Clinical trial NCT04513652
Conditions: Anesthesia, Local
Keywords: AG-920 articaine septocaine Intravitreal Injection
Sponsor: American Genomics, LLC
Start date: 2020-09-03; Completion date: 2020-12-17
Enrollment: 120 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
This is a Phase 3, randomized, placebo-controlled, double masked, parallel design study in healthy subjects performed in the US. It is designed to evaluate the safety and anesthetic efficacy of AG-920 Sterile Topical Ophthalmic Solution.

Detailed description:
This is a Phase 3, randomized, placebo-controlled, double masked, parallel design study in healthy subjects performed in the US. It is designed to evaluate the safety and anesthetic efficacy of AG-920 Sterile Topical Ophthalmic Solution. In this study, subjects who provide informed consent and fulfill all the inclusion criteria and none of the exclusion criteria will be randomized in a 1:1 ratio to receive a single dose of AG-920 or identical looking placebo into the study eye. Subjects will undergo a conjunctival pinch procedure and the pain associated with the pinch rated.

Arms: arm 1: Articaine Sterile Topical Ophthalmic Solution (AG-920) is a sterile, isotonic, non-preserved aqueous solution containing the active ingredient Articaine HCl 8%, Boric Acid, Mannitol, Sodium Acetate Trihydrate, Glacial Acetic Acid, and Edetate Disodium Dihydrate. The product formulation is adjusted to pH 4.5 to 5.0. Each subject randomized to AG-920 will receive a single dose of 2 drops 30 seconds apart from a single vial into study eye. arm 2: Placebo ophthalmic solution is identical to the active product, with the exception of the active ingredient. Each subject randomized to placebo will receive a single dose of 2 drops 30 seconds apart from a single vial into study eye.
Interventions: intervention 1: AG-920 intervention 2: Placebo
Intervention descriptions: intervention 1: AG-920 Sterile Topical Ophthalmic Solution intervention 2: Placebo Topical Ophthalmic Solution

Protocol documents:
Prot_000.pdf: 






American Genomics, LLC 
Clinical Study Protocol 
Confidential 
 
Protocol AG-920-CS301 Amendment 1 
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Abbreviations and Terms 
 
Abbreviation 
Full text 
AE 
Adverse event 
AG-920 
 sterile topical ophthalmic solution 
BCVA 
Best corrected visual acuity 
BOCF 
Baseline observation carried forward 
CFR 
Code of Federal Regulations 
COVID-19 
Coronavirus Disease 2019 
CRO 
Contract Research Organization 
EDC 
Electronic Data Capture 
ETDRS 
Early Treatment of Diabetic Retinopathy Study 
FDA 
Food and Drug Administration  
GCP 
Good Clinical Practices 
GLP 
Good Laboratory Practices 
GMP 
Good Manufacturing Practices 
HCl 
Hydrochloride 
ICH 
International Conference on Harmonisation 
ICF 
Informed Consent Form 
IEC 
Independent Ethics Committee  
IMP 
Investigational Medical Product 
IOP 
Intraocular pressure 
IRB 
Institutional Review Board 
ITT 
Intent-to-Treat 
LOCF 
Last Observation Carried Forward 
logMAR 
Logarithmic Minimum Angle of Resolution 
MedDRA 
Medical Dictionary for Regulatory Activities 
NSAID 
Non-steroidal anti-inflammatory drug 
OD 
Oculus dexter (Right eye) 
OTC 
Over the Counter 
OS 
Oculus sinister (Left eye) 
PP 
Per protocol 
PPE 
Personal protective equipment 
PT 
Preferred Term 
SAE 
Serious adverse event 
SAP 
Statistical Analysis Plan 
SOC 
System/Order/Class 
SOP 
Standard Operating Procedures 
TEAE 
Treatment emergent adverse event 
UPT 
Urine Pregnancy Test 
US 
United States 
VA 
Visual acuity 
 
 
 


American Genomics, LLC 
Clinical Study Protocol 
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Summary 
Study Number 
AG-920-CS301 
Clinical Phase: 
Phase 3 
Type of study 
Efficacy and Safety 
Name of Investigational product: 
 Sterile Topical Ophthalmic Solution (AG-920) 
Duration of treatment per subject 1 day 
Objectives and Endpoints: 
Primary Objective 
Primary Endpoints 
 
To evaluate anesthetic 
efficacy of AG-920  
The proportion of subjects 
with no pain at 5 minutes. 
 
Secondary Objectives 
Secondary Endpoints 
 
To evaluate how long it 
takes one dose of AG-920 
to anesthetize the eye  
Mean time to no pain score 
(onset)  
 
To evaluate how long one 
dose of AG-920 
anesthetizes the eye 
Mean duration of 
anesthetic effect  
 
To evaluate the safety and 
tolerability of AG-920 
Visual acuity, 
biomicroscopy, AEs, 
TEAEs, SAEs, 
withdrawals due to TEAEs 
Subject Population:  
Healthy volunteers 
Exclusion criteria may be found in Section 4.2.  
Design: 
Randomized, placebo (vehicle) controlled, double-
masked, parallel. 
Visit Schedule: 
Study visits will consist of a Screening Visit, a dosing and 
anesthesia testing visit, and a follow-up visit (by 
telephone) 
Number of Investigational Sites: 
1-2 US sites  
Estimated Total Sample Size: 
120/study; ~60/treatment group 
Plan for Data Analysis:  
With 60 subjects per treatment group and assuming 15% 
response in the vehicle group, this study will have 88% 
power to detect a treatment effect of at least 25% (α=0.05, 
two-sided).  
Investigational/Comparator 
Product(s), Dose and Mode of 
Administration 
Subjects will receive a single dose of IMP in one (study) 
eye.  The IMP will be randomized as either AG-920 or 
Placebo.  The study eye will be randomized as either right 
eye (OD) or left eye (OS).  The single dose will be 
administered by the clinic staff as two drops in study eye 
30 seconds apart.  
 
 
 


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Clinical Study Protocol 
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Protocol AG-920-CS301 Amendment 1 
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TABLE OF CONTENTS 
CONTACT INFORMATION AND PROTOCOL AUTHORIZATION.......................................................... 2 
ABBREVIATIONS AND TERMS ....................................................................................................................... 4 
SUMMARY ............................................................................................................................................................ 5 
1 
INTRODUCTION........................................................................................................................................ 8 
1.1 
Findings from nonclinical and clinical studies ...................................................................................... 8 
1.2 
Potential Risks and benefits to human subjects .................................................................................... 9 
1.3 
Design justification .................................................................................................................................. 9 
2 
OBJECTIVES AND ENDPOINTS ............................................................................................................ 9 
3 
STUDY DESIGN ........................................................................................................................................ 10 
3.1 
Description and schedule of visits and procedures ............................................................................. 10 
3.2 
Measures taken to minimize/avoid bias ............................................................................................... 12 
3.3 
Study medications ................................................................................................................................. 12 
 
IMP Packaging and labeling .......................................................................................................... 12 
 
Storage of study medication .......................................................................................................... 13 
 
Study medication accountability .................................................................................................... 13 
3.4 
Expected duration of subject participation ......................................................................................... 13 
3.5 
Randomization and procedure for breaking the code ....................................................................... 13 
3.6 
Participant and Study completion ....................................................................................................... 14 
 
Completed subject ......................................................................................................................... 14 
 
Non-completing subject/Subject Withdrawal ................................................................................ 14 
 
Discontinuation of the Study ......................................................................................................... 14 
 
Discontinuation of a Clinical Site .................................................................................................. 15 
 
Actions after discontinuation ......................................................................................................... 15 
 
Completed study ............................................................................................................................ 15 
 
Procedure after the completion of the study .................................................................................. 15 
4 
SUBJECT INCLUSION AND EXCLUSION CRITERIA ..................................................................... 15 
4.1 
Subject inclusion criteria ...................................................................................................................... 15 
4.2 
Subject exclusion criteria...................................................................................................................... 16 
4.3 
Subject replacement .............................................................................................................................. 16 
5 
TREATMENT OF SUBJECTS ................................................................................................................ 16 
5.1 
Concomitant medications ..................................................................................................................... 17 
 
Prohibited medications .................................................................................................................. 17 
 
Allowed medications ..................................................................................................................... 17 
5.2 
Female Subjects ..................................................................................................................................... 17 
6 
STUDY PROCEDURES ........................................................................................................................... 18 
6.1 
General Procedures: ............................................................................................................................. 18 
6.2 
Re-screening Procedures: ..................................................................................................................... 18 
6.3 
Visit 1 (Screening) – Day -2 to 0/1 ........................................................................................................ 18 
6.4 
Visit 2 (Randomization/Treatment) – Day 1 ....................................................................................... 18 
 
Pre-dose ......................................................................................................................................... 18 
 
IMP Treatment ............................................................................................................................... 19 
 
Post Treatment and Pinch Test ...................................................................................................... 19 
6.5 
Follow-Up Phone Call (Day 2-5) .......................................................................................................... 20 
6.6 
Unscheduled visits ................................................................................................................................. 20 
6.7 
COVID-19 Pandemic Accommodations .............................................................................................. 20 
7 
ASSESSMENT OF EFFICACY ............................................................................................................... 21 
8 
ASSESSMENT OF SAFETY .................................................................................................................... 21 
8.1 
Specification of safety parameters ....................................................................................................... 21 
8.2 
The methods and timing for assessing, recording, and analyzing safety parameters ..................... 21 


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8.3 
Adverse events ....................................................................................................................................... 21 
 
Adverse Event (AE) definitions ..................................................................................................... 21 
 
Serious Adverse Event (SAE) definitions ...................................................................................... 23 
 
Expedited reporting of Serious and Unexpected Adverse Events.................................................. 25 
 
Follow-up of subjects after adverse events .................................................................................... 25 
9 
STATISTICS .............................................................................................................................................. 26 
9.1 
Primary Hypotheses .............................................................................................................................. 26 
9.2 
Sample Size Considerations .................................................................................................................. 26 
9.3 
Analysis populations ............................................................................................................................. 26 
9.4 
Statistical methods to be employed ...................................................................................................... 26 
 
General Considerations .................................................................................................................. 26 
 
Interim analyses ............................................................................................................................. 27 
 
Analysis of Baseline Data .............................................................................................................. 27 
 
Subject Disposition ........................................................................................................................ 27 
 
Protocol Deviations ....................................................................................................................... 27 
 
Analysis of Efficacy ...................................................................................................................... 27 
 
Analysis of Safety .......................................................................................................................... 28 
 
