Conditions: Mydriasis
Allocation: RANDOMIZED | Intervention model: CROSSOVER
Enrollment: 76
Healthy volunteers: yes

Brief summary:
This study evaluated pupil dilation after administration of fixed combination tropicamide-phenylephrine (TR/PH) vs. placebo. Participants attended 3 visits. At each visit, after baseline measurements, either the study drug or placebo was administered to both eyes, then pupil dilation and safety assessments were performed at specific time intervals.

Detailed description:
Volunteer participants were screened for study eligibility during a Screening Visit and enrolled after signing the study-specific informed consent form. Subjects meeting all inclusion/exclusion criteria were scheduled for 3 treatment visits, which were at least 2 days, but no more than 7 days apart. At each treatment visit, baseline measurements were taken, then either the study drug or placebo was administered to both eyes. Afterwards, efficacy and safety assessments were performed at 20, 35, 50, 65, 80, 120, and 180 minutes. Participants' treatment assignments for each treatment visit were equally randomized using 1 of 2 sequences - ABB or BAA, where A was the study drug and B was the placebo.

Arms:
arm 1: Participants were dosed once at 3 separate clinic visits in accordance with this sequence. Each participant received 2 sprays of the designated treatment in each eye. arm 2: Participants were dosed once at 3 separate clinic visits in accordance with this sequence. Each participant received 2 sprays of the designated treatment in each eye.

Interventions: intervention 1: Tropicamide/Phenylephrine ophthalmic solution (TR/PE) intervention 2: Placebo
Intervention descriptions: intervention 1: Tropicamide 1%/phenylephrine 2.5% ophthalmic solution administered with the Optejet microdose dispenser intervention 2: Eyewash administered with the Optejet microdose dispenser

Protocol document:
ICF_002.pdf: 
EYENOVIA, INC. / Protocol Number EYN-MYD-TP-32 
Page 1 of 16   
 
 
 
«PiFullName» 
Advarra IRB Approved Version 30 Nov 2018 
Revised «PIApprovalDate» 
 
 
INFORMED CONSENT FORM  
FOR 
ADULT SUBJECTS, SUBJECTS WHO TURN AGE OF MAJORITY AND  
PARENTAL/LEGAL GUARDIAN PERMISSION 
 
 
Sponsor / Study Title: 
  
EYENOVIA, INC. / “A Multi-center, Double-masked, Placebo-
controlled, Phase 3 Study of the Safety and Efficacy of Fixed 
Combination Phenylephrine 2.5%-Tropicamide 1% Ophthalmic 
Solution Administered with a Microdose Dispenser for 
Dilation of the Pupil (The MIST-2 Study)” 
 
Protocol Number: 
 
EYN-MYD-TP-32 
 
Principal Investigator: 
(Study Doctor) 
 
«PiFullName» 
 
Telephone: 
 
«IcfPhoneNumber» 
 
Additional Contact(s): 
(Study Staff) 
 
«AdditionalStaffMemberContacts» 
 
Address: 
«PiLocations» 
 
 
 
 
 
INTRODUCTION 
You are invited to take part in a research study. This study is to test phenylephrine 2.5%-
tropicamide 1% ophthalmic solution for dilating the pupil at the time of eye examinations.  
Eyenovia, Inc. is sponsoring this research study. 
 
Please read this form carefully. Take your time to ask the study doctor or study staff as many 
questions about the study as you would like. The study doctor or study staff can explain words or 
information that you do not understand. Reading this form and talking to the study doctor or 
study staff may help you decide whether to take part or not. If you decide to take part in this 
study, you must sign your name at the end of this form and date it.  
 
If you are the parent or legal guardian of a child who may take part in this study, permission 
from you is required.  The assent (agreement) of your child may also be required.  When “you” 
appears in this form, it may refer to you or your child; “we” means the study doctors and study 
staff.   


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«PiFullName» 
Advarra IRB Approved Version 30 Nov 2018 
Revised «PIApprovalDate» 
 
BACKGROUND AND PURPOSE  
You are being asked to take part in this research study as a volunteer.  The study will evaluate an 
investigational drug for dilation of the pupil.  Dilation of the pupil means to increase the size of 
the black circle in the center of the colored part (the iris) of the eye. 
 
The purpose of this research study is to test the safety and effectiveness of the study drug, 
phenylephrine 2.5%-tropicamide 1% ophthalmic solution. 
 
The study drug is a fixed combination of the 2 drugs, phenylephrine 2.5% and tropicamide 1%.  
Each of these drugs is currently marketed as an eyedrop for pupil dilation.  The fixed 
combination study drug is administered as a spray mist, and the amount of drug delivered to the 
eye is much less than the amount of drug contained in an eyedrop. 
 
The fixed combination study drug is “investigational” because the United States Food and Drug 
Administration (FDA) has not approved this formulation and dosage of the drug for sale or for 
use in dilating the pupil.   
 
In this study, the safety and pupil dilation performance of the fixed combination study drug will 
be compared versus a placebo that has no therapeutic effect (a placebo looks like study drug, but 
contains no active drug).  Both the study drug and the placebo will be sprayed as a mist to the 
eye by Eyenovia’s dispenser, which has been designed to deliver precise low-volume drug doses.  
You will receive either study drug or placebo at each study treatment visit. 
 
About 90 subjects will take part in this study.  
 
WHAT WILL HAPPEN DURING THE STUDY 
Your time in this study will last approximately 3 weeks.  You will need to come to the study 
center for 4 scheduled visits. 
 
Screening: 
Before any study-related tests and activities are performed, you will be asked to read and sign 
this consent document. The following screening tests and activities will then be performed to 
determine if you qualify to take part in this study:   
 
• You will be asked to provide information about yourself, including your date of birth, 
gender, race, ethnic origin, and personal data (information). 
• You will be asked about your current overall health and the condition of both eyes.  You 
will also be asked about any previous conditions or treatments you have had.  You must 
be honest in providing information about your eyes and your overall health.  Giving false 
or incomplete information could have serious health consequences. 
• A list of the drugs you are taking now and have taken in the past will be recorded. 
• If you are female and able to become pregnant, you will need to have a urine pregnancy 
test. 
• Your vision in both eyes will be checked. 


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• You will be asked to focus on a target and a bright light will be shone into each eye while 
a special instrument called a pupillometer measures the size of your pupil.  For younger 
pediatric subjects, the pupil diameter may be measured using a ruler or pupil gage. 
• The front part of each eye will be examined using a bright light and a special microscope 
called a slit lamp.  Photographs of your eyes may be taken during this examination.  For 
pediatric subjects who cannot cooperate with a traditional slit lamp examination, a 
portable slit lamp model may be used. 
• Numbing drops will be put in your eyes and your eye pressure will be measured by 
touching an instrument called a tonometer to your cornea (the clear front window of your 
eye).  Because your eyes are numb, you should feel no discomfort during this procedure.    
• Dilating drops will be put in your eyes to make your pupils large.  The study doctor will 
use a magnifying lens and a bright light to examine your retina, which is the back of your 
eye.  The effect of these eye drops will last approximately 4 – 5 hours.  During that time, 
you should not drive a car or operate machinery.  You may also need to wear sunglasses 
outside to reduce light sensitivity. 
 
Note:  Younger pediatric subjects who are unable to perform certain testing may have eye 
pressure and uncorrected visual acuity evaluated using an age-appropriate testing method. 
 
Based on the information gathered from these tests and activities, the study doctor will determine 
if you meet the requirements for receipt of the study drugs to be administered in the study.  If 
you are qualified to continue, you will be scheduled to return for 3 Study Treatment Visits after 
your Screening Visit.  These visits will occur 2-7 days apart and require that you be available for 
3 – 4 hours.   
 
This study will use competitive enrollment.  This means that when the designated number of 
subjects begin drug administration, all further enrollment will be closed.  Therefore, it is possible 
that you could be in the screening phase, ready to begin the study, and be discontinued without 
your consent if the target number of subjects has already begun the study. 
 
Study Treatment:  
The study drugs will be put in your eyes only when you come for your clinic visits.  The study 
drugs will be given as a mist that is sprayed in your eyes by trained study staff.  The mist will be 
sprayed 2 times in each eye over a period of about 5 minutes. 
 
You will receive either the study drug or the placebo at each clinic visit.  At the end of the study, 
you will have received either the placebo twice and the study drug once, or the study drug twice 
and the placebo once.   Neither you nor the person who examines you will know which drug you 
receive at each visit.  In case of an emergency, however, the study doctor can get this 
information. 
 
The following tests and activities will be performed:  
 
At Study Treatment Visit – Day 1:   
• Your vision in both eyes will be checked. 


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• The size of each of your pupils will be measured in the same way as at the Screening 
Visit.  Your pupillary light reflex, which is how quickly your pupil gets smaller in the 
presence of a bright light, will also be evaluated. 
• The front part of each eye will be examined with a slit lamp in the same way as at the 
Screening Visit. 
• Numbing drops will be put into both eyes and your eye pressures will be measured. 
• One of the study drugs will be put into each eye and a video recording of the study drug 
administration may be made.  
• At about 20 minutes, 35 minutes, 50 minutes, 65 minutes, 80 minutes and 120 minutes 
after study treatment, the size of each pupil will be measured.  Your pupillary light reflex 
will also be periodically evaluated.  You will be asked if you are feeling differently since 
receiving study treatment. 
- 
About 65 minutes after study treatment, numbing drops will be put into both eyes and 
your eye pressures will be measured. 
• At about 180 minutes after study treatment, your vision will be checked, the size of your 
pupils will be measured, your pupillary light reflex will be evaluated, and the front part of 
each eye will be examined with a slit lamp.  Photographs of your eyes may also be taken.  
You will be asked if you are feeling differently since receiving the study treatment. 
 
Note:  Younger pediatric subjects who are unable to perform certain testing may have eye 
pressure and uncorrected visual acuity evaluated using an age-appropriate testing method. 
 
At Study Treatment Visit - Days 2 and 3:   
• You will be asked how you are feeling and if you have had any changes in your health or 
drugs you have taken since your last visit. 
• Your vision in both eyes will again be checked. 
• The size of each of your pupils will again be measured in the same way as at the 
Screening Visit.   
• The front part of each eye will again be examined with a slit lamp in the same way as at 
the Screening Visit. 
• Numbing drops will be put into both eyes and your eye pressures will again be measured. 
• One of the study drugs will be put into each eye and a video recording of the study drug 
administration may be made.  
• At approximately 20 minutes, 35 minutes, 50 minutes, 65 minutes, 80 minutes and 120 
minutes after study treatment, the size of each pupil will be measured and your pupillary 
light reflex will also be periodically evaluated.  You will be asked if you are feeling 
differently since receiving study treatment. 
- 
About 65 minutes after study treatments, numbing drops will be put into both eyes 
and your eye pressures will be measured again. 
• At approximately 180 minutes after study treatment, your vision will be checked, the size 
of your pupils will be measured, your pupillary light reflex will be evaluated, and the 
front part of each eye will be examined with a slit lamp.  Photographs of your eyes may 
also be taken.  You will be asked if you are feeling differently since receiving study 
treatment. 
 


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Note:  Younger pediatric subjects who are unable to perform certain testing may have eye 
pressure and uncorrected visual acuity evaluated using an age-appropriate testing method. 
 
After Study Treatment: 
The study drugs will be given to you only during the study and not after the study is over.  If 
your pupil diameter is larger than baseline (screening) at the time of study exit, you may be 
asked if you would be willing to stay at the clinic for additional measurements of your pupil 
during the remainder of the day.  These additional measurements help determine how long the 
study drug works.  You are not required to stay for these additional measurements if you do not 
wish to.  
 
If you have a problem with your eyes related to the study drug and the problem continues after 
the end of the study, you may need to return to the clinic for additional follow-up.   
 
EXPECTATIONS  
If you take part in this study, you will be expected to:  
• Tell the truth about your medical history, current conditions and any prescription or over-
the-counter drugs or supplements you are taking. 
• Tell the study doctor if you have been in a research study during the past 1 month or are 
in another research study now.  You will not be able to enroll in any other clinical study 
of an investigational drug or device until you have completed participation in this study. 
• Go to all your scheduled study visits. 
• Follow all instructions given by the study doctor and study staff. 
• Tell the study doctor or study staff about any changes in your health or the way you feel. 
• If you typically wear contact lenses, on the days the study drug is given, you may not 
wear your contact lenses until after you have completed all study tests and activities.  
Instead, you may need to use your eyeglasses during this time. 
• If you are female and able to become pregnant, you will be asked to use medically 
acceptable birth control to prevent pregnancy. 
 
RISKS, SIDE EFFECTS, AND/OR DISCOMFORTS  
As with any drug, there are risks that side effects may occur.  While each individual drug in the 
fixed combination solution is currently marketed in the United States as eyedrops for dilation of 
the pupil, the United States Food and Drug Administration (FDA) has not approved this 
formulation and dosage of the drug for sale or for use in dilating the pupil.  The fixed 
combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution has not been evaluated by 
the FDA.   
 
After each drug is given, your vision may be blurry.  You should not drive until your vision 
returns to normal.  You may also need to wear sunglasses when outside if you experience light 
sensitivity.  The following side effects have been reported with use of phenylephrine and 
tropicamide in the eye.  You may experience some or all these side effects listed below: 
 
Likely 
• Temporary blurred vision 
• Temporary light sensitivity 


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Advarra IRB Approved Version 30 Nov 2018 
Revised «PIApprovalDate» 
 
• Eye stinging at the time of drug administration 
 
Less Frequent 
• Itching, burning, stinging or foreign body sensation in the eye after drug administration 
• Increased pressure in the eye 
• Dryness in the eye 
• Increased tearing in the eye  
• Rash or allergic skin reaction  
• Blood pressure increase or decrease, which may result in fainting 
• Abnormally fast, slow, or irregular heart beat 
• Headache 
• Dryness of the mouth 
• Nausea and vomiting 
• Unhealthy skin appearance around the eye 
• Excitability, behavioral disturbances, or psychotic reactions 
 
RISKS OF STUDY PROCEDURES 
• Eye Pressure Measurement:  There is a slight risk that you may receive a minor scratch to 
your cornea from the instrument that measures eye pressure.  Minor scratches of this nature 
usually heal within 24-72 hours. 
• Retina Exam:   The eyedrops used to make your pupil large so the study doctor can see your 
retina may cause blurred vision or temporary glare.  The eyedrops could also make your eye 
pressure go up.  You may also have an allergic reaction to these drops. 
 
UNFORESEEN RISKS  
There may be other risks or side effects that are currently unknown.  Allergic reactions can occur 
with any drug.  Some symptoms of allergic reactions are rash, wheezing and difficulty breathing, 
dizziness and fainting, swelling around the mouth, throat or eyes, a fast pulse or sweating. It is 
important to tell the study doctor if you start to experience any new symptoms after the study 
drug is administered.  Your study doctor will monitor you for side effects throughout this study.   
 
Also, there may be unknown risks to a pregnancy, embryo, or fetus if you become pregnant. 
 
BIRTH CONTROL RESTRICTIONS  
Use of the study drug may involve risks to a pregnant woman, an embryo, fetus (unborn baby) or 
nursing infant.  Therefore, if you are pregnant, planning to become pregnant, or are breastfeeding 
a child, you cannot participate in this study. 
 
To reduce the risk of pregnancy, you should use an effective method of birth control while you 
are participating in this study.  Acceptable methods of birth control are intrauterine devices 
(IUDs), hormonal drugs, barrier devices and abstinence.  The Investigator or study staff will 
discuss this with you.   
 


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If you become pregnant while you are participating in this study or within the 4 weeks after the 
last study treatment visit, tell your Investigator or the study staff immediately. The drug will not 
be given again and your participation in this study will be ended.   
 
 
ALTERNATIVES TO PARTICIPATION  
This study is for research purposes only. The only alternative is to not take part in this study. 
 
NEW FINDINGS  
Any new important information that is discovered during the study that may influence your 
willingness to continue participation will be provided to you. 
 
BENEFITS 
This study is for research purposes only.  There is no direct benefit to you from your taking part 
in the study.  Information learned from the study may help other people in the future. 
 
COMPENSATION FOR PARTICIPATION  
You will be paid up to a total of $xx.xx if you complete this study. You will be paid for the visits 
you complete according to the following schedule: 
 
Visit 
Payment 
Screening Visit 
 
Study Treatment Visit - Day 1 
 
Study Treatment Visit – Day 2 
 
Study Treatment Visit – Day 3 
 
Study Treatment Day 3 – Post 180 Minutes (optional) 
 
 
If you do not complete the study for any reason, you will be paid for each study visit you do 
complete.   If you do not complete an entire study visit, you will not receive payment for that 
visit.  You will be paid ___________ ‘after each visit’, ‘annually’, ‘bi-weekly’, etc. 
 
If you have any questions regarding your compensation for participation, please contact the study 
staff.     
 
CONFIDENTIALITY    
Records of your participation in this study will be held confidential except when sharing the 
information is required by law or as described in this informed consent.  The Investigator, the 
Sponsor (or persons working on behalf of the Sponsor), and under certain circumstances, the 
United States FDA and the Institutional Review Board (IRB) will be able to inspect and copy 
confidential study-related records that identify you by name.  This means that absolute 
confidentiality cannot be guaranteed.  If the results of this study are published or presented at 
meetings, you will not be identified. 
 


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A description of this clinical trial will be available on http://www.ClinicalTrials.gov, as required 
by U.S. Law.  This Web site will not include information that can identify you.  At most, the 
Web site will include a summary of the results.  You can search this Web site at any time. 
 
