: 
This is an 8-week, randomized, open-label, multicenter, efficacy and safety study in subjects with FCD. 
The study will be conducted at approximately 20 sites in the USA. Two different dosing regimens (QD and 
BID) of netarsudil will be studied (randomized 1:1) to evaluate their efficacy in reducing or resolving corneal 
edema in subjects with FCD. Subjects randomized to the netarsudil QD dosing regimen will be instructed to 
instill one drop of the Study Artificial Tear in the study eye every morning, and one drop of netarsudil in the 
study eye every evening. Subjects randomized to the netarsudil BID dosing regimen will instill one drop of 
netarsudil in the study eye every morning and evening. 
CCT will be measured by ultrasound pachymetry at approximately the same time of day ± 60 minutes and 
always before noon (local time), at each of the 5 scheduled visits (Screening, Baseline [Day 1], Week 2, 
Week 4, and Week 8/Exit). In order to be eligible to participate in the study, subjects will have been 
diagnosed with visually significant central corneal edema due to FCD, in at least one eye at Screening and 
Baseline, for no longer than 12 months’ duration.  
For analysis purposes, the study eye will be defined as the eligible eye with a Baseline CCT of at least 
600 μm, as assessed by ultrasound pachymetry. If both eyes are eligible at both Screening and Baseline based 
on CCT and enrollment criteria, the study eye will be the eye with the thinner cornea at the Baseline Visit.   
Study Population: 
Approximately forty (40) subjects will be enrolled 
Key Inclusion Criteria: 
•
Adults aged 18 years or older
•
Documented diagnosis of FCD


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• 
Evidence of central corneal edema, in at least one eye, at the Screening and Baseline Visit. The study 
eligible eye(s) should have a CCT of at least 600 μm at both the Screening and Baseline Visit 
• 
Best Corrected Visual Acuity (BCVA) using the Early Treatment of Diabetic Retinopathy Study 
(ETDRS) methodology of 70 to 20 letters at Screening and Baseline in the eligible eye(s) 
 
Key Exclusion Criteria: 
• 
FCD so advanced that, in the opinion of the Investigator, surgery would likely be required in the study 
eligible eye(s) within the study period 
• 
Clinically significant ocular disease (other than FCD) or trauma in the eligible eye(s) which could 
interfere with study interpretation 
• 
History of ocular surgery within 6 months of the Screening Visit, or any prior corneal refractive surgery 
in the eligible eye(s)  
Study Medication, Dose, and Mode of Administration: 
Once Daily Netarsudil Dosing Regimen 
Study Artificial Tear; one drop, in the study eye in the morning  
Netarsudil 0.02% ophthalmic solution; one drop, in the study eye in the evening 
Twice Daily Netarsudil Dosing Regimen 
Netarsudil 0.02% ophthalmic solution; one drop, in the study eye in the morning and in the evening  
Duration of Treatment: 
Eight (8) Weeks 
Efficacy Assessments: 
Primary Endpoint:  
• 
Mean change from baseline in CCT as assessed by ultrasound pachymetry at Week 4 
Secondary Endpoints:  
• 
Mean change from baseline in BCVA at Week 4 
• 
Mean change from baseline in CCT as assessed by ultrasound pachymetry at Week 8 
• 
Mean change from baseline in BCVA at Week 8 
• 
Proportion of subjects who gained ≥15 letters (3 lines) in BCVA at Week 4 
• 
Proportion of subjects who gained ≥15 letters (3 lines) in BCVA at Week 8 
• 
Proportion of subjects with complete resolution of corneal edema at Week 4 
• 
Proportion of subjects with complete resolution of corneal edema at Week 8     
Safety Assessments: 
The safety of netarsudil QD and BID will be evaluated at each visit by: 
• 
Adverse events 
• 
BCVA 
• 
Anterior segment biomicroscopy 
• 
Ophthalmoscopy 
• 
Vital signs (heart rate and blood pressure) 
• 
Intraocular pressure 


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Statistical Methods: 
Continuous study assessments will be summarized by treatment and time point (as applicable) using 
descriptive statistics (n, mean, median, standard deviation, minimum, and maximum). Categorical study 
assessments will be summarized by treatment and visit (as applicable) using frequency counts and 
percentages.  
Hypothesis: 
H01: The mean change from baseline CCT by ultrasound pachymetry in study eyes treated with netarsudil 
QD at Week 4 = 0. 
H11: The mean change from baseline CCT by ultrasound pachymetry in study eyes treated with netarsudil 
QD at Week 4 ≠ 0. 
H02: The mean change from baseline CCT by ultrasound pachymetry in study eyes treated with netarsudil 
BID at Week 4 = 0. 
H12: The mean change from baseline CCT by ultrasound pachymetry in study eyes treated with netarsudil 
BID at Week 4 ≠ 0. 
Analysis of Primary Endpoint: 
The primary comparisons in this trial will be within a treatment group on the mean change from baseline 
CCT to Week 4 for each of netarsudil QD and BID. The primary efficacy endpoint change from baseline 
CCT will be summarized descriptively (n, mean, standard deviation, median, min, and max) and analyzed 
primarily using one-sample t-tests; comparisons between treatment groups will be completed using two-
sample t-tests. Sensitivity analyses will be completed using Wilcoxon signed-rank tests within a treatment 
group, Wilcoxon rank sum tests between treatment groups and an ANCOVA model with terms for baseline 
CCT value and treatment. The least squares mean will be presented for each treatment group from the model 
together with two-sided p-values and 90% and 95% confidence intervals around the change from baseline 
within each treatment group.  Comparisons between treatment groups on the change from baseline CCT will 
also be made including the least squares mean difference and corresponding two-sided p-values and 90% and 
95% confidence intervals. 
The primary analysis will use the modified Intent to Treat (mITT), population with available data per subject. 
Robustness analyses will also be presented based on the multiple imputation methodology under different 
assumptions of missingness (at random and not at random), using last observation carried forward (LOCF), 
and using the per protocol population with available data per subject. 
Intercurrent events will be primary handled in the following manner: 
• 
Discontinuation of study medication and non-optimal compliance will be ignored [treatment policy 
strategy]. 
• 
Withdrawal due to lack of efficacy or adverse events: missing data will be singly imputed as the worst 
within subject observation prior to the intercurrent event [hypothetical strategy]. 
• 
Missing data without withdrawal or withdrawal due to reasons other than lack of efficacy or adverse 
events: missing data will be imputed employing Multiple Imputation (MI) using randomized treatment-
based Markov Chain Monte Carlo (MCMC) methodology to impute non-monotone missing data and 
regression methodology to impute monotone missing data [hypothetical strategy]. 
 


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Safety: 
Adverse events (AEs) will be coded using the MedDRA dictionary. Frequencies and percentages of 
treatment emergent adverse events (TEAEs) will be summarized at the subject level by system organ class 
and preferred term for all TEAEs, treatment related TEAEs, serious TEAEs, and TEAEs causing premature 
discontinuation by treatment group. An AE is treatment emergent if it occurs or worsens after the first dose 
of study medication. Similar summaries will be presented for all TEAEs by maximal severity. Separate 
summaries will be performed for ocular and non-ocular AEs. Other safety endpoints including visual acuity, 
anterior segment biomicroscopy, intraocular pressure, ophthalmoscopy, and vital signs will be summarized 
by treatment group and visit using descriptive statistics. Changes or shifts from baseline will also be 
summarized where appropriate. For assessments performed by eye, study eye and fellow eye will be 
summarized separately. 
Date of Original Approved Protocol (Rev 0): 14 August 2020 
Date of Most Recent Protocol Amendment (Rev 1): 03 February 2021 


 
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SCHEDULE OF VISITS AND PROCEDURES 
Study procedures are recommended to be performed in the sequence specified in the schedule below. 
Table 1 
Schedule of Visits and Procedures 
Visit1 
Screening Visit1 
Baseline Visit1 
Week 21 
Week 41 
Week 8/Exit1 
Visit Window (Days) 
Day -8 to -1 
Day 1 
Day 14 ± 3 
Day 28 ± 3 
Day 56 ± 3 
Visit Sequence Number 
1 
2 
3 
4 
5 
Informed Consent 
X 
 
 
 
 
Inclusion/Exclusion  
X 
X 
 
 
 
V-FUCHS Questionnaire 
 
X 
 
X 
X 
Demography 
X 
 
 
 
 
Medical/Ophthalmic History 
X 
X 
 
 
 
Concomitant Medications 
X 
X 
X 
X 
X 
HR/BP 
X 
X 
 
 
X 
Urine Pregnancy Test2 
 
X 
 
 
X 
Ocular Symptoms/AEs 
OU 
OU 
OU 
OU 
OU 
BCVA (ETDRS protocol) 
OU 
OU 
OU 
OU 
OU 
Biomicroscopy 
OU 
OU 
OU 
OU 
OU 
Ultrasound Pachymetry (CCT) 3 
OU 
OU 
OU 
OU 
OU 
IOP 
OU 
OU 
OU 
OU 
OU 
Dilated Ophthalmoscopy  
OU 
 
 
 
OU 
Undilated Ophthalmoscopy 
OU 
OU 
OU 
Tomography (Pentacam®)4 
OU 
OU 
OU 
OU 
OU 
SE Determination5 and Randomization 
 
X 
 
 
 
Eye Drop Instruction and Demonstration 
 
SE 
 
 
 
Dispense Study Medication 
 
X 
 
X 
 
Abbreviations: AEs = adverse events; BCVA = best corrected visual acuity; BP = blood pressure; CCT = central corneal thickness; ETDRS = Early Treatment Diabetic 
Retinopathy Study; HR = heart rate; IOP = intraocular pressure; OU = both eyes; SE = Study Eye; V-FUCHS = Visual Function and Corneal Health Status 
1 
Subjects should be scheduled at approximately the same time of day ± 60 mins in the morning (ideally at or before 10:00 AM local time) for each visit 
2 
Required only for females of childbearing potential 
3 
Ultrasound pachymetry should be performed at approximately the same time of day ± 60 mins and always before noon local time 
4 
Selected sites only, may be completed prior to ophthalmoscopy at the Screening and Week 8/Exit Visit while the subject is dilating 
5 
Please refer to Section 4.4 for instructions on study eye determination  


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SCHEMA 
Figure 1 
Study Design Schema 
 
* Any subject randomized to netarsudil QD who, in the opinion of the Investigator, has not responded adequately to treatment, may have their netarsudil dosing 
frequency increased to BID for the remainder of the study. 
† Study eye only. AM dosing should occur between 06:00 and 09:00 hours. PM dosing should occur between 19:00 and 22:00 hours.  
‡ Subjects randomized to the netarsudil QD dosing regimen will be instructed to instill one drop of the Study Artificial Tear in their study eye every morning, and 
one drop of netarsudil in their study eye every evening. 


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TABLE OF CONTENTS 
SYNOPSIS ..........................................................................................................................5 
SCHEDULE OF VISITS AND PROCEDURES .............................................................9 
SCHEMA ..........................................................................................................................10 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS ..............................14 
1. 
INTRODUCTION ....................................................................................................16 
2. 
STUDY OBJECTIVES ............................................................................................17 
2.1 
Primary Objective(s) ...........................................................................................17 
2.2 
Secondary Objective(s) ........................................................................................17 
3. 
INVESTIGATIONAL PLAN ..................................................................................17 
3.1 
Overall Study Design and Plan ...........................................................................17 
3.2 
Rationale for Study Design and Control Group ...............................................18 
3.3 
Study Duration .....................................................................................................19 
4. 
STUDY POPULATION SELECTION ...................................................................19 
4.1 
Study Population ..................................................................................................19 
4.2 
Inclusion Criteria .................................................................................................19 
4.3 
Exclusion Criteria ................................................................................................20 
4.4 
Study Eye Determination ....................................................................................21 
5. 
STUDY TREATMENTS ..........................................................................................22 
5.1 
Description of Treatments ...................................................................................22 
5.1.1 
Study Medication ...........................................................................................22 
5.1.2 
Placebo or Control Drug ...............................................................................22 
5.2 
Treatments Administered ...................................................................................22 
5.3 
Selection and Timing of Dose for Each Patient .................................................23 
5.4 
Method of Assigning Patients to Treatment Groups ........................................23 
5.5 
Masking .................................................................................................................23 
5.6 
Concomitant Therapy ..........................................................................................23 
5.6.1 
Prohibited Interventions ...............................................................................23 
5.6.2 
Permitted Interventions.................................................................................24 
5.7 
Restrictions ...........................................................................................................24 
5.7.1 
Prior Therapy .................................................................................................24 
5.7.2 
Fluid and Food Intake ...................................................................................24 
5.7.3 
Subject Activity Restrictions .........................................................................25 
5.8 
Treatment Compliance ........................................................................................25 
5.9 
Packaging and Labeling ......................................................................................25 
5.10 
Storage and Accountability .................................................................................25 
5.11 
Study Medication Retention at Study Site .........................................................26 
5.11.1 
Receipt and Disposition of Study Medication .............................................26 
5.11.2 
Return of Study Medication ..........................................................................26 
6. 
STUDY PROCEDURES ..........................................................................................27 
6.1 
Informed Consent ................................................................................................27 
6.2 
Demographics and Medical/Ophthalmic History .............................................27 
6.3 
Concomitant Medication Assessments ...............................................................27 
6.4 
Vital Signs (HR and BP only) .............................................................................28 


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6.5 
Pregnancy Testing ................................................................................................28 
6.6 
Dispensing Study Medication .............................................................................28 
6.7 
Efficacy Assessments ...........................................................................................29 
6.7.1 
Ultrasound Pachymetry.................................................................................29 
6.7.2 
Best Corrected Visual Acuity (ETDRS Method) ........................................29 
6.7.3 
Biomicroscopy ................................................................................................29 
6.7.4 
V-FUCHS Questionnaire...............................................................................29 
6.7.5 
Tomography (Pentacam®; Selected Sites Only) ..........................................30 
6.8 
Safety Assessments ...............................................................................................30 
6.8.1 
IOP ..................................................................................................................30 
6.8.2 
Ophthalmoscopy ............................................................................................30 
6.9 
Adverse Events Assessments ...............................................................................30 
6.9.1 
Performing Adverse Event (AE) Assessments ............................................30 
6.9.2 
Timing .............................................................................................................32 
6.9.3 
Severity............................................................................................................33 
6.9.4 
Relationship ....................................................................................................33 
6.9.5 
Expectedness ...................................................................................................34 
6.9.6 
Serious Adverse Events, Suspected Serious Adverse Reactions or 
Suspected Unexpected Serious Adverse Reactions .....................................34 
6.9.6.1 
Reporting SAEs, SSARs or SUSARs ......................................................34 
6.9.7 
Pregnancy Reporting .....................................................................................35 
6.10 
Removal of Subjects from the Study or Study Medication ..............................35 
6.10.1 
Completed Subject .........................................................................................35 
6.10.2 
Non-completing Subject ................................................................................35 
6.10.3 
Actions after Discontinuation .......................................................................36 
6.10.4 
Discontinuation of the Entire Study .............................................................36 
6.10.5 
Completed Study ............................................................................................36 
6.11 
Appropriateness of Measurements .....................................................................36 
7. 
STUDY ACTIVITIES ..............................................................................................36 
7.1 
Screening Visit (Day -8 to -1) ..............................................................................37 
7.2 
Treatment Period .................................................................................................37 
7.2.1 
Baseline (Day 1) Procedures .........................................................................37 
7.2.2 
Week 2 (Day 14 ± 3) Procedures ...................................................................38 
7.2.3 
Week 4 (Day 28 ± 3) Procedures ...................................................................39 
7.2.4 
Week 8 (Day 56 ± 3) or Early Termination Procedures .............................39 
7.2.5 
Unscheduled Visits .........................................................................................40 
8. 
QUALITY CONTROL AND ASSURANCE .........................................................40 
9. 
PLANNED STATISTICAL METHODS ................................................................40 
9.1 
General Considerations .......................................................................................40 
9.1.1 
Unit of Analysis ..............................................................................................41 
9.1.2 
Missing Data and Intercurrent Events ........................................................41 
9.1.3 
Multiplicity Considerations...........................................................................41 
9.2 
Hypotheses ............................................................................................................42 
9.3 
Determination of Sample Size .............................................................................42 
9.4 
Analysis Populations ............................................................................................43 


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9.4.1 
Modified Intent-to-Treat (mITT) Population .............................................43 
9.4.2 
Per Protocol (PP) Population ........................................................................43 
9.4.3 
Safety Population ...........................................................................................43 
9.5 
Demographics and Baseline Characteristics .....................................................43 
9.6 
Subject Disposition...............................................................................................43 
9.7 
Primary Endpoint(s) ............................................................................................44 
9.8 
Efficacy Analysis ..................................................................................................44 
9.8.1 
Primary Efficacy Endpoint ...........................................................................44 
9.8.2 
Primary Efficacy Analyses ............................................................................44 
9.8.3 
Secondary Efficacy Endpoints ......................................................................45 
9.8.4 
Secondary Efficacy Analyses ........................................................................45 
9.8.5 
Exploratory Efficacy Endpoints ...................................................................46 
9.8.6 
Exploratory Efficacy Analyses......................................................................46 
9.8.7 
Primary Safety Assessments and Analyses ..................................................46 
10. ADMINISTRATIVE CONSIDERATIONS ...........................................................47 
10.1 
Investigators and Study Administrative Structure ...........................................47 
10.2 
Institutional Review Board (IRB) or Independent Ethics Committee 
(IEC) Approval.....................................................................................................47 
10.3 
Ethical Conduct of the Study ..............................................................................47 
10.4 
Subject Information and Consent ......................................................................48 
10.5 
Subject Confidentiality ........................................................................................48 
10.6 
Study Monitoring .................................................................................................49 
10.7 
IRT ........................................................................................................................50 
10.8 
Case Report Forms and Study Records .............................................................50 
10.9 
Protocol Deviations ..............................................................................................51 
10.10 Access to Source Documentation ........................................................................51 
10.11 Data Generation and Analysis ............................................................................52 
10.12 Retention of Data .................................................................................................52 
10.13 Financial Disclosure .............................................................................................52 
10.14 Publication and Disclosure Policy ......................................................................52 
11. AMENDMENT SUMMARY OF CHANGES ........................................................53 
11.1 
Amendment 1: 03 February 2021 .......................................................................53 
12. REFERENCES .........................................................................................................54 
 
TABLE OF TABLES 
Table 1 
Schedule of Visits and Procedures ............................................................9 
 
TABLE OF FIGURES 
Figure 1 
Study Design Schema ...............................................................................10 
 
 


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LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS 
AE 
Adverse Event 
AESI 
Adverse Event of Special Interest 
BCVA 
Best Corrected Visual Acuity 
BID 
Twice Daily 
BP 
Blood Pressure 
CCT 
Central Corneal Thickness 
CNO 
Certificate of Non-Objection 
CONMED 
Concomitant Medication 
DSMC 
Data and Safety Monitoring Committee 
DSO 
Descemet Stripping Only 
DWEK 
Descemetorhexis Without Endothelial Keratoplasty 
eCRF 
Electronic Case Report Form 
EDC 
Electronic Data Capture 
ETDRS 
Early Treatment Diabetic Retinopathy Study 
FCD 
Fuchs Corneal Dystrophy 
GCP 
Good Clinical Practice 
HR 
Heart Rate 
IB 
Investigator’s Brochure 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonization 
IEC 
Independent Ethics Committee 
IOP 
Intraocular Pressure 
IRB 
Institutional Review Board 


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IRT 
Interactive Response Technology 
kg 
Kilogram 
L 
Liters 
mg 
Milligram 
min 
Minute 
mmHg 
Millimeters Mercury 
QD 
Once Daily 
ROCK 
Rho Kinase 
SAE 
Serious Adverse Event 
SAR 
Serious Adverse Reaction 
SSAR 
Serious Suspected Adverse Reaction 
V-FUCHS 
Visual Function and Corneal Health Status  
WHO 
World Health Organization 
 


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1. INTRODUCTION 
Fuchs Corneal Dystrophy (FCD) is a slowly progressing disease of the cornea that is the 
most common indication for corneal-endothelial transplants worldwide (Gain 2016). 
FCD is characterized by corneal edema and the presence of guttae, which are collagenous 
excrescences of Descemet’s membrane (McLaren 2014). As FCD progresses, 
corneal-endothelial cell degeneration compromises the ability of the cornea to pump out 
excess fluid resulting in corneal edema and clouding (Okumura 2018). Patients with FDC 
typically present with glare, blurry vision, and a hazy cornea. Decreased visual acuity may be 
more pronounced in the morning due to increased corneal hydration that occurs from closed 
eyelids overnight. In later stages of the disease, edema may reach the corneal epithelium 
leading to an irregular surface and painful bullae (Moshirfar 2020).  
Currently, the only definitive treatment option for FCD is surgery, and endothelial 
keratoplasty using donor corneas is the gold standard. A newer surgical procedure, called 
Descemet Stripping Only (DSO) or Descemetorhexis Without Endothelial Keratoplasty 
(DWEK), is available for select patients and does not require donor corneal tissue. 
Despite recent improvements in surgical techniques, there are still many risks associated with 
these procedures including infection, cataract formation, secondary ocular hypertension 
(OHT) from long term steroid use, graft rejection, and treatment failure requiring a second 
surgery (Moshirfar 2020). Therefore, there is a large unmet medical need for pharmacologic 
alternatives to surgery for treating FCD (Koizumi 2014).  
Netarsudil is a novel Rho kinase (ROCK) inhibitor that was approved for marketing as 
Rhopressa® (netarsudil ophthalmic solution) 0.02% by the US FDA in December 2017 for 
the reduction of elevated intraocular pressure in patients with open-angle glaucoma or OHT.  
In addition to lowering intraocular pressure, ROCK inhibitors, such as netarsudil, may also 
have potential benefit for the treatment of corneal endothelial decompensation. Although the 
mechanisms are still being elucidated, preclinical evidence is emerging that ROCK inhibition 
can promote adhesion, enhance proliferation, and inhibit apoptosis of corneal-endothelial 
cells (Okumura 2009, Okumura 2017). Findings from a clinical case series suggest that 
topical administration of the ROCK inhibitor Y-27632 after performing transcorneal freezing 
can improve central corneal edema in patients with FCD (Okumura 2013). In one patient, 
full corneal transparency was achieved for more than 6 years, and original plans for corneal 
transplantation were cancelled (Koizumi 2013, Okumura 2018). An additional two case 
reports of patients with FCD have described improvements in corneal edema and corneal 
endothelial cell counts with the use of netarsudil following DSO (Ploysangam 2019, 
Hirabayashi 2020). Several clinical trials are now ongoing to investigate ROCK inhibition in 
both pre-surgical (NCT04051463) and post-surgical (NCT03575130, NCT03813056, 
NCT03249337, NCT03971357, NCT04250207, NCT04057053) FCD patients. 
Overall, netarsudil is well tolerated, and the most common adverse event is conjunctival 
hyperemia. Several case reports of honeycomb or reticular edema have been observed in 
patients taking netarsudil, both with and without pre-existing corneal edema or a history of 