Adverse Events .............................................................................................................................. 28 
9.5 
Procedure for accounting for missing, unused, or spurious data ...................................................... 28 
9.6 
Procedure for reporting deviations from the statistical plan ............................................................ 28 
9.7 
Data listings ........................................................................................................................................... 28 
10 
QUALITY CONTROL AND QUALITY ASSURANCE ....................................................................... 28 
10.1 
Audit and Inspection ........................................................................................................................ 28 
10.2 
Direct Access to Source Data Documents ....................................................................................... 28 
10.3 
Clinical Monitoring .......................................................................................................................... 29 
10.4 
Data Management and Coding ........................................................................................................ 29 
11 
ETHICAL CONSIDERATIONS AND COMPLIANCE WITH GOOD CLINICAL PRACTICE .... 29 
11.1 
Independent Ethics Committee or Institutional Review Board ................................................... 29 
11.2 
Regulatory Authorities ..................................................................................................................... 30 
11.3 
Ethical Conduct of the Study ........................................................................................................... 30 
11.4 
Written Informed Consent .............................................................................................................. 30 
11.5 
Subject Confidentiality .................................................................................................................... 31 
12 
DATA HANDLING AND RECORD KEEPING .................................................................................... 31 
12.1 
Data quality control and reporting ................................................................................................. 31 
12.2 
Inspection of Records ....................................................................................................................... 31 
12.3 
Records retention ............................................................................................................................. 31 
12.4 
Amendments to the protocol ............................................................................................................ 31 
13 
PUBLICATION ......................................................................................................................................... 32 
14 
REFERENCES ........................................................................................................................................... 32 
14.1 
Published references ......................................................................................................................... 32 
14.2 
Internal references ........................................................................................................................... 32 
 
APPENDIX 1: PROCEDURES............................................................................................................. 33 
APPENDIX 2: SAMPLE SUBJECT CONJUNCTIVAL PINCH ASSESSMENT INSTRUCTION ...................... 37 
APPENDIX 3: SAMPLE SUBJECT PAIN ASSESSMENT WORKSHEET ................................................... 38 
 
List of tables 
TABLE 1: SCHEDULE OF VISITS AND PROCEDURES......................................................................... 11 
TABLE 2: AE ASSESSMENT OF CAUSALITY .................................................................................... 24 
TABLE 3: ASSESSMENT OF SEVERITY (INTENSITY) ......................................................................... 24 
TABLE 4: COMPARISON OF SNELLEN RATIO AND LOGMAR UNITS ................................................. 34 
 






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3 STUDY DESIGN 
Randomized, placebo-controlled, double-masked, parallel. 
3.1 
Description and schedule of visits and procedures 
This is a Phase 3, randomized, placebo-controlled, double-masked, parallel design study in healthy 
subjects performed in the US.  It is designed to evaluate the safety and anesthetic efficacy of one 
dose of 
 Sterile Topical Ophthalmic Solution (AG-920).  In this study, subjects who 
provide informed consent and fulfill all the inclusion criteria and none of the exclusion criteria will 
be randomized in a 1:1 ratio to receive a single dose of AG-920 or identical looking placebo into 
one (study) eye (2 drops 30 seconds apart).  Subjects will undergo a conjunctival pinch procedure 
and the pain associated with the pinch rated.  IMP dosing and conjunctival pinch procedure will 
be performed by the study staff. 
 
A schedule of assessments, including allowable visit windows, is displayed in Table 1.   
 
 




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3.2 
Measures taken to minimize/avoid bias 
The Investigator(s), Sponsor, and the subject will be masked to treatment assignment throughout 
the conduct of the study.  Exceptions to this are limited to one statistician at the Contract Research 
Organization (CRO) who will prepare the randomization code, and three people at the Sponsor 
who will review the batch records and release product.  None of these masked persons will be 
involved in the day to day execution of the study.  The masking will be broken after database lock.    
3.3 
Study medications 
 Sterile Topical Ophthalmic Solution (AG-920) is a sterile, isotonic, non-preserved 
aqueous solution containing the active ingredient 
 HCl 8%, Boric Acid, Mannitol, 
Sodium Acetate Trihydrate, Glacial Acetic Acid, and Edetate Disodium Dihydrate.  The product 
formulation is adjusted to pH 4.5 to 5.0.  
 
Placebo ophthalmic solution is identical to the active product, with the exception of the active 
ingredient.  
 
Figure 2: Investigational Medicinal Product 
 
INVESTIGATIONAL PRODUCT 
PRODUCT NAME 
 Sterile Topical 
Ophthalmic Solution (AG-
920) 
Placebo 
ACTIVE INGREDIENT 
 Hydrochloride 
-- 
INACTIVE 
INGREDIENTS 
 
Boric Acid 
Mannitol 
Sodium Acetate Trihydrate 
Glacial Acetic Acid 
Edetate Disodium Dihydrate 
Water for Injection 
 
Boric Acid 
Mannitol 
Sodium Acetate Trihydrate 
Glacial Acetic Acid 
Edetate Disodium Dihydrate 
Water for Injection 
UNIT DOSE 
0.5 mL blow fill seal vial  
0.5 mL blow fill seal vial  
ROUTE OF 
ADMINISTRATION 
Topical ocular 
Topical ocular 
DOSING REGIMEN 
2 drops 30 seconds apart 
from a single vial into study 
eye 
2 drops 30 seconds apart 
from a single vial into study 
eye 
STORAGE 
REQUIREMENTS 
Store at Room Temperature 
(15-25°C or 59-77°F) 
Store at Room Temperature 
(15-25°C or 59-77°F) 
 
 IMP Packaging and labeling 
Investigational Medical Product (IMP) will be packaged and labeled identically in order to 
maintain the integrity of the double mask.  The appearance of the blow fill seal vials for the AG-
920 and matching placebo dosage forms are indistinguishable.   
 


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The container-closure system to be used for the AG-920 and Placebo in this clinical study is a 0.5 
mL low-density polyethylene (LDPE) resin blow fill seal vial.  There are 5 vials inside an 
aluminum foil pouch.  Each foil pouch will be labeled with an investigational label with the 
following minimal information: the study number, kit number, and storage statement, including a 
statement “Caution – New Drug – Limited by Federal (US) Law to Investigational Use” or 
equivalent.  Each foil pouch will be individually placed into a single-unit carton and a similar 
investigational label will be applied to the carton.  The IMP for each individual treatment 
assignment will be packaged into identical subject kits. 
 Storage of study medication 
The IMP should be stored at room temperature (15-25°C or 59-77°F).  Do not freeze the product.  
Prior to dispensing to the subject, all investigational material must be stored in a secure location 
with strictly limited access documented by signature of authorized persons who may dispense 
investigational materials. 
 Study medication accountability 
Accountability of IMP kits will be conducted by a member of the site and verified by a study 
monitor.  Accountability will be ascertained by performing reconciliation between the number of 
kits sent to the site and the amount used and unused at the time of reconciliation. Site staff will be 
queried about any discrepancies. 
 
IMP kit shipment records will be verified, and accountability performed by comparing the 
shipment inventory sheet to the actual quantity of kits received at the site.  In addition, receipt of 
kits will be confirmed by the study monitor.  Accurate records of receipt and disposition of the kits 
(e.g., dates, quantity, subject number, kits used, kits unused, etc.) must be maintained by the 
Investigator or his/her designee.   
 
At the end of the study and after the monitor has verified kit accountability, all IMP is to be 
returned to American Genomics (or designee) or destroyed at the site and documented per the 
site’s standard process. 
3.4 
Expected duration of subject participation 
Each subject is planned to participate in the study for up to 2 Visits (Screening and 
Treatment/Anesthesia testing) and a Follow-up Phone Call 1-4 days following Visit 2 (Day 2-5).  
3.5 
Randomization and procedure for breaking the code 
A randomization code for allocating the treatments will be prepared by an independent 
biostatistician, who is not involved in the day-to-day conduct of the study.  Subjects will be 
randomized in a 1:1 ratio to receive AG-920 or Placebo. 
 
Treatment assignments will be masked to the Investigator, the clinical study team (Sponsor, 
personnel involved in day to day study management, Monitors, Data Managers, and Statisticians), 
and the subjects.  Only in case of medical emergency or occurrence of adverse events that warrant 
unmasking in the opinion of the investigator, will the treatment assignment(s) be unmasked and 
made available to the Investigator and the Sponsor Safety Officer.  In the absence of medical need, 
the randomization code will not be available to the above personnel until after the study is 
completed and the database is locked. 


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If the Investigator feels it is necessary to unmask a subject’s treatment assignment after an 
emergency situation, the Investigator should contact the Sponsor Safety Officer or designee.  Only 
after consultation with the Sponsor Safety Officer will a decision be made as to whether or not the 
treatment for the subject should be unmasked.  The treatment assignment will be revealed on a 
subject-by-subject basis, thus leaving the masking on the remaining subjects intact.   
 
If there is an emergency situation in which treatment of an adverse event requires immediate 
decoding, and the Investigator is unable to contact the Sponsor Safety Officer, the Investigator 
should treat the subject as if they received active treatment (AG-920) and not placebo.  The 
Investigator should contact the Sponsor immediately and document the AE.   
3.6 
Participant and Study completion 
 Completed subject 
A completed subject is defined as one who completes all Visits (1 and 2) and the Follow Up Phone 
call.  
 Non-completing subject/Subject Withdrawal 
A non-completing subject is defined as one who exits the study by their own volition or at the 
discretion of the Investigator and/or the Sponsor Safety Officer.  Any subject may decide to 
voluntarily withdraw from the study at any time without prejudice and the reason will be 
documented.  In the event that discontinuation of treatment is necessary, the Investigator will make 
every attempt to complete all subsequent safety assessments.  Every attempt should be made to 
keep subjects in the study and to perform the required study procedures, but if this is not possible, 
the subject may be withdrawn.   
 
All reasonable efforts should be made to contact the subject who is lost to follow-up.  These efforts 
must be documented in the subject’s file. 
 