COMPENSATION FOR INJURY   
If you become ill or are injured while you are in the study, get the medical care that you need 
right away. You should tell the healthcare professional treating you that you are taking part in 
this study. If you tell the study staff that you think you have been injured, they will help you get 
the care you need.  
 
If you are injured as a result of taking the study drug(s) or from procedures done for this study, 
the Sponsor will pay for those medical expenses necessary to treat your injury that are not 
covered by your medical insurance or any other third-party coverage.  You will not lose any of 
your legal rights or release the Sponsor, the Investigator, the study staff, or study site from 
liability for their mistakes by signing this consent document. 
 
To pay medical expenses, the Sponsor will need to know some information about you, such as 
your name, date of birth, and Medicare Beneficiary Identifier (MBI). This is because the Sponsor 
must check to see if you receive Medicare and if you do, report the payment it makes to 
Medicare.  
 
COSTS  
There will be no charge to you for your participation in this study.  The study drug, study-related 
activities, and study visits will be provided at no charge to you or your insurance company. 
 
WHOM TO CONTACT ABOUT THIS STUDY 
During the study, if you experience any medical problems, suffer a research-related injury, or 
have questions, concerns, or complaints about the study, please contact the Investigator at the 
telephone number listed on the first page of this consent document.  If you seek emergency care, 
or hospitalization is required, alert the treating physician that you are taking part in this study. 
 
An IRB is an independent committee established to help protect the rights of research 
subjects.  If you have any questions about your rights as a research subject, and/or concerns or 
complaints regarding this research study, contact: 
 
• By mail: 
 
Study Subject Advisor 
Advarra IRB 
6940 Columbia Gateway Drive, Suite 110 
Columbia, MD 21046 
• or call toll free:         877-992-4724 
• or by email:               adviser@advarra.com 
 
Please reference the following number when contacting the Study Subject Adviser: 
Pro00029864. 
 


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VOLUNTARY PARTICIPATION / WITHDRAWAL  
Your decision to take part in this study is voluntary.  You may choose to not take part, or you 
may withdraw from the study for any reason without penalty or loss of benefits to which you are 
otherwise entitled and without any effect on your future medical care.  However, please note that 
FDA requires that any information collected up to the point of your withdrawal cannot be 
removed from the study. 
 
If you are an employee of this study center or a family member of someone who is an employee 
at this center and you decide to take part in this study, you may leave the study at any time for 
any reason without penalty or prejudice. 
 
If you decide not to take part in the study or to withdraw after it has started, this decision will 
have no effect on employee performance evaluations, job advancement or current employment 
status.  No action will be taken against an employee based on information the study doctor 
obtains due to their study participation.   
 
The Investigator or the Sponsor can stop your participation at any time without your consent for 
the following reasons: 
 
• If it appears to be medically harmful to you; 
• If you fail to follow directions for participating in the study; 
• If it is discovered that you do not meet the study requirements; 
• If the study is canceled; or  
• For administrative reasons. 
 
If you leave the study for any reason, the Investigator may ask you to have some end-of-study 
tests for your safety.   
 
Primary Health Care Provider Notification Option 
I consent to having my family doctor or primary health care provider notified by the study center 
of my participation in this study and/or any significant findings related to my health (please 
check yes or no). 
 
 
Yes (If yes, please complete the information below) 
No   
 
Name and address of family 
doctor or primary health care 
provider: 
Name: 
Address: 
 
Telephone and Fax Number: 
Tel: 
Fax: 


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CONSENT   
I have read and understand the information in this informed consent document.  I have had an 
opportunity to ask questions and all my questions have been answered to my satisfaction. I 
voluntarily agree to participate in this study until I decide otherwise.  I do not give up any of my 
legal rights by signing this consent document.  I will receive a copy of this signed and dated 
consent document. 
 
 
____________________________________ 
 
 
 
Subject’s Printed Name  
 
 
____________________________________ 
 
 
_________  
Subject’s Signature 
 
 
 
 
 
 
Date 
 
 
STATEMENT OF PARENTAL / LEGAL GUARDIAN PERMISSION 
 
I have read and understand the information in this informed consent document.  I have had an 
opportunity to ask questions and all of my questions have been answered to my satisfaction. I 
voluntarily agree for my child to participate in this study until I decide otherwise.  I do not give 
up any of my or my child’s legal rights by signing this consent document.  I will receive a copy 
of this signed and dated consent document. 
 
_________________________________ 
 
 
 
____/____/____ 
Signature of Parent/Legal Guardian (if subject is under age of majority) 
 
Date 
 
 
 
_________________________________ 
Printed Name of Parent/Legal Guardian (if subject is under age of majority) 
 
______________________________________________   
Printed Name of Second Parent 
 
______________________________________________     _____/____/_____ 
Second Parent Signature 
 
 
 
 
 
Date 
 
 
 


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Second parent permission not documented, reason (person obtaining consent will initial and 
check all that apply): 
 
Initials of person obtaining consent 
2nd Parent is deceased:     
 
 
              
____________________ 
2nd Parent is unknown:                                     
 
____________________ 
2nd Parent is incompetent:                                  
 
____________________ 
2nd Parent does not have legal custody:          
 
____________________ 
2nd Parent “Not Reasonably Available”:           
 
____________________ 
             2nd Parent Not available by telephone 
             2nd Parent Not available by email        
             2nd Parent Not available by mail 
             2nd Parent Not available by fax 
 
 
____________________________________ 
 
 
 
Printed Name of the Person Conducting the   
 
 
Consent Discussion   
 
_____________________________________ 
 
 
___________ 
Signature of the Person Conducting the  
 
 
 
Date 
 
Consent Discussion   
 
 
 
 
 
 
 
FOR CHILDREN WHO BECOME AGE OF MAJORITY DURING THE STUDY 
I have been told that my parents/legal guardian agreed for me to participate in this research study 
as a minor. I have read and understand the information in this informed consent document.  I 
have had an opportunity to ask questions and all my questions have been answered to my 
satisfaction. I voluntarily agree to continue to participate in this study until I decide otherwise.  I 
do not give up any of my legal rights by signing this consent document.  I will receive a copy of 
this signed and dated consent document. 
 
 
____________________________________                            
Subject’s Printed Name  
 
 
____________________________________                           
_________  
Subject’s Signature                                                                     
Date 
 
 
 
 


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WITNESS SIGNATURE FOR SUBJECTS WHO CANNOT READ   
The study subject has indicated that he/she is unable to read.  The consent document has been 
read to the subject by a member of the study staff, discussed with the subject by a member of the 
study staff, and the subject has been given an opportunity to ask questions of the study staff.    
 
____________________________________ 
Printed Name of Impartial Witness  
 
 
____________________________________ 
 
 
__________ 
Signature of Impartial Witness 
 
 
 
 
Date  
 
 


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 AUTHORIZATION TO USE AND DISCLOSE 
PROTECTED HEALTH INFORMATION 
 
If you decide to be in this study, the study doctor and research team will use and 
share health data about you to conduct the study.  Health data may include: 
• Your name. 
• Address. 
• Phone number. 
• Date of birth. 
• Medical history. 
• Information from your study visits, including all test results.   
 
Health data may come from your study records or from existing records kept by 
your doctor or other health care workers.   
 
For this study, the research team may share health data about you with authorized 
users.  Authorized users may include 
• Representatives of Eyenovia, Inc. 
• Representatives of Advarra IRB (an Institutional Review Board that reviews 
this study). 
• The Food and Drug Administration (FDA) and other US federal and state 
agencies. 
• Government agencies to whom certain diseases (like HIV, hepatitis, and 
STDs) must be reported. 
• Governmental agencies of other countries. 
• Outside individuals and companies, such as laboratories and data storage 
companies, that work with the researchers and sponsor and need to access 
your information to conduct this study. 
• Other research doctors and medical centers participating in this research, if 
applicable.  
 
Your health data will be used to conduct and oversee the research, including for 
instance: 
• To see if the study drug works and is safe. 
• To compare the study drug performance to that of phenylephrine 2.5% and 
tropicamide 1%. 
• For other research activities related to the study drug. 


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Once your health data has been shared with authorized users, it may no longer be 
protected by federal privacy law and could possibly be used or disclosed in ways 
other than those listed here.   
 
Your permission to use and share health data about you will end in 50 years unless 
you revoke it (take it back) sooner.  
 
You may revoke (take back) your permission to use and share health data about 
you at any time by writing to the study doctor at the address listed on the first page 
of this form.  If you do this, you will not be able to stay in this study.  No new 
health data that identifies you will be gathered after your written request is 
received.  However, health data about you that has already been gathered may still 
be used and given to others as described in this form.   
 
Your right to access your health data in the study records will be suspended during 
the study to keep from changing the study results.  When the study is over, you can 
access your study health data. 
 
If you decide not to sign this form, you will not be able to take part in the study. 
 
 
 


EYENOVIA, INC. / Protocol Number EYN-MYD-TP-32 
Page 15 of 16   
 
 
 
«PiFullName» 
Advarra IRB Approved Version 30 Nov 2018 
Revised «PIApprovalDate» 
 
STATEMENT OF AUTHORIZATION 
I have read this form and its contents were explained.  My questions have been 
answered.  I voluntarily agree to allow study staff to collect, use and share my 
health data as specified in this form.  I will receive a signed and dated copy of this 
form for my records.  I am not giving up any of my legal rights by signing this 
form.   
 
 
____________________________________ 
Printed Name of Subject 
 
____________________________________ 
 
__________ 
Signature of Subject 
 
 
 
 
 
Date 
 
STATEMENT OF PARENTAL / LEGAL GUARDIAN PERMISSION 
 
I have read and understand the information in this informed consent document.  I 
have had an opportunity to ask questions and all of my questions have been 
answered to my satisfaction. I voluntarily agree for my child to participate in this 
study until I decide otherwise.  I do not give up any of my or my child’s legal 
rights by signing this consent document.  I will receive a copy of this signed and 
dated consent document. 
 
_________________________________ 
 
 
 
____/____/____ 
Signature of Parent/Legal Guardian 
 
 
 
 
 
Date  
 
(if subject is under age of majority) 
 
_________________________________ 
Printed Name of Parent/Legal Guardian (if subject is under age of majority) 
 
 
____________________________________ 
Printed Name of the Person Obtaining the  
Authorization 
 
____________________________________ 
 
 
__________ 
Signature of the Person Obtaining the  
 
 
 
Date 
Authorization 
 
 


EYENOVIA, INC. / Protocol Number EYN-MYD-TP-32 
Page 16 of 16   
 
 
 
«PiFullName» 
Advarra IRB Approved Version 30 Nov 2018 
Revised «PIApprovalDate» 
 
FOR CHILDREN WHO BECOME AGE OF MAJORITY DURING THE 
STUDY  
I have been told that my parents/legal guardian agreed to the use and disclosure of 
my Protected Health Information as outlined in this document. I have read and 
understand the information in this authorization.  I have had an opportunity to ask 
questions and all my questions have been answered to my satisfaction. I continue 
to authorize the use and disclosure of my Protected Health Information. I will 
receive a copy of this signed and dated authorization. 
 
 
____________________________________                            
Subject’s Printed Name  
 
 
 
____________________________________                           
_________  
Subject’s Signature                                                                        Date 
 
 
 
WITNESS SIGNATURE FOR SUBJECTS WHO CANNOT READ   
The study subject has indicated that he/she is unable to read.  This Authorization 
document has been read to the subject by a member of the study staff, discussed 
with the subject by a member of the study staff, and the subject has been given an 
opportunity to ask questions of the study staff.    
 
 
____________________________________ 
Printed Name of Impartial Witness  
 
 
____________________________________ 
 
__________ 
Signature of Impartial Witness   
 
 
 
Date  
 
 
 

Prot_000.pdf: 
PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
EYENOVIA CLINICAL STUDY PROTOCOL EYN-MYD-TP-32 VERSION C  
 
 
 
CONFIDENTIAL  
20 NOVEMBER 2018 
 
  
 
 
  
 
 
PROTOCOL EYN-MYD-TP-32 
VERSION C 
 
 
A MULTI-CENTER, DOUBLE-MASKED, PLACEBO-CONTROLLED, PHASE 3 STUDY OF 
THE SAFETY AND EFFICACY OF FIXED COMBINATION PHENYLEPHRINE 2.5%-
TROPICAMIDE 1% OPHTHALMIC SOLUTION ADMINISTERED WITH A MICRODOSE 
DISPENSER FOR DILATION OF THE PUPIL 
(THE MIST-2 STUDY) 
 
 
 
 
SPONSOR: 
 
EYENOVIA, INC. 
295 MADISON AVENUE, SUITE 2400 
NEW YORK NY  10017 
USA 
PHONE:  917-289-1117 
 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
EYENOVIA CLINICAL STUDY PROTOCOL EYN-MYD-TP-32 VERSION C  
 
 
 
CONFIDENTIAL - PAGE 2 OF 61 
20 NOVEMBER 2018 
 
TABLE OF CONTENTS 
 
TABLE OF CONTENTS ......................................................................................................................... 2 
LIST OF ABBREVIATIONS .................................................................................................................. 5 
1. PERSONNEL AND FACILITIES ....................................................................................................... 6 
2. STUDY SYNOPSIS ............................................................................................................................. 7 
2.1. Study Objective ............................................................................................................................ 7 
2.2. Study Drugs .................................................................................................................................. 7 
2.3. Study Population .......................................................................................................................... 7 
2.4. Study Design ................................................................................................................................ 7 
2.5. Inclusion Criteria .......................................................................................................................... 8 
2.6. Exclusion Criteria ......................................................................................................................... 8 
2.7. Primary Performance Endpoint .................................................................................................... 9 
2.8. Exploratory Outcomes ................................................................................................................. 9 
2.9. Safety Outcomes .......................................................................................................................... 9 
2.10. Schedule of Visits, Procedures and Evaluations ...................................................................... 10 
2.10.1. Study Medication Administration and Clinical Assessments ........................................... 10 
3. INTRODUCTION AND RATIONALE ............................................................................................ 11 
3.1. Introduction ................................................................................................................................ 11 
3.2. Pupil Dilation ............................................................................................................................. 11 
3.3. Existing Mydriatic Options ........................................................................................................ 12 
3.4. Phenylephrine and Tropicamide Ophthalmic Microdosing Solutions for Pupil Dilation .......... 12 
3.5. Ophthalmic Microdosing ........................................................................................................... 13 
4. INVESTIGATIONAL PRODUCT DESCRIPTION ......................................................................... 16 
4.1. Findings from Non-Clinical and Clinical Studies ...................................................................... 16 
4.2. How Provided............................................................................................................................. 16 
4.3. Route of Administration, Dosage, Regimen, Treatment Period(s) ............................................ 17 
5. STUDY OBJECTIVE ........................................................................................................................ 18 
6. STUDY DESIGN ............................................................................................................................... 18 
7. STUDY OUTCOMES ........................................................................................................................ 19 
7.1. Effectiveness Outcomes ............................................................................................................. 19 
7.2. Safety Outcomes ........................................................................................................................ 19 
8. STUDY POPULATION ..................................................................................................................... 20 
8.1. Inclusion Criteria ........................................................................................................................ 20 
8.2. Exclusion Criteria ....................................................................................................................... 20 
9. STUDY METHODS .......................................................................................................................... 22 
9.1. Subject Informed Consent .......................................................................................................... 22 
9.2. Subject Selection and Screening Procedures ............................................................................. 22 
9.3. Randomization to Study Drug Administration Sequence .......................................................... 22 
9.3.1. Procedure for Unmasking Study Drug Assignment Code .................................................. 23 
9.4. Study Drug Packaging, Labeling and Storage............................................................................ 23 
9.5. Study Drug Administration ........................................................................................................ 24 
9.6. Study Visits and Clinical Assessments ...................................................................................... 24 
9.6.1. Screening Visit (-14 Days to -1 Day) ................................................................................. 24 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
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CONFIDENTIAL - PAGE 3 OF 61 
20 NOVEMBER 2018 
 
9.6.2. Treatment Visit 1 (Day 1) ................................................................................................... 25 
9.6.3. Treatment Visit 2 (Day +3 to Day +8) ................................................................................ 26 
9.6.4. Treatment Visit 3 (Day +5 to Day +15) .............................................................................. 27 
9.4.5. Unscheduled Visits ............................................................................................................. 28 
9.4.6. Missed Visits....................................................................................................................... 28 
9.7. Study Exit ................................................................................................................................... 28 
9.7.1. Subject Termination ............................................................................................................ 28 
9.7.2. Subject Completion ............................................................................................................. 28 
10. ADVERSE EVENTS ....................................................................................................................... 29 
10.1. Adverse Event Definitions ....................................................................................................... 29 
10.1.1. Adverse Event (AE) .......................................................................................................... 29 
10.1.2. Adverse Drug Reaction (ADR) ......................................................................................... 29 
10.1.3. Unexpected Adverse Event or Unexpected Suspected Adverse Reaction ........................ 29 
10.1.4. Suspected Adverse Reaction ............................................................................................. 29 
10.1.5. Life-Threatening Adverse Event or Life-Threatening Suspected Adverse Reaction ....... 30 
10.1.6. Serious Adverse Event (SAE) or Serious Suspected Adverse Reaction ........................... 30 
10.1.7. Serious Ocular Adverse Events ........................................................................................ 30 
10.1.8. Treatment-Emergent Adverse Event (TEAE) ................................................................... 31 
10.2. Adverse Event Assessment and Documentation ...................................................................... 31 
10.2.1. Adverse Event Severity..................................................................................................... 31 
10.2.2. Adverse Event Relationship to Study Drug ...................................................................... 32 
10.3 Adverse Event Reporting .......................................................................................................... 32 
10.3.1. Expedited Adverse Event Reporting ................................................................................. 33 
11. STATISTICAL METHODS............................................................................................................. 34 
11.1. Efficacy Endpoints ................................................................................................................... 34 
11.2. Sample Size .............................................................................................................................. 34 
11.3. Safety Outcomes ...................................................................................................................... 35 
11.4. Analysis Populations ................................................................................................................ 35 
11.4.1. Intent to Treat Population ................................................................................................. 35 
11.4.2. Per Protocol Population .................................................................................................... 35 
11.4.3. Modified Per Protocol Population .................................................................................... 35 
11.4.4. Safety Population .............................................................................................................. 36 
11.5. General Statistical Considerations ........................................................................................... 36 
11.5.1. Analysis of Primary Efficacy ............................................................................................ 36 
11.5.2. Additional Efficacy Analyses ........................................................................................... 37 
11.5.3. Sensitivity Analyses .......................................................................................................... 37 
11.5.4. Interim Analyses ............................................................................................................... 38 
11.5.5. Analysis of Baseline Data ................................................................................................. 38 
12. QUALITY ASSURANCE AND QUALITY CONTROL ............................................................... 39 
12.1. Study Monitoring ..................................................................................................................... 39 
12.2. Study Product Accountability .................................................................................................. 39 
13. ETHICAL AND REGULATORY CONSIDERATIONS ................................................................ 40 
13.1. Institutional Review ................................................................................................................. 40 
13.2. Informed Consent ..................................................................................................................... 40 
13.3. Subject Confidentiality ............................................................................................................. 40 
14. DATA HANDLING AND RECORD KEEPING............................................................................. 41 
14.1. Data Quality Control and Reporting ........................................................................................ 41 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
EYENOVIA CLINICAL STUDY PROTOCOL EYN-MYD-TP-32 VERSION C  
 