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FCD (Fernandez 2018, Liu 2019, Moumneh 2020, Wisely 2020). Typically, reticular or 
honeycomb edema resolves after discontinuation of netarsudil. 
The aim of this study is to evaluate the therapeutic potential of netarsudil as a pharmacologic 
agent to delay or prevent the need for surgery in patients with visually significant edema due 
to FCD. 
2. STUDY OBJECTIVES 
2.1 
Primary Objective(s) 
To evaluate the reduction in central corneal thickness (CCT) in eyes dosed with either 
netarsudil once-daily (QD), or netarsudil twice-daily (BID) for the treatment of corneal 
edema due to Fuchs corneal dystrophy (FCD). 
2.2 
Secondary Objective(s) 
(1) To evaluate the proportion of subjects demonstrating improvement in vision following 
treatment for FCD with netarsudil QD or netarsudil BID. 
(2) To evaluate the safety of netarsudil QD and netarsudil BID in subjects with FCD. 
3. INVESTIGATIONAL PLAN 
3.1 
Overall Study Design and Plan 
This is an 8-week, randomized, open-label, multicenter, efficacy and safety study in subjects 
with FCD. The study will be conducted at approximately 20 sites in the USA. Two different 
dosing regimens (QD and BID) of netarsudil will be investigated for their efficacy in 
reducing or resolving corneal edema in subjects with FCD. Subjects randomized to the 
netarsudil QD dosing regimen will be instructed to instill one drop of the Study Artificial 
Tear in their study eye every morning, and one drop of netarsudil in their study eye every 
evening. Subjects randomized to the netarsudil BID dosing regimen will instill one drop of 
netarsudil in their study eye every morning and evening. 
CCT will be measured by ultrasound pachymetry at approximately the same time of day 
± 60 minutes and always before noon (local time), at each of the 5 scheduled visits 
(Screening, Baseline [Day 1], Week 2, Week 4, and Week 8/Exit). In order to be eligible to 
participate in the study, subjects will have been diagnosed with visually significant central 
corneal edema due to FCD, in at least one eye, for no longer than 12 months’ duration.  
At the Screening Visit, the subject’s diagnosis of FCD will be confirmed and CCT measured 
using an ultrasonic pachymeter. Intraocular pressure (IOP) and Best Corrected Visual Acuity 
(BCVA) will also be collected. 


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At the Baseline Visit (Day 1), the eligibility assessments conducted during Screening will be 
repeated to ensure that the subject meets all criteria for enrollment in the study. The same eye 
must qualify at both the Screening and Baseline Visit. If qualified, the subject will be 
provided with the Visual Function and Corneal Health Status (V-FUCHS) patient reported 
visual disability questionnaire (Wacker 2018). The subject will be randomized (1:1) into the 
study and provided with study medication. Subjects randomized to the netarsudil QD dosing 
regimen will be instructed to instill one drop of the Study Artificial Tear in the study eye 
every morning, and one drop of netarsudil in the study eye every evening. Subjects 
randomized to the netarsudil BID dosing regimen will instill one drop of netarsudil in the 
study eye every morning and evening. The subject will also be trained on appropriate 
techniques for applying eye drops. 
At the Week 2 and Week 4 Visits, the subject will return to the study site for efficacy and 
safety assessments.  
At the Week 4 Visit, any subject randomized to netarsudil QD who, in the opinion of the 
Investigator, has not responded adequately to treatment, may have their netarsudil dosing 
frequency increased to BID for the remainder of the study. For these subjects, the morning 
administration of Study Artificial Tear will be terminated.  
At the Week 8 Visit, a final assessment of efficacy and safety parameters will be conducted, 
and the subject will exit the study.  
For analysis purposes, the study eye will be defined as the eligible eye with a Baseline CCT 
of at least 600 μm, as assessed by ultrasound pachymetry. If both eyes qualify at both the 
Screening and Baseline Visit based on CCT and enrollment criteria, the study eye will be the 
eye with the thinner cornea at the Baseline Visit.   
3.2 
Rationale for Study Design and Control Group 
Current treatment options for FCD include hyperosmotic saline ophthalmic drops/ointments 
and surgical procedures. Hyperosmotic saline can provide temporary symptomatic relief by 
facilitating corneal dehydration. Endothelial keratoplasty using donor corneas is currently the 
gold standard therapeutic option to restore vision in patients with FCD. Although 
improvement of surgical techniques has enabled less invasive treatment and better outcomes, 
corneal transplants require long-term steroid use and are still associated with difficult 
surgical techniques, graft rejection, acute/chronic cell loss, and worldwide donor cornea 
shortages (Okumura 2017). A relatively new non-transplant surgical procedure known as 
Descemet Stripping Only (DSO) or Descemetorhexis Without Endothelial Keratoplasty 
(DWEK) is also now available for a minority of FCD patients. 
A pharmacologic alternative to surgery in patients with FCD would be very beneficial, 
even if pharmacologic treatment only delays the need for surgery.  


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This study is designed to investigate the efficacy and safety of netarsudil for the treatment of 
FCD and to enable optimal study design for further clinical development. The optimal dosing 
regimen will provide maximum reduction in corneal edema due to FCD with minimal 
adverse effects.  
It should be noted that, since FCD is a slowly progressing disease, surgery is rarely urgent, 
and patients are often seen multiple times before undergoing surgery. Therefore, an 
approximate 8 to 9-week delay in surgery for participation in this study is acceptable. 
3.3 
Study Duration 
The expected duration of subject participation is approximately 8 to 9 weeks. This includes a 
screening period of up to 8 days, followed by a treatment period of 8 weeks. 
4. STUDY POPULATION SELECTION 
4.1 
Study Population 
A total of approximately 40 subjects will be enrolled into the study at approximately 20 
investigational sites within the United States. Subjects will be at least 18 years of age with a 
diagnosis of FCD, each of whom meets all inclusion criteria and none of the exclusion 
criteria listed below.  
4.2 
Inclusion Criteria 
Subjects must meet all of the following criteria to enter into the study: 
1. Adults, aged 18 years or older at the Screening Visit, of either sex and any race or 
ethnicity 
2. Documented diagnosis of FCD at the Screening Visit 
3. Evidence of central corneal edema, in at least one eye, at the Screening and Baseline 
Visit. The study eligible eye(s) should have a CCT of at least 600 μm at both Screening 
and Baseline, as assessed by ultrasound pachymetry   
4. Evidence of corneal edema attributed to FCD of <12-months’ duration in the eligible 
eye(s) 
5. Best Corrected Visual Acuity using the Early Treatment of Diabetic Retinopathy Study 
(ETDRS) methodology of 70 to 20 letters at Screening and Baseline in the eligible eye(s) 
6. Presence of central corneal edema deemed by the Investigator to be the chief cause of 
reduced visual acuity in the eligible eye(s) 
7. Willing and able to provide written informed consent to participation in the study prior to 
any and all study-related activities 


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4.3 
Exclusion Criteria 
Subjects meeting any of the following criteria will be excluded from entry into the study: 
Ophthalmic: 
1. FCD so advanced that, in the opinion of the Investigator, surgery would likely be 
required in the study eligible eye(s) within the study period   
2. Concurrent or anticipated need for treatment for FCD, e.g., Muro 128 (2% or 5%) during 
the course of the study in the study eligible eye(s). Provided the subject undergoes a 
washout of Muro drops for at least 24 hours prior to the Baseline Visit, they will not be 
excluded  
3. Clinically-significant ocular disease or trauma which could interfere with the 
interpretation of study results (e.g., uveitis, scarring, any severe retinal disease) in the 
eligible eye(s) 
4. Recent (within 3 months of Screening) or current ocular infection (bacterial, fungal or 
viral) in either eye 
5. Recent (within 3 months of Screening) or current severe ocular inflammation (e.g., severe 
blepharitis, conjunctivitis) in either eye 
6. History of any prior ocular refractive surgery in the study eligible eye(s)  
7. Use of contact lenses in either eye within 7 days prior to the Baseline Visit or planned use 
during the study 
8. History of ocular surgery within 6 months of the Screening Visit, any prior complicated 
cataract surgery resulting in vitreous loss, or any anticipated ocular surgery during the 
course of the study in the study eligible eye(s) 
9. Current treatment for, or a history of severe or difficult to treat (i.e., requiring ≥3 ocular 
hypotensive medications) glaucoma (open- or closed- angle) or OHT. Fixed dose 
combination medications will be considered as two medications 
10. Concurrent use of oral or ophthalmic carbonic anhydrase inhibitors (e.g., acetazolamide 
[Diamox®], dorzolamide [Trusopt®], or brinzolamide [Azopt®]) during the course of the 
study is prohibited. Individuals currently on, or previously treated with, oral carbonic 
anhydrase inhibitors for IOP reduction will be excluded from the study. Individuals 
currently using ophthalmic carbonic anhydrase inhibitor drops must undergo a minimum 
washout period of 5 days prior to the Baseline Visit  
11. Concurrent or anticipated use of topical corticosteroids in the study eligible eye(s) 
12. History of prior treatment with netarsudil (monotherapy or in combination) or any other 
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Systemic: 
13. Any clinically-significant or chronic co-morbidity which, in the opinion of the 
Investigator, could interfere with the subject’s ability to participate in the study and 
comply with the protocol, visit schedule and all assessments (e.g., autoimmune disorder, 
terminal illness, psychiatric or cognitive disorder, severe cardiac, pulmonary,
or neurological condition, advanced diabetes, etc.)
14. Known contraindication or hypersensitivity to any of the treatments, anesthetics or 
diagnostic agents or components thereof which may be used during the study
15. Recent (within 60 days of the Screening Visit) or ongoing participation in any other 
investigational interventional clinical study
16. Concurrent or anticipated use of systemic corticosteroids during the study. Individuals 
currently using topical or other locally acting corticosteroids (with the exception of use in 
the study eligible eye[s]) must have been on a stable dose for at least 30 days prior to the 
Screening Visit, and the dose must not be anticipated to change during the course of the 
study
17. Women of child-bearing potential who are pregnant, nursing, planning a pregnancy or not 
using a medically acceptable form of birth control (to be considered NOT of child-
bearing potential, a woman must be at least one-year post-menopause or three-months 
post-surgical sterilization). All women of child-bearing potential will be required to take a 
urine pregnancy test at the Screening and Week 8/Exit Visits 
Subjects who have transient, fluctuating edema, but otherwise meet all of the inclusion 
criteria and none of the exclusion criteria WILL be allowed into the study. 
4.4 
Study Eye Determination 
The subject must have at least one eye meeting all the inclusion criteria (Section 4.2) and 
none of the exclusion criteria (Section 4.3) at the Screening and Baseline Visit. The study 
eligible eye(s) should have a Screening and Baseline CCT of at least 600 μm, as assessed by 
ultrasound pachymetry. If both eyes qualify at both the Screening and Baseline Visit, 
the study eye will be the eye with the thinner cornea at the Baseline Visit. If both eyes have 
the same CCT, the study eye will be the right eye. Study subjects will be instructed to instill 
the study medication into the study eye only.  


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5. STUDY TREATMENTS 
5.1 
Description of Treatments 
5.1.1 
Study Medication 
Netarsudil is a ROCK inhibitor approved by the US Food and Drug Administration for the 
treatment of OHT and open-angle glaucoma.   
Netarsudil 0.02% ophthalmic solution is supplied as a sterile, isotonic, buffered aqueous 
solution of netarsudil dimesylate with a pH of approximately 5 and an osmolality of 
approximately 295 mOsmol/kg. It is intended for topical application in the eye. Each mL 
contains 0.2 mg of netarsudil (equivalent to 0.28 mg of netarsudil dimesylate). Benzalkonium 
chloride, 0.015%, is added as a preservative. The inactive ingredients are boric acid, 
mannitol, sodium hydroxide to adjust pH, and water for injection. 
5.1.2 
Placebo or Control Drug 
A Study Artificial Tear will be provided to subjects randomized to the QD dosing regimen. 
These subjects will be instructed to instill one drop of the Study Artificial Tear every 
morning.  
The Study Artificial Tear will be REFRESH PLUS® (preservative-free) Lubricant Eye 
Drops. The active ingredient is carboxymethylcellulose sodium (0.5%), and the inactive 
ingredients are calcium chloride, magnesium chloride, potassium chloride, purified water, 
sodium chloride, and sodium lactate. The solution may also contain hydrochloric acid and/or 
sodium hydroxide to adjust pH. 
5.2 
Treatments Administered 
Subjects will be randomized using a 1:1 allocation ratio to one of two treatment groups: 
• Once Daily Netarsudil Dosing Regimen 
Study Artificial Tear; one drop, in the study eye in the morning  
Netarsudil 0.02% ophthalmic solution; one drop, in the study eye in the evening 
• Twice Daily Netarsudil Dosing Regimen 
Netarsudil 0.02% ophthalmic solution; one drop, in the study eye in the morning and 
in the evening 
Doses will be self-administered by the study subjects. If the subject is using any another 
topical ophthalmic drop (e.g., ocular hypotensive medications) at the same time as netarsudil, 
the subject should be instructed to instill netarsudil first, and wait at least 15 minutes before 
instilling any additional drops. 


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For subjects deemed unable to self-administer the doses, a caregiver will be asked to 
administer the medication. All subjects will administer study medication for approximately 
8 weeks. 
5.3 
Selection and Timing of Dose for Each Patient 
The dose and regimens selected for this study have previously been tested in studies of 
netarsudil for the treatment of glaucoma and OHT.   
5.4 
Method of Assigning Patients to Treatment Groups 
Subjects will be assigned to treatment groups through the use of an interactive response 
technology (IRT) system. 
5.5 
Masking 
This is an open-label study. 
5.6 
Concomitant Therapy 
5.6.1 
Prohibited Interventions 
Concomitant use of hypertonic sodium chloride ophthalmic solution (e.g., Muro 128 
[2% or 5%]) during the course of the study is prohibited. Individuals currently using 
hypertonic sodium chloride drops or ointment must undergo a minimum washout period of 
24-hours prior to the Baseline Visit.  
Concomitant use of oral or ophthalmic carbonic anhydrase inhibitors (e.g., acetazolamide 
[Diamox®], dorzolamide [Trusopt®], or brinzolamide [Azopt®]) during the course of the 
study is prohibited. Individuals currently on, or previously treated with, oral carbonic 
anhydrase inhibitors for IOP reduction will be excluded from the study. Individuals currently 
using ophthalmic carbonic anhydrase inhibitor drops must undergo a minimum washout 
period of 5 days prior to the Baseline Visit. 
Concomitant use of systemic corticosteroids is prohibited during the course of the study. 
Individuals currently using topical or other locally acting corticosteroids (with the exception 
of use in the study eligible eye[s]) must have been on a stable dose for the month prior to the 
Baseline Visit, and the dose must not be anticipated to change during the course of the study. 
Contact lens wear in either eye during the course of the study is prohibited. Individuals 
currently wearing contact lenses must undergo a minimum washout period of 7 days prior to 
the Baseline Visit. 
 


 
 
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The use of subject’s own artificial tears will not be permitted. All subjects will be provided 
with a Protocol Approved Artificial Tear for intermittent use, up to four times daily. 
See Section 5.6.2 for more information on the Protocol Approved Artificial Tear.  
5.6.2 
Permitted Interventions  
Therapy considered necessary for the subject’s welfare may be given at the discretion of the 
Investigator. If the use of a specific therapy or intervention is in question, please contact 
Aerie. 
Any systemic medication not itemized as an exclusion in Section 4.3 (i.e., oral carbonic 
anhydrase inhibitors or systemic corticosteroids) will be permitted.  
Any topical ophthalmic medication not itemized as an exclusion in Section 4.3 
(i.e., hypertonic saline, ophthalmic carbonic anhydrase inhibitors, or ophthalmic 
corticosteroids in the study eye) will be permitted. Ocular hypotensive medications will be 
permitted, however, a history of severe or difficult to treat (i.e., requiring ≥3 ocular 
hypotensive medications) glaucoma (open- or closed- angle) or OHT will exclude a subject 
from participation in this study. Fixed dose combination medications will be considered as 
two medications. If multiple topical ophthalmic medications are needed at the same time, 
subjects should be instructed to always instill netarsudil first and wait at least 15 minutes 
before instilling any additional drops. 
The intermittent use of artificial tears will be permitted. However, subjects must use the 
(Aerie supplied) Protocol Approved Artificial Tear (REFRESH PLUS® [preservative-free]). 
The Protocol Approved Artificial Tear may be used, as needed, up to four times daily (QID) 
in either eye. If the subject intends on using the Protocol Approved Artificial Tear at the 
same time as their scheduled netarsudil dose, they should be instructed to instill netarsudil 
first and wait a minimum of 15 minutes before instilling the Protocol Approved Artificial 
Tear. 
Any medication taken during the study between the date of the first dose of randomized 
study medication and the date of the Week 8/Exit Visit or Early Termination Visit should be 
recorded in the eCRF as a concomitant medication.  
5.7 
Restrictions 
5.7.1 
Prior Therapy 
Prior use of netarsudil (monotherapy or in combination) will exclude the patient from 
participation in the study. 
5.7.2 
Fluid and Food Intake 
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5.7.3 
Subject Activity Restrictions 
There are no activity restrictions for subjects participating in this study. 
5.8 
Treatment Compliance 
All subjects will be instructed on the importance of following their assigned dosing regimen. 
Subjects in the once daily netarsudil treatment group will instill one drop of the Study 
Artificial Tear in the study eye every morning, and one drop of netarsudil 0.02% in the study 
eye every evening. Subjects in the twice daily netarsudil treatment group will instill one drop 
of netarsudil 0.02% in the study eye in the morning and evening. At Week 4, any subject 
randomized to netarsudil QD who, in the opinion of the Investigator, has not responded 
adequately to treatment, may have their netarsudil dosing frequency increased to BID for the 
remainder of the study. 
Morning dosing should occur between 06:00 and 09:00 hours. Evening dosing should occur 
between 19:00 and 22:00 hours.  
As no commercial method is readily available for direct single-container monitoring of 
treatment adherence with multi-dose ophthalmic products, no formal measure of treatment 
adherence to netarsudil is planned.  
Subjects should be reminded at all visits to adhere to their assigned dosing regimen. 
5.9 
Packaging and Labeling 
Netarsudil will be supplied in a sterile, clear, multi-dose low density polyethylene (LDPE) 
dropper dose ophthalmic bottle with a white propylene cap. 
The Study Artificial Tear (REFRESH PLUS® [preservative-free]) will be supplied in a box 
containing 30 single use containers.   
The Protocol Approved Artificial Tear (REFRESH PLUS® [preservative-free]) will be 
supplied in a box containing 30 single use containers. Please see Section 5.6.2 for more 
information on the Protocol Approved Artificial Tear.  
5.10 
Storage and Accountability 
The study medication, Study Artificial Tears, and Protocol Approved Artificial Tears must be 
dispensed according to the procedures prescribed in this protocol. Only subjects enrolled in 
the study may receive study medication, in accordance with all the applicable regulatory 
requirements. Only authorized staff is allowed to dispense these medications. 
Under normal conditions of handling and administration, the study medication, Study 
Artificial Tears, and Protocol Approved Artificial Tears are not expected to pose significant 
safety risk to site staff. Adequate precautions must be taken to avoid direct contact with the 
study medication.  