Subjects may be withdrawn from the study for any reason at any time, including but not limited to 
the following reasons: 
• Subject request 
• Use of prohibited concomitant medication/therapy 
• Lost to follow-up 
• Occurrence of AEs that are not compatible with the continuation of the subject in the study, 
in the Investigator’s opinion, or that make it unacceptable to the subject to continue 
• Investigator judgment 
• Sponsor request 
• Randomization code broken prematurely 
• Pregnancy 
The reason for early discontinuation will be collected 
 Discontinuation of the Study 
If the clinical study is prematurely terminated or suspended, the Sponsor or designee will inform 
the Investigators and the regulatory authorities of the termination/suspension and the reason(s) for 
the termination/suspension.  The Investigators should promptly notify their Independent Ethics 


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Committee (IEC) or Institutional Review Board (IRB) of the termination or suspension and of the 
reasons. 
 Discontinuation of a Clinical Site 
The Sponsor reserves the right to close an investigational site or terminate the study in its entirety 
at any time, for reasonable cause.  Reasons for the closure of any investigational site or termination 
of the study may include: 
• Failure to accrue subjects into the study at an acceptable rate. 
• Failure of the Investigator to comply with applicable regulations and Good Clinical 
Practice (GCP) guidelines. 
• Submission of knowingly false information from the research facility to the Sponsor, Food 
and Drug Administration (FDA), or other regulatory authorities. 
• Insufficient adherence to protocol requirements and procedures. 
If the study is prematurely discontinued, all study data must be returned to the Sponsor or designee.  
Additionally, the site must conduct final disposition of all unused IMP in accordance with CRO 
procedures.  Study termination and follow-up will be performed in compliance with the conditions 
set forth in regulatory guidelines. 
 
Financial compensation to the Investigator and/or institution will be in accordance with the 
agreement established between the Investigator and the Sponsor or Sponsor designee. 
 Actions after discontinuation 
All subjects who discontinue IMP due to a report of an AE must be followed-up and provided 
appropriate medical care until their signs and symptoms have remitted, stabilized, determined to 
be chronic, or until abnormal laboratory findings have returned to acceptable or pre-study limits.  
 
For the subject who chooses to withdraw consent or who is non-compliant, every possible effort 
should be made by the Investigator to assure there is a final visit that includes all examinations 
listed for Visit 2.  
 Completed study 
The end of the study is defined as the date the last subject completes the Follow-up Phone Call.   
 Procedure after the completion of the study 
When the study is completed and the site has been closed out, the Investigator will be asked to 
notify the governing IRB.  
4 SUBJECT INCLUSION AND EXCLUSION CRITERIA 
Subjects must meet all of the following inclusion criteria to be eligible for participation in this 
study.  On criteria that involves the eyes, both eyes must qualify.   
4.1 
Subject inclusion criteria 
1. Provide written informed consent prior to any study-related procedures being performed. 
2. Is male or a non-pregnant, non-lactating female aged 18 years or older.  Female subjects of 
childbearing potential must have a negative urine pregnancy test at Screening (Visit 1) in 
order to be eligible for randomization.   


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3. Are willing and able to follow instructions and can be present for the required study visits 
and Follow-up Phone Call for the duration of the study. 
4. Have an Early Treatment of Diabetic Retinopathy Study (ETDRS) best corrected visual 
acuity (BCVA) of 20/200 or better in each eye as assessed by Early Treatment of Diabetic 
Retinopathy Study (ETDRS) at the screening visit.  Note: previous refractive procedures 
allowed. 
5. Have an Intraocular Pressure (IOP) between 7 and 30 mmHg. 
6. Certified as healthy by clinical assessment (detailed medical history) including ocular 
examination. 
7. Verbal communication skills adequate to participate. 
8. Able to tolerate bilateral instillation of Over-The-Counter artificial tear product based on 
investigator judgement. 
4.2 
Subject exclusion criteria 
Subjects who meet any of the following exclusion criteria are ineligible for this study: 
 
1. Have participated in an investigational study (drug or device) within the past 30 days. 
2. Have a contraindication to local anesthetics, 
®, or any component of the IMP. 
3. Have known decreased corneal or conjunctival sensitivity (e.g., sequelae of herpetic eye 
disease, corneal graft) or a diagnosed corneal pathology which might lead to decreased 
sensitivity.  
4. Have had ocular surgery (intraocular, refractive, extraocular muscles, eyelid) or general 
surgery in either eye within the past 90 days (Note: 
 restorative work allowed) 
5. Have had an intravitreal injection in either eye within 14 days of randomization. 
6. Have ocular surface disease requiring punctual plugs. 
7. Evidence of any current ocular inflammation. 
8. Subject who must wear contact lenses on Study Day 1 (Visit 2). 
9. Are currently using, or used within the past 30 days, a systemic opioid or opiate analgesic 
or topical Non-steroidal Anti-Inflammatory Drug (NSAID).  
10. Subject who cannot withhold their intermittent over-the-counter (OTC) artificial tear 
lubricant products for one hour preceding or following study medication.  
11. Any condition, including alcohol or drug dependency, that would limit the subject’s ability 
to comply with the procedures of the protocol (per Investigator’s judgment). 
 
Note: Given the approval status and preclinical safety information on the molecule, and low chance 
of systemic exposure, other than the negative urine pregnancy test, there is no restriction on women 
of child-bearing potential. 
4.3 
Subject replacement 
Discontinued subjects may be replaced by enrollment of additional subjects until approximately 
120 subjects are enrolled. 
5 TREATMENT OF SUBJECTS 
Subjects will receive a single dose of IMP in one (study) eye.  The IMP will be randomized as 
either AG-920 or Placebo.  The study eye will be randomized as either right eye (OD) or left eye 
(OS).  The single dose will be administered by the clinic staff as two drops in study eye 30 seconds 
apart.  


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5.1 
Concomitant medications 
 Prohibited medications 
The list of prohibited medications provided below may not be comprehensive.  Investigators are 
encouraged to consult with the Medical Monitor for a decision about proceeding if they have any 
questions or concerns about any medication. 
 
The following medications are prohibited.  
• systemic opioids or opiate analgesics within 30 days of randomization and during the study 
treatment day 
• topical NSAIDs within 30 days of randomization and during the study treatment day 
• Over-the-counter (OTC) artificial tear lubricant products within one hour preceding or 
following study medication, other than the tolerability test  
 
As noted in the exclusion criteria (Section 4.2), individuals must refrain from contact lens wear on 
the Dosing and Anesthesia Testing Day.   
 Allowed medications 
Other than the agents and/or times noted above, systemic therapy with agents is allowed.  
 
Use of all medications should be documented on the appropriate CRF.  Investigators are 
encouraged to contact the Sponsor for any questions regarding allowed medications.  Judgment of 
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 Sponsor.   
 
All medications which the subject has taken within 30 days prior to screening and during the study 
will be recorded in the CRF.  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 desired.  
The use of routine ophthalmic diagnostic pharmaceutical agents (e.g., fluorescein and local 
anesthetic) will be allowed, and individual documentation not required.   
5.2 
Female Subjects 
Female subjects of childbearing potential must not be pregnant or breastfeeding at Randomization.  
These subjects must have a negative urine pregnancy test at Screening in order to be eligible for 
the study.   
Female subjects who are not of childbearing potential must meet at least one of the following 
criteria: 
• At least 1 year since the last menstrual period 
• Surgically sterile (tubal ligation, bilateral oophorectomy, salpingectomy, or hysterectomy) 
• Congenitally sterile 
• Diagnosed as infertile and not undergoing treatment to reverse infertility 


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6 STUDY PROCEDURES 
6.1 
General Procedures: 
The study will consist of 1 or 2 clinic visits and 1 Follow-up Phone Call.  All ocular assessments 
will be performed on both eyes.  Subjects will be screened for entry (Visit 1) and 
randomized/treated at (Visit 2).  Visit 1 and 2 may be on the same day.  At randomization, subjects 
will receive a single dose of IMP (2 drops 30 seconds apart) in the study eye. Subjects will then 
have a safety Follow-up Phone Call 1-4 days (Days 2-5) following Visit 2. 
6.2 
Re-screening Procedures: 
A subject who is first designated as a screen failure prior to being randomized will be allowed to 
rescreen one additional time 7-30 days later.  
6.3 
Visit 1 (Screening) – Day -2 to 0/1 
Visit 1 may be combined with Visit 2 in order to accommodate clinic logistics.  If both visits are 
combined, Visit 1 will last anywhere from 1-3 hours.  If topical anesthetic drops are used to assess 
IOP, subject must wait at least 1 hour before IMP treatment.  Topical anesthetic drops may cause 
a mild stinging or burning sensation upon instillation. 
 
The following procedures will be performed in the following suggested order at Visit 1.  All ocular 
tests at this and all visits will be performed in both eyes (with the exception of conjunctival 
pinching): 
• Obtain written informed consent to participate before any study-required screening 
assessments have been conducted 
• Collect demographics (sex, year of birth, race, ethnicity) 
• Collect systemic medical history and ocular medical history 
• Review of prior and concomitant medications (including ocular medications) 
• Study technician to instill a drop of an Over-The-Counter artificial tear.  At the opinion of 
the Investigator/Study Technician, only individuals who tolerate this drop, will be enrolled. 
• Perform urine pregnancy test (UPT) on women of childbearing potential (see Appendix 1). 
• Measure Best Corrected Visual Acuity (BCVA) (see Appendix 1) 
• Perform slit lamp biomicroscopy and external eye exam (see Appendix 1) 
• Measure IOP (see Appendix 1) 
• Verify subject eligibility based on Inclusion/Exclusion requirements 
• Schedule subject to return for Visit 2 if Visit 1 and 2 are not performed on the same day.  
6.4 
Visit 2 (Randomization/Treatment) – Day 1 
Visit 2 must occur within 3 days of Visit 1 (Screening) if it is not performed on the same day as 
Visit 1.  As noted in Section 6.3, Visit 1 and Visit 2 may be performed on the same day for clinic 
logistics.  If a topical anesthetic is used to measure IOP at Screening (Visit 1), there must be at 
least a 1-hour interval post-anesthetic drops prior to treatment with IMP.  At Visit 2, subjects are 
expected to be in the clinic for approximately 60 minutes post dose.   
 Pre-dose 
The following procedures will be performed in the following order at Visit 2, if Visit 1 was not 
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7 ASSESSMENT OF EFFICACY1 
Assessments of efficacy will be as follows: 
• Assessment of pain questionnaire following conjunctival pinch. 
The timing of this assessment is provided in Section 6.4.3. 
8 ASSESSMENT OF SAFETY 
8.1 
Specification of safety parameters 
The assessment of safety and tolerability will be evaluated by: 
• BCVA (ETDRS) 
• Biomicroscopy and external eye exam of anterior segment including evaluation of cornea, 
conjunctiva and anterior chamber.  
• AEs (post treatment) 
• AEs will be collected at every visit.  See Section 8.3 for details on collection and reporting of 
AEs. 
 