 
 
CONFIDENTIAL - PAGE 4 OF 61 
20 NOVEMBER 2018 
 
14.2. Data Archiving ......................................................................................................................... 41 
14.3. Records Retention .................................................................................................................... 41 
14.4. Protocol Amendments .............................................................................................................. 41 
15. REFERENCES ................................................................................................................................. 42 
APPENDIX 1:  SCHEDULE OF MEDICATION ADMINISTRATION AND EXAMINATIONS .... 44 
APPENDIX 2:  STUDY MEDICATION PREPARATION PROCEDURE .......................................... 45 
APPENDIX 3:  STUDY MEDICATION ADMINISTRATION PROCEDURE .................................. 47 
APPENDIX 4:  EXAMINATION PROCEDURES ............................................................................... 49 
APPENDIX 5:  SPONSOR AND INVESTIGATOR OBLIGATIONS ................................................. 53 
APPENDIX 6:  WORLD MEDICAL ASSOCIATION DECLARATION OF HELSINKI .................. 56 
 
 
 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
EYENOVIA CLINICAL STUDY PROTOCOL EYN-MYD-TP-32 VERSION C  
 
 
 
CONFIDENTIAL - PAGE 5 OF 61 
20 NOVEMBER 2018 
 
LIST OF ABBREVIATIONS 
 
Abbreviation 
Term 
AC 
Anterior chamber 
AE  
Adverse event 
ADR 
Adverse drug reaction 
AT 
Artificial tears 
BCDVA 
Best-corrected distance visual acuity 
CF 
Count fingers 
CFR 
Code of Federal Regulations 
eCRF 
Electronic case report form 
ETDRS 
Early Treatment Diabetic Retinopathy Study (Chart) 
FDA 
United States Food and Drug Administration 
GCP 
Good Clinical Practice 
HM 
Hand motion 
IB 
Investigator brochure 
ICF 
Informed consent form 
ICH 
International Conference on Harmonization 
IND 
Investigational New Drug 
IOL 
Intraocular lens 
IOP 
Intraocular pressure 
IRB 
Institutional Review Board 
ITT 
Intent-to-treat  
LP 
Light perception 
MiDD 
Eyenovia MicroDose Dispenser 
NLP 
No light perception 
OU 
Both eyes (within this protocol, OU does not connote simultaneous treatment/evaluation) 
PETG 
Polyethylene terephthalate glycol 
PP 
Per-protocol 
SAE 
Serious adverse event 
SD 
Standard deviation 
SLE 
Slit lamp examination 
TEAE 
Treatment-emergent adverse event 
TLR 
Total letters read 
UCDVA 
Uncorrected distance visual acuity 
US 
United States 
VA 
Visual acuity 
WMA 
World Medical Association 
µD 
Microdose 
µL 
Microliter 
NOTE:  The first occurrence of some abbreviations is not spelled out in the document (e.g., units of 
measure). 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
EYENOVIA CLINICAL STUDY PROTOCOL EYN-MYD-TP-32 VERSION C  
 
 
 
CONFIDENTIAL - PAGE 6 OF 61 
20 NOVEMBER 2018 
 
 
PROTOCOL EYN-MYD-TP-32 
VERSION C 
 
 
A MULTI-CENTER, DOUBLE-MASKED, PLACEBO-CONTROLLED, PHASE 3 STUDY OF 
THE SAFETY AND EFFICACY OF FIXED COMBINATION PHENYLEPHRINE 2.5% - 
TROPICAMIDE 1% OPHTHALMIC SOLUTION ADMINISTERED WITH A MICRODOSE 
DISPENSER FOR DILATION OF THE PUPIL 
(THE MIST-2 STUDY) 
 
 
1. PERSONNEL AND FACILITIES 
 
Study Sponsor: 
Eyenovia, Inc. 
295 Madison Avenue, Suite 2400 
New York, NY  10017  USA 
Phone:  917-289-1117 
Email:  admin@eyenoviabio.com 
 
Chief Medical Officer: 
Tsontcho Ianchulev, MD, MPH 
Phone:  617-216-4369 
Email:  tianchul@eyenoviabio.com 
 
Medical Monitor: 
Siddarth Rathi, MD 
Phone: 914-714-8990 
Email:  siddarthrathi@gmail.com 
 
Clinical Study Manager: 
Greg Bennett, Sr. Director, Clinical Operations 
Phone: 650-504-2005 
Email: gbennett@eyenoviabio.com  
 
Clinical Data Manager: 
Lucinda Marinus, Director, Clinical Data Management 
Phone:  949-903-4325 
Email:  lmarinus@eyenoviabio.com 
 
Overall Program Management: 
Ginger Clasby, VP, Clinical Operations 
Phone:  714-269-8838 
Email:  gclasby@eyenoviabio.com  
 
  
 
 
 
 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
EYENOVIA CLINICAL STUDY PROTOCOL EYN-MYD-TP-32 VERSION C  
 
 
 
CONFIDENTIAL - PAGE 7 OF 61 
20 NOVEMBER 2018 
 
2. STUDY SYNOPSIS 
 
2.1. Study Objective    
 
The primary objective of this study is to evaluate the safety and efficacy of Eyenovia’s fixed combination 
of phenylephrine 2.5%-tropicamide 1% ophthalmic solution administered with Eyenovia’s microdose 
dispenser (MiDD) for dilation of the pupil as compared a placebo. 
 
2.2. Study Drugs 
 
• Investigational Product, Dosage and Mode of Administration:  Fixed combination ophthalmic 
solution (phenylephrine 2.5%-tropicamide 1%) administered by Eyenovia’s MiDD. 
 
• Placebo, Dosage and Mode of Administration:  Eyewash solution administered by Eyenovia’s 
MiDD. 
 
2.3. Study Population 
 
Up to 90 volunteer participants will be enrolled at 2 – 3 study sites in the United States (US) and a 
minimum of 65 subjects will be randomized to study drug administration at Treatment Visit 1 in order to 
complete follow-up on 54 subjects.   
 
2.4. Study Design   
 
This trial is a multicenter, double-masked, placebo-controlled, cross-over superiority study evaluating 
Eyenovia’s fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution versus placebo.   
 
Volunteer participants will be screened for study eligibility during a Screening Visit and enrolled after 
signing the study-specific informed consent form (ICF).  Subjects meeting all inclusion/exclusion criteria 
will be scheduled for 3 treatment visits, which must occur at least 2 days, but no more than 7 days apart.  
A 2 sequence, 3 period crossover design will be used.  At each of the three treatment visits, baseline 
measurements will be taken, then either the investigational drug or the placebo will be administered to 
both eyes (OU) in two separate instances, approximately 5 minutes apart.  Only 1 drug will be 
administered per treatment visit, and drug administration will occur in a randomized order using one of 
two sequences - ABB and BAA, where A is the Eyenovia fixed combination and B is the placebo control. 
Afterwards, efficacy and safety assessments will be performed.   
 
This study will be double-masked.  There will be no differences in the presentation of the investigational 
drug or placebo administered and all study personnel conducting ophthalmic assessments will be masked 
to study drug assignment.  Study drug administration will be performed by at least 3 different trained 
personnel during the study.  Personnel who administer study drug on a given day will not be allowed to 
perform post-study drug administration ophthalmic assessments on that day. 
 
 
 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
EYENOVIA CLINICAL STUDY PROTOCOL EYN-MYD-TP-32 VERSION C  
 
 
 
CONFIDENTIAL - PAGE 8 OF 61 
20 NOVEMBER 2018 
 
2.5. Inclusion Criteria 
 
Subjects eligible for study participation must meet each criterion listed below.  Ocular criteria must be 
met for both eyes. 
  
1. Male or female of any age. 
2. Female subjects must be either pre-menarche, 1-year postmenopausal, surgically sterilized, 
or, if of childbearing potential, have a negative urine pregnancy test during the Screening 
Visit and agree to use an acceptable form of contraception throughout the study.  Acceptable 
methods include the use of at least one of the following: intrauterine (intrauterine device), 
hormonal (oral, injection, patch, implant, ring), barrier with spermicide (condom, 
diaphragm), or abstinence. 
3. Ability to provide signed written consent prior to participation in any study-related 
procedures.  Subjects under the age required for informed consent must have the ability to 
provide additional written assent and/or parental consent, as required by the reviewing 
Institutional Review Board (IRB). 
4. Ability to return for all study treatment visits. 
5. Photopic screening pupil diameter ≤ 3.5 mm in each eye. 
 
2.6. Exclusion Criteria 
 
Subjects with any of the following diseases, surgeries or conditions are ineligible for study participation.  
Subjects may not participate if either eye meets any of the ocular exclusion criteria. 
 
1. Pregnancy or lactation. 
2. Allergy to phenylephrine hydrochloride. 
3. Allergy to tropicamide. 
4. Allergy to benzalkonium chloride. 
5. History of benign prostatic hyperplasia. 
6. Use of benzodiazepines, monoamine oxidase inhibitors, tricyclic antidepressants, 
anticonvulsants, cholinergic drug  at screening or anticipated during the study period.  
7. Participation in any study of an investigational drug or device within 30 days prior to the 
Screening Visit, or at any time during the study period. 
8. History of closed-angle glaucoma. 
9. Anatomically narrow anterior chamber angles (Van Herrick grade ≤ 2 in either eye). 
10. Ocular surgery or laser treatment of any kind prior to the Screening Visit. 
11. History of chronic or acute uveitis. 
12. History of traumatic iritis or hyphema. 
13. History of traumatic mydriasis or angle recession. 
14. History of heterochromia. 
15. Irregularly-shaped pupil secondary to ocular trauma or congenital defect. 
16. History of neurogenic pupil disorder (e.g., Horner’s syndrome, third cranial nerve palsy, 
Adie’s pupil, Argyl Robertson syndrome, etc.). 
17. History of anterior chamber intraocular lens (IOL) or iris-fixated IOL. 
18. History of iris surgery of any kind (e.g., iridotomy, iridectomy, coreoplasty). 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
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CONFIDENTIAL - PAGE 9 OF 61 
20 NOVEMBER 2018 
 
19. History of iris atrophy. 
20. History of iris – cornea apposition/touch. 
21. Unwilling or unable to discontinue use of contact lenses at treatment visits. 
22. Current active eye disease for which topical or systemic ophthalmic medication is necessary, 
except for dry eye disease managed using artificial tears (AT).   AT’s must be 
discontinued on the day of each treatment visit. 
23. Presence of a severe/serious ocular condition, or any other unstable medical condition that, in 
the Investigator’s opinion, may preclude study treatment and/or follow-up. 
 
2.7. Primary Performance Endpoint 
 
The primary performance endpoint is mean change in pupil diameter at 35 minutes versus baseline, as 
measured by digital pupillometry in highly photopic conditions.  The highly photopic condition will be 
established using a fully-charged transilluminator (muscle light) at the brightest setting.  
 
2.8. Exploratory Outcomes 
 
• 
Proportion of eyes achieving pupil diameter of 6.0 mm or greater at 35 minutes 
• 
Proportion of eyes achieving pupil diameter of 7.0 mm or greater at 35 minutes 
• 
Mean change in pupil diameter at other timepoints (20, 50, 65, 80, 120, and 180 minutes) 
• 
Distribution of pupil diameters at 20, 35, 50, 65, 80, 120, and 180 minutes 
• 
Time from baseline to maximal pupil dilation 
 
These exploratory outcomes will also be measured using digital pupillometry in highly photopic 
conditions. 
 
2.9. Safety Outcomes 
 
• 
Slit-lamp examination findings 
• 
Occurrence of AEs 
• 
Intraocular pressure (IOP) measured at 65 minutes post-administration 
• 
Visual acuity changes 
 
 
 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
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CONFIDENTIAL - PAGE 10 OF 61 
20 NOVEMBER 2018 
 
2.10. Schedule of Visits, Procedures and Evaluations 
 
Study visits include a Screening Visit, which must occur between 1 and 14 days prior to treatment; 
followed by Treatment Day 1, Treatment Day 2, and Treatment Day 3.  Treatment visits must occur at 
least 2 days and not more than 7 days apart.  The study visit schedule is presented in flow chart form in 
Figure 1. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Figure 1:  Study Visit Flow Chart 
 
 
2.10.1. Study Medication Administration and Clinical Assessments 
 
Study medication administration and clinical assessments will be performed at study visits as shown in 
APPENDIX 1:  SCHEDULE OF MEDICATION ADMINISTRATION AND EXAMINATIONS. 
Screening 
Visit 
Treatment Day 1 
Randomization 
Investigational drug or placebo administered 
Efficacy & Safety assessments 
Treatment Day 2 
Investigational drug or placebo administered 
Efficacy & Safety assessments 
 
Treatment Day 3 
Investigational drug or placebo administered 
Efficacy & Safety Assessments 
 
-14 Days to 1 Day prior to Treatment Day 1 
2 – 7 Days after Screening Visit 
2 – 7 Days after Treatment Day 1 
2 – 7 Days after Treatment Day 2 
Study Exit 
Fails to meet 
inclusion/exclusion criteria 
Meets inclusion/exclusion criteria 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
EYENOVIA CLINICAL STUDY PROTOCOL EYN-MYD-TP-32 VERSION C  
 
 
 
CONFIDENTIAL - PAGE 11 OF 61 
20 NOVEMBER 2018 
 
3. INTRODUCTION AND RATIONALE  
  
3.1. Introduction 
 
Eyenovia, the Sponsor of this study, is a specialty pharmaceutical company pursuing development of a 
variety of ocular topical medications, including a pupil dilation medication with a fixed combination 
formulation of phenylephrine 2.5% and tropicamide 1%, for microdose delivery.  Eyenovia’s drug 
formulations are administered using a proprietary system called the Microdose Delivery Dispenser 
(MiDD), which is designed to accurately deliver precise low-volume doses of topical ocular 
pharmaceuticals.  
 
Phenylephrine is an α1-adrenegic receptor sympathetic agonist that has been used for more than 70 years 
to dilate the pupil for diagnostic, therapeutic, and surgical procedures. Systemically, phenylephrine, when 
administered topically to the eye at higher doses, is known to cause serious cardiovascular reactions 
including ventricular arrhythmias, myocardial infarction, and elevated blood pressure in some patients1,2. 
These risks are especially relevant when more mydriasis is required, and phenylephrine 10% is used, 
rather than the lower 2.5% concentration.   Tropicamide is a synthetic derivative of tropic acid that 
became available for ocular use in 19593.  It is an anticholinergic mydriatic agent that acts by blocking the 
iris sphincter muscle and the ciliary muscle responses to acetylcholine.  As an anticholinergic related to 
atropine and scopolamine, systemically absorbed tropicamide in higher doses has the potential to cause 
rare but potentially serious adverse events, particularly in the pediatric population.  
 
Eyenovia’s formulation (fixed combination phenylephrine 2.5%-tropicamide 1%) is a proprietary 
investigational product that utilizes microdosing technology to facilitate accurate topical ocular delivery 
of a controlled quantity of mydriatic agents in a precise manner to maximize therapeutic control and 
minimize systemic absorption and loss to washout. 
 
3.2. Pupil Dilation 
 
A variety of ophthalmic evaluations require pupil dilation to provide an unobstructed view of the lens, 
retina, and optic nerve. The degree of pupil dilation required depends on the procedure being performed.   
For routine diagnostic procedures, and in conditions where short term pupil dilation is desired, the goal is 
to achieve adequate mydriasis for visualization with minimal cycloplegic effect. 
 
 
 
 
1 Vaajanen A, Vapaatalo H. A single drop in the eye - Effects on the whole body? Open Ophthalmol J. 
2017;11:305‒314. doi: 10.2174/1874364101711010305 
2 Farkouh A, Frigo P, Czejka M. Systemic side effects of eye drops: a pharmacokinetic perspective. Clin 
Ophthalmol. 2016;10:2433‒2441. 
3 Merrill D L, Goldber, B, Zavell S. bis-Tropamide, A new parasympatholytic. Curr Ther Res. 1960;12:43‒50. 
 