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The study medication, Study Artificial Tears, and Protocol Approved Artificial Tears will be 
stored in a secure area under the appropriate physical conditions for the product. Access to 
the study medication, Study Artificial Tears, and Protocol Approved Artificial Tears will be 
limited to authorized site staff only. The study medication and artificial tears will be stored as 
directed on the label. Netarsudil should be stored refrigerated (2°C to 8°C / 36°F to 46°F) 
and protected from light. Study Artificial Tears and Protocol Approved Artificial Tears will 
be stored between 15°C and 30°C (59°F and 86°F). Temperature of the study medication 
storage locations at the site is to be monitored using a calibrated monitoring device and 
documented. 
At time of dispensing, the subject will be instructed to store the bottle(s) as directed on the 
label. Once the netarsudil bottle is opened, the product must always be stored refrigerated 
and protected from light. It is recommended that the product is stored in the carton provided.  
5.11 
Study Medication Retention at Study Site 
5.11.1 
Receipt and Disposition of Study Medication 
Study medication and artificial tears (Study Artificial Tears and Protocol Approved Artificial 
Tears) will be shipped to the Investigator’s site from a central depot. A study staff member at 
the Investigator’s site will verify study medication shipment records by comparing the 
shipping documentation accompanying the study medication to the study medication 
received at the Investigator’s site. If a discrepancy is noted, the appropriate individual at the 
Sponsor or designee must be notified immediately. The responsible person (e.g., study 
coordinator) at the Investigator’s institution has to account for all used, partially used, and 
unused study medication and artificial tears. The responsible person will also maintain the 
study medication accountability records. 
5.11.2 
Return of Study Medication 
When the site is closed, the study is completed or is terminated by the Sponsor; all study 
material including used and unused study medication and artificial tears will be returned to 
the Sponsor’s designee. All study medication accounting procedures must be completed 
before the study is considered to be concluded. The responsible person at the Investigator’s 
institution has to account for all used, partially used and unused study medication and 
artificial tears. The monitor will complete a study medication returns form or equivalent that 
will be signed by the Investigator or designee prior to returning the used and unused study 
medication and artificial tears to the Sponsor’s designee. 
 


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6. STUDY PROCEDURES 
6.1 
Informed Consent 
Prior to any study procedures in the treatment period, the study will be discussed with each 
subject. Subjects wishing to participate must give written informed consent. The verbal 
explanation of the study will cover all the elements specified in the written information 
provided for the subject. The Investigator will inform the subject of the aims, methods, 
anticipated benefits and potential hazards of the study, including any discomfort it may 
entail. The subject must be given every opportunity to clarify any points he/she does not 
understand and, if necessary, may ask for more information. At the end of the interview, the 
subject should be given time to reflect. Subjects and/or legally authorized representative then 
will be required to sign and date the informed consent form. The completion of this process 
should be documented in the source documents.  
The informed consent form must have received approval/favorable review by a properly 
constituted Institutional Review Board/Independent Ethics Committee (IRB/IEC) prior to 
use. A copy of the signed and dated consent document will be given to each subject. 
The original signed and dated informed consent document must be maintained in the study 
files at the Investigator’s site. 
The Investigator or staff is responsible for ensuring that no subject undergoes any study 
related examination or activity before the subject has given written informed consent. 
It should be emphasized that the subject is at liberty to withdraw consent to participate at any 
time, without penalty or loss of benefits to which the subject is otherwise entitled. Subjects 
who refuse to give, or withdraw, written informed consent may not be included or continued 
in this study, and should be notified that discontinuation from the study will not impact on 
their subsequent care. 
6.2 
Demographics and Medical/Ophthalmic History 
Demographic data will be collected and recorded at the Screening Visit. Significant, relevant 
(to the condition under investigation) medical/ophthalmic history will also be collected at 
Screening and Baseline. Any current underlying medical conditions, including those that 
began within the last 30 days and which may have resolved before screening, must be 
recorded. In addition, prior treatments for FCD should be recorded. Any medications the 
subject took but discontinued within the 30 days prior to screening also will be recorded as 
part of the medical history. 
6.3 
Concomitant Medication Assessments 
Use of any medication, prescription or over-the-counter (OTC), should be recorded at the 
Screening Visit, and captured on the appropriate eCRF, and the indication noted as part of 
the medical history. Treatments that are permitted to continue throughout the duration of the 
study will be recorded as concomitant medications at all subsequent visits. Judgment of 


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continued study participation by the subject, and inclusion of this subject’s subsequent visits 
in the safety and efficacy analysis will be made by the Investigator. 
All medications which the subject has taken within 30 days prior to screening and during the 
study will be recorded in the eCRF. The name of the drug, dose, route of administration, 
duration of treatment and indication will be recorded for each medication. For combination 
products (e.g., CONTAC®), the brand name is required. For non-combination products, the 
generic name is preferred. The use of routine ophthalmic diagnostic pharmaceutical agents 
(e.g., fluorescein) will be allowed, and individual documentation not required. Any change in 
dosing parameters should also be recorded in the eCRF. 
6.4 
Vital Signs (HR and BP only) 
Subject heart rate and blood pressure will be measured at the Screening, Baseline, and 
Week 8/Exit Visits. Heart rate will be determined only once during these study visits by the 
method described in the Study Procedure Manual. 
Blood pressure will be measured once for each subject after the subject heart rate has been 
determined, with the subject in a sitting position. A mechanical or digital 
sphygmomanometer may be used, but effort should be made to use the same instrument and 
the same arm of the subject for each reading. Please refer to the Study Procedure Manual for 
more details. 
6.5 
Pregnancy Testing 
A urine pregnancy test will be conducted at the Baseline Visit and the Week 8/Exit Visit for 
all females of child-bearing potential. A female of child-bearing potential is defined as an 
adult woman unless she is 1-year post-menopausal or 3 months post-surgical sterilization. 
Any female who is pregnant, nursing, planning a pregnancy, or not using a medically 
acceptable form of birth control will be excluded from the study. All females of childbearing 
potential must have a negative pregnancy test result at the Baseline Visit to be enrolled and 
must not intend to become pregnant during the study. 
6.6 
Dispensing Study Medication 
Study staff responsible for dispensing the study medication will be listed on the Delegation 
of Responsibilities Log. When a subject meets all criteria for selection and has completed all 
screening assessments, the subject will be assigned to a treatment group according to the 
IRT. 
Study medication, Study Artificial Tears (for subjects randomized to the QD dosing 
regimen), and Protocol Approved Artificial Tears will be dispensed at the Baseline Visit. 
Additional study medication, Study Artificial Tears (for subjects randomized to the QD 
dosing regimen), and Protocol Approved Artificial Tears (if needed) will be dispensed at the 
Week 4 Visit.   


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The responsible study staff will account for used and unused study medication and artificial 
tears by maintaining a study medication accountability log. 
6.7 
Efficacy Assessments 
6.7.1 
Ultrasound Pachymetry 
CCT will be measured at each visit at approximately the same time of day ± 60 minutes and 
always before noon (local time). Every effort should be made to use the same instrument for 
each reading. For more details, please refer to the Study Procedure Manual. 
6.7.2 
Best Corrected Visual Acuity (ETDRS Method) 
At each visit, BCVA should be measured using the ETDRS method, following manifest 
refraction, and prior to the slit lamp examination. Please refer to the BCVA Procedure 
Manual for the BCVA Worksheet and more details.  
6.7.3 
Biomicroscopy  
External magnification and biomicroscopy will be performed using a slit-lamp. 
Magnification will be consistent with standard clinical practice. The subject will be seated.  
The following will be examined:  
• Eyelid 
• Cornea  
• Conjunctiva 
• Anterior Chamber 
• Iris 
• Lens 
For more details, please refer to the Study Procedure Manual. 
6.7.4 
V-FUCHS Questionnaire 
The Visual Function and Corneal Health Status (V-FUCHS) patient reported visual disability 
questionnaire will be administered at the Baseline, Week 4, and Week 8 Visits 
(Wacker 2018). Subjects who are not fluent in the English language may be exempt from 
completing this questionnaire. Please refer to the Study Procedure Manual for the full 
15-item questionnaire and additional details.  


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6.7.5 
Tomography (Pentacam®; Selected Sites Only) 
Tomography assessments using a Pentacam® will be performed at selected sites only. 
Please refer to the Study Procedure Manual for more details.   
6.8 
Safety Assessments  
6.8.1 
IOP 
At each visit, IOP will be measured using an Applanation Tonometer. 
6.8.2 
Ophthalmoscopy  
Indirect ophthalmoscopy will be performed with pupil dilation at the Screening and 
Week 8/Exit Visits and without dilation at all other visits. The Investigator will make 
observations of the vitreous, retina, macula, choroid and optic nerve. Please refer to the Study 
Procedure Manual for more details. 
6.9 
Adverse Events Assessments 
6.9.1 
Performing Adverse Event (AE) Assessments 
Qualified study staff responsible for assessing AEs will be listed on the Site Authorization 
and Delegation Log. This includes assessment of AE severity and relationship to the study 
medication. AE information may be volunteered by the subject or solicited by study 
personnel through non-leading questions. 
All AEs occurring during the study, regardless of the assumption of causal relationship, must 
be documented on the respective electronic case report form (eCRF). Adverse events should 
be documented from the time the subject signs the informed consent until the subject’s last 
study visit. If an event occurs during the washout period (prior to subject enrollment and the 
administration of study medication), it should be recorded as an AE. Any medical condition 
present prior to administration of the study medication which remains unchanged or 
improved should not be recorded as an AE at subsequent visits. 
If a subject has an ongoing AE at the time of study completion, the ongoing AE must be 
followed-up and provided appropriate medical care until the event has resolved or stabilized, 
the subject is lost to follow-up, or there is other resolution to the event. 
Documentation of AEs/adverse reactions includes start date and stop date, severity, action(s) 
taken, seriousness and outcome. 
Investigators are also asked to note all observations of new or worsening conjunctival 
hyperemia on the biomicroscopy eCRF as well as on the study AE form if deemed necessary 
(per Investigator discretion). 


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The following definitions of terms apply to this section: 
• Adverse event (AE). Any untoward medical occurrence associated with the use of a drug 
in humans, whether or not considered drug related. 
• Suspected adverse reaction (SAR). Any AE for which there is a reasonable possibility 
that the drug caused the AE. For the purposes of IND safety reporting, ‘‘reasonable 
possibility’’ means there is evidence to suggest a causal relationship between the drug 
and the AE. Suspected adverse reaction implies a lesser degree of certainty about 
causality than adverse reaction, which means any AE caused by a drug. 
• Life-threatening AE or life-threatening SAR. An AE or SAR is considered 
“life-threatening” if, in the view of either the Investigator or Sponsor, its occurrence 
places the patient at immediate risk of death. It does not include an AE or SAR that, 
had it occurred in a more severe form, might have caused death. 
• Serious adverse event (SAE) or serious suspected adverse reaction (SSAR). An AE or 
SAR is considered ‘‘serious’’ if, in the view of either the Investigator or Sponsor, 
it results in any of the following outcomes: Death, a life-threatening AE, patient 
hospitalization or prolongation of existing hospitalization, a persistent or significant 
incapacity or substantial disruption of the ability to conduct normal life functions, or a 
congenital anomaly/birth defect. Important medical events that may not result in death, 
be life-threatening, or require hospitalization may be considered serious when, based 
upon appropriate medical judgment, they may jeopardize the patient and may require 
medical or surgical intervention to prevent one of the outcomes listed in this definition. 
Examples of such medical events include allergic bronchospasm requiring intensive 
treatment in an emergency room or at home, blood dyscrasias or convulsions that do not 
result in patient hospitalization, or the development of drug dependency or drug abuse. 
• Unexpected AE or unexpected SAR. An AE or SAR is considered ‘‘unexpected’’ if it is 
not listed in the Investigator’s Brochure (IB), the Package Insert, or is not listed at the 
specificity or severity that has been observed. For example, under this definition, hepatic 
necrosis would be unexpected (by virtue of greater severity) if the IB referred only to 
elevated hepatic enzymes or hepatitis. Similarly, cerebral thromboembolism and cerebral 
vasculitis would be unexpected (by virtue of greater specificity) if the IB listed only 
cerebral vascular accidents. “Unexpected,” as used in this definition, also refers to AEs or 
SARs that are mentioned in the IB as occurring with a class of drugs or as anticipated 
from the pharmacological properties of the drug, but are not specifically mentioned as 
occurring with the particular drug under investigation. 


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6.9.2 
Timing 
The AEs occurring during the study must be documented, regardless of the assumption of a 
causal relationship. AEs should be documented from the time the subject signs and dates the 
patient consent form until subject participation in the study has been completed. If a subject 
has one or more ongoing AEs at the time of study completion, the subject should be followed 
and provided appropriate medical care until the event is resolved or stabilized, the subject is 
lost to follow-up, or there is other resolution to the event. These follow-up visits will be 
documented. 
When recording an AE, the following information should be provided on the study AE 
eCRF: 
1. Action Taken with Study Medication: 
• None 
• Study Medication Interrupted 
• Study Medication Discontinued 
2. Other Action Taken: 
• None 
• Non-Drug Therapy 
• New OTC or Rx Drug Added 
• Hospitalized less than 24 hours 
• Hospitalized greater than or equal to 24 hours  
3. Outcome of an adverse event is coded as:  
• Fatal  
• Not Recovered/Not Resolved  
• Recovered/Resolved  
• Recovered/Resolved with sequelae  
• Recovering/Resolving  
• Unknown/Lost to follow-up 


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6.9.3 
Severity 
Severity of an AE is defined as a qualitative assessment of the level of discomfort or the 
degree of intensity of an AE as determined by the Investigator or reported to him/her by the 
subject. The assessment of severity is made irrespective of study medication relationship or 
seriousness of the event and should be evaluated according to the following scale: 
1 = Mild: present and noticeable, but not distressing, and no disruption of normal daily 
activities. 
2 = Moderate: bothersome, discomfort sufficient to possibly reduce or affect normal daily 
activity. 
3 = Severe: incapacitating, with inability to work or perform normal daily activity. 
A change in increased severity for a reported AE will require a stop date for the previous 
severity and a new start and stop date for the new severity. For example, a change in severity 
may go from mild to severe or from moderate to severe. In both cases, the start and stop dates 
should be recorded. 
Note: A severe AE is not the same as a serious AE. Seriousness of an AE (NOT severity) 
serves as a guide for defining regulatory reporting obligations (see Section 6.9.6 for further 
information on serious AEs [SAEs]). 
6.9.4 
Relationship 
The study medication relationship for each AE/adverse reaction should be determined by the 
Investigator using these explanations: 
• Not Related: The event is clearly related to other factors such as subject’s clinical 
condition; therapeutic interventions, concomitant disease or therapy administered to the 
subject and does not follow a known response pattern to the product. 
• Unlikely Related: The event is most probably caused by other etiologies such as 
participant’s underlying condition, therapeutic intervention, or concomitant therapy; or 
the delay between administration and the onset of the AE is incompatible with a causal 
relationship. Therefore, there is not a reasonable possibility that the AE was caused by 
the study medication. 
• Possibly Related: The event follows a reasonable, temporal sequence from the time of 
study medication administration and/or follows a known response pattern to the study 
medication, but could have been produced by other factors such as the subject’s clinical 
state, therapeutic interventions or concomitant therapy administered to the subject. 
• Related: The event follows a reasonable, temporal sequence from the time of study 
medication administration and/or follows a known response pattern to the study 


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medication and cannot be reasonably explained by other factors such as subject’s clinical 
state, therapeutic interventions or concomitant therapy administered to the subject, and 
either occurs immediately following study medication administration, or improves on 
stopping the study medication, or reappears on repeat exposure, or there is a positive 
reaction at the application site. 
6.9.5 
Expectedness 
An AE or suspected adverse reaction is considered “unexpected” if it is not listed in the IB or 
is not listed at the specificity or severity that has been observed. “Unexpected,” as used in 
this definition, also refers to adverse events or suspected adverse reactions that are mentioned 
in the IB as occurring with this class of drugs or as anticipated from the pharmacological 
properties of netarsudil, and are not specifically mentioned as occurring with the study 
medication. The AEs and adverse reactions that are both unexpected and serious should be 
reported in an expedited fashion to the Sponsor (see Section 6.9.6 for further details). 
6.9.6 
Serious Adverse Events, Suspected Serious Adverse Reactions or Suspected 
Unexpected Serious Adverse Reactions 
6.9.6.1 
Reporting SAEs, SSARs or SUSARs 
An Investigator must immediately (i.e., within 24 hours) report any SAE, SSAR or SUSAR 
to the Sponsor using the SAE report form, whether or not considered medication-related, 
including those listed in the IB, and must include an assessment of whether there is a 
reasonable possibility that the medication caused the event. In addition, in the case of 
immediately life-threatening AEs or AEs with fatal outcome, or SUSARs, the Investigator 
must inform the Sponsor by phone within 24 hours of observation or occurrence of the SAE. 
The Investigator must report any SAE, SSAR, or SUSAR that occurs during the course of the 
study or within 4 weeks of last study visit. In case of incomplete information, the Investigator 
must provide follow-up information as soon as possible, again using the SAE report form. 
SAEs, SSARs, and SUSARs must be reported to the IRB/IEC according to the IRB/IEC 
requirements. 
The contact information of the study Safety Mailbox is as follows: 
Covance PSS 
SAEintake@covance.com 
Fax: 1-888-887-8097 
The contact information of the study Medical Monitors is as follows: 
Richard A. Lewis, MD  
Phone: (916) 768-7703  
Email: rlewis@aeriepharma.com 
 
 


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6.9.7 
Pregnancy Reporting 
Pregnancies occurring in subjects in this study, occurring up to the subject’s last study visit, 
must be reported and followed to outcome. Information on pregnant partners will not be 
collected. 
While pregnancy itself is not considered to be an AE or SAE, pregnancy reports are tracked 
by Covance and the study Medical Monitor. Premature terminations including miscarriage, 
spontaneous abortion, or elective termination of a pregnancy for medical reasons will be 
reported as an SAE. Other pregnancy complications should be reported as SAEs, if they meet 
serious criteria. Should a pregnancy result in a congenital anomaly or birth defect, a separate 
SAE report must be submitted for the baby. Furthermore, all neonatal deaths that occur 
within 30 days of the birth should be reported as SAEs, without regard to causality. 
The Investigator must complete the pregnancy report form and email the form to the study 
Safety Mailbox within 1 business day of knowledge of the pregnancy. Following delivery or 
termination of pregnancy, the updated or follow-up pregnancy report form is to be completed 
and submitted by fax or email to the Safety Mailbox for the study. 
6.10 
Removal of Subjects from the Study or Study Medication 
Subjects may be discontinued from the study at any time for any reason. Participation is 
entirely voluntary, and only possible if the subject has signed informed consent. Consent may 
be withdrawn at any time. 
6.10.1 
Completed Subject 
A completed subject is defined as one who completes all planned study treatments and visits. 
6.10.2 
Non-completing Subject 
A non-completing subject is defined as one who exits the study by their own volition or at 
the discretion of the Investigator, the Medical Monitor, and/or an Aerie Safety 
Officer/designee. Any subject may decide to voluntarily withdraw from the study at any time 
without prejudice. If discontinuation of treatment is necessary, the Investigator will make 
every attempt to complete all subsequent safety assessments listed for the Week 8 / Early 
Termination Visit. 
The subject may also be discontinued from the study for the following reasons:  
• 
AEs (including intercurrent diseases reported by the subject or observed by the 
Investigator with documentation on the CRF)  
• 
Withdrawal of Consent  
• 
Non-compliance (e.g., non-adherence to scheduled follow-up visits or use of study 
treatment)  


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• 
Lost to Follow-up  
• 
Pregnancy  
• 
Investigator Decision  
• 
Protocol Deviation  
• 
Death  
• 
Other  
6.10.3 
Actions after Discontinuation 
All subjects who discontinue study treatment due to a report of an AE must be followed and 
provided appropriate medical care until their signs and symptoms have remitted or stabilized.  
For subjects who choose to withdraw consent or who are discontinued for non-compliance 
prior to completing the study, every possible effort should be made by the Investigator to 
assure there is a final visit that includes all examinations listed for the Week 8 / Early 
Termination Visit. 
6.10.4 
Discontinuation of the Entire Study 
The entire study may be discontinued at a given site (by the Investigator or Aerie /Aerie 
representative) or at all sites by Aerie. Prompt, written notice of reasonable cause to all other 
relevant parties (Aerie or Investigator) is required. Prompt notice to the IRB and to 
regulatory authorities is also required. 
6.10.5 
Completed Study  
The study is completed when the planned enrollment has been completed, and all the 
enrolled subjects have completed the study. An Aerie representative will be in 
communication with the investigational sites regarding enrollment. 
6.11 
Appropriateness of Measurements 
The ophthalmic and systemic measures included in this study are consistent with standard of 
care. 
7. STUDY ACTIVITIES 
A detailed Schedule of Visits and Procedures is provided in Table 1. Subjects should be 
scheduled at approximately the same time of day ± 60 mins in the morning (ideally at or 
before 10:00 local time) for each visit to ensure that ultrasound pachymetry is completed 
before noon (local time). 