Screening only: 
• Intraocular pressure  
• Urine pregnancy test for females of childbearing potential 
8.2 
The methods and timing for assessing, recording, and analyzing safety parameters 
Methods may be found in Table 1 and Appendix 1.   
8.3 
Adverse events2 
All AEs occurring after the first dose of IMP and throughout the remainder of the study period 
(Follow-up Phone Call) will be considered TEAEs and must be documented on the relevant 
Electronic Data Capture (EDC) pages and in the source documents.  Whenever possible, the 
diagnosis, if available, and not the symptoms should be reported as the AE.  AEs and Serious 
Adverse Events (SAEs) will be coded according to the Medical Dictionary for Regulatory 
Activities (MedDRA).  
 
Documentation of adverse events/adverse reactions includes start date and stop date, severity, 
action(s) taken, seriousness and outcome.   
 Adverse Event (AE) definitions 
An AE is any untoward medical occurrence in a subject or clinical investigation subject, temporally 
associated with the use of a medicinal product, whether or not considered related to the medicinal 
product 
 
                                                 
 
2 Note: This section is referenced to 21 CFR 312.32 (IND safety reports), updated as per “Investigational New Drug 
Safety Reporting Requirements for Human Drug and Biological Products and Safety Reporting Requirements for 
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Note: An AE can therefore be any untoward or unfavorable medical occurrence in a human subject, 
including any abnormal sign (for example, abnormal physical exam or laboratory finding), 
symptom, or disease, temporally associated with the subject’s participation in the research, 
whether or not considered related to the subject’s participation in the research. 
 
Any untoward or unfavorable medical occurrence in a human subject, including any abnormal sign 
(for example, abnormal physical exam or laboratory finding), symptom, or disease, temporally 
associated with the subject’s participation in the research, whether or not considered related to the 
subject’s participation in the research will be documented.  
 
Events meeting the definition of an AE include: 
• Exacerbation of a chronic or intermittent pre-existing condition including either an increase 
in frequency and/or intensity of the condition 
• New conditions detected or diagnosed after study drug administration even though it may 
have been present prior to the start of the study 
• Signs, symptoms, or clinical sequelae of a suspected interaction 
• Signs, symptoms, or clinical sequelae of a suspected overdose of either study drug or a 
concomitant medication (overdose per se will not be reported as an AE/SAE). 
 
Events that do not meet the definition of an AE include: 
• Medical or surgical procedure (e.g., endoscopy, appendectomy); the condition that leads to 
the procedure is an AE 
• Situations where an untoward medical occurrence did not occur (social and/or convenience 
admission to a hospital) 
• Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present or 
detected at the start of the study that do not significantly worsen 
• The disease/disorder being studied, or expected progression, signs, or symptoms of the 
disease/disorder being studied, unless rescue medication or other medical treatment is 
required. 
 
8.3.1.1 Life-threatening adverse event or life-threatening suspected adverse reaction.   
An adverse event or suspected adverse reaction is considered “life-threatening” if, in the view of 
either the Investigator or Sponsor, its occurrence places the subject at immediate risk of death.  It 
does not include an adverse event or suspected adverse reaction that, had it occurred in a more 
severe form, might have caused death.   
8.3.1.2 Suspected adverse reaction 
Suspected adverse reaction means any adverse event for which there is a reasonable possibility 
that the drug caused the adverse event.  For the purposes of IND safety reporting, ‘‘reasonable 
possibility’’ means there is evidence to suggest a causal relationship between the drug and the 
adverse event.  Suspected adverse reaction implies a lesser degree of certainty about causality than 
adverse reaction, which means any adverse event caused by a drug.   
8.3.1.3 Unexpected adverse event or unexpected suspected adverse reaction.   
An adverse event or suspected adverse reaction is considered ‘‘unexpected’’ if it is not listed in 
the Investigator brochure or is not listed at the specificity or severity that has been observed; or, if 


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an investigator brochure is not required or available, is not consistent with the risk information 
described in the general investigational plan or elsewhere in the current application, as amended.  
For example, under this definition, hepatic necrosis would be unexpected (by virtue of greater 
severity) if the investigator brochure referred only to elevated hepatic enzymes or hepatitis.  
Similarly, cerebral thromboembolism and cerebral vasculitis would be unexpected (by virtue of 
greater specificity) if the Investigator brochure listed only cerebral vascular accidents.  
“Unexpected,” as used in this definition, also refers to adverse events or suspected adverse 
reactions that are mentioned in the Investigator brochure 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. 
 
Note: Any medical condition present prior to administration of the masked study medication which 
remains unchanged or improved should not be recorded as an adverse event at subsequent visits. 
 Serious Adverse Event (SAE) definitions 
An SAE is an AE that:  
• Results in death. 
• Is life-threatening (an AE is life-threatening if the subject was at immediate risk of death 
from the event as it occurred, i.e., it does not include a reaction that might have caused 
death if it had occurred in a more serious form). 
• Requires or prolongs inpatient hospitalization.  Complications occurring during 
hospitalization are AEs or SAEs if they cause prolongation of the current hospitalization.  
Hospitalization or prolonged hospitalization for elective treatment of a pre-existing non 
worsening condition is not, however, considered an AE.   
• Results in persistent or significant disability/incapacity.  (An AE is incapacitating or 
disabling if it results in a substantial and/or permanent disruption of the subject’s ability to 
carry out normal life functions). 
• Is a congenital anomaly/birth defect. 
• Is an important medical event that may jeopardize the subject or may require medical 
intervention to prevent one of the outcomes listed above. 
 
Medical and scientific judgment is required to decide if prompt notification is required in situations 
other than those defined for SAEs above.  This can include an AE that suggests any significant 
hazard, contraindication, adverse event, or precaution that may be associated with the use of the 
IMP. 
8.3.2.1 Recording an AE and/or SAE including Onset Date, End Date  
When an AE/SAE occurs, it is the responsibility of the investigator to review all documentation 
(eg, hospital progress notes, laboratory reports, and diagnostics reports) related to the event. The 
investigator will then record all relevant AE/SAE information in the CRF. 
If an AE starts but does not end before the final visit, the Investigator must make a reasonable 
effort to establish the outcome and the end date of the AE.  If this is not possible (e.g., because the 
AE is still ongoing) or the subject is lost to follow-up, there will be no end date for the AE and the 
status will be recorded as “ongoing.”  For all AEs that resolve, resolve with sequelae, or have a 
fatal outcome, an end date must be provided. 


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The investigator will attempt to establish a diagnosis of the event based on signs, symptoms, and/or 
other clinical information. Whenever possible, the diagnosis (not the individual signs/symptoms) 
will be documented as the AE/SAE. 
8.3.2.2 Assessment of Causality 
Every effort will be made by the Investigator to assess the relationship of the AE, if any, to the 
IMP.  Causality should be assessed as unrelated or related using the categories defined in Table 2.   
 
Table 2: AE Assessment of Causality 
Unrelated: 
• Clinical event with an incompatible time relationship to IMP 
administration, and that could be explained by underlying disease or other 
drugs or chemicals or is incontrovertibly not related to the IMP, or 
• Clinical event whose time relationship to IMP makes a causal connection 
improbable, but that could plausibly be explained by underlying disease 
or other drugs or chemicals. 
Related: 
• Clinical event with a reasonable time relationship to IMP, but that could 
also be explained by concurrent disease or other drugs or chemicals, or 
• Clinical event with a reasonable time relationship to IMP and is unlikely 
to be attributed to concurrent disease or other drugs or chemicals, or 
• Clinical event with plausible time relationship to IMP, and that cannot be 
explained by concurrent disease or other drugs or chemicals. 
Abbreviations:  AE=Adverse Event; IMP=investigational medicinal product 
8.3.2.3 Assessment of Severity (Intensity) 
The severity of an adverse event is defined as a qualitative assessment of the level of discomfort 
of an adverse event as is determined by the Investigator or reported to him/her by the subject.  The 
assessment of intensity is made irrespective of study medication relationship or seriousness of the 
event.  The investigator will make an assessment of intensity for each AE and SAE reported during 
the study and assign it to 1 of the following categories: and should be evaluated according to the 
scale in Table 3. 
 
Table 3: Assessment of Severity (Intensity) 
Mild 
An event that is easily tolerated by the participant, causing minimal discomfort 
and not interfering with everyday activities. 
Moderate 
An event that causes sufficiently discomfort and interferes with normal 
everyday activities. 
Severe 
An event that prevents normal everyday activities. An AE that is assessed as 
severe should not be confused with a SAE. Severe is a category utilized for 
rating the intensity of an event; and both AEs and SAEs can be assessed as 
severe. 
 
Please note: the term “severe” is used to describe the intensity (severity, see above) of an 
event/reaction; the event/reaction itself may be of relatively minor medical significance (such as 
severe headache).  This is not the same as a “Serious” Adverse Event, which is based on 
subject/event outcome or action criteria usually associated with events that pose a threat to the 




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of the last visit, efforts will be made to follow up until the adverse event/adverse reaction is 
resolved or stabilized, the subject is lost to follow-up, or there is other resolution to the event.  
9 Statistics 
9.1 
Primary Hypotheses 
H01: 
The percentage of subjects with no pain within 5 minutes is NOT different between 
subjects treated with AG-920 and Placebo.  
 
H11: 
The percentage of subjects with no pain within 5 minutes is different between 
subjects treated with AG-920 and Placebo.  
9.2 
Sample Size Considerations 
Efficacy:   
With 60 subjects per treatment group and assuming 15% response in the vehicle group, this study 
will have 88% power to detect a treatment effect of at least 25% between AG-920 and vehicle 
(α=0.05, two-sided).  
9.3 
Analysis populations 
Randomized Population: The randomized population will include all subjects who were 
randomized to treatment.  Baseline (screening) variables and demographic characteristics will be 
summarized for this population. 
 
Intent-to-Treat Population (ITT): The ITT population will include all randomized subjects who 
have received at least one dose (2 drops) of study medication.  This population will be the primary 
population for the efficacy analyses and all efficacy variables will be summarized using this set of 
subjects.  The ITT population will include subjects as randomized. 
 
Per-protocol population (PP): The PP population is a subset of the ITT population, which will 
include those subjects (and their visits) who do not have major protocol violations likely to 
seriously affect the primary outcome of the study as judged by a masked evaluation prior to the 
unmasking of the study treatment.  This population will be the secondary population for efficacy 
analyses and all efficacy variables will be summarized using this set of subjects.  If the PP and ITT 
populations are exactly the same and there are no missing data, then additional efficacy analyses 
on the PP population will not be performed.  The PP population will include subjects as treated. 
 
Safety Population: The safety population will include all randomized subjects who have received 
at least one dose (2 drops) of study medication.  This population will be used to summarize the 
safety variables and will summarize subjects as treated.   
9.4 
Statistical methods to be employed 
 General Considerations 
All continuous study assessments will be summarized by treatment and time point (as applicable) 
using descriptive statistics (n, mean, median, standard deviation, minimum, and maximum).  All 
categorical study assessments will be summarized by treatment and time point (as applicable) 
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 Analysis of Safety 
Slit lamp biomicroscopy and external eye exam measures will be summarized at each measured 
time point using discrete summary statistics. 
 