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CONFIDENTIAL - PAGE 12 OF 61 
20 NOVEMBER 2018 
 
3.3. Existing Mydriatic Options 
 
Dilation of the pupil is currently produced with parasympathetic anticholinergics such as tropicamide, 
cyclopentolate, homatropine, and atropine; or with sympathetic agonists (sympathomimetics) such as 
phenylephrine4. 
 
Parasympathetic antagonists block acetylcholine activation of muscarinic receptors, thereby paralyzing 
the iris sphincter muscle. The result is an enlarged pupil; however, at the same time, the ciliary muscle 
that regulates visual accommodation can be paralyzed, resulting in blurry vision that can persist for hours. 
Additionally, rare but serious systemic side effects have been observed after ocular administration of 
anticholinergics such as tropicamide, particularly in infants and children5,6. 
 
Sympathetic agonists produce dilation by stimulating the iris dilator muscle and constricting ophthalmic 
blood vessels. Although phenylephrine does not induce blurry vision, at lower concentrations; e.g., 2.5%, 
it generally does not dilate the pupil as well as parasympathetic agonists. Where full mydriasis is required, 
higher concentrations of phenylephrine; e.g., 10%, are required.  Higher concentration phenylephrine, 
however, has been known to cause cardiovascular effects after systemic absorption, which include 
hypertension, tachycardia, and more rarely, arrhythmia and stroke, especially in patients with pre-existing 
heart conditions7.  
 
To maximize dilation efficacy and minimize side effects, most ophthalmologists employ a combination of 
both classes of mydriatic agents.  
 
3.4. Phenylephrine and Tropicamide Ophthalmic Microdosing Solutions for Pupil Dilation 
 
Eyenovia’s MiDD is designed to deliver precise microvolumes of topical ophthalmic medications using a 
piezo-generated microdroplet spray. When the dispenser is actuated, a precisely calibrated single 
microdose is delivered.  It is anticipated that microdosing of ophthalmic drugs will enhance drug 
bioavailability and efficacy while minimizing systemic absorption and any associated side effects. 
 
Phenylephrine ophthalmic solution is currently marketed with 2.5% and 10% concentrations (Paragon 
BioTeck; Akorn, Inc.) for dilation of the pupil.  Tropicamide ophthalmic solution is also approved for 
eyedrop delivery at 0.5% and 1.0% concentrations (Alcon Laboratories; Bausch & Lomb; Akorn, Inc.).  
Eyenovia’s fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution formulation uses 
US Food and Drug Administration (FDA)-approved formulations of phenylephrine 2.5% and tropicamide 
1.0%.  The drug is administered in approximately 8 µL microdoses using the MiDD.   Eyedrops 
 
4 Grob SR, Gonzalez LA, Daly MK. Management of mydriasis and pain in cataract and intraocular lens surgery: 
review of current medications and future directions. Clin Ophthalmol. 2014;8:1281‒1289. doi: 
10.2147/OPTH.S47569 
5 Vaajanen A, Vapaatalo H. A single drop in the eye - Effects on the whole body? Open Ophthalmol J. 
2017;11:305‒314. doi: 10.2174/1874364101711010305 
6 Farkouh A, Frigo P, Czejka M. Systemic side effects of eye drops: a pharmacokinetic perspective. Clin Ophthalmol. 
2016;10:2433‒2441. 
7 Ibid. 


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20 NOVEMBER 2018 
 
containing a combination of phenylephrine and tropicamide are only available outside of the US (Mydrin-
P, Japan; Phenyltrope, Canada); no similar drugs are formulated for microdosing. 
 
Ocular microdosing with the MiDD has been evaluated in humans using phenylephrine 2.5%, 
phenylephrine 10%, and tropicamide 1.0% solutions, individually.   
 
3.5. Ophthalmic Microdosing 
 
The rationale for Eyenovia’s fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution 
development is the desire to optimize mydriasis while reducing systemic side effects, improving local 
tolerability, and decreasing the amount of medication lost to drainage from increased tear turnover rate.  
 
The average tear volume of an adult eye is 6−7 μL8,9. The estimated maximum volume of eyedrops that 
the cul de sac can contain, albeit momentarily, is approximately 30 μL; this is reduced to 10 µL with 
normal blinking10.  The typical eyedropper (either unit dose or multidose), however, provides drops that 
range from 25 μL to > 50 μL. Even among bottles of the same size and shape, eyedrop sizes vary 
significantly depending on the angle of the bottle, the amount of ophthalmic solution remaining, and even 
among manufacturers11.  
 
Much of the eyedrop volume overflows into the nasolacrimal canal where the active drug product 
becomes available systemically. Ocular drugs that are absorbed by the nasolacrimal mucosa mimic 
intravenous injection delivery insofar as they are not susceptible to first-pass hepatic metabolism. 
Additionally, ocular medication in swallowed nasolacrimal secretions is theoretically available for 
absorption in the gastrointestinal tract. Taken together, only a small fraction of the applied medication is 
absorbed directly into the eye, while there remain multiple opportunities for systemic absorption12. This 
can result in significant safety concerns, especially with drugs like phenylephrine and tropicamide that are 
known to produce dose-dependent systemic side effects13,14.  Additionally, excess drug in the eye may 
cause ocular surface toxicity and tolerability issues. 
 
Instillation of eyedrops stimulates lacrimation and can increase the tear turnover rate more than 4-fold, 
which additionally dilutes the drug product administered15.  If the eyedrop causes discomfort, the loss rate 
 
8 Mishima S, Gasset A, Klyce SD Jr, Baum JL. Determination of tear volume and tear flow. Invest. Ophthalmol. 
5(3), 264‒276 (1966). 
9 Scherz W, Doane MG, Dohlman CH. Tear volume in normal eyes and keratoconjunctivitis sicca. Albrecht Von 
Graefes Arch. Klin. Exp. Ophthalmol. 1974;192(2):141‒150. 
10 Shell JW. Pharmacokinetics of topically applied ophthalmic drugs. Surv Ophthalmol. 1982;26(4):207-18. 
11Mammo ZN, Flanagan JG, James DF, Trope GE.Generic versus brand-name North American topical glaucoma 
drops. Can J Ophthalmol. 2012;47(1):55‒61. doi: 10.1016/j.jcjo.2011.12.004. 
12 Urtti A, Salminen L. Minimizing systemic absorption of topically administered ophthalmic drugs. Surv. 
Ophthalmol. 37(6), 435−456 (1993). 
13Vaajanen A, Vapaatalo H. A single drop in the eye - Effects on the whole body? Open Ophthalmol J. 
2017;11:305‒314. doi: 10.2174/1874364101711010305 
14 Farkouh A, Frigo P, Czejka M. Systemic side effects of eye drops: a pharmacokinetic perspective. Clin Ophthalmol. 
2016;10:2433‒2441. 
15 Jordan A, Baum J. Basic tear flow. Does it exist? Ophthalmology. 1980 Sep;87(9):920‒930. 


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20 NOVEMBER 2018 
 
can be even higher16.  Approximately 80% of a medication instilled as an eyedrop is lost to drainage 
during the first 15‒30 seconds17. 
 
Microdosed mydriatic medications have been demonstrated to provide similar or superior efficacy 
compared to traditional eyedropper systems. Microdosing can also reduce systemic absorption and side 
effects, improve patient comfort, and minimize drug dilution and spillage. Studies comparing microdosed 
phenylephrine and cyclopentolate to traditional eyedrops (26 ‒ 30 µL drop size) in premature babies18 
and in full-term infants19 have shown equivalent pupil dilation with drop sizes ranging from 5 ‒ 8 µL20,21 
while reducing systemic levels by more than 50%22. 
 
Tropicamide 1.0% spray application caused similar pupil dilation as the same solution delivered as an 
eyedrop, but with less discomfort23 .  The main reason microdosing has not become the accepted method 
for topical ocular dosing of ophthalmic medication is that no system has been developed that can 
consistently deliver accurate microdoses of these medications. 
 
As shown in Figure 1, the Eyenovia MiDD is designed to deliver a finely controlled microdroplet spray of 
phenylephrine 2.5%-tropicamide 1% ophthalmic solution with precisely defined volume, velocity, and 
geometry.24 These characteristics facilitate high-precision, piezo-generated medication delivery that 
gently coats the ocular surface. 
 
 
16 Farris RL, Stuchell RN, Mandell ID. Basal and reflex human tear analysis. I Physical measurements: osmolarity, 
basal volumes, and reflex flow rate. Am Acad Ophthal 1981;88:852. 
17 Lallemand F, Daull P, Benita S, Buggage R, Garrigue JS. Successfully improving ocular drug delivery using the 
cationic nanoemulsion, novasorb. J Drug Deliv. 2012;2012:604204. 
18 Wheatcroft S, Sharma A, McAllister J. Reduction in mydriatic drop size in premature infants. Br J Ophthalmol. 
1993;77(6):364-5. 
19 Lynch MG, Brown RH, Goode SM, Schoenwald RD, Chien DS. Reduction of phenylephrine drop size in infants 
achieves equal dilation with decreased systemic absorption. Arch Ophthalmol. 1987;105(10):1364-5. 
20 Wheatcroft S, Sharma A, McAllister J. Reduction in mydriatic drop size in premature infants. Br J Ophthalmol. 
1993;77(6):364-5. 
21 Lynch MG, Brown RH, Goode SM, Schoenwald RD, Chien DS. Reduction of phenylephrine drop size in infants 
achieves equal dilation with decreased systemic absorption. Arch Ophthalmol. 1987;105(10):1364-5. 
22 Ibid. 
23Akman A, Aydin P. Comparison of mydriatic efficacy of spray application and drop instillation of tropicamide 
1%. Eye (Lond). 1999;13(5):653‒655. 
24 Ianchulev T, Chayet A, Kahook M, Packer M, Pasquale L, Weinreb RN. Pharmacodynamic profile of mydriatic 
agents delivered by ocular piezo-ejection microdosing compared with conventional eyedropper. Ther Deliv. 
2016;7(11):751-760. 
 


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Figure 1: Microdroplet Distribution Pattern from the MiDD 
Image analysis of high-speed photographs of the microdroplet plume quantitatively confirms precise 
medication microdroplet delivery. Shown are representative side (A) and cross-sectional (B) views of 
the droplet dispersal pattern. Color-coding indicates the percentage of the total medication dose 
present at different distances from the nozzle-to-target dead- center axis. The cross-sectional pattern on 
the right was captured 3 cm from the MiDD nozzle. 
 
 
 


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4. INVESTIGATIONAL PRODUCT DESCRIPTION 
 
This study will evaluate the safety and efficacy of topical ocular microdose administration of a fixed 
combination of phenylephrine 2.5%-tropicamide 1% ophthalmic solution for mydriasis in routine 
diagnostic procedures and in conditions where short-term pupil dilation is desired. 
 
4.1. Findings from Non-Clinical and Clinical Studies  
 
Detailed information on nonclinical and clinical studies of fixed combination phenylephrine 2.5%-
tropicamide 1% ophthalmic solution is provided in the Investigator Brochure (EYN-MYD-TP-3-
1/32). 
4.2. How Provided 
Eyenovia’s fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution is contained 
sterile in the Eyenovia MiDD, which is designed to accurately deliver precise low-volume doses of 
topical ocular pharmaceutical ingredients.  As shown in Figure 2, the MiDD is composed of a “cartridge 
assembly” (shown in white in the the image below) and the base unit (shown in gray in the image below).  
 
 
 
 
Figure 2: Eyenovia MiDD Front (L) and Back (R) View 
 
Figure 3 shows a diagram of the MiDD base unit and cartridge assembly.  The cartridge assembly is the 
primary container closure system, which contains a vial filled with sterile fixed combination 
phenylephrine 2.5% - tropicamide 1% ophthalmic solution.  At the time of initial MiDD actuation, the 
seal on the vial is punctured to allow the drug to flow to the MiDD delivery system.  The cartridge 
assembly also contains a piezo-delivery system designed to drive delivery of the drug through the Spray 
Nozzle in discrete pulses repeated in a discrete cycle of duration ≤ 100 milliseconds, which is the average 
involuntary blink response time. 
 
Dose Button 
Load Button 
MiDD Base Unit 
MiDD Cartridge 
Spray Nozzle 


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Figure 3:  Diagram of the MiDD Base Unit (L) and Cartridge Assemblies (R) 
 
Prior to initial MiDD use, fixed combination phenylephrine 2.5%-tropicamide 1% solution is aseptically 
transferred from the 2R vial in which it was received into the MiDD vial, the MiDD vial is inserted into 
the cartridge assembly, the cartridge and base are clipped together, and the seal on the vial containing the 
drug is punctured by depressing the MiDD Load Button.  Puncture of this seal creates a fluid path for the 
drug.  Depression and release of the Load Button 5 times primes the system for drug delivery.  At the time 
of dosing, the Load Button is again pressed and released to activate the piezo element to create the aerosol 
containing the drug dose.  When the Dose Button is pressed, approximately 8 µL of the drug is delivered 
through the Spray Nozzle to the cornea.  
4.3. Route of Administration, Dosage, Regimen, Treatment Period(s) 
The study medication will be administered in accordance with the treatment assignment schedule on a 
single day in the clinic by trained clinical study staff.  Two sprays of fixed combination phenylephrine 
2.5%-tropicamide 1% ophthalmic solution will be topically administered within a 5-minute period to 
both eyes.  The concentration of the active ingredients selected for this study is based on the standard 
regimen used for mydriasis where short term pupil dilation is required. The single-use regimen and 1-
day treatment period is selected based on the medication’s intended use for in-office mydriasis.  
 
 
 
 
Load Button 
Vial 
Cartridge Assembly 
Base Unit Assembly  
 
(Electrical mechanical 
components in top diagram) 
Spray Nozzle 


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5. STUDY OBJECTIVE 
 
The primary objective of this study is to evaluate the safety and efficacy of Eyenovia’s fixed combination 
of phenylephrine 2.5%-tropicamide 1% ophthalmic solution administered with the MiDD for dilation of 
the pupil as compared to 
 placebo. 
 
 
6. STUDY DESIGN 
 
This trial is a multicenter, double-masked, placebo-controlled, cross-over superiority study evaluating 
Eyenovia’s fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution versus placebo.   
 
Volunteer participants from 2 – 3 US-based investigational sites will be screened for study eligibility 
during a Screening Visit and enrolled after signing the study-specific informed consent form (ICF).  
Subjects meeting all inclusion/exclusion criteria will be scheduled for 3 treatment visits, which must 
occur at least 2 days, but no more than 7 days apart.  A 2 sequence, 3 period crossover design will be 
used.  At each of the three treatment visits, baseline measurements will be taken, then either the 
investigational drug or the placebo will be administered to both eyes (OU) in two separate instances, 
approximately 5 minutes apart.  Only 1 investigational product will be administered per treatment visit, 
and product administration will occur in a randomized order using one of two sequences - ABB and BAA, 
where A is the Eyenovia fixed combination and B is the placebo control. Afterwards, efficacy and safety 
assessments will be performed at specific time intervals.   
 
This study will be double-masked.  There will be no differences in the presentation of the investigational 
drug or placebo administered and all study personnel conducting ophthalmic assessments will be masked 
to study drug assignment.  Study drug administration will be performed by at least 3 different trained 
personnel during the study. Personnel who administer study drug on a given day will not be allowed to 
perform post-study drug administration ophthalmic assessments on that day.  
 
 
 
 


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7. STUDY OUTCOMES 
 
7.1. Effectiveness Outcomes 
 
The primary performance endpoint is mean change in pupil diameter at 35 minutes from the time of first 
drug dose versus baseline, as measured by digital pupillometry in highly photopic conditions.  The highly 
photopic condition will be established using a fully-charged transilluminator (muscle light) at the 
brightest setting.  The difference in mean change in pupil diameter will be compared between Eyenovia’s 
fixed combination phenylephrine 2.5%-tropicamide 1% and placebo.   
 
The study’s sample size will provide 95% power to detect a 0.5 mm difference between drugs assuming a 
standard deviation of 1.0 mm using a paired t-test with a two-sided significance level of 0.05. 
 
Additional exploratory outcomes (also to be measured in highly photopic conditions) include: 
 
• 
Proportion of eyes achieving pupil diameter of 6.0 mm or greater at 35 minutes 
• 
Proportion of eyes achieving pupil diameter of 7.0 mm or greater at 35 minutes 
• 
Mean change in pupil diameter at other timepoints (20, 50, 65, 80, 120, and 180 minutes) 
• 
Distribution of pupil diameters at 20, 35, 50, 65, 80, 120, and 180 minutes 
• 
Time from baseline to maximal pupil dilation  
 
7.2. Safety Outcomes 
 
A safety analysis will be conducted to compare fixed combination of phenylephrine 2.5%-tropicamide 1% 
µD against the placebo control for the following outcomes: 
 
• 
Slit-lamp examination (SLE) findings 
• 
Occurrence of AEs 
• 
IOP measured at 65 minutes post-administration 
• 
Visual acuity changes 
 
 
 
 


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8. STUDY POPULATION   
 
Volunteer participants will be considered enrolled after signing the study-specific ICF(s) and screened for 
study eligibility during a Screening Visit.  Subjects meeting all inclusion/exclusion criteria will be 
scheduled for 3 treatment visits.  It is assumed that up to 20% of subjects (n = 10) will fail to complete the 
study visits; therefore, to have 54 subjects evaluable for the primary efficacy analysis, a minimum of 65 
subjects will be randomized to  study drug administration at Treatment Visit 1 (Day 1).  To account for 
possible screen failures, the study will enroll up to 90 subjects. 
 