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7.1 
Screening Visit (Day -8 to -1) 
• Informed consent 
• Inclusion and exclusion criteria 
• Demographics 
• Medical, ophthalmic, and surgical history 
• Prior or concomitant medication review 
• Vital signs (heart rate and blood pressure only) 
• Ocular symptoms and AE review for both eyes 
• BCVA using the ETDRS method and following manifest refraction for both eyes 
• Anterior segment biomicroscopic examination of both eyes 
• CCT will be measured by ultrasound pachymetry in both eyes  
o Note: Ultrasound pachymetry should be performed at approximately the same time of 
day for all visits (± 60 minutes) and always before noon (local time) 
• IOP in both eyes 
• Dilated Ophthalmoscopy in both eyes 
• Tomography (Pentacam®) for both eyes (at selected sites only) – may be performed prior 
to ophthalmoscopy while the subject is dilating 
7.2 
Treatment Period 
7.2.1 
Baseline (Day 1) Procedures 
• Inclusion and exclusion criteria 
• Visual Function and Corneal Health Status (V-FUCHS) patient reported visual disability 
questionnaire (Wacker 2018) 
• Medical, ophthalmic, and surgical history 
• Concomitant medication review 
• Vital signs (heart rate and blood pressure only) 


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• Urine pregnancy test (WOCBP only) 
• Ocular symptoms and AE review for both eyes 
• BCVA using the ETDRS method and following manifest refraction for both eyes 
• Anterior segment biomicroscopic examination of both eyes 
• CCT will be measured by ultrasound pachymetry in both eyes 
o Note: Ultrasound pachymetry should be performed at approximately the same time of 
day for all visits (± 60 minutes) and always before noon (local time) 
• IOP in both eyes 
• Ophthalmoscopy without dilation in both eyes 
• Tomography (Pentacam®) for both eyes (at selected sites only) 
• Study eye determination (See Section 4.4) 
• Randomization 
• Eye drop instruction and demonstration for the study eye 
• Dispensing of study medication, Study Artificial Tears (if randomized to the QD dosing 
arm), and Protocol Approved Artificial Tears 
7.2.2 
Week 2 (Day 14 ± 3) Procedures 
• Concomitant medication review 
• Ocular symptoms and AE review for both eyes 
• BCVA using the ETDRS method and following manifest refraction for both eyes 
• Anterior segment biomicroscopic examination of both eyes 
• CCT will be measured by ultrasound pachymetry in both eyes 
o Note: Ultrasound pachymetry should be performed at approximately the same time of 
day for all visits (± 60 minutes) and always before noon (local time) 
• IOP in both eyes 
• Ophthalmoscopy without dilation in both eyes 
• Tomography (Pentacam®) for both eyes (at selected sites only) 


 
 
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7.2.3 
Week 4 (Day 28 ± 3) Procedures 
 Visual Function and Corneal Health Status (V-FUCHS) patient reported visual disability 
questionnaire (Wacker 2018) 
 Concomitant medication review 
 Ocular symptoms and AE review for both eyes 
 BCVA using the ETDRS method and following manifest refraction for both eyes 
 Anterior segment biomicroscopic examination of both eyes 
 CCT will be measured by ultrasound pachymetry in both eyes  
o Note: Ultrasound pachymetry should be performed at approximately the same time of 
day for all visits (± 60 minutes) and always before noon (local time) 
 IOP in both eyes 
 Ophthalmoscopy without dilation in both eyes 
 Tomography (Pentacam®) for both eyes (at selected sites only)  
 Dispensing of study medication, Study Artificial Tears (if randomized to the QD dosing 
arm), and Protocol Approved Artificial Tears (if needed) 
o Note: At the Week 4 Visit, any subject randomized to netarsudil QD who, in the 
opinion of the Investigator, has not responded adequately to treatment, may have their 
netarsudil dosing frequency increased to BID for the remainder of the study. 
The subject should be re-trained on their new dosing regimen, if applicable.  
7.2.4 
Week 8 (Day 56 ± 3) or Early Termination Procedures 
 Visual Function and Corneal Health Status (V-FUCHS) patient reported visual disability 
questionnaire (Wacker 2018) 
 Concomitant medication review 
 Vital signs (heart rate and blood pressure) 
 Urine pregnancy test (WOCBP only) 
 Ocular symptoms and AE review for both eyes 
 BCVA using the ETDRS method and following manifest refraction for both eyes 


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• Anterior segment biomicroscopic examination of both eyes 
• CCT will be measured by ultrasound pachymetry in both eyes 
o Note: Ultrasound pachymetry should be performed at approximately the same time of 
day for all visits (± 60 minutes) and always before noon (local time) 
• IOP in both eyes 
• Dilated ophthalmoscopy in both eyes 
• Tomography (Pentacam®) for both eyes (at selected sites only) – may be performed prior 
to ophthalmoscopy while the subject is dilating 
The Week 8 Visit will be the final visit, and the subject should be thanked for their 
participation in the study. 
7.2.5 
Unscheduled Visits 
An unscheduled visit may be any visit to the Investigator other than the specific visits 
specified in the protocol as possibly required for the subject’s ophthalmic condition. 
The Investigator will perform all procedures necessary to evaluate the study participant at 
these visits and record any AEs in the CRF. 
8. QUALITY CONTROL AND ASSURANCE 
The progress of the study will be monitored by on-site, written, and telephone 
communications between personnel at the Investigator’s site and the Study Monitor. 
The Investigator will allow the Sponsor or designee to inspect all CRFs, subject records 
(source documents), signed consent forms, study medication records (receipt, storage, 
preparation, and disposition), and regulatory files related to this study. 
9. PLANNED STATISTICAL METHODS 
9.1 
General Considerations 
Continuous study assessments will be summarized by treatment and time point 
(as applicable) using descriptive statistics (n, mean, median, standard deviation, minimum, 
and maximum). Categorical study assessments will be summarized by treatment and visit 
(as applicable) using frequency counts and percentages. Hypothesis testing, unless otherwise 
indicated, will be performed at a 2-sided 0.10 significance level. Where applicable, 2-sided 
90% and 95% CIs will be reported. All p-values will be displayed to four decimal places, 
with p-values less than 0.0001 presented as '<0.0001' and p-values greater than 0.9999 
presented as ‘>0.9999’. Difference between netarsudil QD and netarsudil BID will be 
calculated as netarsudil QD – netarsudil BID. 


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All study data will be listed by treatment, subject and time point (as applicable). 
Statistical methods will be more fully described in a separate Statistical Analysis Plan. 
9.1.1 
Unit of Analysis 
Safety endpoints will be analyzed for both eyes. For efficacy endpoints, the primary unit of 
analysis will be the study eye defined based on CCT of at least 600 μm evaluated by 
ultrasound pachymetry. If both eyes qualify based on CCT and enrollment criteria, the study 
eye will be the eye with the thinner cornea at the baseline.  Qualified fellow eyes may also be 
summarized. 
9.1.2 
Missing Data and Intercurrent Events 
The primary analysis will be completed with available data per subject from the 
mITT population. Robustness analyses will also be presented based on the multiple 
imputation methodology under different assumptions of missingness and intercurrent events 
(at random and not at random), using last observation carried forward (LOCF), and using the 
per protocol population with available data per subject. 
Intercurrent events will be primary handled in the following manner: 
1. Discontinuation of study medication and non-optimal compliance will be ignored, values 
measured after discontinuation of study medication or regardless of compliance will be 
used in the analyses [treatment policy strategy]. 
2. Withdrawal due to lack of efficacy or adverse events: missing data will be singly imputed 
as the worst within subject observation prior to the intercurrent event [hypothetical 
strategy]. 
3. Missing data without withdrawal or withdrawal due to reasons other than lack of efficacy 
or adverse events: missing data will be imputed employing Multiple Imputation (MI) 
using randomized treatment-based Markov Chain Monte Carlo (MCMC) methodology to 
impute non-monotone missing data and regression methodology to impute monotone 
missing data [hypothetical strategy]. 
9.1.3 
Multiplicity Considerations 
Adjustments for multiplicity will not be made in this early phase trial.   
 
 


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9.2 
Hypotheses 
H01: The mean change from baseline CCT by ultrasound pachymetry in study eyes treated 
with netarsudil QD at Week 4 = 0. 
H11: The mean change from baseline CCT by ultrasound pachymetry in study eyes treated 
with netarsudil QD at Week 4 ≠ 0. 
H02: The mean change from baseline CCT by ultrasound pachymetry in study eyes treated 
with netarsudil BID at Week 4 = 0. 
H12: The mean change from baseline CCT by ultrasound pachymetry in study eyes treated 
with netarsudil BID at Week 4 ≠ 0. 
9.3 
Determination of Sample Size 
This study is not powered to detect a pre-stated efficacy signal, but rather will be used to 
inform the design and power for any possible future studies.  With a sample size of up-to 
20 subjects per treatment group, the precision of the point estimate of mean change from 
baseline in CCT, as measured by the half-width of the two-sided 90% confidence interval 
(CI), will be approximately 1/2.7 of the standard deviation (SD) of the change from baseline, 
yielding 90% confidence that the true mean change from baseline is within ± SD/2.7 µm of 
the observed mean change from baseline.  That is, if the SD of the change from baseline is 
60 µm, then the half-width of the two-sided 90% CI would be 60 µm x 1/2.7 = 22.2 µm, 
yielding 90% confidence that the true mean change from baseline is within ± 22.2 µm of the 
observed mean change from baseline.  
With a sample size of up-to 20 subjects within a treatment group, the study will have 80% 
power to reject H01 or H02 and in favor of H11 or H12 respectively and demonstrate a 
statistically significant mean change from baseline, assuming the true effect size 
(mean change / SD) is 0.577 or larger (e.g., assuming the true mean change from baseline is 
34.6 µm and the standard deviation is 60 µm), a one sample t-test and a two-sided alpha 
= 0.10. 
With a sample size of up-to 20 subjects per treatment group, the study will have 95% 
confidence of ruling out AEs with true incidence rates of 13.9% or higher within each 
treatment group.  That is, with up-to 20 subjects in a treatment group, if an AE of a specific 
type is not observed, then with 95% confidence, the true incidence rate of that adverse event 
is less than 13.9%. 
Similarly, with a sample size of up-to 40 subjects combined over treatment groups, the study 
will have 95% confidence of ruling out AEs with true incidence rates of 7.2% or higher 
within each treatment group.  That is, with up-to 40 subjects in a treatment group, if an AE of 
a specific type is not observed, then with 95% confidence, the true incidence rate of that 
adverse event is less than 7.2%. 


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9.4 
Analysis Populations 
9.4.1 
Modified Intent-to-Treat (mITT) Population 
The mITT population will include all randomized subjects who have received at least one 
dose of study medication. This population will be the primary population for efficacy 
analyses and will be used to summarize a subset of efficacy variables and will summarize 
subjects as randomized. 
9.4.2 
Per Protocol (PP) Population 
The PP population is a subset of the mITT population, which will include those subjects 
(and their visits) who do not have major protocol deviations likely to seriously affect the 
primary outcome of the study. This population will be the secondary population for efficacy 
analyses and will be used to summarize a subset of efficacy variables. If the PP and mITT 
populations are exactly the same, then additional efficacy analyses on the PP population will 
not be performed. The PP population will summarize subjects as treated. 
9.4.3 
Safety Population 
The safety population will include all randomized subjects who have received at least one 
dose of investigational product. This population will be used to summarize safety variables 
and will summarize subjects as treated. 
9.5 
Demographics and Baseline Characteristics 
Demographic and baseline characteristics such as age, gender, race, and ethnicity will be 
summarized and listed. Medical history, history of ocular surgery and procedures will also be 
summarized and listed. 
9.6 
Subject Disposition 
Subject disposition will be presented in terms of the numbers and percentages of subjects 
who completed the study and discontinued from the study. Disposition will be summarized 
by treatment group and for all subjects. 
The number of randomized subjects in each of the analysis populations (mITT, PP, and 
Safety) will be displayed by treatment. 
The number and percentage of subjects who prematurely discontinue from the study and the 
reasons for study discontinuation will be summarized by treatment group for all subjects. 


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9.7 
Primary Endpoint(s) 
9.8 
Efficacy Analysis 
9.8.1 
Primary Efficacy Endpoint 
The primary efficacy endpoint is the mean change from baseline in mean CCT as assessed by 
ultrasound pachymetry at Week 4. 
9.8.2 
Primary Efficacy Analyses 
The primary comparisons in this trial will be within a treatment group on the mean change 
from baseline CCT to Week 4/Day 28 (Visit 4) for each of netarsudil QD and BID. 
The primary efficacy endpoint change from baseline CCT will be summarized descriptively 
(n, mean, standard deviation, median, min, and max) and analyzed primarily using one-
sample t-tests; comparisons between treatment groups will be completed using two-sample 
t-tests. Sensitivity analyses will be completed using Wilcoxon signed-rank tests within a 
treatment group, Wilcoxon rank sum tests between treatment groups and an ANCOVA 
model with terms for baseline CCT value, treatment. The least squares mean will be 
presented for each treatment group from the model together with two-sided p-values and 90 
and 95% confidence intervals around the change from baseline within each treatment group. 
Comparisons between treatment groups on the change from baseline CCT will also be made 
including the least squares mean difference and corresponding two-sided p-values and 90 and 
95% confidence intervals. 
The primary analysis will use the mITT population with available data per subject. 
Robustness analyses will also be presented based on the multiple imputation methodology 
under different assumptions of missingness and intercurrent events (at random and not at 
random), using last observation carried forward (LOCF), and using the per protocol 
population with available data per subject. 
Intercurrent events will be primary handled in the following manner: 
1. Discontinuation of study medication and non-optimal compliance will be ignored, values 
measured after discontinuation of study medication or regardless of compliance will be 
used in the analyses [treatment policy strategy]. 
2. Withdrawal due to lack of efficacy or adverse events: missing data will be singly imputed 
as the worst within subject observation prior to the intercurrent event [hypothetical 
strategy]. 
3. Missing data without withdrawal or withdrawal due to reasons other than lack of efficacy 
or adverse events: missing data will be imputed employing Multiple Imputation (MI) 
using randomized treatment-based Markov Chain Monte Carlo (MCMC) methodology to 
impute non-monotone missing data and regression methodology to impute monotone 
missing data [hypothetical strategy]. 


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9.8.3 
Secondary Efficacy Endpoints 
Secondary endpoints include:  
• Mean change from baseline in BCVA at Week 4 
• Mean change from baseline in CCT as assessed by ultrasound pachymetry at Week 8  
• Mean change from baseline in BCVA at Week 8 
• Proportion of subjects who gained ≥15 letters (3 lines) in BCVA at Week 4 
• Proportion of subjects who gained ≥15 letters (3 lines) in BCVA at Week 8 
• Proportion of subjects with complete resolution of corneal edema at Week 4 
• Proportion of subjects with complete resolution of corneal edema at Week 8 
9.8.4 
Secondary Efficacy Analyses 
The secondary efficacy analyses will be conducted in the mITT population with available 
data per subject. Descriptive statistics will be presented by treatment group. Summarization 
and analysis of the mean change from baseline in BCVA at Weeks 4 and 8 and mean change 
from baseline in CCT by ultrasound pachymetry at Week 8 will be completed using a similar 
strategy as for the primary endpoints.   
Testing of the proportion of subjects who gained ≥15 (3 lines) in BCVA at Weeks 4 and 8 
(separately) and the proportion of subjects with complete resolution of corneal edema at 
Weeks 4 and 8 (separately) will be completed using logistic regression with fixed effects of 
treatment and corresponding baseline measure.  
The adjusted odds ratios and marginal proportions and difference in proportions along with 
corresponding two-sided 95% Confidence Intervals (CIs) and p-values will be presented.   
Treatment comparisons will also be made using Pearson’s chi-squared test or Fisher’s exact 
test if any of the cell counts are less than five as a sensitivity analysis. 
The primary analyses of the secondary endpoints will use the mITT population with available 
data per subject. Robustness analyses may also be presented based on the multiple imputation 
methodology under different assumptions of missingness and intercurrent events (at random 
and not at random), using last observation carried forward (LOCF), and using the per 
protocol population with available data per subject. 
 
 


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9.8.5 
Exploratory Efficacy Endpoints 
Exploratory endpoints include:  
• Proportion of subjects who gained ≥15 letters (3 lines) in BCVA at Week 2 
• Proportion of subjects who gained ≥10 letters (2 lines) in BCVA at Weeks 2, 4, and 8 
• Proportion of subjects who gained ≥5 letters (1 line) in BCVA at Weeks 2, 4, and 8 
• Mean change from baseline in CCT as assessed by Pentacam® at Week 4 (selected sites 
only) 
• Mean change from baseline in V-FUCHS glare factor score at Week 4 
9.8.6 
Exploratory Efficacy Analyses 
Exploratory endpoints will be analyzed in a similar manner as the secondary endpoints.  
9.8.7 
Primary Safety Assessments and Analyses 
Adverse events (AEs) will be coded using the MedDRA dictionary. Frequencies and 
percentages of treatment-emergent adverse events (TEAEs) will be summarized at the 
subject level by system organ class and preferred term for all TEAEs, treatment related 
TEAEs, serious TEAEs, and TEAEs causing premature treatment discontinuation by 
treatment group. An AE is treatment emergent if it occurs or worsens after the first dose of 
study medication. Similar summaries will be presented for all TEAEs by maximal severity. 
Separate summaries will be performed for ocular and non-ocular AEs.  
Best corrected visual acuity and intraocular pressure data for both the study eye and the 
fellow eye will be summarized for each measured visit and for change from baseline to each 
post-treatment visit using the continuous summary statistics.  
Anterior segment biomicroscopy and ophthalmoscopy measures will be summarized at each 
measured visit using discrete summary statistics. Additionally for anterior segment 
biomicroscopy, discrete summaries will be provided by region, finding, visit, time point, and 
eye (study eye and fellow eye) for the number of subjects with at least one severity grade 
increase from baseline and for the number of subjects judged to be clinically significant by 
the Investigator. Shift tables from baseline may also be summarized. 
Vital signs will be summarized at each measured visit and for change from baseline to each 
measured visit using continuous summary statistics by treatment group and visit. 
 
 


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10. ADMINISTRATIVE CONSIDERATIONS 
10.1 
Investigators and Study Administrative Structure 
The Principal Investigator is responsible for all site medical-related decisions. The qualified 
Sponsor Medical Monitor is responsible for the safe conduct of this study. The contact 
information of the Sponsor Medical Monitor is as follows: 
 
Richard A. Lewis, MD  
Phone: (916) 768-7703  
Email: rlewis@aeriepharma.com 
10.2 
Institutional Review Board (IRB) or Independent Ethics Committee (IEC) 
Approval 
This protocol, materials used to recruit subjects, and materials used to document consent 
must be approved by the IRB/IEC prior to initiation of the study. The name and address of 
each reviewing IRB/ IEC will be documented in the Trial Master File. Written IRB/IEC 
approval must adequately identify the protocol and informed consent. In addition to 
approving the protocol, the IRB/IEC must also approve the Subject Information and Consent 
Form, as well as any advertising tools that will be used for the study. Written approval also 
must indicate whether approval was granted based on full committee review or expedited 
review. Copies of all approved materials, all correspondence with the IRB/IEC and written 
approval from the IRB/IEC must be made available to the Sponsor, prior to the start of 
subject enrollment into the study. The Investigator will report promptly to the IRB/IEC any 
new information that may adversely affect the safety of the subjects or the conduct of the 
study. The Investigator will submit written summaries of the study to the IRB/IEC as 
required. On completion of the study the IRB/IEC will be notified that the study has ended. 
10.3 
Ethical Conduct of the Study 
The study will be conducted according to this clinical protocol and will be governed by the 
following directives and guidelines: 
• US CFR, Title 21 
• ICH – Consolidated Good Clinical Practices Guideline (E6) 
• Standard Operating Procedures (SOPs) of the Sponsor and any vendors participating in 
the conduct of the study 
• The ethical principles that have their origin in the Declaration of Helsinki 


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10.4 
Subject Information and Consent 
Written informed consent will be obtained from each subject before any subject specific 
procedures are initiated. A copy of the signed and dated consent document will be given to 
each subject. The original signed and dated informed consent document must be maintained 
in the study files at the Investigator’s site. 
The Investigator is responsible for ensuring that no subject is subject to any study-related 
examination or activity before that subject has given informed consent. The subject must give 
written consent after the receipt of detailed information. The verbal explanation will cover all 
the elements specified in the written information provided for the subject. 
It should be emphasized that the subject is at liberty to withdraw consent to participate at any 
time, without penalty or loss of benefits to which the subject is otherwise entitled. 
Subjects who refuse to give, or withdraw, written informed consent may not be included or 
continued in this study, but this will not impact on their subsequent care. 
The Investigator will inform the subject of the aims, methods, anticipated benefits and 
potential hazards of the study, including any discomfort it may entail. The subject must be 
given every opportunity to clarify any points he/she does not understand and if necessary, ask 
for more information. At the end of the interview, the subject may be given time to reflect if 
this is required, or if the subject requests more time. Subjects and/or legal guardian will be 
required to sign and date the informed consent form. 
Signed informed consent must be attained prior to the conductance of any study procedures. 
10.5 
Subject Confidentiality 
The Investigator and his/her staff will maintain all personal subject data collected and 
processed for the purposes of this study using adequate precautions to ensure confidentiality, 
in accordance with local, state and federal laws and regulations. 
Monitors, auditors and other authorized representatives of the Sponsor, the IRB/IEC 
approving this study, and government regulatory authorities (e.g., FDA) may be granted 
direct access to the study subject’s original medical and study records for verification of the 
data or clinical study procedures. Access to this information will be permitted to 
representatives of the aforementioned organizations to the extent permitted by law. 
A report of this study’s results may be published or sent to the appropriate health authorities 
in any country in which the study medication may ultimately be marketed, but subject 
identities will not be disclosed in these documents. 