Visual Acuity data will be summarized at each time point using both continuous summaries 
(Logarithmic Minimum Angle of Resolution, logMAR), including change from baseline, and 
discrete summaries, including change from baseline in the number of lines and the proportion of 
subjects with a worsening of ≥ 3 lines from baseline.   
 Adverse Events 
Verbatim descriptions of AEs will be mapped to MedDRA thesaurus terms and be presented in a 
data listing.  Treatment emergent AEs, those that occur after the first dose of study medication, 
will be summarized by treatment group using frequency counts and percentages for each system 
organ class (SOC) and preferred term (PT) within each SOC.  Summaries will be presented 
separately for ocular and non-ocular AEs.  These summaries will also be presented by the 
relationship to the Investigational Medical Product (related, unrelated) and by severity of the AE 
(mild, moderate, severe).  Fisher’s exact test will be used to test the difference in proportions of 
subjects with each AE between treatment groups (SOC and PT).   
9.5 
Procedure for accounting for missing, unused, or spurious data 
A minimal amount of missing data is expected in this study because of the short duration and the 
subjects being in the clinic on Day 1.  For the ITT population, any missing data on Day 1 will be 
imputed using the method of last observation carried forward (LOCF), where the closest non-
missing value prior to the missing value will be carried forward and imputed for the missing value.  
The PP and Safety populations will be based on observed data only (without imputation).  Any 
missing, unused, or spurious data will be noted and explained in the final statistical report. 
9.6 
Procedure for reporting deviations from the statistical plan 
Any deviations from the statistical plan will be described and a justification given in the final 
statistical report. 
9.7 
Data listings 
Data listings will be prepared for all data on the database. 
10 QUALITY CONTROL AND QUALITY ASSURANCE 
10.1 Audit and Inspection 
Study centers and study documentation may be subject to Quality Assurance audits during the 
study by the Sponsor or its designee.  In addition, inspections may be conducted by regulatory 
authorities at their discretion. 
10.2 Direct Access to Source Data Documents 
Authorized representatives of the Sponsor, a regulatory authority, an IEC or IRB may visit the site 
to perform audits or inspections, including source data verification.  The purpose of a Sponsor 
audit or inspection is to systematically and independently examine all study-related activities and 
documents to determine whether these activities were conducted, and data were recorded, 
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regulatory requirements.  The Investigator should contact the Sponsor immediately if contacted by 
a regulatory agency about an inspection. 
10.3 Clinical Monitoring 
Data for each subject will be recorded in a source document and in EDC for each subject who 
signs an informed consent form (ICF) and is administered IMP. 
 
In accordance with current GCP and International Conference of Harmonisation (ICH) guidelines, 
the study monitor will carry out source document verification at regular intervals to ensure that the 
data collected in EDC are accurate and reliable.  
 
The Investigator must permit the monitor, the IEC/IRB, the Sponsor’s internal auditors, and 
representatives from regulatory authorities’ direct access to all study-related documents and 
pertinent hospital or medical records for confirmation of data contained within EDC.   
Due to the COVID-19 Pandemic, remote monitoring visits may replace some of the onsite 
monitoring visits.  It will still be the responsibility of the monitor to discuss the study and any 
issues with the Principal Investigator via telephone or videoconference.  
10.4 Data Management and Coding 
The CRO will be responsible for data management per their SOPs.  This will include setting up a 
relevant database and data transfer mechanisms, along with appropriate validation of data and 
resolution of queries.  Data generated within this clinical study will be handled according to the 
relevant standard operating procedures of the data management and biostatistics departments of 
the CRO. 
 
All data for subjects who sign an ICF and receive a dose of IMP will be recorded via EDC.  
Subjects who are screened but found ineligible for the study and who do not receive IMP will be 
considered screen failures.  The reason for exclusion from the study will be recorded.   
 
Study centers will enter data directly into the EDC via a secure internet connection.  Data entered 
into EDC must be verifiable against source documents at the study center.  Data recorded directly 
into EDC, if any, will be identified and the EDC will be considered the source document.  Any 
changes to the data entered into the EDC system will be recorded in the audit trail and will be FDA 
Code of Federal Regulations (CFR) 21 Part 11 compliant. 
 
MedDRA will be used to code AEs.  Medications will be coded by WHODrug.  Missing or 
inconsistent data will be noted within the EDC system and queried with the Investigator for 
clarification.  Subsequent modifications to the database will be documented. 
11 ETHICAL CONSIDERATIONS AND COMPLIANCE WITH GOOD 
CLINICAL PRACTICE 
11.1 Independent Ethics Committee or Institutional Review Board 
Before initiation of the study at each study center, the protocol, the ICF, other written material 
given to the subject, and any other relevant study documentation will be submitted to the 
appropriate IEC/IRB.  Written approval of the study and all relevant study information must be 
obtained before the study center can be initiated or the IMP released to the Investigator.  Any 


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necessary extensions or renewals of IEC/IRB approval must be obtained for changes to the study 
such as amendments to the protocol, the ICF or other study documentation. The written approval 
of the IEC/IRB together with the approved ICF must be filed in the study files. 
The Investigator will report promptly to the IEC/IRB 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 status to the IEC/IRB as required.  On completion of the study, the IEC/IRB 
will be notified that the study has ended. 
11.2 Regulatory Authorities 
Relevant study documentation will be submitted to the regulatory authorities of the participating 
countries, according to local/national requirements, for review and approval before the beginning 
of the study.  On completion of the study, the regulatory authorities will be notified that the study 
has ended. 
11.3 Ethical Conduct of the Study 
The Investigator(s) and all parties involved in this study should conduct the study in adherence to 
the ethical principles based on the Declaration of Helsinki, GCP, ICH guidelines, and the 
applicable national and local laws and regulatory requirements. 
11.4 Written Informed Consent 
The process of obtaining informed consent must be in accordance with applicable regulatory 
requirement(s) and must adhere to GCP. 
 
The Principal Investigator(s) at each center will ensure that the subject is given full and adequate 
oral and written information about the nature, purpose, possible risk, inconveniences, and potential 
benefit of the study.  Subjects must also be notified that they are free to discontinue from the study 
at any time.  The subject should be given the opportunity to ask questions and allowed time to 
consider the information provided.  The subject should be given every opportunity to ask for 
clarification of any points s/he does not understand and, if necessary, ask for more information.  
The subject will be required to sign and date the ICF.  After signatures are obtained, the ICF will 
be kept and archived by the Investigator in the Investigator’s study file.  A signed and dated copy 
of the subject ICF will be provided to the subject.  The Principal Investigator(s) must maintain the 
original, signed ICF. 
 
The Investigator is responsible for ensuring that no subject undergoes any study-related 
examination or activity before the subject has given written informed consent to participate in the 
study. 
 
It should be emphasized that the subject may decline to participate in the study and may withdraw 
from the study at any time without consequences for their further care or penalty or loss of benefits 
to which the subject is otherwise entitled.  
 
If new information becomes available that may be relevant to the willingness of the subject to 
continue in the study, a new ICF will be approved by the IEC(s)/IRB(s) (and regulatory authorities, 
if required). The subject will be informed about this new information and reconsent will be 
obtained. 


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11.5 Subject Confidentiality 
Monitors, auditors, and other authorized agents of the Sponsor and/or its designee, the 
IEC(s)/IRB(s) approving this research, as well as that of any other applicable agency, will be 
granted direct access to the study subjects’ original medical records for verification of clinical 
study procedures and/or data, without violating the confidentiality of the subjects to the extent 
permitted by the law and regulations.  In any presentations of the results of this study or in 
publications, the subjects’ identity will remain confidential. 
12 DATA HANDLING AND RECORD KEEPING 
All procedures for the handling and analysis of data will be conducted using good computing 
practices meeting ICH and U.S. Food and Drug Administration (FDA) guidelines for the handling 
and analysis of data for clinical trials. 
12.1 Data quality control and reporting 
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.   
12.2 Inspection of Records 
American Genomics or designee will be allowed to conduct site visits to the investigation facilities 
for the purpose of monitoring any aspect of the study.  The Investigator agrees to allow the monitor 
to inspect the drug storage area, IMP stocks, drug accountability records, subject charts and study 
source documents, and other records pertaining to study conduct. 
12.3 Records retention 
Essential documents are those documents that individually and collectively permit evaluation of 
the study and quality of the data produced.  After completion of the study, all documents and data 
relating to the study will be kept in an orderly manner by the Investigator in a secure study file.  
This file will be available for inspection by the Sponsor or its representatives.  Essential documents 
should be retained for 2 years after the final marketing approval in an ICH region or for at least 2 
years from the discontinuation of clinical development of the investigational product.  It is the 
responsibility of the Sponsor or its representatives to inform the study center when these 
documents no longer need to be retained.  The Investigator must contact the Sponsor before 
destroying any study related documentation.  In addition, all subject medical records and other 
source documentation will be kept for the maximum time permitted by the hospital, institution, or 
medical practice. 
12.4 Amendments to the protocol 
Modifications of the signed protocol are only possible by approved protocol amendments and with 
the agreement of all responsible persons.  The procedure for approval of a protocol amendment is 
identical to that for approval of the protocol.  The ethics committee must be informed of all 
protocol amendments and should be asked for its opinion as to whether a full re-evaluation of the 
ethical aspects of the study is necessary by the committee.  This should be fully documented.   
 


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The investigator must not implement any deviation from or change to the protocol, without 
discussion with, and agreement by American Genomics, LLC and prior review and documented 
approval/favorable opinion of the amendment from the relevant ethics committee, except where it 
is necessary to eliminate an immediate hazard to study subjects, or where the change(s) involves 
only logistical or administrative aspects of the study (e.g., change in monitor(s), change of 
telephone number(s)).   
 