8.1. Inclusion Criteria 
 
Subjects eligible for study participation must meet each criterion listed below.  Ocular criteria must be 
met for both eyes. 
  
1. Male or female of any age. 
2. Female subjects must be either pre-menarche, 1-year postmenopausal, surgically sterilized, 
or, if of childbearing potential, have a negative urine pregnancy test during the Screening 
Visit and agree to use an acceptable form of contraception throughout the study.  Acceptable 
methods include the use of at least one of the following: intrauterine (intrauterine device), 
hormonal (oral, injection, patch, implant, ring), barrier with spermicide (condom, 
diaphragm), or abstinence. 
3. Ability to provide signed written consent prior to participation in any study-related 
procedures.  Subjects under the age required for informed consent must have the ability to 
provide additional written assent and/or parental consent, as required by the reviewing IRB. 
4. Ability to return for all study treatment visits. 
5. Photopic screening pupil diameter ≤ 3.5 mm in each eye. 
 
8.2. Exclusion Criteria 
 
Subjects with any of the following diseases, surgeries or conditions are ineligible for study participation.  
Subjects may not participate if either eye meets any of the ocular exclusion criteria. 
 
1. Pregnancy or lactation. 
2. Allergy to phenylephrine hydrochloride. 
3. Allergy to tropicamide. 
4. Allergy to benzalkonium chloride. 
5. History of benign prostatic hyperplasia. 
6. Use of benzodiazepines, monoamine oxidase inhibitors, tricyclic antidepressants, 
anticonvulsants at screening or anticipated during the study period.  
7. Participation in any study of an investigational drug or device within 30 days prior to the 
Screening Visit, or at any time during the study period. 
8. History of closed-angle glaucoma. 
9. Anatomically narrow anterior chamber angles (Van Herrick grade ≤ 2 in either eye). 
10. Ocular surgery or laser treatment of any kind prior to the Screening Visit. 
11. History of chronic or acute uveitis. 


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12. History of traumatic iritis or hyphema. 
13. History of traumatic mydriasis or angle recession. 
14. History of heterochromia. 
15. Irregularly-shaped pupil secondary to ocular trauma or congenital defect. 
16. History of neurogenic pupil disorder (e.g., Horner’s syndrome, third cranial nerve palsy, 
Adie’s pupil, Argyl Robertson syndrome, etc.). 
17. History of anterior chamber intraocular lens (IOL) or iris-fixated IOL. 
18. History of iris surgery of any kind (e.g., iridotomy, iridectomy, coreoplasty). 
19. History of iris atrophy. 
20. History of iris – cornea apposition/touch. 
21. Unwilling or unable to discontinue use of contact lenses at treatment visits. 
22. Current active eye disease for which topical or systemic ophthalmic medication is necessary, 
except for dry eye disease managed using artificial tears (AT).   AT’s must be discontinued on 
the day of each treatment visit. 
23. Presence of a severe/serious ocular condition, or any other unstable medical condition that, in 
the Investigator’s opinion, may preclude study treatment and/or follow-up. 
 
 
 
 


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9. STUDY METHODS 
 
9.1. Subject Informed Consent 
 
The IRB-approved study ICF must be used for administration of informed consent.  Dependent on IRB 
requirements for subjects under 18 years of age, an IRB-approved study assent form may be supplemental 
to the study ICF.   
 
The investigator or designee will explain the study purpose, procedures and responsibilities to the 
potential participant and provide sufficient opportunity to ask questions, while allowing adequate time for 
consideration of the information provided.  Written consent (and assent, if applicable) for study 
participation must be present prior to initiation of any study-specific procedure; subjects will be 
considered enrolled in the study upon their (and/or their parent’s/guardian’s, if applicable) signature on 
the consent form(s).  It is the responsibility of the Investigator to complete the informed consent process, 
maintain a copy of the signed consent/assent forms in the subject’s medical records, and provide each 
subject with a copy of their fully-executed consent documents. 
 
9.2. Subject Selection and Screening Procedures 
 
Study participants will be recruited from the Investigator’s patient population, referrals, or other outreach 
methods.  Written recruitment materials directed to potential study participants must be approved by the 
IRB.   
 
Where possible, candidate participants may be pre-screened via review of their medical charts to evaluate 
potential eligibility based on study inclusion/exclusion criteria.  During the informed consent discussion, 
the potential participant’s willingness and ability to meet the follow-up requirements will be evaluated.  
Those who elect to sign the study consent form(s) will be considered enrolled in the study and given a 
study identification code.   At/after the time of enrollment, the subject will be evaluated at the Screening 
Visit for study eligibility based on inclusion/exclusion requirements established in Sections 8.1 and 8.2.  
Subjects who fail to qualify for the study will be considered “screen failures” and exited from the study.  
Subjects who successfully complete the Screening Visit will be scheduled for their next visit at which the 
first study medication will be administered.   
 
9.3. Randomization to Study Drug Administration Sequence  
 
Each eligible subject will be randomly assigned to receive one of 2 sequences (ABB or BAA) of study 
drug administration where either investigational drug or placebo will be administered in both eyes at each 
of the 3 treatment visits.  After Treatment Visit 3, each study subject should have received either 2 
treatments with the investigational product or 2 treatments with the placebo. The study drug 
administration schedule is equally distributed across the 2 randomization sequences.   Subject drug 
administration assignments will be prepared using a computer-generated randomization scheme by an 
independent statistician who is not involved in the day-to-day conduct of the study.  The  drug assignment 
sequence (Study Drug ABB or BAA) will be provided in paper format contained in a sealed envelope for 
each subject who randomized to treatment. 
 


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The study drug administered will be masked to the study subject, the investigator, and study staff 
administering performing clinical assessments.  The Sponsor (or designee) involved in day-to-day study 
management will also be masked to study drug assignments.  
 
9.3.1. Procedure for Unmasking Study Drug Assignment Code  
 
Unmasking of study drug assignments can only occur in the event of a medical emergency or occurrence 
of an AE that, in the opinion of the investigator, warrants such action.  In the absence of medical need, the 
study drug assignment code will not be available to the investigator, site staff, or Sponsor (or designee) 
representative involved in day-to-day study management until study completion and database lock. 
 
In the event an investigator considers unmasking of a subject’s study drug assignment to be necessary, 
he/she must first contact the study Medical Monitor (or designee) to discuss the case.  Only after 
consultation with the study Medical Monitor will the decision be made regarding unmasking. Study drug 
assignments will be revealed only for the subjects approved for unmasking, thus leaving masking on 
remaining subjects intact. 
 
9.4. Study Drug Packaging, Labeling and Storage 
 
The investigational product and placebo, each delivered in microdose form using the Eyenovia MiDD, 
will be evaluated in this study.  Each product must be aseptically transferred by a licensed pharmacist 
affiliated with the study site from the container in which they are received to the sterile vial provided with 
the MiDD cartridge.  After aseptic transfer, the filled vial is re-inserted into the MiDD cartridge. 
 
To maintain masking of the study drug assignments for staff administering investigational product and/or 
performing clinical evaluations, study inventory storage and management will be performed by a 
pharmacy associated with the study site.  The Sponsor, or designee, will ship study drug kits to each site 
pharmacy.  Each kit will be labeled with the study protocol number, study drug code, product storage 
requirements, and the Caution – New Drug – Limited by Federal (US) Law to Investigational Use label.  
 
Instructions regarding study medication preparation are included in  
APPENDIX 2:  STUDY MEDICATION PREPARATION PROCEDURE.  Each designated site 
pharmacist will be trained and certified by the Sponsor, or designee, prior to study medication 
preparation. 
 
Prior to use, study medication must be stored in a secure area.  To maintain product stability, store at 20° 
to 25° C (68° to 77° F).  Protect product from light and excessive heat.   
 
New MiDDs must be prepared by the pharmacy for use by site personnel on each treatment day.  To 
maintain masking of site personnel, the MiDD containing each study drug will be labeled with the 
appropriate study randomization code, pharmacy preparation date and “use by” date, then forwarded to 
site personnel for use.  The date on which the MiDD is used will be added by site personnel.  At the end 
of the treatment day, each MiDD provided will be returned to the pharmacy for storage. Please note that 
each MiDD prepared must be used on the day of preparation; the MiDD may not be re-used on 
subsequent days.   


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9.5. Study Drug Administration 
 
Site personnel designated to administer study medication will be trained by the Sponsor on proper use of 
the MiDD prior to study initiation.  A single MiDD should be used for study drug administration to all 
subjects whose study drug assignment corresponds to the dispenser code on that treatment day. At each 
treatment visit, after collection of baseline BCDVA, IOP and SLE data, study medication will be 
administered by designated site personnel in accordance with the subject’s assigned sequence.  Subjects 
will be administered 2 doses of study drug in each eye.  After the initial dosing of each eye, 
approximately 5 minutes should elapse before the second dose administration.  Study drug administration 
procedures are further detailed in APPENDIX 3:  STUDY MEDICATION ADMINISTRATION 
PROCEDURE. 
 
9.6. Study Visits and Clinical Assessments 
 
Study participants will be evaluated at a Screening Visit.  Subjects who qualify for further study 
participation will be scheduled for visits on Treatment Day 1, Treatment Day 2 and Treatment Day 3 at 
which the assigned study drug is administered and examinations are performed.  Data collected from each 
visit will be documented in the source records and recorded on study electronic Case Report Forms 
(eCRF).   
 
The timing and frequency of assessments/procedures to be performed at each visit is outlined in 
APPENDIX 1:  SCHEDULE OF MEDICATION ADMINISTRATION AND EXAMINATIONS.  
Methodology for study examinations is presented in APPENDIX 4:  EXAMINATION 
PROCEDURES. 
 
9.6.1. Screening Visit (-14 Days to -1 Day) 
 
The Screening Visit must occur no more than 14 days prior and no less than 1 day prior to Treatment 
Visit 1.  At the Screening Visit, the following activities will be performed:  
 
• 
Documentation of study informed consent/assent using IRB-approved consent form(s) 
• 
Collection of demographic data (gender, age, ocular comorbidities) 
• 
Collection of medical/ocular history and medication use data 
• 
Administration of urine pregnancy test, if applicable 
• 
BCDVA25 – OU 
• 
Pupil diameter measurement – OU 
• 
Slit lamp biomicroscopy including Van Herrick angle assessment – OU 
• 
Dilated Fundus Exam – OU  
• 
IOP measurement - OU   
• 
Determination of study eligibility 
 
25 Pediatric subjects who are unable to perform ETDRS testing must have uncorrected visual acuity evaluated using 
an age-appropriate acuity testing method.   
 


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9.6.2. Treatment Visit 1 (Day 1) 
 
Subjects will return to the site at least 1 day and no more than 14 days after the Screening Visit.  The 
subject will be queried regarding the occurrence of any adverse events since the time of the Screening 
Visit.  The subject will be assigned to his/her study drug randomization schedule based on iris color 
cohort assignment (dark irises are either black or brown in color, while light irises are all other colors), 
and the following baseline measurements will be taken prior to study drug administration: 
  
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
BCDVA26 – OU 
• 
Slit lamp biomicroscopy – OU 
• 
IOP measurement - OU   
 
The first study drug in the drug administration sequence will be administered OU, then administered 
again approximately 5 minutes later OU.  The completion of the first administration of medications will 
be considered Time 0 (T0).  The following assessments will be done at the timepoints specified below: 
 
Timepoint 
Clinical Assessment (to be performed in the specified order) 
T0 + 20 minutes (+/- 2 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 35 minutes (+/- 5 minutes) 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
AE assessment 
T0 + 50 minutes (+/- 5 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 65 minutes (+/- 5 minutes) 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
 IOP measurement – OU 
T0 + 80 minutes (+/-10 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 120 minutes (+/-10 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 180 minutes (+/-10 minutes) 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
BCDVA23 – OU 
• 
Slit lamp biomicroscopy – OU 
• 
AE assessment 
 
 
 
 
26 Pediatric subjects who are unable to perform ETDRS testing must have uncorrected visual acuity evaluated using 
an age-appropriate acuity testing method.   


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9.6.3. Treatment Visit 2 (Day +3 to Day +8) 
 
Subjects will return to the site.  Subjects will be queried regarding the occurrence of any adverse events 
since the previous visit and the following baseline measurements will be taken prior to study drug 
administration: 
 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
BCDVA27 – OU 
• 
Slit lamp biomicroscopy – OU 
• 
IOP measurement - OU   
 
The second study drug in the drug administration sequence will be administered OU, then administered 
again approximately 5 minutes later OU.  The completion of the first administration of medications will 
be considered Time 0 (T0).  The following assessments will be done at the timepoints specified below: 
 
Timepoint 
Clinical Assessment (to be performed in the specified order) 
T0 + 20 minutes (+/- 2 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 35 minutes (+/- 5 minutes) 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
AE assessment 
T0 + 50 minutes (+/- 5 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 65 minutes (+/- 5 minutes) 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
 IOP measurement – OU 
T0 + 80 minutes (+/-10 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 120 minutes (+/-10 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 180 minutes (+/-10 minutes) 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
BCDVA23 – OU 
• 
Slit lamp biomicroscopy – OU 
• 
AE assessment 
 
 
 
 
 
27 Pediatric subjects who are unable to perform ETDRS testing must have uncorrected visual acuity evaluated using 
an age-appropriate acuity testing method.   
 


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9.6.4. Treatment Visit 3 (Day +5 to Day +15) 
 
Subjects will return to the site.  Subjects will be queried regarding the occurrence of any adverse events 
since the previous visit and the following baseline measurements will be taken prior to study drug 
administration: 
 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
BCDVA28 – OU 
• 
Slit lamp biomicroscopy – OU 
• 
IOP measurement - OU   
 
The third study drug in the drug administration sequence will be administered OU, then administered 
again approximately 5 minutes later OU.  The completion of the first administration of medications will 
be considered T0.  The following assessments will be done at the timepoints specified below: 
 
Timepoint 
Clinical Assessment (to be performed in the specified order) 
T0 + 20 minutes (+/- 2 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 35 minutes (+/- 5 minutes) 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
AE assessment 
T0 + 50 minutes (+/- 5 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 65 minutes (+/- 5 minutes) 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
 IOP measurement – OU 
T0 + 80 minutes (+/-10 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 120 minutes (+/-10 minutes) 
• 
Pupil diameter measurement – OU 
T0 + 180 minutes (+/-10 minutes) 
• 
Pupil diameter measurement – OU 
• 
Pupillary light reflex - OU 
• 
BCDVA25 – OU 
• 
Slit lamp biomicroscopy – OU 
• 
AE assessment 
 
If applicable, a urine pregnancy test will be administered at this visit and, upon completion of the T0 + 
180-minute assessment, the subject will have completed the study.29 
 
 
 
28 Pediatric subjects who are unable to perform ETDRS testing must have uncorrected visual acuity evaluated using 
an age-appropriate acuity testing method.  
29 Subjects whose pupil diameter is larger than baseline at the time of study exit may be followed at hourly intervals 
up to 6 hours after medication administration to gather additional data regarding the duration of study treatment 
effect.  
 
 


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9.4.5. Unscheduled Visits 
 
Unscheduled visits are those which are not required by the study protocol, but which occur due to an 
ocular intervention or a subject complaint regarding their eye.  No specific testing is required at 
unscheduled visits; rather, the investigator and/or qualified study staff will perform the procedures 
necessary to treat/evaluate the subject at these visits.  Clinical data from these visits will be recorded on 
the relevant Unscheduled Visit eCRF.    
 
9.4.6. Missed Visits 
 
Subjects who miss a scheduled visit should be contacted by site personnel to encourage the subject to 
return to clinic as soon as possible after the missed visit.  In the event a subject does not return for 
multiple consecutive examinations, site personnel must make a minimum of 3 documented attempts via 
telephone, email, or regular mail to contact the subject.  If the subject does not reply to any of these 
attempts, site personnel must send a letter by certified mail (with request for notification of delivery 
receipt) to the subject.  If a subject is non-responsive to each of these follow-up attempts, he/she will be 
terminated from the study and considered lost to follow-up. 
 
9.7. Study Exit 
 
9.7.1. Subject Termination 
 
Subjects may be terminated from the study due to: 
 
• 
Failure to meet protocol eligibility criteria  
• 
Investigator decision that termination is medically indicated 
• 
Voluntary withdrawal from the study 
• 
Loss to follow-up 
• 
Other administrative reasons (e.g., study terminated by Sponsor, technical problems, non-
compliance with study medication administration or other study requirements, etc.) 
 
Subjects who terminate due to the occurrence of an AE will be followed until resolution or stabilization of 
the event. 
 
Subjects who exit the study due to failure to meet eligibility criteria may be replaced.  Subjects who exit 
the study prematurely for other reasons will not be replaced.  The Sponsor (or designee) should be 
promptly notified of a subject’s early termination, and information regarding the termination (e.g., date 
and reason) should be documented in the source records and on the Study Exit eCRF. 
 
9.7.2. Subject Completion 
 
Subjects will be considered to have completed the study if they did not terminate study participation prior 
to completion of primary endpoint assessment at Treatment Visit 3. 
 


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10. ADVERSE EVENTS 
 
Throughout the course of the study, all efforts must be made by the investigator to remain alert to possible 
AEs or untoward findings.  All AEs must be assessed for severity and relationship to the study drug, and 
the investigator must take all appropriate and necessary therapeutic measures required for AE resolution.  
Adverse events must be evaluated until resolution or, if the AE is assessed as chronic, until stable.   
 
10.1. Adverse Event Definitions 
 
Most of the AE definitions provided are published in either CFR 312.32 – IND Safety Reporting or 
ICH E6 – Good Clinical Practice:  Consolidated Guidance.  Because of the differing sources for these 
definitions, some terminology; e.g., “adverse event”, “adverse reaction”, and “adverse experience”, 
differs.  For this study, these terms should be considered equivalent. 
 