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10.6 
Study Monitoring 
Clinical research associates hired or contracted by the Sponsor will be responsible for 
monitoring the study sites and study activities. Clinical research associates will contact and 
visit the Investigator regularly. The actual frequency of monitoring visits depends on subject 
enrollment and on study site performance. Among others, the following items will be 
reviewed: 
• Study progress 
• Compliance with the protocol 
• Completion of eCRFs 
• Dispensing, storage, and accountability of study medication 
• Source data verification 
• AE and SAE reporting 
• Essential documents contained within the Investigator’s site file 
For source data verification (i.e., comparison of eCRF entries with subject records), data will 
be 100% source verified and will include at a minimum: 
• Subject identification 
• Informed consent (procedure, signature, and date) 
• Selection criteria 
• Primary efficacy and safety parameters (i.e., AEs) 
Member(s) of the Sponsor or their designee, in the role of Clinical Research Associate, will 
meet with the Investigator prior to the initiation of the study in order to assess the adequacy 
of the Investigator's subject population, facilities, and equipment, and to familiarize the 
Investigator with the protocol. 
The Clinical Research Associate will subsequently meet with the Investigator after several of 
the subjects have initiated the study in order to ensure that the subjects are being properly 
selected, that adequate supplies for the study have been provided and that the assignment of 
medication is properly recorded. In addition, the Clinical Research Associate will verify that 
the Investigator follows the approved protocol and all approved amendments, if any, by 
reviewing the Investigator’s regulatory documents, source documents, ICFs, and eCRFs of 
study subjects. 


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The Clinical Research Associate will meet with the Investigator when all subjects have 
completed the Final Visit of the study, in order to collect unused study medications and 
unused supplies and materials. 
Interim monitoring visits and telephone consultations will be done by the Clinical Research 
Associate as necessary, to ensure the proper progression and documentation of the study. 
10.7 
IRT 
Interactive response technology (IRT) activities will be performed as described in the IRT 
User Manual. 
10.8 
Case Report Forms and Study Records 
The initial point of entry of study data should be the subject source documentation. 
The location and nature of the source documentation for all data collected in the study will be 
identified in the study files at the Investigator’s site. In cases where no source documents will 
be used (i.e., data will be recorded directly into the eCRF without first being recorded on 
another document, such as a flowsheet, laboratory report, or other typical form of data 
reporting for later transcription to the eCRF), the original data will be included in the eCRF. 
Source document information should be legible. Recorded data should only be corrected by 
drawing a single line through the incorrect entry and writing the revision next to the 
corrected data. The person who has made the correction should place his or her initials as 
well as the date of the correction next to the correction. Data may not be obliterated by 
erasure, redaction, or with correction fluid. 
Study data will be transcribed and recorded via an electronic data capture (EDC) system as 
eCRFs. Security and authorization procedures consistent with the EDC system must be used. 
At each subject visit, the appropriate eCRFs must be completed. Whenever an eCRF is used, 
be sure to provide all information requested including subject identification number and 
initials, name or number of Investigator, date(s), etc. All applicable questions should be 
answered, and all data requested should be provided. Those areas that require a response but 
are not filled in correctly are considered incomplete or erroneous entries and will have to be 
corrected. 
Each authorized study staff member will receive a unique access account in order to use the 
EDC system. Access accounts will not be shared among study staff. Authorized users will 
make entries and/or changes to eCRFs via a secure internet access. Each completed set of 
eCRFs will be reviewed by the Investigator who will then electronically sign and date the 
eCRF confirming that data for the subjects are complete and accurate. 
The study records must include a copy of each Investigator's CV and medical license, and 
statement of Investigator qualifications. The name of each Sub-Investigator working under 
the supervision of the Investigator is also required to be filed in the study records. 
In addition, each eCRF, subject charts/source documents, IB, protocol, protocol amendments, 


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correspondence with the Sponsor/designee and the IRB/IEC, study medication storage, 
receipts, returns and dispensing records, Delegation of Responsibilities Log, site training 
records, records of site monitoring, any unmasking documentation, AE and SAE reporting, 
IRB/IEC approvals, advertisements, written information provided to subjects, and subject 
completed ICFs will be included in the study records. 
If the Investigator moves, withdraws from an investigation, or retires, the responsibility for 
maintaining the records may be transferred to another person (e.g., Sponsor, other 
Investigator) who will accept the responsibility. Notice of this transfer, including written 
acceptance, must be made to and agreed upon by the Sponsor. 
10.9 
Protocol Deviations 
Per ICH E6 (GCP) R2 Section 4.5.1 the Investigator/institution should conduct the trial in 
compliance with the protocol agreed with the sponsor and, if required, by the Regulatory 
Authority and which was given approval/favorable opinion by IRB/IEC. 
Protocol waivers or deviations from the protocol inclusion and exclusion criteria are not 
allowed because they can potentially jeopardize the scientific integrity of the study regulatory 
acceptability or subject safety. Therefore, adherence to the criteria as specified in the 
protocol is essential. 
The site will contact the Sponsor for clarification of inclusion and/or exclusion criteria as 
needed prior to enrollment of the study subject. The Sponsor will document clarification 
requests and responses or their representative. If a subject does not meet all the inclusion and 
exclusion criteria during screening, that subject may not be enrolled into the study. 
If a protocol deviation is identified by the Investigator or through site monitoring activities an 
immediate submission to the IRB/IEC may be required e.g., 24 or 48 hours (as per IRB 
guidelines). The Sponsor will assess any protocol deviation and decide whether any of these 
non-compliances should be reported to the relevant competent authority as a serious breach 
of GCP and the protocol. If per the relevant competent authorities’ requirements, the protocol 
deviation is not required to be reported immediately but is still required to be notified to the 
IRB/IEC, the specific protocol deviation will be added to the annual progress report. 
The Sponsor will review, designate, and/or approve all protocol deviations prior to the 
database lock. 
10.10 Access to Source Documentation 
The Investigator will permit study-related monitoring visits, audits, IRB/IEC review, 
and regulatory inspection(s) by providing direct access to source data and documents. 
The monitor and the head of the medical institution (where applicable) agrees to allow the 
monitor direct access to all relevant documents and to allocate their time and the time of their 
staff to monitor to discuss findings and any issues. 


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Sponsor/designee will monitor the study to ensure: 
• Data are authentic, accurate and complete. 
• Safety and rights of the subjects are being protected. 
• Study is conducted in accordance with the currently approved protocol and any other 
study agreements, ICH GCP and all applicable regulatory requirements. 
10.11 Data Generation and Analysis 
After data have been entered into the study database, a system of computerized data 
validation checks will be implemented and applied to the database. Query reports pertaining 
to data omissions and discrepancies will be forwarded to the clinical Investigator and the 
Sponsor for resolution. The study database will be updated by the clinical Investigator or 
their staff, in accordance with the resolved query reports. All changes to the study database 
will be documented. 
10.12 Retention of Data 
The Investigator’s site will retain all records related to the study in compliance with ICH 
Good Clinical Practices Guidelines E6 (R2) Section 4.9.4. 
Archived versions of the database will be saved by the Sponsor consistent with ICH Good 
Clinical Practices Guidelines E6 (R2) Section 5.5.11, complying with whichever of the 
requirements is longer. The Sponsor will notify the Investigator when study records should 
be destroyed. 
10.13 Financial Disclosure 
The Principal Investigator and Sub-Investigators (as listed on Form FDA 1572) will provide 
financial disclosure information prior to participation in the study. The Principal Investigator 
and any Sub-Investigators will notify the Sponsor promptly of any required revision to their 
financial disclosure status during the term of this study, annually, or at the end of the study 
(if applicable) and 1-year post-study completion. The Principal Investigator and Sub-
Investigators will provide updated financial disclosure information upon the Sponsor’s 
written request following completion of the study. 
10.14 Publication and Disclosure Policy 
Aerie Pharmaceuticals, as the Sponsor, has proprietary interest in the study. Authorship and 
manuscript composition will reflect joint cooperation between multiple Investigators and 
sites and Aerie Pharmaceuticals personnel. For studies with multiple centers, no individual 
publications will be allowed prior to completion of the final report of the multicenter study 
except as agreed with Aerie Pharmaceuticals. 
 
 


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11. AMENDMENT SUMMARY OF CHANGES 
11.1 
Amendment 1: 03 February 2021 
Section(s) 
Original 
Amendment 1 
Rationale 
4.3 Exclusion 
Criteria 
2. Concurrent or anticipated need 
for treatment for FCD, e.g., Muro 
128 (2% or 5%) during the course 
of the study 
2. Concurrent or anticipated need for 
treatment for FCD, e.g., Muro 128 
(2% or 5%) during the course of the 
study in the study eligible eye(s) 
Reduce 
medication 
restrictions for 
non-study eye 
4.3 Exclusion 
Criteria 
11. Concurrent or anticipated use 
of topical corticosteroids applied 
to the ocular surface or in the 
vicinity of the eyes (including 
intranasal corticosteroids and 
topical corticosteroids applied to 
the face) 
11. Concurrent or anticipated use of 
topical corticosteroids in the study 
eligible eye(s) 
Reduce 
medication 
restrictions for 
non-study eye 
4.3 Exclusion 
Criteria 
12. History of prior treatment with 
netarsudil (monotherapy or in 
combination) 
12. History of prior treatment with 
netarsudil (monotherapy or in 
combination) or any other Rho Kinase 
inhibitor 
Clarify 
exclusion 
criteria 
4.3 Exclusion 
Criteria and 
5.6.1 Prohibited 
Interventions 
16. Concurrent or anticipated use 
of corticosteroids during the study 
(with the exception of 
intraarticular corticosteroids and 
topical corticosteroids applied 
anywhere other than the eyes or 
face – see Exclusion #10). 
Individuals currently using 
corticosteroids must have been on 
a stable dose for at least 30 days 
prior to the Screening Visit, and 
the dose must not be anticipated to 
change during the course of the 
study 
16. Concurrent or anticipated use of 
systemic corticosteroids during the 
study. Individuals currently using 
topical or other locally acting 
corticosteroids (with the exception 
of use in the study eligible eye[s]) 
must have been on a stable dose for at 
least 30 days prior to the Screening 
Visit, and the dose must not be 
anticipated to change during the 
course of the study 
 
Reduce 
medication 
restrictions 
6.3 
Concomitant 
Medication 
Assessments 
However, medications used as part 
of the intravitreal injection process 
other than topical or 
subconjunctival anesthetic and 
povidone iodine swabs (e.g., pre-
or post-injection antibiotics or 
IOP-lowering medications) should 
be recorded on the eCRF. 
[removed] 
This statement 
is not necessary 
for this study 
6.7.4 V-FUCHS 
Questionnaire 
N/A 
Subjects who are not fluent in the 
English language may be exempt from 
completing this questionnaire. 
Clarification - 
There are 
currently no 
validated 
translations of 
the V-FUCHS 
questionnaire 


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6.9.1 
Performing 
Adverse Event 
(AE) 
Assessments 
Investigators are asked to use the 
verbatim term “conjunctival 
hyperemia” on the study AE form 
to describe observations of 
conjunctival redness if the ocular 
redness observation is increased 
from baseline observations and 
clinically meaningful. 
Investigators are also asked to note 
all observations of new or 
worsening conjunctival hyperemia 
on the biomicroscopy eCRF as 
well as on the study AE form. 
Investigators are also asked to note all 
observations of new or worsening 
conjunctival hyperemia on the 
biomicroscopy eCRF as well as on the 
study AE form if deemed necessary 
(per Investigator discretion). 
Clarification 
6.9.6.1 
Reporting 
SAEs, SSARs 
or SUSARs 
N/A 
The contact information of the study 
Safety Mailbox is as follows: 
Covance PSS 
SAEintake@covance.com 
Fax: 1-888-887-8097 
Adding to / 
updating safety 
reporting 
information 
6.9.7 Pregnancy 
Reporting 
While pregnancy itself is not 
considered to be an AE or SAE, 
pregnancy reports are tracked by 
the study Medical Monitor.  
The Investigator must complete 
the pregnancy report form and 
email the form to the study 
Medical Monitor within 1 business 
day of knowledge of the 
pregnancy. Following delivery or 
termination of pregnancy, the 
pregnancy report form is to be 
completed and submitted by fax or 
email to the Medical Monitor for 
the study. 
While pregnancy itself is not 
considered to be an AE or SAE, 
pregnancy reports are tracked by 
Covance and the study Medical 
Monitor… 
The Investigator must complete the 
pregnancy report form and email the 
form to the study Safety Mailbox 
within 1 business day of knowledge of 
the pregnancy. Following delivery or 
termination of pregnancy, the 
updated or follow-up pregnancy 
report form is to be completed and 
submitted by fax or email to the 
Safety Mailbox for the study. 
Adding to / 
updating safety 
reporting 
information 
9.1.1 Unit of 
Analysis 
Qualified fellow eyes will also be 
summarized. 
Qualified fellow eyes may also be 
summarized. 
Updating plan 
for potential 
posthoc 
analysis 
 
12. REFERENCES 
Fernandez MM. Reticular epithelial edema in edematous corneas treated with netarsudil. 
Ophthalmology. 2018;125;1709. 
Gain P, Jullienne R, He Z, Aldossary M, Acquart S, Cognasse F, Thuret G. Global Survey of 
Corneal Transplantation and Eye Banking. JAMA Ophthalmol. 2016 Feb;134(2):167-73. 
Hirabayashi KE, et al. Descemet stripping only for a chronic Descemet detachment after 
cataract surgery. Cornea. 2020;39:379–381. 


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Koizumi N, Okumura N, Ueno M, Nakagawa H, Hamuro J, Kinoshita S. Rho-associated 
kinase inhibitor eye drop treatment as a possible medical treatment for Fuchs corneal 
dystrophy. Cornea 2013; 32(8): 1167-70.  
Koizumi N, Okumura N, Ueno M, Kinoshita S. New therapeutic modality for corneal 
endothelial disease using Rho-associated kinase inhibitor eye drops. Cornea. 2014;33 Suppl 
11:S25‐S31.  
Liu KC, Gupta D. Netarsudil-associated reticular corneal epithelial edema with raised 
intraocular pressure. Ophthalmol Glaucoma. 2019;2(3):166. 
McLaren JW, Bachman LA, Kane KM, Patel SV. Objective assessment of the corneal 
endothelium in Fuchs' endothelial dystrophy. Invest Ophthalmol Vis Sci. 2014;55(2):1184‐
1190. Published 2014 Feb 26.  
Moshirfar M, Somani AN, Vaidyanathan U, et al. Fuchs Endothelial Dystrophy (FED) 
[Updated 2020 May 12]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls 
Publishing; 2020 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK545248/ 
Moumneh K, Sheybani A, Fellman RL, Godfrey DG, Grover DS. Reticular Corneal Edema 
or Corneal Honeycombing in Eyes Treated With Netarsudil: A Case Series. J Glaucoma. 
2020;29(7):607-610. [ePub ahead of print] 
Okumura N, Ueno M, Koizumi N, et al. Enhancement on primate corneal endothelial cell 
survival in vitro by a ROCK inhibitor. Invest Ophthalmol Vis Sci 2009; 50(8): 3680-7.  
Okumura N, Koizumi N, Kay EP, et al. The ROCK inhibitor eye drop accelerates corneal 
endothelium wound healing. Invest Ophthalmol Vis Sci 2013; 54(4): 2493-502.  
Okumura N, Kinoshita S, Koizumi N. Application of Rho Kinase Inhibitors for the 
Treatment of Corneal Endothelial Diseases. J Ophthalmol. 2017;2017:2646904. 
Okumura N, Hayashi R, Koizumi N. Perspective of Future Potent Therapies for Fuchs 
Endothelial Corneal Dystrophy. Open Ophthalmol J. 2018;12:154-163. 
Ploysangam P, Patel SP. A Case Report Illustrating the Postoperative Course of 
Descemetorhexis without Endothelial Keratoplasty with Topical Netarsudil Therapy. Case 
Rep Ophthalmol Med. 2019;2019:6139026. 
Wacker K, Baratz KH, Bourne WM, Patel SV. Patient-Reported Visual Disability in Fuchs' 
Endothelial Corneal Dystrophy Measured by the Visual Function and Corneal Health Status 
Instrument. Ophthalmology. 2018;125(12):1854-1861.  
Wisely CE, Liu KC, Gupta D, Carlson AN, Asrani SG, Kim T. Reticular Bullous Epithelial 
Edema in Corneas Treated with Netarsudil: A Case Series [published online ahead of print, 
2020 Apr 11]. Am J Ophthalmol. 2020;S0002-9394(20)30169-0. [ePub ahead of print] 

SAP_001.pdf: 
Statistical Analysis Plan 
 
 
A Randomized, Open-Label, Parallel-Group Study to Evaluate 
the Safety and Efficacy of Two Dosing Regimens of Netarsudil 
Ophthalmic Solution in Patients with Corneal Edema Due to 
Fuchs Corneal Dystrophy 
 
 
NCT 04498169 
 
 
SAP v1.0 02 September 2021 


 
STATISTICAL ANALYSIS PLAN  
 
A Randomized, Open-Label, Parallel-Group Study to Evaluate the Safety and 
Efficacy of Two Dosing Regimens of Netarsudil Ophthalmic Solution in Patients 
with Corneal Edema Due to Fuchs Corneal Dystrophy 
 
 
Sponsor:   
 
Aerie Pharmaceuticals, Inc. 
 
 
 
 
 
 
Protocol Number: 
AR-13324-CS210 
 
 
Author:   
 
Yan Liu  
 
 
 
Senior Statistician 
Statistics & Data Corporation 
 
Date:    
 
02 September 2021 
Version:  
 
1.0 
 


 
Page 2 of 34 
 
A Randomized, Open-Label, Parallel-Group Study to Evaluate the Safety and Efficacy of Two 
Dosing Regimens of Netarsudil Ophthalmic Solution in Patients with Corneal Edema Due to Fuchs 
Corneal Dystrophy 
 
Protocol Number: 
AR-13324-CS210 
SAP Version:   
1.0 
SAP Date:   
 
02 September 2021 
 
Statistical Analysis Plan Approval 
 
 
Prepared by: 
 
 
 
 
 
 
 
 
 
 
 
 
Yan Liu, MS 
 
 
 
 
 
 
Date 
 
 
Senior Biostatistician 
 
 
Statistics & Data Corporation 
 
 
Reviewed by:  
 
 
 
 
 
 
 
_____________ 
Kalyani Kothapalli, MS
Date
 
 
Senior Director, Biostatistics 
 
 
Statistics & Data Corporation 
 
 
Approved by:  
 
 
 
 
 
 
 
 
 
 
 
Amber Lewis, PharmD, PSM 
 
 
 
 
Date 
Manager, Clinical Development 
 
 
Aerie Pharmaceuticals, Inc.  
 
 
Approved by:  
 
 
 
 
 
 
 
 
 
 
 
Michelle Senchyna, PhD 
 
 
 
 
Date 
Vice President, Clinical Development and Medical Affairs 
 
 
Aerie Pharmaceuticals, Inc.  
 
s, PharmD, PSM
nical Development
	
	
hapalli, MS
	

chyna, PhD
S



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Table of Contents 
1. 
Introduction............................................................................................................................... 7 
2. 
Study Objectives ...................................................................................................................... 7 
2.1 Efficacy ..................................................................................................................................... 7 
2.1.1 
Primary Endpoint .............................................................................................................. 7 
2.1.2 
Secondary Endpoints ....................................................................................................... 7 
2.1.3 
Exploratory Endpoints ...................................................................................................... 8 
2.2 Safety ....................................................................................................................................... 8 
2.3 Statistical Hypotheses .............................................................................................................. 8 
3. 
Study Design and Procedures ................................................................................................. 8 
3.1 General Study Design and Plan ............................................................................................... 8 
3.2 Schedule of Visits and Assessments ..................................................................................... 10 
4. 
Study Treatments ................................................................................................................... 11 
4.1 Treatment Administered ......................................................................................................... 11 
4.2 Method of Assigning Patients to Treatment Groups .............................................................. 11 
4.3 Masking and Unmasking ........................................................................................................ 11 
5. 
Sample Size and Power Considerations ................................................................................ 11 
6. 
Data Preparation .................................................................................................................... 12 
6.1 Input Data ............................................................................................................................... 12 
6.2 Output Data ............................................................................................................................ 12 
7. 
Analysis Populations .............................................................................................................. 12 
7.1 Modified Intent-to-Treat (mITT) Population ............................................................................ 13 
7.2 Per Protocol (PP) Population ................................................................................................. 13 
7.3 Safety Population ................................................................................................................... 13 
8. 
General Statistical Considerations ......................................................................................... 13 
8.1 Unit of Analysis....................................................................................................................... 13 
8.2 Missing or Inconclusive Data Handling .................................................................................. 13 
8.2.1 
Assessment Windows .................................................................................................... 13 
8.2.2 
Missing Data Handling for Efficacy Variables ................................................................ 13 
8.2.3 
Missing data handling for partial missing dates ............................................................. 14 
8.3 Definition of Baseline ............................................................................................................. 15 
8.4 Data Analysis Conventions .................................................................................................... 15 
8.5 Adjustments for Multiplicity ..................................................................................................... 16 
9. 
Disposition of Subjects ........................................................................................................... 16 
10. 
Demographic and Pretreatment Variables ............................................................................. 16 
10.1 Demographic Variables .......................................................................................................... 16 
11. 
Medical History and Concomitant Medications ...................................................................... 17 