Protocol amendments will be submitted to the appropriate authority(ies) as required by the 
applicable regulatory requirement(s).   
13 PUBLICATION 
The Sponsor must review and approve any results of the study or abstracts for professional 
meetings prepared by the Investigator(s).  Published data must not compromise the objectives of 
the study. Data from individual study centers in multicenter studies must not be published 
separately. 
14 REFERENCES 
14.1 Published references 
• Bailey IL, Lovie JE. New design principles for visual acuity letter charts. Am J Optom 
Physiol Opt 1976;53:740-5. 
• Ferris FL, Freidlin V, Kassoff A, et al. Relative letter and position difficulty on visual 
acuity charts from the Early Treatment Diabetic Retinopathy Study. Am J Ophthalmol 
1993;116:735-40. 
• 
® package insert, 2018. 
• Sherwood MB, Craven ER, Chou C, DuBiner HB, Batoosingh AL, Schiffman RM, et al. 
Twice-daily 0.2% brimonidine-0.5% timolol fixed-combination therapy vs monotherapy 
with timolol or brimonidine in patients with glaucoma or ocular hypertension: a 12-month 
randomized trial. Arch Ophthalmol. 2006;124(9):1230-8. 
• FDA Guidance on Conduct of Clinical Trials of Medicinal Products during COVID-19 
Public Health Emergency, March 2020; https://www.fda.gov/media/136238/download. 
14.2 Internal references 
• American Genomics, LLC, 
 Sterile Topical Ophthalmic Solution Investigator’s 
Brochure, 2020.  
 


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Appendix 1: Procedures 
Procedures: Best Corrected Visual Acuity 
Introduction: Best corrected visual acuity (Distance) will be measured at Visit 1 and at the end of 
Visit 2.  Visual acuity will be measured using Bailey Lovie charts, ETDRS charts, or their 
equivalents.  Accepted charts are those designed according to the following principles described 
by Bailey and Lovie (1976), Ferris et al (1993) and the National Academy of Science-National 
Research Council (NAS-NRC) Committee on Vision 1980 (1980): 1) letters of equal legibility; 2) 
combine the letters so that each line is of approximately equal difficulty (as described by Ferris et 
al, 1993); 3) present five letters at each acuity level; 4) space rows by the height of the smaller 
letter 5) space letters by the width of same-sized letters and 6) use a logarithmic progression of 
letter size from logMAR (Minimum Angle of Resolution to base 10) -0.3 (20/10) to 1.0 (20/200).   
 
Rationale: Best corrected acuity taken at follow-up visits as a measure of ocular function.  Best 
corrected visual acuity will be measured at screening and frequently throughout the study.   
 
Procedure: Distance visual acuity must be assessed using an Early Treatment of Diabetic 
Retinopathy Study (ETDRS) or equivalent chart.  Visual acuity testing should precede intraocular 
pressure measurement, the administration of topical anesthetic agents, or any examination 
requiring contact with the anterior segment.   
 
Distance visual acuity will be measured with best correction.   
 
The visual acuity chart may be either retro-illuminated (“back-lit”), or reflectance illuminated.  If 
the latter, then the illumination must be checked at regular intervals to be consistent with ETDRS 
guidelines.  Standard charts for a distance from subject to chart of 10 feet to 20 feet must be used.  
Ideally, the subject should be seated.  The right eye should be tested first.  Sites are directed to 
refer to the instructions on the commercial ETDRS charts.  If there is any question, contact the 
Sponsor’s monitor.   
 
The subject should attempt to read each letter, line by line, left to right, beginning with line 1 at 
the top of the chart (20/200 line).  The subjects should be told that the chart has letters only, no 
numbers.  If the subject reads a number, he or she should be reminded that the chart contains no 
numbers, and the examiner should then request a letter in lieu of the number.  The subjects should 
be asked to read slowly, about one letter per second, so as to achieve the best identification of each 
letter.  He/she is not to proceed to the next letter until he/she has given a definite response. 
 
A maximum effort should be made to identify each letter on the chart.  When the subject says 
he/she cannot read a letter, he/she should be encouraged to guess.  If the subject identified two 
letters (e.g., A or B), he/she should be asked to choose one letter and, if necessary, to guess.  When 
it becomes evident that no further meaningful readings can be made despite encouragement to read 
or guess, the examiner should stop the testing for that eye.  However, all letters on the last line 
should be attempted as letter difficulties vary and the last letter may be the only one read correctly.  
The number of letters missed or read incorrectly should be noted.  Repeat with left eye.   
 
In order to provide standardized and well-controlled assessment of visual acuity during the study, 
all visual acuity assessments for a subject must be performed consistently (e.g., the same lighting 
conditions, viewing distance, etc.) during the entire study.   


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15 September 2020 
 
 
The number of letters missed is multiplied by 0.02 and added to the baseline value to determine 
the logMAR visual acuity.  Baseline is defined as the last line for which the subject reads at least 
one letter.   
 
logMAR units VA = Baseline value + (n x 0.02) 
 
If refraction was used at the screening or baseline, then this refraction should be used in the 
phoropter or trial frame for subsequent examinations.  A repeated refraction is not required at these 
subsequent examinations.   
 
A priori, a change of three lines (ETDRS) in visual acuity in either eye is considered clinically 
significant.   
 
For comparison with Snellen, a chart of the Snellen ratio that best corresponds with LogMAR is 
provided in Table 4.  This is provided for convenience only.  Visual Acuity is not to be measured 
by Snellen Acuity charts in this study. 
 
Table 4: Comparison of Snellen ratio and logMAR units 
Snellen ratio 
logMAR 
20/200 
+1.0 
20/160 
+0.9 
20/125 
+0.8 
20/100 
+0.7 
20/80 
+0.6 
20/62.5 
+0.5 
20/50 
+0.4 
20/40 
+0.3 
20/32 
+0.2 
20/25 
+0.1 
20/20 
+0.0 
20/16 
-0.1 
20/12.5 
-0.2 
20/10 
-0.3 
 
Procedures: Measurement of intraocular pressure: 
Local anesthetic may be applied in order to facilitate IOP measurements with Tonopen® or 
Goldmann Tonometer.   
 
If a Goldmann tonometer is used, then two consecutive IOP measurements of each eye are to be 
obtained. If the 2 measurements differ by more than 2 mm Hg, a third measurement should be 
obtained.  IOP will be analyzed as the mean of 2 measurements or as the median of 3 measurements 
(Sherwood et al 2006). 
 
If local anesthetic is applied in order to facilitate IOP measurements, there must be at least a 1-
hour interval post-anesthetic drops prior to treatment with IMP.  Subjects should be reminded that 


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Clinical Study Protocol 
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Protocol AG-920-CS301 Amendment 1 
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15 September 2020 
 
topical local anesthetics may cause a mild stinging or burning sensation upon instillation.  This 
should not be considered as “pain” when conjunctival pinching is conducted.  
 
If a Goldmann Tonometer is used, it must be regularly calibrated per manufactures 
recommendations.  
 
Procedures: Biomicroscopy and External Eye Exam 
The subject will be seated while being examined. 
 
External examination and biomicroscopy will be performed using a slit lamp.  Magnification will 
be consistent with standard clinical practice. 
 
The clinician will examine and grade the eyelid.  Observations will be documented on the 
appropriate CRF.  The clinician will examine the conjunctiva, cornea, anterior chamber, iris, pupil 
and lens of the eye with the aid of a slit lamp, which is a table-mounted binocular microscope.   
 
The Sponsor recommends that the same examiner should conduct all biomicroscopy examinations 
at each time point and at each visit for a given subject. 
 
Biomicroscopic grading will be done as follows: 
 
LID 
Erythema 
None (0)= 
Normal, without any redness, or less than mild 
Mild (+1)= 
A low grade flushed reddish color 
Moderate (+2)= 
Diffused redness encompassing the entire lid margin 
Severe (+3)= 
Deep diffused reddish color of lid margins and superior or inferior eyelid 
 
Edema 
None (0)= 
Normal, no swelling of the lid tissue, or less than mild 
Mild (+1)= 
Slight diffuse swelling above normal 
Moderate (+2)= 
General swelling 
Severe (+3)= 
Extensive swelling of the eyelid(s), with or without eversion of upper and/or 
lower lids. 
 
CONJUNCTIVA  
Hyperemia 
None (0)=  
Normal. Appears white with a small number of conjunctival blood vessels easily 
observed.  
Mild (+1)= 
Prominent, pinkish-red color of both the bulbar and palpebral conjunctiva; 
Moderate (+2)= 
Bright, scarlet red color of the bulbar and palpebral conjunctiva 
Severe (+3)= 
“Beefy Red” with petechiae --- Dark red bulbar and palpebral conjunctiva with 
evidence of subconjunctival hemorrhage   
 
Edema 
None (0)= 
Normal, no swelling of the conjunctiva or less than mild 
Mild (+1)= 
Slight diffuse or regional swelling of the conjunctiva 
Moderate (+2)= 
General swelling of the conjunctiva 
Severe (+3)= 
Extensive swelling of the conjunctiva 
 
CORNEA 
Edema 
None (0)= 
Transparent and clear or less than mild 


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Protocol AG-920-CS301 Amendment 1 
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Mild (+1) = 
Dull glassy appearance 
Moderate (+2)= 
Dull glassy appearance of epithelium with large number of vacuoles 
Severe (+3) = 
Stromal edema, localized or diffuse, with stromal striae 
 
 
ANTERIOR CHAMBER 
Cells 
None (0)= 
No cells seen or less than mild 
Mild (+ 1) = 
+ cells 
Moderate (+2) = 
++ cells 
Severe (+3) = 
+++ cells 
Hypopyon (+4)= 
++++ cells, Hypopyon Formation (indicate size of hypopyon) 
 
 
Flare 
None (0)= 
No Tyndall effect or less than mild 
Mild (+1) = 
Tyndall beam in the anterior chamber has a mild intensity 
Moderate (+2) = 
Tyndall beam in the anterior chamber is of strong intensity 
Severe (+3) = 
Tyndall beam is very intense.  The aqueous has a white, milky appearance 
 
Procedures: Urine  
Urine pregnancy tests will be performed on all women of childbearing potential at Visit 1 (prior 
to treatment).   
 