10.1.1. Adverse Event (AE) 
 
Any untoward medical occurrence associated with the use of a drug in humans, whether or not considered 
drug-related.30    
 
10.1.2. Adverse Drug Reaction (ADR) 
 
Any noxious and unintended response to a medicinal product administered at any dose.  A reasonable 
possibility must exist that the adverse reaction is related to the medicinal product administered.31 
 
10.1.3. Unexpected Adverse Event or Unexpected Suspected Adverse Reaction  
 
An AE or suspected adverse reaction is considered "unexpected" if it is not listed in the Investigator 
Brochure (IB) or is not listed at the specificity or severity that has been observed; or, if an IB 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.  "Unexpected," as used in this definition, also 
refers to AEs or suspected adverse reactions that are mentioned in the IB as occurring with a class of 
drugs or as anticipated from the pharmacological properties of the drug but are not specifically mentioned 
as occurring with the particular drug under investigation.32 
 
10.1.4.  Suspected Adverse Reaction 
 
Any AE for which there is a reasonable possibility that the drug caused the event. For the purposes of 
Investigational New Drug (IND) safety reporting, "reasonable possibility" means there is evidence to 
 
30 CFR 312.23 – IND Safety Reporting, 
https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=312.32 
31 ICH E6 – Good Clinical Practice:  Consolidated Guidance, Glossary, p 1. 
32 32 CFR 312.23 – IND Safety Reporting, 
https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=312.32 
 


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suggest a causal relationship between the study drug and the AE. Suspected adverse reaction implies a 
lesser degree of certainty about causality than adverse reaction, which means any AE caused by a drug.33  
 
10.1.5. Life-Threatening Adverse Event or Life-Threatening Suspected Adverse Reaction 
 
An AE 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.  Life-threatening 
AEs do not include AEs or suspected adverse reactions that, had they occurred in a more severe form, 
might have caused death.34 
 
10.1.6. Serious Adverse Event (SAE) or Serious Suspected Adverse Reaction 
 
An AE or suspected adverse reaction that, in the view of either the Investigator or Sponsor, results in any 
of the following outcomes:  
 
• 
Death 
• 
Is life-threatening35  
• 
Requires in-patient hospitalization or prolongation of existing hospitalization 
• 
Persistent or significant disability/incapacity, meaning there is a substantial disruption of the 
subject’s ability to conduct normal life functions 
• 
A congenital anomaly or birth defect 
 
Important medical events that may not result in death, be life-threatening, or require hospitalization may 
be considered serious when, based upon appropriate medical judgment, they may jeopardize the subject 
and may require medical or surgical intervention to prevent one of the outcomes listed in this 
definition.36  
 
10.1.7. Serious Ocular Adverse Events 
 
Serious ocular AEs include, but are not limited to: 
 
• 
A decrease in BCDVA of ≥ 30 total letters read (TLR) or an increase of ≥ 0.6 logMar from 
the most recent previous measurement of BCDVA 
• 
A decrease in BCDVA to light perception (LP) or worse that lasts > 1 hour 
• 
An ocular event that, in the opinion of the investigator, requires medical or surgical 
intervention to prevent permanent loss of sight 
• 
Severe intraocular inflammation; e.g., 4+ anterior chamber cell/flare or 4+ vitritis 
 
33 CFR 312.23 – IND Safety Reporting, 
https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=312.32 
34 CFR 312.23 – IND Safety Reporting, 
https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=312.32 
35 A life-threatening SAE is any AE or adverse reaction that places the subject, in the view of the Investigator, at 
immediate risk of death from the reaction as it occurred (i.e., it does not include a reaction that, had it occurred in a 
more severe form, might have caused death). 
36 CFR 312.23 – IND Safety Reporting, 
https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=312.32 
 


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• 
Corneal decompensation 
• 
Retinal tear or detachment 
• 
Central or branch retinal vein occlusion 
 
10.1.8. Treatment-Emergent Adverse Event (TEAE) 
 
An AE not present prior to initiation of therapy, or an already present event that worsens either in 
intensity or frequency following initiation of therapy.  
 
10.2. Adverse Event Assessment and Documentation 
 
All AEs that occur from the time the subject receives the first dose of study drug37 until study 
participation has been completed (e.g., TEAEs) must be documented.  Any medical condition present 
prior to the first study drug dose that remains unchanged or improves will not be recorded as an AE; 
however, a worsening of the condition after dosing with the study drug, will be considered an AE.   
 
Adverse events may be determined by evaluating the following in relation to the study subject: 
 
• 
Observed or volunteered problems 
• 
Complaints 
• 
Physical signs and symptoms  
• 
The occurrence of a medical condition during the study, which was absent at baseline 
• 
The worsening of a baseline medical condition during the study 
 
Treatment-emergent AEs, regardless of causal relationship, must be assessed by the investigator and 
recorded in the source documentation and the appropriate eCRF.  Each AE must be described as ocular or 
non-ocular along with the following information: date of onset, date of resolution, severity, frequency of 
the event (single episode, intermittent, continuous), action taken (none, medical and/or surgical), 
relationship to study drug, and seriousness criteria.  Any medication necessary for the treatment of an AE 
must be recorded on the Concomitant Medication eCRF and any surgical intervention necessary for AE 
treatment must be recorded on the Concomitant Procedure eCRF.  If more than one distinct AE occurs, 
each event should be recorded separately. 
 
Adverse events will be documented beginning at the time of onset, and documentation must continue 
until recovery is noted.  Events that are ongoing at the time of study exit must be followed until resolution 
or stabilization. 
 
10.2.1. Adverse Event Severity 
 
Adverse event severity will be assessed by the investigator using the following definitions: 
 
 
37 Events that occur between the Screening Visit and initial administration of study drug should be recorded as 
Medical History, and not as an AE.   
 


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Mild 
Subject is aware of sign or symptom, but it is easily tolerated 
Moderate 
Subject experiences discomfort enough to cause interference with 
usual activity 
Severe 
AE is incapacitating to subject, causing inability to work or do usual 
activity 
 
10.2.2. Adverse Event Relationship to Study Drug 
 
The relationship of the AE to study drug will be assessed by the investigator using the following 
definitions: 
 
Not Related 
Evidence exists that the AE has a cause other than the study 
drug (e.g. pre-existing condition or underlying disease, 
intercurrent illness, or concomitant medication) and does not 
meet any other criteria listed.  
  
Possibly Related 
A temporal relationship exists between event onset and 
administration of study drug.  Although the AE may appear 
unlikely to be related to the study drug, it cannot be ruled out 
with certainty; and/or the event cannot be readily explained by 
the patient’s clinical state or concomitant therapies.  
 
Probably Related 
A temporal relationship exists between the event onset and 
administration of study drug; it appears with some degree of 
certainty to be related based on known therapeutic and 
pharmacologic actions of the study drug.  It cannot be readily 
explained by the patient’s clinical state or concomitant 
therapies. 
 
Definitely Related 
Strong evidence exists that the study drug caused the AE.  
There is a temporal relationship between the event onset and 
administration of the study drug.  There is strong therapeutic 
and pharmacologic evidence that the event was caused by the 
study drug.  The patient’s clinical state and concomitant 
therapies have been ruled out as a cause.  
 
10.3 Adverse Event Reporting 
 
Any treatment-emergent AE occurring in the study must be reported on the AE eCRF.  Expedited 
reporting of an AE is required if the event is considered “serious”, regardless of relationship to the study 
drug or whether the event is expected or unexpected. 
 
 
 


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10.3.1. Expedited Adverse Event Reporting 
 
Any SAE that occurs during the study, regardless of relationship to the study drug, or whether the event is 
expected or unexpected, must be reported to the Sponsor, or the Sponsor’s designee, within 24 hours of 
the investigator’s becoming aware of the event.   
 
Reports should be made by completing the SAE eCRF.  Information provided on the SAE eCRF should 
be supplemented with hospitalization records, death certificate, clinic notes from specialists evaluating the 
subject’s condition, etc. as applicable for the event.  The urgency for reporting SAEs is 3-fold:  
 
• 
to facilitate discussion [and implementation, if necessary] by the Sponsor and the Investigator of 
appropriate follow-up measures;  
• 
To facilitate Investigator reporting of unanticipated problems involving risk to human subjects to 
the IRB/EC; and  
• 
To enable the Sponsor to fulfill the reporting requirements to the appropriate regulatory authority. 
 
 
 
 


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11. STATISTICAL METHODS 
 
The primary objective of this study is to evaluate the safety and efficacy of Eyenovia’s fixed combination 
of phenylephrine 2.5%-tropicamide 1% ophthalmic solution administered with the MiDD for dilation of 
the pupil as compared to the individual components. 
 
11.1. Efficacy Endpoints 
 
Primary Efficacy Endpoint: An efficacy analysis will be conducted to compare fixed combination of 
phenylephrine 2.5%-tropicamide 1% microdose ophthalmic solution against placebo with the following 
endpoint: 
 
The primary performance endpoint is mean change per visit in pupil diameter at 35 minutes versus 
baseline, as measured by digital pupillometry in highly photopic conditions.  The highly photopic 
condition will be established using a fully-charged transilluminator (muscle light) at the brightest setting.  
 
Additional Efficacy Analyses: 
• 
Proportion of eyes achieving pupil diameter of 6.0 mm or greater at 35 minutes 
• 
Proportion of eyes achieving pupil diameter of 7.0 mm or greater at 35 minutes 
• 
Mean change in pupil diameter at other timepoints (20, 50, 65, 80, 120, and 180 minutes) 
• 
Distribution of pupil diameters at 20, 35, 50, 65, 80, 120, and 180 minutes 
• 
Time from baseline to maximal pupil dilation 
 
These additional efficacy analyses will also be measured using digital pupillometry in highly photopic 
conditions. 
 
11.2. Sample Size 
 
Up to 90 subjects who have provided informed consent for study participation will be enrolled in the 
study and a minimum of 65 subjects will be randomized to study drug administration at Treatment Visit 1 
with the goal of having 54 subjects evaluable for the primary efficacy endpoint. A sample size of 27 
subjects per sequence, 54 total (the study has two sequences, ABB and BAA where A is the Eyenovia 
fixed combination drug, and B is the placebo control), will have 95% power to detect a difference in 
means of 0.5 mm, assuming a standard deviation of differences of 1.0 mm, using a paired t-test with a 
0.05 two-sided significance level. 
 
Power calculations are provided for additional exploratory outcomes below on an informational basis.  
 
When the sample size is 54, a two-sided 95% exact confidence interval for a single proportion will have a 
half-width of 13.9% or less.  
• 
For evaluation of the proportion of eyes achieving pupil size of 6.0 mm or greater at 35 minutes: 
if the observed proportion in the fixed combination treatment is 70.4% (38/54), then the exact 
95% confidence interval for the proportion will be [56.4%, 82.0%], with a half-width of 12.8%. 


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• 
For evaluation of the proportion of eyes achieving pupil size of 7.0 mm or greater at 35 minutes: 
if the observed proportion in the fixed combination treatment is 40.7% (22/54), then the exact 
95% confidence interval for the proportion will be [27.6% 55.0%], with a half-width of 13.7%. 
 
11.3. Safety Outcomes 
 
A safety analysis will be conducted to compare fixed combination of phenylephrine 2.5%-tropicamide 1% 
microdose against placebo for the following outcomes: 
• 
Slit-lamp examination findings 
• 
Occurrence of AEs 
• 
IOP measured at 65 minutes post-administration* 
• 
Visual acuity changes* 
 
*IOP and VA line listings will be provided for pediatric subjects because of the variable testing methods 
appropriate for this sub-group  
 
11.4. Analysis Populations 
 
Subjects who are missing efficacy assessments at any of the time points or are otherwise unevaluable for 
efficacy considerations will not be replaced.  
 
11.4.1. Intent to Treat Population 
 
The intent to treat (ITT) population will consist of all randomized subjects who received a dose of study 
drug.   Subjects will be analyzed according to the planned sequence of drugs. 
 
11.4.2. Per Protocol Population 
 
The per protocol (PP) population will consist of all ITT subjects who completed all planned assessments 
without major protocol violations.  Subjects to be excluded from the PP population will be selected prior 
to unmasking drug assignment.  Subjects will be analyzed according to the sequence of study drugs 
received. 
 
11.4.3. Modified Per Protocol Population 
 
The modified per protocol (MPP) population is defined similarly to the PP population, except that 
classification will be made for each subject at the individual treatment visit level, rather than at the subject 
level. Unlike the PP population, subjects who only complete 1 or 2 of the 3 treatment visits may be 
included.  
 
The MPP  will consist of all ITT subjects who completed all planned assessments without major protocol 
violations for the given treatment visit. Subjects to be excluded from the MPP population will be selected 
prior to unmasking treatment 
 


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11.4.4. Safety Population 
 
The safety population will consist of all ITT subjects.  Subjects will be analyzed according to the 
sequence of study drugs received 
 
11.5. General Statistical Considerations 
 
Primary efficacy analyses will be conducted on the PP population. Safety analyses will be performed 
using the safety analysis set. All primary analyses will be performed in both eyes.  The pupil diameter 
measurement at each corresponding time point after administration will be compared between study 
drugs. 
 
Descriptive statistics will be used to summarize continuous outcomes (number of subjects [N], mean, 
standard deviation or standard error of the mean, median, maximum, and minimum) and categorical 
variables, i.e. race, sex, iris color (frequency and percentage) at each assessment time point.  
 
The ABB/BAA crossover design used in this study allows for within-subject (i.e. within-eye) estimates of 
both drug effects (A and B) and carryover (AA, AB, BA, BB).  Thus, there is no need to include random 
effects for subject or eye in the analysis model, we will use fixed effects ANOVA to analyze these data 
(Jones and Kenward, 2015, section 5.2, 5.3.1) and tests will be based on within-eye variances. A SAS 
REPEATED statement will be used to adjust for correlation between fellow eyes. 
 
This study has both a “pre-randomization” covariate (iris color) and a “period-dependent” covariate 
(baseline pupil measurement) as discussed in Jones and Kenward (2015,  section 5.4.4).  We are assuming 
that the time between visits is sufficiently long that the baseline pupil measurement at each visit is 
unaffected by carryover. 
 
Unless explicitly stated otherwise, all statistical tests and confidence intervals will be two-sided, alpha = 
0.05. 
 
11.5.1. Analysis of Primary Efficacy 
 
All time points for pupil diameter will be analyzed separately; however, only the 35-minute time point 
will support statistical inference.  Success is defined as a statistically significant drug effect, with the 
fixed combination better (greater deltaPD) than placebo. 
 
The null and alternative hypotheses for comparison of Eyenovia’s fixed combination phenylephrine 
2.5%-tropicamide 1% ophthalmic solution with placebo, are defined as follows: 
 
• 
H0: The mean change per visit in pupil diameter at 35 minutes versus baseline, as measured by 
digital pupillometry in highly photopic conditions, is not different between Eyenovia’s fixed 
combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution and placebo  


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• 
HA: The mean change per visit in pupil diameter at 35 minutes versus baseline, as measured by 
digital pupillometry in highly photopic conditions, is different between Eyenovia’s fixed 
combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution and placebo 
 
Comparisons among drugs on mean pupil diameter will be based on a fixed-effects analysis of variance 
model.  The model will contain an effect due to subject, eye, subject x eye, period (1, 2, or 3), direct effect 
of drug, first order carryover, baseline pupil diameter, and iris color (dark vs light).  The SAS PROC 
MIXED procedure will be used to fit the ANOVA model.  Hypothesis tests will be based on confidence 
intervals produced by the SAS LSMEANS option for the difference between treatment groups. Details are 
provided in the statistical analysis plan. 
 
The SAS PROC MIXED procedure uses all available data to produce maximum likelihood parameter 
estimates, in such a setting the missing observations are ignorable if data are missing at random (MAR).   
 
11.5.2. Additional Efficacy Analyses 
 
The proportion of eyes achieving pupil size of 6.0 mm or greater and 7.0 mm or greater at 35 minutes 
post-dose will be summarized by eye and by treatment group with descriptive statistics.  
 
For each of these proportions, pairwise comparisons of the fixed combination of phenylephrine 2.5%-
tropicamide 1% microdose ophthalmic solution administered against placebo will be conducted with a 
generalized estimating equation (GEE) model for binomial outcome, allowing for correlation between 
eyes.   
 
The SAS PROC GENMOD procedure will be used to fit the model, using the same fixed effects as the 
primary analysis.  Once again, comparisons between treatment groups will be based on SAS LSMEANS 
for the difference between drug and placebo.  Details are provided in the statistical analysis plan. 
 
In addition, a cumulative proportion of responders analysis (CPRA) will be created to graphically display 
the proportions of subjects with a pupil size at 35 minutes post-dose over the entire range of possible 
pupil size cut-offs (Farrar 2006), with separate lines for each treatment and eye combination.  
 
The distribution of time to maximum pupil diameter will be described by Kaplan-Meier plots.   
 
11.5.3. Sensitivity Analyses 
 
The primary efficacy analysis described above will be rerun using the MPP population as a sensitivity 
analysis.  
 
As a further sensitivity analysis, multiple imputation based on reference-based multiple imputation will be 
used to impute missing data at the 35-minute time point based on a missing not at random (MNAR) 
assumption. Since the PP population contains only subjects with non-missing values, this analysis will be 
conducted using the ITT population, which may contain subjects who have only partially available 
information. Placebo treatment will serve as reference group for this approach. Each subject will be 


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CONFIDENTIAL - PAGE 38 OF 61 
20 NOVEMBER 2018 
 
included for all three treatment visits for this approach, regardless of how many treatment visits were 
actually completed.  Details are provided in the statistical analysis plan. 
 