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11.1 Medical History ....................................................................................................................... 17 
11.2 Concomitant Medications ....................................................................................................... 17 
12. 
Efficacy Analyses ................................................................................................................... 18 
12.1 Efficacy Assessments ............................................................................................................ 18 
12.1.1 
Central Corneal Thickness by Ultrasound Pachymetry ............................................. 18 
12.1.2 
Best Corrected Visual Acuity by ETDRS Method ...................................................... 18 
12.1.3 
Biomicroscopy ............................................................................................................ 18 
12.1.4 
Tomography (Pentacam®; Selected Sites Only) ........................................................ 18 
12.1.5 
V-FUCHS Questionnaire ............................................................................................ 19 
12.2 Primary Efficacy Analysis ....................................................................................................... 19 
12.3 Secondary Efficacy Analyses ................................................................................................. 25 
12.4 Exploratory Efficacy Analyses ................................................................................................ 26 
12.5 Ad-hoc Efficacy Analyses ....................................................................................................... 27 
13. 
Safety Analyses ..................................................................................................................... 27 
13.1 Adverse Events ...................................................................................................................... 27 
13.2 Best Corrected Visual Acuity .................................................................................................. 29 
13.3 Biomicroscopy ........................................................................................................................ 29 
13.4 Intraocular Pressure ............................................................................................................... 30 
13.5 Ophthalmoscopy .................................................................................................................... 30 
13.6 Vital Signs .............................................................................................................................. 30 
14. 
Interim Analyses ..................................................................................................................... 30 
15. 
Changes from Protocol-Stated Analyses ............................................................................... 30 
16. 
References ............................................................................................................................. 31 
17. 
Revision History ..................................................................................................................... 31 
18. 
Tables ..................................................................................................................................... 31 
19. 
Listings ................................................................................................................................... 33 
20. 
Figures ................................................................................................................................... 34 
 
 
 


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List of Abbreviations 
AD 
Analysis Dataset 
AE 
Adverse Event 
ANCOVA 
Analysis of Covariance 
ATC 
Anatomical Therapeutic Chemical 
BCVA 
Best Corrected Visual Acuity 
BID 
Twice Daily 
CCT 
Central Corneal Thickness 
CI 
Confidence Interval 
CRF 
Case Report Form 
CS 
Clinically Significant 
eCRF 
Electronic Case Report Form 
EDC 
Electronic Data Capture 
ETDRS 
Early Treatment of Diabetic Retinopathy Study 
FCD 
Fuchs Corneal Dystrophy 
FCS 
Fully Conditional Specification 
ICH 
International Conference on Harmonisation 
ICT 
Inferior Corneal Thickness 
IOP 
Intraocular Pressure 
IRT 
Interactive Response Technology 
LOCF 
Last Observation Carried Forward 
logMAR 
Logarithm of the Minimum Angle of Resolution 
LS 
Least Squares 
mITT 
Modified Intent-to-Treat 
MedDRA 
Medical Dictionary for Regulatory Activities 
MI 
Multiple Imputation 
NCS 
Not Clinically Significant 
NCT 
Nasal Corneal Thickness 
ODO 
Observed Data Only 
PDF 
Portable Document Format 
PP 
Per Protocol 
PT 
Preferred Term 
QD 
Once Daily 
RTF 
Rich Text Format 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SCT 
Superior Corneal Thickness 
SD 
Standard Deviation 
SDC 
Statistics & Data Corporation 
SE 
Standard Error 
SOC 
System Organ Class 
TCT 
Temporal Corneal Thickness 
TEAE 
Treatment-Emergent Adverse Event 
V-FUCHS 
Visual Function and Corneal Health Status 


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WHODrug  
World Health Organization Drug Dictionary 
 
 
 


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1. Introduction 
The purpose of this statistical analysis plan (SAP) is to describe the planned analyses and reporting for 
protocol AR-13324-CS210 (Amendment 1) dated 03FEB2021. 
This SAP is being written with due consideration of the recommendations outlined in the most recent 
International Conference on Harmonisation (ICH) E9 Guideline entitled Guidance for Industry: Statistical 
Principles for Clinical Trials and the most recent ICH E3 Guideline, entitled Guidance for Industry: Structure 
and Content of Clinical Study Reports. 
This SAP describes the data that will be analyzed and the subject characteristics, efficacy, and safety 
assessments that will be evaluated. This SAP provides details of the specific statistical methods that will 
be used. The statistical analysis methods presented in this document will supersede the statistical analysis 
methods described in the clinical protocol. If additional analyses are required to supplement the planned 
analyses described in this SAP, they may be completed and will be identified in the clinical study report. 
2. Study Objectives 
The primary objective of the study is:  
• 
To evaluate the reduction in central corneal thickness (CCT) in eyes dosed with either Netarsudil 
Ophthalmic Solution 0.02% (netarsudil) once-daily (QD) or netarsudil twice-daily (BID) for the 
treatment of corneal edema due to Fuchs corneal dystrophy (FCD). 
The secondary objectives of the study are:  
1. To evaluate the proportion of subjects demonstrating improvement in vision following treatment for 
FCD with netarsudil QD or netarsudil BID; and 
2. To evaluate the safety of netarsudil QD and netarsudil BID in subjects with FCD. 
2.1 
Efficacy  
Efficacy endpoints are described below. The primary comparisons in this trial will be within a treatment 
group on the mean change from baseline. The primary comparisons for proportion of subjects achieving a 
specified endpoint will be between treatment groups.  
2.1.1 
PRIMARY ENDPOINT 
• 
Mean change from baseline in CCT as assessed by ultrasound pachymetry at Week 4 
2.1.2 
SECONDARY ENDPOINTS 
• 
Mean change from baseline in CCT as assessed by ultrasound pachymetry at Week 8 
• 
Mean change from baseline in best corrected visual acuity (BCVA) at Weeks 4, and 8 
• 
Proportion of subjects who gained ≥15 letters (3 lines) in BCVA at Weeks 4, and 8 
• 
Proportion of subjects with complete resolution of corneal edema at Weeks 4, and 8 


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2.1.3 
EXPLORATORY ENDPOINTS 
• 
Proportion of subjects who gained ≥15 letters (3 lines) in BCVA  at Week 2 
• 
Proportion of subjects who gained ≥10 letters (2 lines) in BCVA at Weeks 2, 4, and 8 
• 
Proportion of subjects who gained ≥5 letters (1 line) in BCVA at Weeks 2, 4, and 8 
• 
Mean change from baseline in CCT as assessed by Pentacam® at Week 4 (selected sites 
only) 
• 
Mean change from baseline in visual function and corneal health status (V-FUCHS) glare and 
diurnal variation (Glare Factor) score at Week 4 
• 
Proportion of subjects with complete resolution of epithelial edema at Weeks 4, and 8 
2.2 
Safety 
The safety of netarsudil QD and BID will be evaluated by: 
• 
Adverse events (AEs) 
• 
BCVA 
• 
Anterior segment biomicroscopy 
• 
Intraocular pressure (IOP)  
• 
Ophthalmoscopy 
• 
Vital signs (heart rate and blood pressure) 
2.3 
Statistical Hypotheses 
The null and alternative hypotheses, based on the primary variables, are as follows: 
H01: The mean change from baseline in CCT by ultrasound pachymetry in study eyes 
treated with netarsudil QD at Week 4 = 0. 
H11: The mean change from baseline in CCT by ultrasound pachymetry in study eyes 
treated with netarsudil QD at Week 4 ≠ 0. 
H02: The mean change from baseline in CCT by ultrasound pachymetry in study eyes 
treated with netarsudil BID at Week 4 = 0. 
H12: The mean change from baseline in CCT by ultrasound pachymetry in study eyes 
treated with netarsudil BID at Week 4 ≠ 0. 
3. Study Design and Procedures 
3.1 
General Study Design and Plan 
This is an 8-week, randomized, open-label, multicenter, efficacy and safety study in subjects with FCD. 
The study will be conducted at approximately 20 sites in the USA. Two different dosing regimens (QD and 
BID) of netarsudil will be investigated for their efficacy in reducing or resolving corneal edema in subjects 
with FCD. Subjects randomized to the netarsudil QD dosing regimen will be instructed to instill one drop 
of the Study Artificial Tear in their study eye every morning, and one drop of netarsudil in their study eye 


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every evening. Subjects randomized to the netarsudil BID dosing regimen will instill one drop of netarsudil 
in their study eye every morning and evening. 
CCT will be measured by ultrasound pachymetry at approximately the same time of day ± 60 minutes and 
always before noon (local time), at each of the 5 scheduled visits (Screening, Baseline [Day 1], Week 2, 
Week 4, and Week 8/Exit). In order to be eligible to participate in the study, subjects will have been 
diagnosed with visually significant central corneal edema due to FCD, in at least one eye, for no longer 
than 12 months’ duration. 
At the Screening visit, the subject’s diagnosis of FCD will be confirmed and CCT measured using an 
ultrasonic pachymeter. IOP and BCVA will also be collected. 
At the Baseline visit (Day 1), the eligibility assessments conducted during Screening will be repeated to 
ensure that the subject meets all criteria for enrollment in the study. The same eye must qualify at both 
the Screening and Baseline visits. If qualified, the subject will be provided with the V-FUCHS patient 
reported visual disability questionnaire (Wacker 2018). The subject will be randomized (1:1) into the study 
and provided with study medication. The subject will also be trained on appropriate techniques for 
applying eye drops.  
At the Week 2 and Week 4 visits, the subject will return to the study site for efficacy and safety 
assessments. 
At the Week 4 visit, any subject randomized to netarsudil QD who, in the opinion of the Investigator, has 
not responded adequately to treatment, may have their netarsudil dosing frequency increased to BID for 
the remainder of the study. For these subjects, the morning administration of Study Artificial Tear will be 
terminated. 
At the Week 8 visit, a final assessment of efficacy and safety parameters will be conducted, and the 
subject will exit the study. 
Scheduled study visits (Screening, Baseline, Week 2, Week 4, and Week 8/Exit) will be referred to in all 
tables and listings to enable reviewers to understand the assessment timing without referring to the protocol 
visit schedule. Figure 1 shows the scheduled study visits and visit window (days) for each study visit. There 
is no Day 0, Day 1 is the day of study eye determination and randomization, on which the subjects get their 
first study medication dispensation. All other study days will be labeled relative to Day 1. For event dates 
on or after Day 1, study day will be calculated as [Date of Event] – [Date of First Dose] + 1. For event dates 
before Day 1, study day will be calculated as [Date of Event] – [Date of First Dose]. The first dose of study 
medication will not be captured on the electronic case report form (eCRF) and therefore will be assumed 
to be same as the first study medication dispensation date.  
 
 


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3.2 
Schedule of Visits and Assessments 
The schedule of visits and assessments along with study design schema is provided below. 
Figure 1. Schedule of Visits and Procedures 
 
Figure 2. Study Design Schema 
 
 


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4. Study Treatments 
4.1 
Treatment Administered 
Subjects will be randomized using a 1:1 allocation ratio to one of two treatment groups: 
• 
Netarsudil QD Dosing Regimen 
Study Artificial Tear: one drop in the study eye in the morning 
Netarsudil 0.02% ophthalmic solution: one drop in the study eye in the evening 
• 
Netarsudil BID Dosing Regimen 
Netarsudil 0.02% ophthalmic solution: one drop in the study eye in the morning and in the 
evening 
Doses will be self-administered by the study subjects. If the subject is using any another topical 
ophthalmic drop (e.g., ocular hypotensive medications) at the same time as netarsudil, the subject should 
be instructed to instill netarsudil first, and wait at least 15 minutes before instilling any additional drops. 
For subjects deemed unable to self-administer the doses, a caregiver will be asked to administer the 
medication. All subjects will administer study medication for approximately 8 weeks. 
4.2 
Method of Assigning Patients to Treatment Groups 
Subjects will be assigned to treatment groups through the use of an interactive response technology (IRT) 
system. Subjects will be identified by subject ID (xxx-xxx) with 3 digits for site ID followed by 3 digits for 
subject number in all datasets and listings for this study. 
4.3 
Masking and Unmasking 
This is an open-label study, therefore masking and unmasking procedures do not apply. 
5. Sample Size and Power Considerations 
This study is not powered to detect a pre-stated efficacy signal, but rather will be used to inform the 
design and power for any possible future studies. With a sample size of up-to 20 subjects per treatment 
group, the precision of the point estimate of mean change from baseline in CCT, as measured by the half-
width of the two-sided 90% confidence interval (CI), will be approximately 1/2.7 of the standard deviation 
(SD) of the change from baseline, yielding 90% confidence that the true mean change from baseline is 
within ± SD/2.7 µm of the observed mean change from baseline. That is, if the SD of the change from 
baseline is 60 µm, then the half-width of the two-sided 90% CI would be 60 µm x 1/2.7 = 22.2 µm, yielding 
90% confidence that the true mean change from baseline is within ± 22.2 µm of the observed mean 
change from baseline. 
With a sample size of up-to 20 subjects within a treatment group, the study will have 80% power to reject 
H01 or H02 and in favor of H11 or H12 respectively and demonstrate a statistically significant mean change 
from baseline, assuming the true effect size (mean change / SD) is 0.577 or larger (e.g. assuming the 
true mean change from baseline is 34.6 µm and the SD is 60 µm), a one sample t-test and a two-sided 
alpha = 0.10. 


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With a sample size of up-to 20 subjects per treatment group, the study will have 95% confidence of ruling 
out AEs with true incidence rates of 13.9% or higher within each treatment group. That is, with up-to 20 
subjects in a treatment group, if an AE of a specific type is not observed, then with 95% confidence, the 
true incidence rate of that adverse event is less than 13.9%. 
Similarly, with a sample size of up-to 40 subjects combined over treatment groups, the study will have 
95% confidence of ruling out AEs with true incidence rates of 7.2% or higher  within each treatment 
group. That is, with up-to 40 subjects in a treatment group, if an AE of a specific type is not observed, 
then with 95% confidence, the true incidence rate of that AE is less than 7.2%.  
6. Data Preparation 
6.1 
Input Data 
Study data will primarily be recorded on the electronic case report form (eCRF) supplied by Statistics & 
Data Corporation (SDC) using electronic data capture (EDC) system, iMednetTM. The only external data 
source is the site level protocol deviations that will be collected separately at site and will be transferred to 
SDC from Aerie. It will be combined with the protocol deviations in EDC by SDC for the final deviation 
classification by Aerie.  
When all prerequisites for database lock have been met, the database will be locked. Any changes to the 
database after data have been locked can only be made with the approval of the Sponsor in consultation 
with SDC.  
Final analysis will be carried out after the following have occurred: 
• 
Database lock has occurred, including receipt of all final versions of external vendor data, with 
written authorization provided by appropriate SDC and Sponsor personnel 
• 
Protocol deviations have been identified and status defined (major/minor deviations) 
• 
Analysis populations have been determined 
6.2 
Output Data 
Data from EDC will be transferred to SDC Biostatistics and will then be mapped to analysis datasets (ADs). 
Raw data will be used to create the subject listings along with ADs as needed, while all tables will be based 
on the ADs. 
7. Analysis Populations 
A total of approximately 40 subjects will be enrolled into the study at approximately 20 investigational 
sites within the United States. Subjects will be at least 18 years of age with a diagnosis of FCD, each of 
whom should meet all of the inclusion criteria and none of the exclusion criteria. 


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7.1 
Modified Intent-to-Treat (mITT) Population 
The mITT population will include all randomized subjects who have received at least one dose of study 
medication. This population will be the primary population for efficacy analyses and will be used to 
summarize a subset of efficacy variables and will summarize subjects as randomized. 
7.2 
Per Protocol (PP) Population 
The PP population is a subset of the mITT population, which will include those subjects (and their visits) 
who do not have major protocol deviations likely to seriously affect the primary outcome of the study. This 
population will be the secondary population for efficacy analyses and will be used to summarize a subset 
of efficacy variables. If the PP and mITT populations are exactly the same, then additional efficacy analyses 
on the PP population will not be performed. The PP population will summarize subjects as treated.  
7.3 
Safety Population 
The safety population will include all randomized subjects who have received at least one dose of 
investigational product. This population will be used to summarize safety variables and will summarize 
subjects as treated. 
8. General Statistical Considerations 
8.1 
Unit of Analysis 
Safety endpoints will be analyzed for both eyes. For efficacy endpoints, the primary unit of analysis will be 
the study eye defined based on CCT of at least 600 µm evaluated by ultrasound pachymetry. If both eyes 
qualify based on CCT and enrollment criteria, the study eye will be the eye with the thinner cornea at the 
baseline. If both eyes have the same CCT, the study eye will be the right eye. Qualified fellow eyes will also 
be summarized. A qualified fellow eye for a given subject is a fellow eye that also met all of the inclusion 
criteria and none of the exclusion criteria at the Screening and Baseline Visits.  
8.2 
Missing or Inconclusive Data Handling 
8.2.1 
ASSESSMENT WINDOWS 
Assessment window mapping will be applied to both efficacy and safety outcomes for table summaries 
using observed data only (ODO). Unscheduled or early termination visits that are within the protocol 
specified window of a scheduled visit will be mapped to that visit to replace the missing scheduled visit. 
Missing data handling for efficacy variables as described in Section 8.2.2 will be applied after assessment 
window mapping. Both the actual visit and windowed visit will be displayed in the listings. 
8.2.2 
MISSING DATA HANDLING FOR EFFICACY VARIABLES 
The primary analysis will be completed with available data per subject from the mITT population. 
Robustness analyses will also be presented based on the multiple imputation (MI) methodology under 
different assumptions of missingness and intercurrent events, using last observation carried forward 
(LOCF), and using the PP population with available data per subject. Imputation using MI or LOCF will 


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only be performed on study eye missing data at post-baseline scheduled visits. No imputation will be 
performed for missing fellow eye data and unscheduled post-baseline visits.  
Intercurrent events will be primary handled in the following manner: 
1. Discontinuation of study medication and non-optimal compliance will be ignored, values measured 
after discontinuation of study medication or regardless of compliance will be used in the analyses 
[treatment policy strategy]. 
2. Withdrawal due to lack of efficacy or AEs: missing data will be singly imputed as the worst within 
subject observation prior to the intercurrent event [hypothetical strategy]. For efficacy variable CCT, 
the worst is the maximum within subject observations. In addition, the worst observation should 
come from post-baseline scheduled and unscheduled visits prior to the intercurrent event.  
3. Missing data without withdrawal or withdrawal due to reasons other than lack of efficacy or AEs: 
missing data will be imputed employing MI fully conditional specification (FCS) regression method.  
For the LOCF imputation, only the observations from post-baseline visits including scheduled, 
unscheduled, and early termination visits for a given subject will be carried forward.  
8.2.3 
MISSING DATA HANDLING FOR PARTIAL MISSING DATES 
In general, there will be no imputation of missing dates other than for partial or missing dates where 
complete dates are required to flag data as treatment-emergent or concomitant with treatment. 
Partial/missing start and end dates for AEs and concomitant medications will be imputed as follows: 
Partial/missing start date:  
• 
Dates with missing day only will be imputed as the 1st of the month unless the month and year are 
same as the month and year of first dose of study medication, in which case missing day will be 
imputed as the first dose day of study medication.  
• 
Dates with both day and month missing will be imputed as 1 Jan unless the year is same as the 
year of first dose of study medication, in which case missing day and month will be imputed as the 
first dose day and month of study medication. 
• 
Completely missing dates will be imputed as the first dose date of study medication unless the end 
date is on or before the first dose date of study medication, in which case missing date will be 
imputed as 1 Jan of the same year as the end date. 
Partial/missing end date:  
• 
Dates with missing day only will be imputed as the last day of the month unless the month and year 
are the same as the month and year of the last dose of study medication, in which case missing 
day will be imputed as the last dose day of study medication.  


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• 
Dates with both day and month missing will be imputed as 31 Dec unless the year is same as the 
year of the last dose of study medication, in which case missing day and month will be imputed as 
the last dose day and month of study medication.  
• 
If the ongoing flag is missing or “Yes” then the date will not be imputed unless death date is 
available, in which case the missing date will be imputed as the death date. If ongoing is “No” then 
the missing end date will be imputed as the last dose date. 
• 
If the imputed date is after the date of death, then the end date will be set equal to the date of 
death.  
The original dates will be displayed in data listings and the imputed dates will be used in derivations only 
(study day, treatment-emergence status, prior or concomitant medication etc). 
8.3 
Definition of Baseline 
Baseline is defined as the last measurement (including measurement from unscheduled visits) prior to the 
first dose of study medication. The date of first dose of study medication is assumed to be same as the first 
study medication dispensation date. 
8.4 
Data Analysis Conventions 
All data analysis will be performed by SDC. Statistical programming and analyses will be performed using 
SAS® Version 9.4 or higher. Output will be provided in rich text format (RTF) for tables and portable 
document format (PDF) for tables, and listings using landscape orientation. Unless otherwise specified, all 
study data for randomized subjects will be listed by treatment group, subject number, visit, and parameter 
as applicable. Only a disposition listing will be created for screen failure subjects. Summaries will be 
presented by treatment group and visit as applicable.  
Continuous study assessments will be summarized by treatment group and visit as applicable using 
descriptive statistics (n, mean, median, SD, minimum, and maximum). Minima and maxima will be 
reported with the same precision as the raw values; means and medians will be presented to one 
additional decimal place than reported in the raw values. Standard deviations will be presented to two 
additional decimal places than reported in the raw values. Categorical study assessments will be 
summarized by treatment group and visit as applicable using discrete descriptive statistics (frequency 
counts and percentages). All percentages will be rounded to one decimal place (ie, XX.X%). Change from 
baseline will be calculated as follow-up visit – baseline visit. 
Hypothesis testing, unless otherwise indicated, will be performed at a 2-sided with a significant level of 
0.10 (α = 0.1). Where applicable, 2-sided 90% and 95% confidence intervals (CIs) will be reported. All p-
values will be displayed to four decimal places, with p-values less than 0.0001 presented as ”<0.0001” 
and p-values greater than 0.9999 presented as ”>0.9999”. All odds ratios less than 0.01 will be presented 
as “<0.01” and all odds ratio greater than 999.99 will be presented as “>999.99”. Difference between 
netarsudil QD and netarsudil BID will be calculated as netarsudil QD – netarsudil BID. 