 
 






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Clinical Study Protocol 
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Clinical Study Protocol 
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Protocol AG-920-CS301 Amendment 1 
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15 September 2020 
 
Appendix 4: Investigator Signature Page 
 
Protocol Title: 
A Randomized, Double-Masked, Vehicle-Controlled, Parallel 
Evaluation of the Local Anesthetic Effect of 
 Sterile 
Topical Ophthalmic Solution 
Protocol Number: 
AG-920-CS301 
Confidentiality and Current Good Clinical Practice Compliance Statement 
I, the undersigned, have reviewed this protocol (and amendments), including appendices, and I 
will conduct the study as described in compliance with this protocol (and amendments), GCP, and 
relevant ICH guidelines. 
Once the protocol has been approved by the IEC/IRB, I will not modify this protocol without 
obtaining prior approval of American Genomics, LLC and of the IEC/IRB.  I will submit the 
protocol amendments and/or any ICF modifications to American Genomics, LLC and IEC/IRB, 
and approval will be obtained before any amendments are implemented. 
I understand that all information obtained during the conduct of the study with regard to the 
subjects’ state of health will be regarded as confidential.  No subjects’ name will be disclosed.  All 
subjects will be identified by assigned numbers on all EDC, laboratory samples, or source 
documents forwarded to American Genomics, LLC.  Clinical information may be reviewed by 
American Genomics, LLC or its agents or regulatory agencies.  Agreement must be obtained from 
the subject before disclosure of subject information to a third party. 
Information developed in this clinical study may be disclosed by American Genomics, LLC, to 
other clinical Investigators, regulatory agencies, or other health authority or government agencies 
as required. 
Investigator Signature 
Date 
Printed Name 
 
Institution 
 
 

SAP_001.pdf: 








American Genomics, LLC 
Statistical Analysis Plan 
Confidential 
 
Protocol AG-920-CS301 
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Table of Contents 
List of Abbreviations .............................................................................................................................. 9 
1. 
Administrative Structure ........................................................................................................... 10 
1.1. 
Study Sponsor ....................................................................................................................... 10 
1.2. 
Introduction and Scope ......................................................................................................... 10 
1.3. 
Responsibility of Data Management Team ........................................................................... 10 
1.4. 
Responsibility of SAS Programmer or Statistician ............................................................... 10 
2. 
Study Objectives and Endpoints ............................................................................................... 10 
2.1. 
Study Objectives ................................................................................................................... 10 
2.2. 
Endpoints .............................................................................................................................. 11 
3. 
Study Design ............................................................................................................................. 11 
3.1. 
Overall................................................................................................................................... 11 
3.2. 
Sample Size ........................................................................................................................... 11 
3.3. 
Investigational Products ........................................................................................................ 11 
3.4. 
Randomization ...................................................................................................................... 12 
3.5. 
Post Treatment and Pinch Test .............................................................................................. 12 
3.6. 
Study Schedule ...................................................................................................................... 13 
4. 
Analysis Sets ............................................................................................................................. 14 
4.1. 
Randomized Population ........................................................................................................ 14 
4.2. 
Intent-to-Treat (ITT) Population ........................................................................................... 14 
4.3. 
Per-Protocol (PP) Population ................................................................................................ 14 
4.4. 
Safety Population .................................................................................................................. 14 
5. 
General Aspects of Statistical Analysis .................................................................................... 14 
5.1. 
Presentation of Summaries and Analyses ............................................................................. 14 
5.2. 
Precision of Display .............................................................................................................. 15 
5.3. 
Strategy of Pooling the Data Across Centers ........................................................................ 15 
5.4. 
Coding Dictionaries .............................................................................................................. 15 
5.5. 
Handling of Missing/Partial Dates for Adverse Events ........................................................ 15 
5.6. 
Handling of Data from Unscheduled Visits .......................................................................... 15 
6. 
Subject Disposition ................................................................................................................... 16 
7. 
Demographic and Baseline Measurements ............................................................................... 16 
7.1. 
Demographics ....................................................................................................................... 16 
7.2. 
Inclusion/Exclusion Criteria ................................................................................................. 16 
7.3. 
Protocol Deviations ............................................................................................................... 16 
7.4. 
Medical History .................................................................................................................... 17 
7.5. 
Prior and Concomitant Medication ....................................................................................... 17 
7.6. 
Investigational Product Administration ................................................................................ 17 
8. 
Evaluation of Efficacy .............................................................................................................. 17 
8.1. 
Primary Endpoint .................................................................................................................. 18 
8.2. 
Secondary Endpoints ............................................................................................................ 18 
8.3. 
Interim Analyses ................................................................................................................... 19 
9. 
Evaluation of Safety .................................................................................................................. 19 
9.1. 
Adverse Events ..................................................................................................................... 19 
9.2. 
Other Safety Parameters........................................................................................................ 21 








American Genomics, LLC 
Statistical Analysis Plan 
Confidential 
 
Protocol AG-920-CS301 
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List of Abbreviations 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
ADaM 
Analysis Data Model 
AE 
Adverse Event 
ATC 
Anatomical Therapeutic Chemical 
BCVA 
Best Corrected Visual Acuity 
CI 
Confidence Interval 
CRF 
Case Report Form 
CSR 
Clinical Study Report 
DDD 
Defined Daily Dose 
EDC 
Electronic Data Capture 
GCP 
Good Clinical Practice 
ICH 
International Conference on Harmonization 
IMP 
Investigational Medicinal Product 
IOP 
Intraocular Pressure 
ITT 
Intent to Treat 
LOCF 
Last Observation Carried Forward 
MedDRA 
Medical Dictionary for Regulatory Activities 
OD 
Right Eye 
OS 
Left Eye 
OTC 
Over The Counter 
PP 
Per Protocol 
PT 
Preferred Term 
SAE  
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SD 
Standard Deviation 
SDTM 
Study Data Tabulation Model 
SOC 
System Organ Class 
SOP 
Standard Operating Procedure 
TEAE 
Treatment Emergent Adverse Event 
TLFs 
Tables, Listings, and Figures 
WHOCC 
World Health Organization Collaborating 
Center 


American Genomics, LLC 
Statistical Analysis Plan 
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Protocol AG-920-CS301 
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1. Administrative Structure 
1.1. Study Sponsor 
 
 
 
1.2. Introduction and Scope 
This Statistical Analysis Plan (SAP) provides detailed statistical methodology for the analysis of 
data from protocol number AG-920-CS301, Version 1.0 (09-Jun-2020) that will be presented in 
the Clinical Study Report (CSR). The variables and methods described in this plan supersede those 
described in the protocol. Any deviations from the analyses described in the final SAP will be 
noted in the CSR. 
 
The SAP may also serve as a reference for the creation of any outputs required outside of the CSR, 
e.g., abstracts, posters, presentations, manuscripts and management updates. Data used for these 
analyses will have a status aligned to the database lock guidance. 
 
The statistical analyses will be done in accordance with the ICH-E9 guidelines “Statistical 
Principles for Clinical Trials” and applicable local guidelines. 
1.3. Responsibility of Data Management Team 
The responsibility of providing clean data for the statistical analysis lies with TAB Clinical Data 
Management team. The Data Management team has designed the eCRF in the Castor EDC system, 
version 2020.2.8, in order to perform data capture, data review and data cleaning including a query 
management process. The data review, cleaning and locking are described in detail in the Data 
Management Plan and the related SOPs. 
1.4. Responsibility of SAS Programmer or Statistician 
SAS programmers or statisticians will be responsible for programming the statistical tables, 
listings and figures (TLFs), as well as the ADaM/STDM datasets as per the approved SAP for the 
study clinical data. 
2. Study Objectives and Endpoints 
2.1. Study Objectives 
Primary Objective: 
• To evaluate anesthetic efficacy of AG-920 
Secondary Objectives: 
• To evaluate how long it takes one dose of AG-920 to anesthetize the eye 
• To evaluate how long one dose of AG-920 anesthetizes the eye 
• To evaluate the safety and tolerability of AG-920 








 
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4. 
Analysis Sets 
4.1. Randomized Population 
The randomized population will include all subjects who are randomized to treatment. Baseline 
(screening) variables and demographic characteristics will be summarized for this population. 
4.2. Intent-to-Treat (ITT) Population 
The ITT population will include all randomized subjects who have received at least one dose (drop) 
of study medication. This population will be the primary population for the efficacy analyses and 
all efficacy variables will be summarized using this set of subjects. The ITT population will include 
all subjects as randomized, even if there are subjects who are given the wrong study medication. 
4.3. Per-Protocol (PP) Population 
The PP population is a subset of the ITT population, which will include those subjects (and their 
visits) who do not have major protocol violations likely to seriously affect the primary outcome of 
the study as judged by a masked evaluation prior to the unmasking of the study treatment. This 
population will be the secondary population for the efficacy analyses and all efficacy variables 
will be summarized using this set of subjects. If the PP and ITT populations are exactly the same 
and there are no missing data, then additional efficacy analyses on the PP population will not be 
performed. The PP population will include subjects as treated, so if there are subjects who are 
given the wrong study medication, then they will be included in the treatment group based on what 
they received. 
4.4. Safety Population 
The Safety population will include all randomized subjects who have received at least one dose 
(drop) of study medication. This population will be used to summarize the safety variables and 
will include all subjects as treated. 
5. General Aspects of Statistical Analysis 
All TLFs will be independently checked for consistency, integrity and in accordance with the 
applicable SOP(s). The shells for the TLFs are presented in a separate document. 
5.1. Presentation of Summaries and Analyses 
In general, continuous data will be summarized by descriptive statistics, i.e., n, mean, standard 
deviation (SD), median, minimum and maximum values, and categorical data will be summarized 
using counts (N: number of subjects per treatment group, n: number of subjects with non-missing 
values) and percentages. Percentages will be suppressed when the count is zero, however the 
category will still be displayed. The denominator for all percentages will be the number of subjects 
per treatment in the analysis set of interest (%: n/N*100), unless otherwise indicated. 
 


 
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In general, and unless specified otherwise, all data summaries will be sorted by the parameter 
under consideration and then by visit. Listings will be sorted as per subject ID, the parameter under 
consideration and then by visit. 
 
Unless otherwise stated, baseline will be defined as the last value of the assessment recorded prior 
to first study drug administration. 
5.2. Precision of Display 
Means and medians will be rounded off to one decimal place more than the reported data values. 
Standard deviations will be rounded off to two decimal places more than the reported data values. 
Minimum and maximum values will be of the same precision as that of the reported data. 
 
The precision of display for the variables will be up to one decimal place for percentage values, 
e.g., xx (xx.x), where the value inside the parentheses represents the percentage value. Counts (n) 
will be displayed as integers. 
 
All P values will be reported based on two-sided significance tests and all P values less than or 
equal to 0.05 (≤0.05) will be considered statistically significant. P values will be reported to four 
decimal places or as <0.0001. 
5.3. Strategy of Pooling the Data Across Centers 
Not applicable for this single-site study. 
5.4. Coding Dictionaries 
The Medical Dictionary for Regulatory Activities (MedDRA) version 23.0 or higher will be used 
to code the adverse events and medical history. The World Health Organization Collaborating 
Center (WHOCC) Anatomical Therapeutic Chemical/Defined Daily Dose (ATC/DDD) Index 
2020 classification system will be used to code the concomitant medications. 
5.5. Handling of Missing/Partial Dates for Adverse Events 
Because of the short duration of the study and only a single dose of study drug being given, missing 
or partial start or end dates for AEs are not expected. Therefore, the duration of most AEs should 
be able to be calculated. If there is a missing or partial start or end date for an AE, then the duration 
of that event will be missing. 
 