11.5.4. Interim Analyses 
 
No interim analyses are planned for this study. 
 
11.5.5. Analysis of Baseline Data 
 
No separate analyses of baseline data are planned for this study. 
 
 
 
 


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20 NOVEMBER 2018 
 
12. QUALITY ASSURANCE AND QUALITY CONTROL 
 
The Sponsor or designee will perform quality control and quality assurance checks for this study.  Before 
initiation of study enrollment, the following documents will be reviewed with the investigator, site study 
staff, and pharmacy:  
 
• 
Study protocol 
• 
Study medication preparation procedures 
• 
Study medication administration procedures 
• 
Electronic Case Report Forms (eCRF) and procedures for completion 
• 
Informed consent process 
• 
Adverse event (AE) reporting procedures 
 
12.1. Study Monitoring 
 
During the study, the Sponsor or designee will perform periodic visits to study sites. During these visits, 
information recorded on the study eCRFs will be verified against source documents to confirm data 
capture completeness, accuracy and logical consistency.  Study documents will be reviewed to confirm 
protocol compliance and adherence to IRB and Sponsor-specified reporting requirements, and product 
accountability will be checked.   
 
12.2. Study Product Accountability 
 
Accurate records of each MiDD base unit and cartridge shipped and medication administered must be 
maintained by the Sponsor, or designee, and each study investigator.  Each of the 2 study medications 
will be provided to the site pharmacy in a kit labeled with the medication code.  Study kits will be 
accompanied by a shipper listing the number of kits provided and the contents of each kit.  Pharmacy 
personnel must review the shipper at the time of kit receipt and notify the Sponsor, or designee, of any 
discrepancies.  The Sponsor, or designee, will provide a study kit accountability log to the pharmacy for 
documentation of kits received, kits used for site product administration, and kits returned or discarded.    
 
The study investigator is responsible for documenting each MiDD used, the date of use, the identifiers for 
subjects who received study drug with the MiDD, and the number of sprays administered to each subject 
eye.    
 
MiDD product accountability will be performed by reconciling all dispensers provided to the pharmacy 
with dispensers used for study drug administration, subjects for whom the dispenser was used, and 
dispensers returned to the Sponsor or designee. Additionally, for the placebo product, each commercial 
bottle provided will be reconciled with vials prepared by the pharmacy, the associated dispensers used in 
study drug administration, subjects for whom the dispenser was used, and the dispenser returned to the 
Sponsor or designee.  
 
 


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13. ETHICAL AND REGULATORY CONSIDERATIONS 
 
This protocol was designed and will be conducted, recorded, and reported in compliance with the 
principles of Good Clinical Practice (GCP)38.  Sponsor and Investigator responsibilities associated with 
adherence to GCP and CFR 312 are specified in APPENDIX 5:  SPONSOR AND INVESTIGATOR 
OBLIGATIONS.  The study will be performed in accordance with the recommendations guiding 
physicians in biomedical research involving human subjects adopted by the 18th World Medical 
Assembly, Helsinki, Finland, 1964 and later amendments (see APPENDIX 6:  WORLD MEDICAL 
ASSOCIATION DECLARATION OF HELSINKI). 
 
13.1. Institutional Review 
 
Before enrollment of study subjects, this protocol and the study-specific consent/assent form(s) must be 
reviewed and approved by an IRB operating in accordance with 21 CFR Part 50.  Any changes to the 
study protocol or consent forms must be approved by the IRB prior to implementation.  Materials for 
study patient recruitment and study-specific written materials provided to the subject must also be 
approved by the IRB prior to use.   
 
Ongoing study progress reports will be submitted to the IRB at least annually, and more frequently if 
specified by the IRB.  Reports of safety events and any protocol deviations that affect the safety and 
welfare of a study subject will be submitted to the IRB in accordance with FDA and IRB requirements. 
 
13.2. Informed Consent 
 
The study consent form has been developed in compliance with 21 CFR Part 50.25.  Study consent 
form(s) must be submitted to and approved by the IRB prior to implementation.  Any IRB-requested 
modifications to the consent form must remain in compliance with 21 CFR Part 50.25. 
 
As discussed in Section 9.1, each participant must be provided with a copy of the IRB-approved consent 
form(s) for their review, and the participants written approval(s) must be provided prior to initiation of 
study procedures.   
 
13.3. Subject Confidentiality 
 
The Investigator will maintain the confidentiality of the identity of subjects enrolled in the study and the 
information contained in the study records.  The records will be made available as required for review by 
FDA and a reviewing IRB; however, to the extent possible, the subject’s identity will not be disclosed.  
 
38 Good Clinical Practice: Integrated Addendum to ICH E6(R1) Guidance for Industry, March 2018. 


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20 NOVEMBER 2018 
 
14. DATA HANDLING AND RECORD KEEPING 
 
Procedures for the handling and analysis of data will be conducted using good computing practices 
meeting ICH and US FDA guidelines for the handling and analysis of data for clinical trials. 
 
14.1. Data Quality Control and Reporting 
 
After data have been entered into the study database, a system of computerized data validation checks 
will be applied and written queries pertaining to data omissions and discrepancies will be forwarded to 
study sites for resolution. Study staff will update the database as appropriate to resolve queries 
generated.  All changes to the study database will be documented. 
 
14.2. Data Archiving 
 
Archived versions of the database will be saved by the Sponsor or designee consistent with ICH GCP 
Guidelines, complying with whichever of the requirements is longer. The Sponsor will notify the 
investigator when documents should be returned. 
 
14.3. Records Retention 
 
The Investigator’s site will retain all records related to the study in compliance with ICH GCP 
Guidelines. 
 
14.4. Protocol Amendments 
 
Modifications to the approved protocol are only possible using 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 IRB 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.  This should be fully documented. 
 
The investigator must not implement any deviation from or change to the protocol, without 
discussion with, and agreement by the Sponsor and prior review and documented approval/favorable 
opinion of the amendment from the relevant IRB, except where it is necessary to eliminate an 
immediate hazard to study patients, 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). 
 
 


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20 NOVEMBER 2018 
 
15. REFERENCES 
 
• 
Akman A, Aydin P. Comparison of mydriatic efficacy of spray application and drop instillation of 
tropicamide 1%. Eye (Lond). 1999;13(5):653‒655. 
• 
CFR 312.23 – IND Safety Reporting, 
https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/CFRSearch.cfm?fr=312.32 
• 
Farkouh A, Frigo P, Czejka M. Systemic side effects of eye drops: a pharmacokinetic perspective. 
Clin Ophthalmol. 2016;10:2433‒2441. 
• 
Farris RL, Stuchell RN, Mandell ID. Basal and reflex human tear analysis. I Physical measurements: 
osmolarity, basal volumes, and reflex flow rate. Am Acad Ophthal 1981;88:852. 
• 
Good Clinical Practice: Integrated Addendum to ICH E6(R1) Guidance for Industry, March 2018. 
• 
Grob SR, GonzalezGonzalez LA, Daly MK. Management of mydriasis and pain in cataract and 
intraocular lens surgery: review of current medications and future directions. Clin Ophthalmol. 
2014;8:1281‒1289. doi: 10.2147/OPTH.S47569 
• 
Ianchulev T, Chayet A, Kahook M, Packer M, Pasquale L, Weinreb RN. Pharmacodynamic profile of 
mydriatic agents delivered by ocular piezo-ejection microdosing compared with conventional 
eyedropper. Ther Deliv. 2016;7(11):751-760. 
• 
Jordan A, Baum J. Basic tear flow. Does it exist? Ophthalmology. 1980 Sep;87(9):920‒
930.Lallemand F, Daull P, Benita S, Buggage R, Garrigue JS. Successfully improving ocular drug 
delivery using the cationic nanoemulsion, novasorb. J Drug Deliv. 2012;2012:604204. 
• 
Lallemand F, Daull P, Benita S, Buggage R, Garrigue JS. Successfully improving ocular drug 
delivery using the cationic nanoemulsion, novasorb. J Drug Deliv. 2012;2012:604204. 
• 
Lynch MG, Brown RH, Goode SM, Schoenwald RD, Chien DS. Reduction of phenylephrine drop 
size in infants achieves equal dilation with decreased systemic absorption. Arch Ophthalmol. 
1987;105(10):1364-5. 
• 
Madsen JT, Andersen KE. Phenylephrine is a frequent cause of periorbital allergic contact dermatitis. 
Contact Dermatitis. 2015;73(1):64-5. doi: 10.1111/cod.12394. 
• 
Mammo ZN, Flanagan JG, James DF, Trope GE.Generic versus brand-name North American topical 
glaucoma drops. Can J Ophthalmol. 2012;47(1):55‒61. doi: 10.1016/j.jcjo.2011.12.004. 
• 
Merrill D L, Goldber, B, Zavell S. bis-Tropamide, A new parasympatholytic. Curr Ther Res. 
1960;12:43‒50. 
• 
Mishima S, Gasset A, Klyce SD Jr, Baum JL. Determination of tear volume and tear flow. Invest. 
Ophthalmol. 5(3), 264‒276 (1966). 
• 
Petursson G, Cole R, Hanna C. Treatment of Glaucoma Using Minidrops of Clonidine. Arch 
Ophthalmol. 1984;102(8):1180-1181. 
• 
Ratkowsky, Evans and Alldredge, 1993. Cross-over experiments: design, analysis, and application  
• 
Scherz W, Doane MG, Dohlman CH. Tear volume in normal eyes and keratoconjunctivitis sicca. 
Albrecht Von Graefes Arch. Klin. Exp. Ophthalmol. 1974;192(2):141‒150. 
• 
Shell JW. Pharmacokinetics of topically applied ophthalmic drugs. Surv Ophthalmol. 
1982;26(4):207-18. 


PHENYLEPHRINE 2.5%-TROPICAMIDE 1% MICRODOSE OPHTHALMIC SOLUTION  
EYENOVIA CLINICAL STUDY PROTOCOL EYN-MYD-TP-32 VERSION C  
 
 
 
CONFIDENTIAL - PAGE 43 OF 61 
20 NOVEMBER 2018 
 
• 
Urtti A, Salminen L. Minimizing systemic absorption of topically administered ophthalmic drugs. 
Surv. Ophthalmol. 37(6), 435−456 (1993). 
• 
Vaajanen A, Vapaatalo H. A single drop in the eye - Effects on the whole body? Open Ophthalmol J. 
2017;11:305‒314. doi: 10.2174/1874364101711010305 
• 
Wheatcroft S, Sharma A, McAllister J. Reduction in mydriatic drop size in premature infants. Br J 
Ophthalmol. 1993;77(6):364-5. 
 


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APPENDIX 1:  SCHEDULE OF MEDICATION ADMINISTRATION AND EXAMINATIONS 
 
Assessment/Procedure 
Screening 
Visit 
(Day -14 to 
Day -1) 
Treatment  
Visit 1 (Day 1)1, Visit 21 and Visit 31 
Baseline0 
Time 02 
Time 1 
T0+20 min 
(± 2 min) 
Time 2 
T0+35 min 
(± 5 min) 
Time 3 
T0+50 min 
(± 5 min) 
Time 4 
T0+65 min 
(± 5 min) 
Time 5 
T0+80 min 
(± 10 min) 
Time 6 
T0+120 min 
(± 10 min) 
Time 7 
T0+180 min 
(± 10 min) 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
Medical history 
X 
 
 
 
 
 
 
 
 
 
Ocular history 
X 
 
 
 
 
 
 
 
 
 
Prior/concomitant medication use 
X 
 
 
 
 
 
 
 
 
 
Urine pregnancy test3 
X 
X3 
Manifest refraction (OU) 
X 
 
 
 
 
 
 
 
 
 
BCDVA (OU)4 
X 
X 
 
 
 
 
 
 
 
X 
Study drug administration sequence 
determination (OU) 
 
X 
 
 
 
 
 
 
 
 
Study drug administration (OU) 
 
 
X 
 
 
 
 
 
 
 
Slit lamp biomicroscopy (OU)5 
X 
X 
 
 
 
 
 
 
 
X 
Van Herrick Angle Assessment (OU)6 
X 
 
 
 
 
 
 
 
 
 
IOP (OU)7 
X 
X 
 
 
 
 
X 
 
 
 
Pupil diameter assessment (OU)8 
X 
X 
 
X 
X 
X 
X 
X 
X 
X 
Pupillary light reflex (OU) 
 
X 
 
 
X 
 
X 
 
 
X 
Study eligibility determination 
X 
 
 
 
 
 
 
 
 
 
Dilated fundus exam (OU) 
X 
 
 
 
 
 
 
 
 
 
AE assessment 
 
X 
 
 
X 
 
 
 
 
X 
0 Baseline refers to evaluations made at each Treatment Day prior to study medication administration. 
1 Treatment Visit 1, 2 and 3 must be separated by at least 2 days; but may be up to 7 days apart to allow for scheduling flexibility. 
2 Time 0 starts at the point the first of two study medication doses is administered in second eye of the study subject 
3 A urine pregnancy test will be conducted in females of childbearing potential at the Screening Visit and at the last visit (Treatment Visit 3). The test may be administered at any time during 
the Screening and Treatment Day 3 Visits.  If the subject terminates study participation prior to Treatment Visit 3, this test must be administered promptly at the time of termination. 
4 BCDVA to be measured using ETDRS methods.  For younger pediatric subjects, UCDVA may be measured using age-appropriate methods per investigator’s usual practice. 
5 For younger pediatric subjects who cannot cooperate with a traditional SLE, a portable slit lamp model may be used, if necessary. 
6 Performed as part of slit lamp examination. 
7 IOP to be measured using Goldmann Applanation tonometry. For younger pediatric subjects, IOP may be measured using age-appropriate methods per investigator’s usual practice. 
8 Performed using Neuroptics pupillometer – VIP 300.  For younger pediatric subjects for whom the pupillometer cannot be successfully used, a ruler or pupil gauge may be used.  At 
Treatment Visit 3 - Time 7, subjects whose pupil diameter is larger than baseline may be followed at hourly intervals up to 6 hours after medication administration to gather additional data 
regarding the duration of study treatment effect.  
 


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APPENDIX 2:  STUDY MEDICATION PREPARATION PROCEDURE  
 
To maintain masking of the drug assignments for study staff performing clinical evaluations, study 
medication preparation and inventory management will be performed by a pharmacy associated with 
the study site.  The Sponsor, or designee, will ship the following study drug kits to each site’s 
pharmacy: 
 
Kit Components 
• 2 x MiDD base units 
• 2 x MiDD cartridges containing empty 2 mL vial (sterile) + loose adhesive label to be 
applied to cartridge 
• 2 x Site transport boxes 
• 2 x 10cc syringes (sterile) 
• 2 x 21g needles (sterile) 
• 2 x 2 mL injection vials of phenylephrine 2.5%-tropicamide 1% ophthalmic solution for 
sterile fill 
• 1 x Micro USB charging cable/connector plug 
 
• 2 x MiDD base units 
• 2 x MiDD cartridges containing empty 2 mL vial (sterile) + loose adhesive label to be 
applied to cartridge 
• 2 x Site transport boxes 
• 2 x 10cc syringes (sterile) 
• 2 x 21g needles (sterile) 
• 2 x 4mL bottles of eyewash solution for sterile fill (commercial container) 
• 1 x Micro USB charging cable/connector plug 
 
 
Each kit will be labeled with the following information: 
• 
Study protocol number 
• 
Study drug code (A or B) 
• 
Statement that kit contains either phenylephrine 2.5%-tropicamide 1% ophthalmic solution or 
eyewash solution 
• 
Product storage requirement 
• 
Caution – New Drug – Limited by Federal (US) Law to Investigational Use 
 
The MiDD base unit and cartridge are supplied in separate boxes within each study drug kit.  The 
MiDD base unit is contained in a polyethylene terephthalate glycol (PETG) plastic tray inside the box.  
The MiDD cartridge, which includes the empty vial into which the drug is transferred, is supplied 
sterile in a PETG plastic tray with a peel lid.   Prior to initial MiDD use, the relevant study ophthalmic 
solution must be aseptically transferred from the container in which it is received  into the MiDD vial.  


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The vial is then inserted into the MiDD cartridge assembly, the cartridge and base are clipped together, 
and the seal on the vial containing the drug is punctured by depressing the MiDD Load Button.  
Puncture of this seal creates a fluid path for the drug.  Depression and release of the Load Button 5 
times primes the system for drug delivery.  At the time of dosing, the Load Button is again pressed and 
released to activate the piezo element to create the aerosol containing the dose.  When the Dose Button 
is pressed, approximately 8 µL of the ophthalmic solution is delivered through the Spray Nozzle to the 
cornea.  
 
The site study pharmacist will be trained and certified by the Sponsor, or designee, regarding aseptic 
drug transfer and MiDD cartridge assembly processes prior to their performance.   
 
New study drug MiDDs must be prepared for each treatment day.  MiDD cartridges containing each of 
the 2 study drugs and labeled with the appropriate study drug code, pharmacy preparation date and “use 
by date” date will be forwarded with the accompanying base unit by the pharmacy to site personnel for 
use.  Site personnel will assemble the cartridge and base unit for each study drug to be used that day 
and the “date used” for the MiDD will be added to the product preparation/use label by site personnel.  
At the end of the treatment day, each MiDD provided will be returned to the pharmacy for storage. 
Please note that each MiDD prepared must be used on the day of preparation; the MiDD may not 
be re-used on subsequent days. 
 