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8.5 
Adjustments for Multiplicity 
Adjustments for multiplicity will not be made in this early phase trial. 
9. Disposition of Subjects 
Disposition will be presented in terms of numbers and percentage of subjects by treatment group and for 
all subjects. Percentages will be calculated using treated subjects as the denominator unless otherwise 
specified.  
The numbers and percentages of subjects who were screened, enrolled, screen failed, and treated will be 
presented. The reasons for screen failure will be displayed with the percentages calculated using total 
number of screen failures as the denominator. The screen failure reason will include eligibility criteria not 
met, lost to follow up, investigator decision, enrollment complete, withdrawal of consent, and other. The 
number of subjects with COVID-19 related screen failure will be identified. 
The number and percentages of subjects in each analysis population (mITT, PP, and Safety) will be 
presented. 
The number and percentage of subjects who completed the study or prematurely discontinued from the 
study will be presented. The reasons for study discontinuation will include AE, withdrawal of consent, non-
compliant, lost to follow up, lack of efficacy, disallowed concurrent medication, investigator decision, 
protocol deviation, and other. Percentages will be calculated using the number of discontinued subjects as 
the denominator. The number of subjects with COVID-19 related study discontinuation will be identified. 
The number and percentage of subjects with any deviation, major deviation, and minor deviation will be 
presented. The protocol deviations that will be reviewed include informed consent, inclusion/exclusion,  test 
article/study drug instillation and assignment at site, improper protocol procedures at site, site’s failure to 
report serious adverse event (SAE)/AE, visit out of window, subjects’ use of prohibited concomitant 
medication, subjects’ failure to follow instructions and other. The number of subjects with COVID-19 related 
protocol deviations will be identified.  
In addition, subject listings for randomization schedule, disposition (for all randomized subjects and for 
screen failed subjects), protocol deviations, COVID-19 related protocol deviations, and analysis populations 
will be provided. 
10. Demographic and Pretreatment Variables 
10.1 Demographic Variables 
The demographic variables including age, sex, race, and ethnicity will be summarized for mITT population.  
Age (years) will be summarized by treatment group and for all subjects, using continuous descriptive 
statistics. In addition, age will be categorized into four age groups, <18, 18–65, 66–75, >75, and 
summarized using counts and percentages.  


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Sex, race, and ethnicity will be summarized by treatment group and for all subjects using discrete 
descriptive statistics. Subjects who record more than one race will be grouped into a single category 
denoted as multi-racial. 
Subject listings will be provided for demographic variables, childbearing potential for female subjects, and 
pregnancy tests for female subjects with childbearing potential.  
11. Medical History and Concomitant Medications 
11.1 Medical History 
Medical and ophthalmic history will be obtained at the Screening and Baseline visit and coded using Medical 
Dictionary for Regulatory Activities (MedDRA) Version 23.0. 
Medical history will be summarized using discrete descriptive statistics and presented by treatment group 
and for all subjects at the subject level by System Organ Class (SOC) and Preferred Term (PT) using the 
mITT population. Ocular and non-ocular medical history will be summarized separately. If a subject reports 
the same PT multiple times within the same SOC, that PT will only be reported once within that SOC. If a 
subject reports multiple conditions within the same SOC, that SOC will only be reported once. In the 
summaries, SOCs are listed in alphabetical order; PTs within a SOC are listed in order of descending 
frequency across all subjects. 
Subject listings will be provided for ocular medical history, ocular surgery and laser procedure, and non-
ocular medical history. 
11.2 Concomitant Medications 
Prior and concomitant medications will be coded using World Health Organization Drug Dictionary 
(WHODrug) Global (B3, March 2020) and summarized to the therapeutic drug class (Anatomical 
Therapeutic Chemical [ATC] 4 classification) and preferred name. If the ATC 4 classification is not provided, 
the next highest classification that is provided in the coding dictionary will be used. The preferred name will 
be defined as the active ingredient; if the active ingredient is not provided or includes more than two 
ingredients (e.g., multivitamins), then the drug name will be summarized as the preferred name.  
Concomitant medications are defined as those medications listed as having been taken (1) prior to initiation 
of study medication administration and continuing for any period of time following the first administration of 
study medication or (2) at any time following the first administration of study medication. 
Prior and concomitant medications will be summarized using the mITT population. Medications will be 
tabulated for each treatment group using frequencies and percentages. Subjects may have more than 1 
medication per ATC text. At each level of subject summarization, a subject will be counted once if he/she 
reports 1 or more medications. Percentages will be based on the number of subjects in each treatment 
group. In the summaries, ATC classes are listed in alphabetical order and preferred names within an ATC 
class are listed in order of descending frequency across all subjects.  
A subject listing of prior and concomitant medications will be provided. 


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12. Efficacy Analyses 
Efficacy assessments include CCT by ultrasound pachymetry, BCVA using Early Treatment of Diabetic 
Retinopathy Study (ETDRS) method, tomography (Pentacam®) and V-FUCHS questionnaire that are 
described in Section 12.1. All collected efficacy  measurements will be presented in subject listings. Efficacy 
analyses for primary, secondary, and exploratory endpoints will be produced in summary tables as specified 
in Sections 12.2, 12.3, and 12.4.  
12.1 Efficacy Assessments 
12.1.1 CENTRAL CORNEAL THICKNESS BY ULTRASOUND PACHYMETRY 
CCT will be measured at all scheduled visits for both eyes at approximately the same time of day ± 60 
minutes, always before noon (local time) and always performed before IOP measurements. Every effort 
should be made to ensure that the same individual performs the measurement and uses the same device 
for a given subject at each visit. Five consecutive measurements of CCT will be measured for each 
subject, for each eye at each visit. The mean CCT and SD of the five readings will be recorded, and the 
mean will be analyzed for efficacy. 
12.1.2 BEST CORRECTED VISUAL ACUITY BY ETDRS METHOD 
The BCVA assessment using the ETDRS method will be performed at all scheduled visits for both eyes 
following manifest refraction, and prior to the slit lamp examination. BCVA will be analyzed for efficacy and 
presented as letter score and logarithm of the minimum angle of resolution (logMAR), calculated as: (85 – 
letter core)/50. 
12.1.3 BIOMICROSCOPY 
An external magnification and biomicroscopy will be performed using a slit-lamp. Magnification will be 
consistent with standard clinical practice. The subject will be seated. Examinations of the conjunctiva 
(hyperemia and edema), cornea (staining, stromal edema, and epithelial edema), anterior chamber (cells 
and flare), eyelid (erythema and edema), iris/pupil, and lens (lens status and lens opacity for phakic only) 
will be performed at all scheduled visits for both eyes. The result scoring values are None (0), Mild (+1),  
Moderate (+2), Severe (+3), and Hypopyon (+4 for anterior chamber cells only). Positive findings will be 
graded as being clinically significant (CS) and being non-clinically significant (NCS). A “No positive findings” 
result as collected on the CRF is considered a score value of 0 and NCS. Resolution of corneal edema (a 
grade of 0 for both stromal edema and epithelial edema) and resolution of epithelial edema (a grade of 0 
for epithelial edema) will be identified and analyzed for efficacy. 
12.1.4 TOMOGRAPHY (PENTACAM®; SELECTED SITES ONLY) 
Tomography assessments using a Pentacam® will be performed at selected sites only for both eyes at all 
scheduled visits. Measurements of CCT, superior corneal thickness (SCT), temporal corneal thickness 
(TCT), nasal corneal thickness (NCT), and inferior corneal thickness (ICT) will be collected and analyzed 
for efficacy.  


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12.1.5 V-FUCHS QUESTIONNAIRE 
Patient-reported V-FUCHS questionnaire will be administered at the Baseline, Week 4, and Week 8 visits 
(Wacker 2018). Raw questionnaire data will be processed and be converted to Rasch-based scores 
including visual acuity factor mean score and SD and glare and diurnal variation (Glare Factor) mean score 
and SD. Rasch-based factor mean score will be analyzed for efficacy. 
12.2 Primary Efficacy Analysis 
The primary comparisons in this trial will be within a treatment group on the mean change from baseline 
CCT by ultrasound pachymetry at Week 4/Day 28 (visit 4) for each of netarsudil QD and BID. The 
summary will be provided for the study eye. The primary efficacy endpoint change from baseline in CCT 
will be summarized using continuous descriptive statistics and analyzed primarily using one-sample t-
tests by treatment group; comparisons between treatment groups (QD – BID) will be completed using 
two-sample t-tests. Sensitivity analyses will be completed using Wilcoxon signed-rank tests within a 
treatment group, Wilcoxon rank sum tests between treatment groups and an analysis of covariance 
(ANCOVA) model with terms for baseline CCT value and treatment.  
The mean change from baseline in CCT for each treatment group along with 2-sided p-values and 90% 
and 95% CIs around the change from baseline within each treatment group for the mean from one-
sample t-test will be presented. The mean difference in change from baseline between treatment groups 
along with 2-sided p-values and 90% and 95% CIs for the mean differences between treatment groups 
from two-sample t-test will be presented. P-values from Wilcoxon signed-rank test within each treatment 
group, and Wilcoxon rank-sum test between treatment groups will be presented. The least squares (LS) 
mean for each treatment group along with 2-sided p-values and 2-sided 90% and 95% CIs around the 
change from baseline within each treatment group will be presented from the ANCOVA model. 
Comparisons between treatment groups on the change from baseline in CCT will also be made including 
the LS mean difference and the corresponding 2-sided p-values and 90% and 95% CIs. 
Example SAS® code for the primary efficacy analysis on change from baseline (CHG_CCT): 
One-sample t-test within a treatment: 
proc ttest data = indata; 
var CHG_CCT; 
by treatment; 
run; 
Wilcoxon signed-rank test within a treatment: 
proc univariate data = indata; 
var CHG_CCT; 
by treatment; 
run; 
 
Two-sample t-test for comparison between treatments: 
proc ttest data = indata; 
class treatment; 
var CHG_CCT; 


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run; 
 
Wilcoxon rank sum test for comparison between treatments: 
proc npar1way data = indata wilcoxon; 
class treatment; 
var CHG_CCT; 
run; 
 
ANCOVA model for modeling the treatment effect: 
proc mixed data = indata; 
class treatment; 
model CHG_CCT = treatment baseline_CCT; 
lsmeans treatment / cl pdiff; 
run; 
The primary analysis will use the mITT population with available data per subject at eye level (ie, ODO). 
Robustness analyses (details provided in in Section 8.2.1) will also be presented based on the MI 
methodology under different assumptions of missingness and intercurrent events (where missing data or 
withdrawal due to lack of efficacy or AEs will be imputed using FCS regression method and missing data 
for all other reasons will be imputed using worst within subject observation prior to the intercurrent event).  
Missing data handling using MI under different assumptions of missingness and intercurrent events are 
described in steps along with example SAS® code for the primary analyses one-sample t-test and two-
sample t-test. 
Step 1: Imputation step 
The following SAS® code will be employed to produce twenty “complete” (imputed) datasets using MI 
FCS methodology: 
proc mi data = indata (where (seye = eye)) seed = 999 out = outdata1 
nimpute = 20; 
class treatment; 
var treatment baseline_CCT aval2 aval4 aval8; 
fcs; 
run; 
where 
• indata is the name of the input dataset  
• outdata1 is the name of the output dataset; 
• treatment is the treatment group variable; 
• baseline_CCT is the baseline mean CCT value; and 
• aval2, aval4, and aval8 are the mean CCT values at Week 2, Week 4, and Week 8, 
respectively.  
Step 2. Finalization of imputed datasets from MI under different assumptions of missingness and 
intercurrent events 


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After obtaining twenty “complete” datasets, for subjects with withdrawal due to lack of efficacy or AEs, the 
imputed missing values in each dataset will be replaced by the worst within subject observation from 
post-baseline visits (including both scheduled and unscheduled visits) prior to the intercurrent event. After 
replacement, change from baseline mean CCT (CHG_CCT) will be calculated all post-baseline visits in 
each dataset. 
Step 3. Analysis step 
For each of the “complete” datasets from step 2, the following SAS® code will be employed to run the 
primary analyses.  
One-sample t-test 
proc sort data = outdata1; 
by visit treatment _imputation_; 
run; 
ods output statistics = outdata2; 
proc ttest data = outdata1 alpha = 0.05; 
by visit treatment _imputation_;  
var chg_cct; 
run; 
Two-sample t-test  
<data outdata1; 
set outdata1; 
if treatment = “Netarsudil QD” then treatment1 = “1”; 
else treatment1 = “2”; 
treatment = treatment1; 
drop treatment1; 
run;> 
proc sort data = outdata1; 
by visit _imputation_; 
run; 
ods output statistics = outdata2; 
proc ttest data = outdata1 alpha = 0.05; 
by visit _imputation_;  
class treatment; 
var chg_cct; 
run; 
Step 4. Pooling step 
SAS® procedure MIANALYZE will then be used to combine results from step 3 across imputations to 
obtain final estimates for inference.  
One-sample t-test 
ods output ParameterEstimates = outdata3; 
proc mianalyze data = outdata2; 
by visit treatment; 
modeleffects mean; 
stderr stderr; 
run; 
Two-sample t-test 


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proc sort data = outdata2; 
by visit class; 
run; 
ods output ParameterEstimates = outdata3; 
proc mianalyze data = outdata2; 
by visit class; 
modeleffects mean; 
stderr stderr; 
run; 
Additional example SAS® code to be employed in MI step 3 and 4 for other sensitivity analyses for the 
primary efficacy endpoints. 
ANCOVA model  
<data outdata11; 
set outdata1; 
if treatment = “Netarsudil QD” then treatment1 = “1”; 
else treatment1 = “2”; 
treatment = treatment1; 
drop treatment1; 
run;> 
proc sort data = outdata11; 
by visit _imputation_; 
run; 
ods output lsmeans = lsm diffs = diff; 
proc mixed data = outdata11; 
by visit _imputation_;  
class treatment; 
model CHG_CCT = treatment baseline_CCT / solution ; 
lsmeans treatment / pdiff cl alpha = 0.05; 
lsmeans treatment / pdiff cl alpha = 0.1; 
run; 
proc sort data = lsm; 
by visit treatment alpha; 
run; 
ods output ParameterEstimates = outdata2; 
proc mianalyze data = lsm; 
by visit treatment alpha; 
modeleffects estimate; 
stderr stderr; 
run; 
 
proc sort data = diff; 
by visit treatment alpha; 
run; 
ods output ParameterEstimates = outdata3; 
proc mianalyze data = diff; 
by visit treatment alpha; 
modeleffects estimate; 
stderr stderr; 
run; 
Wilcoxon signed-rank test  
**Read the imputed dataset for study eye; 
data eff2; 
set outdata1; 


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chg=aval-baseline; 
achg=abs(chg); 
run; 
 
**ignore zero values; 
proc rank data=eff2 (where=(chg ne 0))  ties=mean out=rank; 
by _imputation_ visit treatment; 
var achg; 
ranks rchg; 
run; 
 
proc sort data=rank; 
by _imputation_ visit treatment chg; 
run; 
** distinguish negative and positive values; 
data sign; 
set rank; 
by _imputation_ visit treatment chg; 
if chg <0 then type="Neg"; 
else if chg >0 then type="Pos"; 
run; 
 
proc sort data=sign; 
by _imputation_ visit treatment type rchg; 
run; 
** calculate sum of positive and negative ranks separately; 
data sign2; 
set sign; 
by  _imputation_ visit treatment type rchg; 
if first.type then cum_sum=rchg; 
else cum_sum+rchg; 
if last.type; 
keep _imputation_ visit treatment type cum_sum; 
run; 
 
**Calculate Wilcoxon location parameter; 
proc transpose data=sign2 out=sign3; 
by  _imputation_ visit treatment; 
var cum_sum; 
id type; 
run; 
 
** Calculate number of observations for each treatment; 
data sign4; 
set sign3; 
pos1=pos; 
neg1=neg; 
if pos= . then pos1 = 0; 
if neg= . then neg1 = 0; 
pos = pos1; 
neg = neg1;  
stat=abs(neg-pos)/2; 
keep _imputation_ visit treatment stat; 
drop neg1 pos1; 
run; 
 
proc sort data=sign4; 


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by _imputation_; 
run; 
** Calculate number of observations for each treatment; 
Proc freq data=rank; 
tables visit*treatment*achg/out=counts; 
tables visit*treatment/out=freq (rename=(count=n)); 
by _imputation_; 
run; 
 
data cfactor; 
set counts (where=(count>1));  
ci=count*(count-1)*(count+1); 
run; 
** calculate correction factor; 
data cfactor2; 
set cfactor; 
by _imputation_ visit treatment; 
if first._imputation_ then cum_c=ci; 
else cum_c+ci; 
if last.treatment; 
run; 
 
** Perform Wilcoxon test; 
data final; 
merge freq (keep=_imputation_ visit treatment n) 
cfactor2(keep=_imputation_  visit treatment cum_c) sign4; 
by _imputation_ visit treatment; 
v=(n*(n+1)*(2*n+1)-cum_c/2)/24; 
tderr=sqrt((n*v-stat**2)/(n-1)); 
est=stat/stderr; 
p=2*(1-probt(test,n-1)); 
run; 
 
**verify location parameter calculated manually is same as the one from 
SAS procedure; 
ods output  TestsForLocation=location(where=(test="Signed Rank")); 
proc univariate data = eff2; 
var CHG; 
by _imputation_ visit treatment; 
run; 
proc sort data=final; 
by visit treatment; 
run; 
 
**combine all results; 
ods output ParameterEstimates = outdata3; 
proc mianalyze data = final; 
by visit treatment; 
modeleffects stat; 
stderr stderr; 
run; 
 
Wilcoxon rank sum test 
<data outdata1; 
set outdata1; 
if treatment = “Netarsudil QD” then treatment1 = “1”; 


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else treatment1 = “2”; 
treatment = treatment1; 
drop treatment1; 
run;> 
proc sort data = outdata1; 
by visit _imputation_; 
run; 
ods output WilcoxonScores = outdata2 (where=(class=“1”)); 
proc npar1way data = outdata1 wilcoxon; 
by visit _imputation_; 
class treatment; 
var chg_cct; 
run; 
 
data outdata2; 
set outdata2; 
by visit _imputation_; 
mean = (sumofscores – expectedsum); 
if mean > 0 then mean1 = mean – 0.5; 
else mean1 = mean + 0.5;  
mean = mean1; 
stderr = stddevofsum; 
drop mean1; 
run; 
ods output ParameterEstimates = outdata3; 
proc mianalyze data = outdata2; 
by visit; 
modeleffects mean; 
stderr stderr; 
run; 
Robustness analysis will also be performed on mITT population using LOCF imputation as well as using 
PP population ODO (see Section 8.2.1 for additional imputation details).  
12.3 Secondary Efficacy Analyses 
The secondary efficacy analyses will be conducted on the mITT population ODO. Descriptive statistics 
will be presented by treatment group for both study eye and qualified fellow eye. Summarization and 
analysis of the mean change from baseline in BCVA at Weeks 4 and 8 and mean change from baseline in 
mean CCT by ultrasound pachymetry at Week 8 will be completed using a similar strategy as for the 
primary endpoints described in Section 12.2. 
Testing of the proportion of subjects who gained ≥15 letters (3 lines) in BCVA at Weeks 4 and 8 
(separately) and testing of the proportion of subjects with complete resolution of corneal edema at Weeks 
4 and 8 (separately), will be completed using logistic regression with fixed effects of treatment and 
corresponding baseline measure. The adjusted odds ratios, marginal proportion of each treatment group, 
and difference in marginal proportions along with corresponding two-sided 95% CIs and p-values will be 
presented. When none of the subjects in both treatment groups achieve a gain of 3 lines in BCVA, no 
inferential statistics for the testing of the proportions will be produced. Treatment comparisons will also be 
made using Pearson’s chi-squared test or Fisher’s exact test if any of the cell counts are less than five as 
a sensitivity analysis. 