Missing or partial dates will be listed in the subject data listing as they appear in the database. 
5.6. Handling of Data from Unscheduled Visits 
Data from unscheduled visits will only be displayed in subject data listings and will not be used in 
calculating summary statistics. However, if AEs occur or are reported during an unscheduled visit 
then those will be summarized. 


 
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6. Subject Disposition 
The following subject disposition details will be summarized by treatment group and overall and 
listed to include the number and percentage of subjects: 
• Informed Consent signed 
• Screened 
• Screened Failure 
• Dosed or did not initiate treatment 
• Completed the study as per protocol 
• Discontinued study 
• Withdrew from the study 
• Included in each analysis set 
These data will also be listed. Individual reasons for withdrawal will be presented in listings for 
all enrolled subjects. 
7. Demographic and Baseline Measurements 
7.1. Demographics 
The following demographic variables and subject characteristics will be summarized for the 
enrolled (randomized) population (based upon screening visit data): 
• Age (Completed years) 
• Sex 
• Ethnicity 
• Race 
• Iris color 
Age will be summarized using descriptive statistics (n, mean, SD, median, minimum and 
maximum), and sex, ethnicity, race and iris color will be summarized using counts and 
percentages. All summary data will be presented by treatment group and overall. A subject-wise 
data listing of all demographic characteristics will also be presented. 
7.2. Inclusion/Exclusion Criteria 
The inclusion and exclusion criteria will be listed together with the overall eligibility for each 
subject. 
7.3. Protocol Deviations 
Protocol deviations are considered any deviation from the clinical study protocol relating to a 
subject, and include the following: 
• Inclusion/exclusion criteria deviations. 
• Dosing deviations (e.g., incorrect treatment received or administered to the incorrect eye). 


 
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• Time window deviations for safety assessments. 
• Subjects receiving prohibited concomitant medications. 
• Other procedural and study conduct deviations recorded by the site on the protocol 
deviation log. 
All protocol deviations will be reviewed and finalized prior to database lock in order to define the 
per-protocol analysis population for the study. 
 
All protocol deviations will be listed by subject. 
7.4. Medical History 
Medical history will be coded using MedDRA version 23.0 or higher. All medical history events 
will be listed by system organ class (SOC) and preferred term (PT) and will be summarized by 
treatment group and overall using the Safety population. 
7.5. Prior and Concomitant Medication 
All medications which the subject has taken within 30 days prior to screening and during the study 
will be recorded on the CRF. Concomitant medications are defined as those with a start date on or 
after the date of first IMP use and those with a start date prior to the first IMP use but with an end 
date on or after the date of first IMP use. If the medication dates are incomplete and it is not clear 
whether the medication was concomitant, it will be assumed to be concomitant. 
 
Concomitant medication drug names will be coded using the WHOCC ATC/DDD Index 2020. All 
concomitant medications will be listed by reported generic name and coded medication term, 
indication, dose, dose unit, frequency, route of administration, start date and end date or ‘ongoing’ 
and duration of use. 
7.6. Investigational Product Administration 
Listings will be provided for the date of administration, kit number, study eye, time of each drop 
administration, treatment given, etc. for each enrolled subject. 
8. Evaluation of Efficacy 
The primary analysis population for efficacy will be the ITT population as defined in Section 4.2, 
and the secondary analysis population for efficacy will be the PP population as defined in Section 
4.3. The PP population will be used to determine the robustness of the results, and all efficacy 
variables will be analyzed using both sets of subjects. 
 
A minimal amount of missing data is expected in this study because of the short duration and the 
subjects being in the clinic on Day 1. For the ITT population, any missing efficacy data on Day 1 
will be imputed using the method of last observation carried forward (LOCF), where the closest 
non-missing value prior to the missing value will be carried forward and imputed for the missing 
value. The PP population will be based on observed data only (without imputation). Any missing, 
unused, or spurious data will be noted and explained in the final clinical study report. 






 
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For the following cases of a partial or missing start date or time, the adverse event will be 
considered treatment emergent: 
• The event started on Day 1 but has a missing start time 
• The event has a partial start date and the month and/or year are the same as the month 
and/or year of Visit 2 or later 
• The start date is missing 
Severity of event 
For AEs, the following guidelines will be used to describe severity: 
 
Mild 
An event that is easily tolerated by the participant, causing minimal 
discomfort and not interfering with everyday activities. 
Moderate 
An event that causes sufficient discomfort and interferes with normal 
everyday activities. 
Severe 
An event that prevents normal everyday activities. An AE that is assessed as 
severe should not be confused with a SAE. Severe is a category utilized for 
rating the intensity of an event; and both AEs and SAEs can be assessed as 
severe. 
 
Relationship of event to study drug 
All AEs will have their relationship to study drug assessed by the Principal Investigator (PI) who 
will examine and evaluate the subject based on temporal relationship and his/her clinical judgment. 
The degree of certainty about causality will be graded using the following two categories: 
 
Unrelated 
• Clinical event with an incompatible time relationship to IMP 
administration, and that could be explained by underlying disease or other 
drugs or chemicals or is incontrovertibly not related to the IMP, or 
• Clinical event whose time relationship to IMP makes a causal connection 
improbable, but that could plausibly be explained by underlying disease or 
other drugs or chemicals. 
Related 
• Clinical event with a reasonable time relationship to IMP, but that could 
also be explained by concurrent disease or other drugs or chemicals, or 
• Clinical event with a reasonable time relationship to IMP and is unlikely to 
be attributed to concurrent disease or other drugs or chemicals, or 
• Clinical event with plausible time relationship to IMP, and that cannot be 
explained by concurrent disease or other drugs or chemicals. 
Abbreviations: IMP=investigational medicinal product 
 


 
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If the PI is unable to determine the relationship between the adverse event and study drug 
administration, such a case will be classified as related to study drug administration, and the whole 
episode will be assessed in terms of expedited reporting to the regulatory authorities until the PI 
determines the final causality. 
 
In general, events will be summarized by counts and percentages. 
 
The following information will be summarized for TEAEs. All summaries will be presented by 
treatment group and overall: 
• Overall Summary of TEAEs. 
• TEAEs by SOC and preferred term (PT). 
• TEAEs by SOC, PT, and causality. 
• TEAEs by SOC, PT, and severity. 
• TEAEs by SOC, PT, and verbatim terms. 
• Listing of Serious AEs. 
• Listing of AEs leading to discontinuation. 
Also, a listing of all adverse events will be provided and will include the reported term, PT, SOC, 
start and end dates, seriousness, frequency of occurrence, severity, action taken regarding 
investigational drug, outcome, and relationship to investigational drug. 
 
For each AE and SAE, the frequency of the event in the two arms will be summarized. The 
incidence of each event will be calculated as the percentage of subjects experiencing a particular 
AE, out of the total number of subjects at risk (Safety population) in that treatment arm and overall. 
9.2. Other Safety Parameters 
Best corrected visual acuity and slit lamp biomicroscopy will be measured at Visit 1 (screening) 
and at the end of Visit 2. BCVA will be measured in logMAR units and results will be summarized 
using descriptive statistics for the Visit 1 and Visit 2 values and for the changes from Visit 1 to 
Visit 2. Results will be presented separately for the study eye and non-study eye. 
 
The biomicroscopy grading scales are given in Appendix 1 of the protocol; the following 
assessments will be done on four anterior locations of each eye: 
• Lid (erythema and edema) 
• Conjunctiva (hyperemia and edema) 
• Cornea (edema) 
• Anterior chamber (cells and flare) 
The data for each assessment will be summarized using counts and percentages of eyes within 
each category and will be summarized for the Visit 1 and Visit 2 values and for the changes from 
Visit 1 to Visit 2. Results will be presented separately for the study eye and non-study eye. 


 
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10. Software Used for Analysis 
SAS Software Version 9.4 or higher (SAS Institute Inc., Cary, NC), will be used for the statistical 
analysis and statistical reporting. 
11. Changes from the Protocol 
The variables and methods described in this plan will supersede those described in the protocol. 
 
12. Requirements and Specifications (Mock-shells) for Programming and 
Presentation of Tables, Listings and Figures (TLFs) 
This document is intended to be used only as a guide in the development of TLFs and may be 
adjusted to meet study and Sponsor specific needs. 
The specifications for the TLFs must follow the description in the associated Statistical Analysis 
Plan (SAP). 
The following details the requirements and specifications for the programming and presentation 
of the TLFs, as set out in the TLF shells. 
• 
A separate RTF document will be created for each TLF. 
• 
All TLFs will be produced in a landscape format, as far as is feasible. 
• 
The margin, page size specifications are stipulated below and will used for the presentation 
of all TLFs: 
Item: 
Specification: 
Page Layout 
Landscape 
Paper Size 
 8.5 x 11 inch 
• 
The standard font size and font type are "9 point", "Courier New" for all TLFs. 
• 
Every page of each output will have a header indicating the Sponsor name and protocol 
number (to be presented as set out in the example below). 
• 
Every page of each output will contain a footer indicating the program name and the date 
and time when the output was produced (to be presented as set out in the example below). 
• 
Every page of each output will contain a page number written in the format "Page X of Y" 
(to be presented as set out in the example below). 
• 
The following treatment group labels will be used for all TLFs in the following order:  
 Treatment A: AG-920 
 Treatment B: Placebo 


 
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• 
The ordering of visits should be chronological (unless otherwise specified): 
• 
Footnotes should be ordered as follows: 
1. Non-standard abbreviations separated by semi-colons 
2. n = Number of subjects … N = Total number of subjects … % = Percentage of 
subjects. 
3. Definitions. 
4. Footnotes pertaining to statistical methodology 
5. Version of the medical coding dictionary used for programming 
6. Study-specific footnotes further clarifying data presentation 
• 
Each of the footnote types enumerated above will flow continuously instead of starting on 
a new line and end with a period. 
• 
Footnotes are left justified. 
• Where applicable, each table should provide the listing supporting the table. 
• All listings should be sorted by site name/country, subject identifier, visit date/event date 
(in the case of multiple observations per visit date/event date, the observations should be 
sorted alphabetically within visit date/event date). 
 An example of country/subject identifier field: SGP/4000101 
• Calendar dates in the listings will be displayed in the format DD-MMM-YYYY, e.g. 15-
SEP-2015.  Where dates are partial, only the partial data will be listed. 
• Where a listing or table has been planned, but no data meet the criteria, then a single line 
stating ‘No data meet the criteria for this report’ will be provided in the output. 
• 
All tables will be decimal aligned as specified in the study SAP 
For all outputs containing calculated changes from baseline, the reference point will be specified 
in a footnote.