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Example SAS® code for analyzing categorical outcomes (“Y” when desired criterion is met, “N” if not)  
Logistic regression for modeling the treatments effect on outcome: 
proc logistic data = indata descending alpha = 0.05;  
class treatment (ref = ‘BID’) / param=glm; 
model  outcome = treatment baseline_BCVA;  
lsmeans treatment / diff oddsratio cl exp; 
ods output ParameterEstimates = pe; 
run; 
 
In the logistic model for the probability of outcome = Y, the following call to “Margins” (a SAS macro) 
estimates the treatment predictive margins and their difference (the margin effect) and reverses the 
direction of the difference (QD – BID rather than default BID – QD): 
%Margins(data = indata, 
            class = treatment, 
            response = outcome,  
            roptions = event='Y', 
            model = treatment baseline_BCVA, 
            dist = binomial, 
            margins = treatment,  
            options = cl diff reverse) 
 
Chi-Squared test / Fisher’s Exact test for comparison between treatments: 
proc freq data=indata;  
tables outcome*treatment / chisq fisher; 
weight count/zeros; 
run; 
Robustness analysis for secondary efficacy endpoints will be performed using PP population ODO. In the 
summaries using PP population ODO, subjects whose dosing regimen was increased from QD to BID 
starting from Week 4 will be reported separately from the subjects who did not change regimens 
throughout the study duration under a group called “Netarsudil QD to BID” for the Week 8 visit. The 
baseline value for these subjects will remain the same. At Week 8 visit, comparisons for efficacy 
endpoints between subjects treated under QD and subjects who switched from QD to BID will be 
performed as appropriate. However, a high degree of caution should be exerted when interpreting the 
results as sample sizes might be smaller. 
12.4 Exploratory Efficacy Analyses 
The exploratory efficacy analyses will be conducted in the mITT population with available data per 
subject. Descriptive statistics will be presented by treatment group and for study eye and qualified fellow 
eye. Testing of the proportion of subjects who gained ≥15 letters (3 lines) in BCVA at Week 2, testing of 
the proportion of subjects who gained ≥10 letters (2 lines) and ≥5 letters (1 line) in BCVA at all scheduled 
visits (ie, Weeks 2, 4, and 8), and testing of the proportion of subjects with complete resolution of 
epithelial edema (a grade 0 for epithelial edema assessed by biomicroscopy) at all scheduled visits (ie, 
Weeks 2, 4, and 8) for those who have epithelial edema at Baseline visit (a grade > 1 for epithelial edema 
assessed by biomicroscopy) will be completed in a similar manner as the secondary endpoints. If there 


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are less than 5 subjects who have epithelial edema at Baseline visit within each treatment group, no 
inferential statistics will be produced. Summarization and analysis of the mean change from baseline in 
corneal thickness including CCT, SCT, TCT, NCT, and ICT as assessed by Pentacam® at all scheduled 
visits (ie, Weeks 2, 4, and 8), and mean change from baseline in V-FUCHS factor score including glare 
and diurnal variation (Glare Factor) mean score and visual acuity factor mean score at all scheduled visits 
(ie, Weeks 2, 4, and 8) will be completed using a similar strategy as for the continuous secondary 
endpoints.  
Robust analysis will be performed on the exploratory efficacy endpoints, the proportion of subjects who 
gained ≥15 letters [3 lines], ≥10 letters [2 lines], or ≥5 letters [1 line] in BCVA using PP population ODO in 
similar manner as secondary endpoints. No imputation will be performed on other exploratory efficacy 
endpoints. 
12.5 Ad-hoc Efficacy Analyses 
If PP population is not exactly the same as mITT population, ad-hoc efficacy tables using mITT population 
ODO reported as treated may be produced in the same format as efficacy tables using PP population ODO. 
In the ad-hoc tables, subjects whose dosing regimen was increased from QD to BID starting from Week 4 
will be reported under the group “Netarsudil QD to BID” at Week 8 visit. ” If PP population is exactly the 
same as mITT population, no additional analyses will be performed.  
13. Safety Analyses 
All safety analyses will be conducted using the safety population. For visit-based safety summaries 
(BCVA, biomicroscopy, IOP, and vital signs), subjects who switched dose group from QD to BID starting 
from Week 4 will be reported under treatment group BID at Week 8 visit. The baseline value for these 
subjects will remain the same.   
13.1 Adverse Events 
An AE is defined as any untoward medical occurrence associated with the use of a drug in humans, whether 
or not the event is considered drug-related. An AE can therefore be any unfavorable and unintended sign 
(eg, an abnormal laboratory finding), symptom, or disease occurring after the first dose of study drug, 
without any judgment about causality. Any pre-existing medical condition that worsens after first 
administration of the study drug will also be considered a new AE. The AE reporting period starts from the 
time the subject signs the informed consent until the subject’s last study visit. All AEs will be coded using 
MedDRA Version 23.0.  
A treatment-emergent adverse event (TEAE) is defined as any event that occurs or worsens on or after the 
first dose of study medication. Adverse events recorded on the eCRF which began prior to treatment and 
remained unchanged or improved will not be included in the summary tables but will be included in the AE 
data listings. For all AE summary tables, a third group other than QD and BID dose group will be presented 
to capture the AEs for those subjects who switched the treatment dose from QD to BID starting from Week 


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4. The AE that began on or after the switch will be considered treatment emergent to the BID treatment; 
The AE that began before the switch and continued after the switch will be considered treatment emergent 
to the QD treatment only; if the AE worsened on or after the switch, the new AE (one with higher severity) 
would become a TEAE for the BID dose and the original AE would be treatment emergent to the QD dose. 
An overall summary will be presented that includes the number of events and the number and percentage 
of subjects who experienced at least one TEAE, by treatment group. This summary will also include 
breakdowns of TEAEs further categorized as ocular (study eye and fellow eye separately with TEAEs 
occurred on both eyes counted towards study eye once and fellow eye once) or non-ocular, TEAEs by 
maximum severity, TEAEs by maximum relationship to study medication, TEAE by expectedness, serious 
TEAEs, TEAEs leading to study medication discontinuation, TEAEs leading to death.  
Additional summaries of TEAEs will be provided showing the number and percentage of subjects who 
experienced at least one TEAE by treatment group. Non-ocular TEAEs will be summarized using discrete 
summary statistics and presented by treatment group at the subject level by SOC and PT. Ocular TEAEs 
will be summarized similarly at the subject level for study eyes and fellow eyes, separately. If a subject 
reports the same PT multiple times within the same SOC, that PT will only be reported once within that 
SOC. As with the PT, if a subject reports multiple conditions within the same SOC, that SOC will only be 
reported once. In the summary, SOCs and PTs within each SOC will be listed in ascending alphabetical 
order. 
Separate summaries will be provided for the following categories of AEs:  
• 
Ocular TEAEs in the study eye and fellow eye  
• 
Non-ocular TEAEs  
• 
Ocular TEAEs related or possibly related to study medication 
• 
Non-Ocular TEAEs related or possibly related to study medication 
• 
Serious TEAEs 
• 
Serious TEAEs related or possibly related to study medication 
Severity of an AE is defined as a qualitative assessment of the level of discomfort or the degree of 
intensity of an AE as determined by the Investigator or reported to him/her by the subject. The 
assessment of severity is made irrespective of study medication relationship or seriousness of the event 
and should be evaluated according to the following scale: 
• Mild: Present and noticeable, but not distressing, and no disruption of normal daily activities. 
• Moderate: Bothersome, discomfort sufficient to possibly reduce or affect normal daily activity. 
• Severe: Incapacitating, with inability to work or perform normal daily activity. 
A change in increased severity for a reported AE will require a stop date for the previous severity and a 
new start and stop date for the new severity. For example, a change in severity may go from mild to severe 
or from moderate to severe. In both cases, the start and stop dates should be recorded.  


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The relationship of each AE to the study drug should be determined by the Investigator using these 
explanations: 
• Not Related: The event is clearly related to other factors such as subject’s clinical condition; 
therapeutic interventions, concomitant disease or therapy administered to the subject and does not 
follow a known response pattern to the product. 
• Unlikely Related: The event is most probably caused by other etiologies such as participant’s 
underlying condition, therapeutic intervention, or concomitant therapy; or the delay between 
administration and the onset of the AE is incompatible with a causal relationship. Therefore, there is 
not a reasonable possibility that the AE was caused by the study medication. 
• Possibly Related: The event follows a reasonable, temporal sequence from the time of study 
medication administration and/or follows a known response pattern to the study medication, but could 
have been produced by other factors such as the subject’s clinical state, therapeutic interventions or 
concomitant therapy administered to the subject. 
• Related: The event follows a reasonable, temporal sequence from the time of study medication 
administration and/or follows a known response pattern to the study medication and cannot be 
reasonably explained by other factors such as subject’s clinical state, therapeutic interventions or 
concomitant therapy administered to the subject, and either occurs immediately following study 
medication administration, or improves on stopping the study medication, or reappears on repeat 
exposure, or there is a positive reaction at the application site. 
Summaries of TEAEs by maximum severity will be presented for ocular TEAEs and non-ocular TEAEs 
separately. The number of subjects with any TEAEs (along with percentages) will be tabulated by SOC and 
PT within each SOC by treatment group. To count the number of subjects with any TEAEs, if a subject has 
multiple TEAEs coded to the same PT within the same SOC, the subject will be counted once under the 
maximum severity. 
Individual subject listing will be provided for all AEs, SAEs, AEs leading to study medication discontinuation, 
AEs leading to death, AEs related to COVID-19 , and SAEs related to COVID-19. 
13.2 Best Corrected Visual Acuity 
BCVA testing using ETDRS will be performed at all scheduled visits for both eyes. BCVA will be presented 
as letter score and logMAR. The observed and change from baseline values will be summarized for each 
eye (study eye and fellow eye) using continuous summary statistics by visit for each treatment group. A 
subject listing of visual acuity will also be provided.  
13.3 Biomicroscopy 
An external magnification and biomicroscopy will be performed using a slit-lamp at all scheduled visits for 
both eyes as described in Section 12.1.3.  


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A shift table of score values will also be provided comparing each follow-up visit to the baseline. Additionally, 
discrete summaries will be provided by region, finding, visit, and eye (study eye and fellow eye) for the 
number of subjects with at least one severity grade increase from baseline and for the number of subjects 
judged to be clinically significant by the Investigator. A subject listing of biomicroscopy will also be provided. 
13.4 Intraocular Pressure 
IOP will be collected at all scheduled visits for both eyes using an Applanation Tonometer. Two or three 
results (when IOP difference between test 1 and test 2 is more than 2 mmHg in one or both eyes) will be 
collected for both eyes. Mean IOP (mean when two results are collected or median IOP when three results 
are collected) will be calculated and summarized. The mean IOP and change from baseline mean IOP will 
be summarized for each eye (study eye and fellow eye) using continuous summary statistics by visit for 
each treatment group. A subject listing of IOP will also be provided. 
13.5 Ophthalmoscopy 
A dilated ophthalmoscopy at Screening visit and Week 8 and an undilated ophthalmoscopy at Baseline visit 
and Weeks 2 and 4 of the retina, macula, choroid, optic nerve, and vitreous humour will be performed. The 
results will be graded as normal, abnormal CS, or abnormal NCS.  
A shift table will also be provided comparing each follow-up visit to baseline. A subject listing of 
ophthalmoscopy will also be provided. 
13.6 Vital Signs 
Vital signs (heart rate, systolic sitting blood pressure, diastolic sitting blood pressure) will be collected at 
Screening, Baseline and Week 8. The results will be summarized by visit for each treatment group, using 
continuous summary statistics, including change from baseline. A subject listing of the vital signs results 
will also be provided. 
14. Interim Analyses 
No interim analysis is planned.  
15. Changes from Protocol-Stated Analyses 
1. Assessment window mapping will be applied for missing values at scheduled visits in the table 
summaries using ODO.  
2. Multiple imputations by FCS regression method instead of protocol-stated monotone regression 
will be employed to handle missing data without withdrawal or withdrawal due to reasons other 
than lack of efficacy or adverse events. FCS is a valid statistical method that has greater flexibility 
than monotone regression method.   
3. Additional exploratory efficacy endpoints are added to look at proportion of subjects with 
complete resolution of epithelial edema at week 4 and 8 among subjects who have epithelial 
edema at baseline.   


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16. References 
Wacker K, Baratz KH, Bourne WM, Patel SV. Patient-Reported Visual Disability in Fuchs' 
Endothelial Corneal Dystrophy Measured by the Visual Function and Corneal Health Status 
Instrument. Ophthalmology. 2018;125(12):1854-1861. 
17. Revision History 
Documentation of revision to the SAP will commence after approval of the final version 1.0. 
18. Tables 
Tables that will be included in the topline delivery are shown in boldface font.  
Table Number 
Title 
Population 
Table 14.1.1 
Subject Disposition 
 
Table 14.1.2 
Demographics  
Modified Intent-to-Treat 
Population 
Table 14.1.3.1 
Ocular Medical History 
Modified Intent-to-Treat 
Population 
Table 14.1.3.2 
Non-Ocular Medical History  
Modified Intent-to-Treat 
Population 
Table 14.1.4.1 
Ocular Prior and Concomitant Medications  
Modified Intent-to-Treat 
Population 
Table 14.1.4.2 
Non-Ocular Prior and Concomitant Medications  
Modified Intent-to-Treat 
Population 
Table 14.2.1.1 
Change from Baseline in Central Corneal Thickness (µm) as 
Assessed by Ultrasound Pachymetry at Week 4 (Study Eye)  
Modified Intent-to-Treat 
Population (Observed Data 
Only) 
Table 14.2.1.2 
Change from Baseline in Central Corneal Thickness (µm) as 
Assessed by Ultrasound Pachymetry at Week 4 (Study Eye) 
Modified Intent-to-Treat 
Population (Multiple Imputation) 
Table 14.2.1.3 
Change from Baseline in Central Corneal Thickness (µm) as 
Assessed by Ultrasound Pachymetry at Week 4 (Study Eye) 
Modified Intent-to-Treat 
Population (LOCF) 
Table 14.2.1.4 
Change from Baseline in Central Corneal Thickness (µm) as 
Assessed by Ultrasound Pachymetry at Week 4 (Study Eye) 
Per Protocol Population 
(Observed Data Only) 
Table 14.2.1.5 
Change from Baseline in Central Corneal Thickness (µm) as 
Assessed by Ultrasound Pachymetry by Visit 
Modified Intent-to-Treat 
Population (Observed Data 
Only) 
Table 14.2.1.6 
Change from Baseline in Central Corneal Thickness (µm) as 
Assessed by Ultrasound Pachymetry by Visit 
Per Protocol Population 
(Observed Data Only) 
Table 14.2.1.7 
Central Corneal Thickness (µm) as Assessed by Ultrasound 
Pachymetry by Visit 
Modified Intent-to-Treat 
Population (Observed Data 
Only) 


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Table 14.2.2.1 
Change from Baseline in Best Corrected Visual Acuity using 
ETDRS Method by Visit  
Modified Intent-to-Treat 
Population (Observed Data 
Only) 
Table 14.2.2.2 
Change from Baseline in Best Corrected Visual Acuity using ETDRS 
Method by Visit  
Per Protocol Population 
(Observed Data Only) 
Table 14.2.2.3 
Best Corrected Visual Acuity using ETDRS Method by Visit 
Modified Intent-to-Treat 
Population (Observed Data 
Only) 
Table 14.2.3.1.1 
Proportion of Subjects Who Gained 15 Letters or More in BCVA 
using ETDRS Method by Visit  
Modified Intent-to-Treat 
Population (Observed Data 
Only) 
Table 14.2.3.1.2 
Proportion of Subjects Who Gained 15 Letters or More in BCVA 
using ETDRS Method by Visit  
Per Protocol Population 
(Observed Data Only) 
Table 14.2.3.2.1 
Proportion of Subjects Who Gained 10 Letters or More in BCVA 
using ETDRS Method by Visit  
Modified Intent-to-Treat 
Population (Observed Data 
Only) 
Table 14.2.3.2.2 
Proportion of Subjects Who Gained 10 Letters or More in BCVA 
using ETDRS Method by Visit  
Per Protocol Population 
(Observed Data Only) 
Table 14.2.3.3.1 
Proportion of Subjects Who Gained 5 Letters or More in BCVA 
using ETDRS Method by Visit  
Modified Intent-to-Treat 
Population (Observed Data 
Only) 
Table 14.2.3.3.2 
Proportion of Subjects Who Gained 5 Letters or More in BCVA using 
ETDRS Method by Visit  
Per Protocol Population 
(Observed Data Only) 
Table 14.2.4.1 
Proportion of Subjects with Complete Resolution of Corneal 
Edema by Visit 
Modified Intent-to-Treat 
Population (Observed Data 
Only) 
Table 14.2.4.2 
Proportion of Subjects with Complete Resolution of Corneal Edema 
by Visit 
Per Protocol Population 
(Observed Data Only) 
Table 14.2.5 
Change from Baseline in Central Corneal Thickness (µm) as 
Assessed by Pentacam by Visit 
Modified Intent-to-Treat 
Population (Observed Data Only) 
Table 14.2.6 
Change from Baseline in V-FUCHS Factor Score by Visit 
Modified Intent-to-Treat 
Population (Observed Data Only) 
Table 14.2.7 
Proportion of Subjects with Complete Resolution of Epithelial Edema 
by Visit 
Modified Intent-to-Treat 
Population (Observed Data Only) 
Table 14.3.1 
Overall Summary of Treatment-Emergent Adverse Events by 
Treatment Group  
Safety Population 
Table 14.3.2.1 
Ocular Treatment-Emergent Adverse Events by System Organ 
Class and Preferred Term 
Safety Population 
Table 14.3.2.2 
Non-Ocular Treatment-Emergent Adverse Events by System Organ 
Class and Preferred Term  
Safety Population 


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Table 14.3.2.3 
Ocular Treatment-Emergent Adverse Events Related or 
Possibly Related to Study Medication by System Organ Class 
and Preferred Term  
Safety Population 
Table 14.3.2.4 
Non-Ocular Treatment-Emergent Adverse Events Related or 
Possibly Related to Study Medication by System Organ Class and 
Preferred Term  
Safety Population 
Table 14.3.2.5 
Serious Treatment-Emergent Adverse Events by System Organ 
Class and Preferred Term  
Safety Population 
Table 14.3.2.6 
Serious Treatment-Emergent Adverse Events Related or Possibly 
Related to Study Medication by System Organ Class and Preferred 
Term 
Safety Population 
Table 14.3.2.7 
Ocular Treatment-Emergent Adverse Events by System Organ 
Class, Preferred Term, and Maximum Severity 
Safety Population 
Table 14.3.2.8 
Non-Ocular Treatment-Emergent Adverse Events by System Organ 
Class, Preferred Term, and Maximum Severity 
Safety Population 
Table 14.3.3.1 
Best Corrected Visual Acuity (BCVA) Using ETDRS Method by 
Treatment and Visit 
Safety Population 
Table 14.3.3.2.1.1 
Shifts in Biomicroscopy from Baseline by Treatment and Visit – 
Study Eye  
Safety Population 
Table 14.3.3.2.1.2 
Shifts in Biomicroscopy from Baseline by Treatment and Visit – 
Fellow Eye  
Safety Population 
Table 14.3.3.2.2 
Number and Percentage of Subjects with at Least a One Severity 
Grade Increase from Baseline Biomicroscopy Findings 
Safety Population 
Table 14.3.3.2.3 
Number and Percentage of Subjects with Biomicroscopy Findings 
Judged Clinically Significant by the Investigator  
Safety Population 
Table 14.3.3.3 
Mean Intraocular Pressure (IOP) by Treatment and Visit  
Safety Population 
Table 14.3.3.4.1 
Shifts in Ophthalmoscopy from Baseline by Treatment and Visit - 
Study Eye 
Safety Population 
Table 14.3.3.4.2 
Shifts in Ophthalmoscopy from Baseline by Treatment and Visit - 
Fellow Eye 
Safety Population 
Table 14.3.3.5 
Vital Signs by Treatment and Visit  
Safety Population 
19. Listings 
Listing Number 
Title 
Population 
Listing 16.1.7 
Randomization Schedule  
All Randomized Subjects 
Listing 16.2.1.1 
Subject Disposition  
All Randomized Subjects 
Listing 16.2.1.2 
Subject Disposition 
Screen Failed Subjects 
Listing 16.2.2 
Protocol Deviations  
All Randomized Subjects 
Listing 16.2.3 
Analysis Populations  
All Randomized Subjects 


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Listing 16.2.4.1 
Demographics  
All Randomized Subjects 
Listing 16.2.4.2 
Childbearing Potential  
All Randomized Female Subjects 
Only 
Listing 16.2.4.3 
Ocular Medical History  
All Randomized Subjects 
Listing 16.2.4.4 
Non-Ocular Medical History 
All Randomized Subjects 
Listing 16.2.4.5 
Ocular Surgery Laser Procedure  
All Randomized Subjects 
Listing 16.2.4.6 
Prior and Concomitant Medications  
All Randomized Subjects 
Listing 16.2.5 
Study Drug and Artificial Tears Assignment  
All Randomized Subjects 
Listing 16.2.6.1 
Central Corneal Thickness (CCT) by Ultrasound Pachymetry  
All Randomized Subjects 
Listing 16.2.6.2 
Best Corrected Visual Acuity by ETDRS  
All Randomized Subjects 
Listing 16.2.6.3 
Biomicroscopy  
All Randomized Subjects 
Listing 16.2.6.4 
Tomography  
All Randomized Subjects 
Listing 16.2.6.5.1 
Visual Function and Corneal Health Status (V-FUCHS) – Visual 
Acuity Factor  
All Randomized Subjects 
Listing 16.2.6.5.2 
Visual Function and Corneal Health Status (V-FUCHS) – Glare 
Factor   
All Randomized Subjects 
Listing 16.2.7.1 
All Adverse Events 
All Randomized Subjects 
Listing 16.2.7.2 
Serious Adverse Events  
All Randomized Subjects 
Listing 16.2.7.3 
Adverse Events Leading to Study Medication Discontinuation  
All Randomized Subjects 
Listing 16.2.7.4 
Adverse Events Leading to Death   
All Randomized Subjects 
Listing 16.2.8 
Pregnancy Test  
All Randomized Female Subjects 
with Childbearing Potential 
Listing 16.2.9 
Intraocular Pressure (IOP)  
All Randomized Subjects 
Listing 16.2.10 
Ophthalmoscopy  
All Randomized Subjects 
Listing 16.2.11 
Vital Signs  
All Randomized Subjects 
20. Figures 
Figure Number 
Title 
Population 
Figure 14.2.1 
Line Plot of Mean Change from Baseline in CCT Assessed by 
Ultrasound Pachymetry (Study Eye) 
Modified Intent-to-Treat Population 
Figure 14.2.2 
Line Plot of Mean Change from Baseline in BCVA Assessed by 
ETDRS (Study Eye) 
Modified Intent-to-Treat Population