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APPENDIX 3:  STUDY MEDICATION ADMINISTRATION PROCEDURE  
 
This study will be double-masked so that there are no differences in presentation (Eyenovia’s fixed 
combination mydriasis agent is formulated as a multi-dose microdroplet spray and the placebo is also 
administrated as a microdroplet spray). Site personnel designated to administer study medication will be 
trained by the Sponsor on proper use of the MiDD prior to study medication administration.  In order to 
reduce the potential for operational bias, study personnel who administer study medication on a given 
day will not be allowed to perform post-study drug administration ophthalmic assessments on that day.  
Study drug must be administered in location unattended by examining staff so that examiners are not 
aware of which drug the subject received. 
 
For each treatment day, one or more new MiDD base units and cartridges containing the study drug to 
be administered will be provided by the pharmacy for use.  Each MiDD will appear the same except for 
a label indicating the study drug code.  Study site personnel will assemble the cartridge and base unit, 
then add the study drug administration day to the MiDD label and prime the system for use by 
depressing and releasing the Load Button, then depressing and releasing the Dose Button.  This 
sequence must be performed 5 times to complete the priming process.   
 
 
 
 
Figure 1: Eyenovia MiDD Front (L) and Back (R) View 
 
At the time of study medication administration, the following steps should be followed sequentially: 
 
1. Load MiDD dose by depressing the LOAD BUTTON once. 
2. Hold MiDD such that that the thumb and 3rd finger are positioned on each side of the cartridge 
and index finger is positioned on DOSE BUTTON at the back of the MiDD.  Note:  Holding 
MiDD with fingers around the front of the device may affect dispenser alignment with the eye, 
resulting in spray administration outside of the targeted corneal area.   
3. Begin to position the MiDD over the patient’s eye such that the SPRAY NOZZLE is facing the 
cornea and the DOSE BUTTON is facing the medication administrator.  While positioning, it is 
Dose Button 
Load Button 
MiDD Base Unit 
MiDD Cartridge 
Spray Nozzle 


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20 NOVEMBER 2018 
 
 
suggested to use the little finger of the hand holding the MiDD to open the lower lid and the 
thumb of the opposite hand to hold the upper lid open.  
4. Lower the MiDD toward the eye such that it is almost touching the patient’s brow and cheek.  
The MiDD may be rotated slightly to avoid interference that might affect stability during spray 
administration; however, it is important that the SPRAY NOZZLE be aligned directly toward 
the patient’s cornea. 
5. Ask patient to first look up, then look at the blue light around the SPRAY NOZZLE 
6. When blue light is centered on the patient’s iris, depress, then quickly release DOSE BUTTON 
to administer medication.  Visually confirm medication delivery to the cornea.   
7. If medication delivery missed the cornea, withdraw MiDD from eye, then reload by depressing 
the LOAD BUTTON once and repeat Steps 1-6. 
8. Repeat steps 1-7 for the contralateral eye. 
 
 
 
. 
 
 


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APPENDIX 4:  EXAMINATION PROCEDURES 
 
All study personnel conducting ophthalmic assessments will be masked to the study drug administered.  
 
1. Visual Acuity 
 
The evaluation of VA should be performed before the subject’s pupils have been dilated.  Distance VA 
measurements should be obtained at a testing distance of 4 meters by a physician or trained technician 
using ETDRS lighted charts.  Chart background luminance must be between 80 cd/m2 and 160 cd/m2, 
inclusive (85 cd/m2 is recommended).    
  
Adult subjects’ BCDVA will be determined using subjective manifest refraction. 
 
The subject should be seated so that the distance from the subject's eyes to the ETDRS chart (Chart 1 
for right eye and Chart 2 for left eye) is 4 meters (157.5 inches).  The middle line of letters to 
approximately one-third of the distance from the top line of letters should be at eye level for the subject.  
The subject should be asked to start reading letters at the top line of the chart.   Document the number 
of letters correctly read the line.  Move to the next line down if 4 or more letters are read correctly and 
document the number of letters correctly read on this line.  Continue moving down the lines 
(documenting the number of letters read for each line) until the subject reads 2 or more letters 
incorrectly, then stop.  Note:  If the total number of letters read (TLR) correctly is less than 20 letters, 
position the subject 1 meter from the chart and add +0.75 D sphere, then resume testing in the same 
manner.  If the subject is unable to read letters correctly at both 4.0 meters and 1.0 meters, then the 
subject should be tested to determine if vision is count fingers (CF), hand motion (HM), light 
perception (LP), or no light perception (NLP).  All visual acuities will be expressed in logMar units. 
 
For younger pediatric subjects, uncorrected distance visual acuity (UCDVA) should be measured using 
age-appropriate methods per the Investigator’s usual practice. 
 
2. Pupil Diameter Measurement 
 
Pupil diameter should be measured prior to additional study-specified examinations using the 
Neuroptics VIP 300 pupillometer; the same pupillometer should be used for each evaluation on any 
study visit day for a given subject.  The subject should be asked to focus straight ahead at a target 
placed at 3 meters to avoid accommodation.  A fully-charged transilluminator, or muscle light, will be 
shone into one eye while the pupillometer is used to measure the pupil diameter in the fellow eye.  After 
a brief rest period, the process will be repeated to measure pupil diameter in the other eye.   
 
For younger pediatric subjects in whom the Neuroptics pupillometer cannot be used to obtain a Time 0 
measurement, the pupil diameter may be measured using a ruler or pupil gage using the instructions that 
follow.  Only a single measurement method can be used on any study visit day for a given subject. 
 
 
 


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a. Pupil Diameter Determination by Ruler or Pupil Gauge 
 
For younger pediatric subjects in whom the Neuroptics pupillometer cannot be used to obtain a 
Time 0 measurement, the pupil diameter may be measured using a ruler or pupil gage using the 
instructions that follow.  A ruler or pupil gauge requires special care to ensure the measurement 
is accurate. Begin by placing the ruler as close as possible alongside the eye, making certain not 
to tilt or bend the device. The subject should be asked to focus straight ahead at a target placed 
at 3 meters to avoid accommodation.  A fully-charged transilluminator, or muscle light, will be 
shone into one eye while the pupil diameter in the fellow eye is measured. For optimal 
accuracy, take the measurement at the patient’s eye level and repeat each measurement 2-3 
times to ensure the result is consistent.  The pupil diameter recorded should be either the mean 
(if 2 measurements are taken) or the median (if 3 measurements are taken) values. 
 
3. Pupillary Light Reflex 
 
Pupillary light reflex will be evaluated using a fully-charged transillumator, or muscle light.  The 
response will be recorded on a scale ranging from 0 (non-responsive) to 3 (brisk). 
 
4. Intraocular Pressure Measurement   
 
After completion of slit lamp biomicroscopy, local anesthetic will be applied to facilitate IOP 
measurements with the Goldmann Applanation Tonometer. Recent tonometer calibration must be 
documented. 
 
Two consecutive IOP measurements of each eye must be obtained. If the 2 measurements differ by 
more than 2 mmHg, a third measurement must be obtained. IOP will be analyzed as the mean of these 
2 measurements, or as the median of the 3 measurements. 
 
For pediatric subjects, IOP should be measured using age-appropriate methods per the Investigator’s 
usual practice. 
 
5. Slit Lamp Biomicroscopy 
 
The eyelids, conjunctiva, sclera, cornea, anterior chamber, lens, iris and anterior vitreous of the eye will 
be examined with the aid of a table-mounted binocular microscope, called a slit lamp.  The patient will 
be seated during this examination and fluorescein dye should be instilled into the ocular cul-de-sac to 
facilitate the examination.  For pediatric subjects who cannot cooperate with a traditional SLE, a 
portable slit lamp model may be used according to the Investigator’s usual practice.  Grading will be 
assigned as follows: 
 
 
 


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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 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, with/without eversion of upper 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 conjunctival 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 
 
SCLERA 
Normal 
Abnormal 
 
CORNEA 
Edema 
None (0) 
 
Transparent and clear, or less than mild 
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 
 
Staining 
None (0) 
 
No fluorescein staining of epithelium, or less than mild 
Mild (+1) 
 
Slight punctate fluorescein staining 
Moderate (+2) 
Regionally dense coalescent fluorescein staining 
Severe (+3) 
Marked fluorescein staining with immediate stromal leakage as a result of epithelial 
loss 
 
 
 


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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 hypopyon size) 
Flare 
None (0) 
 
No Tyndall effect, or less than mild 
Mild (+1) 
 
Tyndall beam in the AC has mild intensity 
Moderate (+2) 
Tyndall beam in the AC is of strong intensity 
Severe (+3)  
Tyndall beam is very intensive.  Aqueous has a white, milky appearance 
 
IRIS 
Normal 
Abnormal 
 
LENS 
Lens Status 
Phakic* 
 
*Indicate Lens Opacity Grade: 
 
None (0) 
None present, or less than mild 
 
 
 
 
 
 
Mild (+1) 
Subtle 
 
 
 
 
 
 
Moderate (+2) 
Moderate 
 
 
 
 
 
 
Severe (+3) 
Dense 
Pseudophakic 
Aphakic 
 
ANTERIOR VITREOUS 
Normal 
Abnormal 
 
 
6. Dilated Fundus Examination 
 
Using ophthalmoscopy, the fundus will be examined and evaluated by the Investigator to be either 
normal or abnormal at the Screening Visit.   
 
 


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APPENDIX 5:  SPONSOR AND INVESTIGATOR OBLIGATIONS  
 
SPONSOR OBLIGATIONS 
 
The Sponsor is committed to: 
 
1. 
Complying with all applicable health authority regulations governing the conduct of clinical 
research studies, including the US FDA. 
2. 
Protecting the rights, health, safety and welfare of study subjects. 
3. 
Informing clinical investigators of any new information about the study which may affect the 
health, safety or welfare of the subjects, or may influence their decision to continue 
participation in the study. 
4. 
Providing clinical investigators with the study protocol, and CRFs on which to document the 
study evaluation variables for each subject entered into the study. 
5. 
Providing the statistical analysis and study report writing resources necessary to complete 
reporting of study results. 
6. 
Ensuring equity of consideration among all investigators in multicenter studies in all matters of 
publications, meeting presentations, etc. 
7. 
Certifying that IRB/EC approval of the protocol and Investigator Agreement will be completed 
prior to treatment at an investigational site.  
 
The Sponsor shall have the right to terminate the study at any time by written notice to the investigator.  
Without limiting this right, the Sponsor will normally only terminate the study under the following 
circumstances: 
 
• 
If severe and/or SAEs associated with the study medication in human and/or animal studies 
indicate discontinuation of the study 
• 
If the Sponsor wishes to discontinue the study for commercial reasons 
• 
If the Sponsor has reasons to believe that the study cannot be satisfactorily completed due to 
insufficient patient enrollment, or an insufficient number of participating study sites identified 
within a reasonable time frame. 
 
 
 


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INVESTIGATOR OBLIGATIONS 
 
Each Investigator must be a licensed physician who has completed a residency or preceptorship in 
ophthalmology.  The Investigators have the following responsibilities: 
 
1. Subject Selection 
The Investigator is responsible for assuring that all subjects entering the study conform to the 
patient selection criteria. 
 
2. Informed Consent 
The Investigator is responsible for fully reviewing the nature of the study, the possible risks, and 
alternative treatments with prospective patients prior to their enrollment in the study.  The 
Investigator is responsible for obtaining written informed consent in compliance with 21 CFR 50 
for each patient, prior to enrollment in the trial.  A copy of the signed Informed Consent Form will 
be maintained in the patient's medical record. 
 
3. Institutional Review Board/Ethics Committee Approval 
The Investigator must obtain approval for participation in this protocol from the IRB/EC for the 
institution at which the procedure will be performed prior to entering any patients in the study.  The 
ICF to be used will also be submitted to the IRB/EC for approval prior to initiation of the study.  
Assurance that IRB/EC approval of the study protocol and ICF has been obtained will be provided 
to the Sponsor, or Sponsor designee, prior to initiation of the study. 
 
4. Subject Evaluations and Data Reporting 
The Investigator is responsible for complying with the requirements of the study protocol and any 
amendment or clarification as published by the Sponsor, or Sponsor designee.  Patient evaluations 
will be performed as described in the study protocol.  All information generated by the patient 
evaluation will be recorded using eCRFs with access provided by the Sponsor or Sponsor designee.     
 
Investigator(s) will not deviate from the study protocol without prior approval of the Sponsor, or 
Sponsor designee, unless the protection of health, safety or welfare of study subjects requires 
prompt action. 
 
5. Record Retention 
The Investigator shall maintain all patient records for whichever of the following periods is 
shortest: 
a. A period of 2 years after the date on which the FDA approves the marketing of the drug for the 
purpose that was the subject of the study. 
b. A period of 5 years after the date on which the results of the study are submitted to the FDA in 
support of the marketing of the drug for the purpose that was the subject of the study. 
 
6. Investigational Material Accountability 
The Investigator must maintain accurate records of the receipt of all investigational material 
shipped by the Sponsor, or Sponsor designee, including the date, and identification numbers of the 


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product received.  In addition, accurate records must be kept on the amount and date that 
investigational material was dispensed or returned to the Sponsor.  The Investigator must assure 
that study supplies be dispensed only to subjects enrolled in the study per study inclusion / 
exclusion criteria and under the direct supervision of the Investigator or his/her Co-Investigators. 
 
Records of all investigational supplies received, used and returned must be kept by the Investigator.  
All unused investigational supplies must be returned to the Sponsor, or Sponsor designee, as soon 
as practical upon completion of the trial.  Investigational material accounting procedures must be 
completed before the study is considered terminated. 
 
 


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APPENDIX 6:  WORLD MEDICAL ASSOCIATION DECLARATION OF HELSINKI 
 
Ethical Principles for Medical Research Involving Human Subjects 
 
Adopted by the 18th WMA General Assembly, Helsinki, Finland, June 1964, and amended by the: 
29th WMA General Assembly, Tokyo, Japan, October 1975 
35th WMA General Assembly, Venice, Italy, October 1983 
41st WMA General Assembly, Hong Kong, September 1989 
48th WMA General Assembly, Somerset West, Republic of South Africa, October 1996 
52nd WMA General Assembly, Edinburgh, Scotland, October 2000 
53th WMA General Assembly, Washington 2002 (Note of Clarification on paragraph 29 added) 
55th WMA General Assembly, Tokyo 2004 (Note of Clarification on Paragraph 30 added) 
59th WMA General Assembly, Seoul, October 2008 
 
A.  INTRODUCTION 
 
1.  The World Medical Association (WMA) has developed the Declaration of Helsinki as a statement 
of ethical principles for medical research involving human subjects, including research on 
identifiable human material and data. The Declaration is intended to be read as a whole and each of 
its constituent paragraphs should not be applied without consideration of all other relevant 
paragraphs. 
 
2.  Although the Declaration is addressed primarily to physicians, the WMA encourages other 
participants in medical research involving human subjects to adopt these principles. 
 
3.  It is the duty of the physician to promote and safeguard the health of patients, including those who 
are involved in medical research. The physician's knowledge and conscience are dedicated to the 
fulfillment of this duty. 
 
4.  The Declaration of Geneva of the WMA binds the physician with the words, “The health of my 
patient will be my first consideration,” and the International Code of Medical Ethics declares that, 
“A physician shall act in the patient's best interest when providing medical care.” 
 
5.  Medical progress is based on research that ultimately must include studies involving human 
subjects. Populations that are underrepresented in medical research should be provided appropriate 
access to participation in research. 
 
6.  In medical research involving human subjects, the well-being of the individual research subject 
must take precedence over all other interests. 
 
7.  The primary purpose of medical research involving human subjects is to understand the causes, 
development and effects of diseases and improve preventive, diagnostic and therapeutic 
interventions (methods, procedures and treatments). Even the best current interventions must be 


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evaluated continually through research for their safety, effectiveness, efficiency, accessibility and 
quality. 
 
8.  In medical practice and in medical research, most interventions involve risks and 
 
burdens. 
 
9.  Medical research is subject to ethical standards that promote respect for all human subjects and 
protect their health and rights. Some research populations are particularly vulnerable and need 
special protection. These include those who cannot give or refuse consent for themselves and those 
who may be vulnerable to coercion or undue influence. 
 
10. Physicians should consider the ethical, legal and regulatory norms and standards for research 
involving human subjects in their own countries as well as applicable international norms and 
standards. No national or international ethical, legal or regulatory requirement should reduce or 
eliminate any of the protections for research subjects set forth in this Declaration. 
 
B.  PRINCIPLES FOR ALL MEDICAL RESEARCH 
 
11. It is the duty of physicians who participate in medical research to protect the life, health, dignity, 
integrity, right to self-determination, privacy, and confidentiality of personal information of 
research subjects. 
 
12. Medical research involving human subjects must conform to generally accepted scientific 
principles, be based on a thorough knowledge of the scientific literature, other relevant sources of 
information, and adequate laboratory and, as appropriate, animal experimentation. The welfare of 
animals used for research must be respected. 
 
13.  Appropriate caution must be exercised in the conduct of medical research that may harm the 
environment. 
 
14. The design and performance of each research study involving human subjects must be clearly 
described in a research protocol. The protocol should contain a statement of the ethical 
considerations involved and should indicate how the principles in this Declaration have been 
addressed. The protocol should include information regarding funding, institutional affiliations, 
other potential conflicts of interest, incentives for subjects and provisions for treating and/or 
compensating subjects who are harmed as a consequence of participation in the research study. The 
protocol should describe arrangements for post-study access by study subjects to interventions 
identified as beneficial in the study or access to other appropriate care or benefits. 
 
15. The research protocol must be submitted for consideration, comment, guidance and approval to a 
research ethics committee before the study begins. This committee must be independent of the 
researcher, the sponsor and any other undue influence. It must take into consideration the laws and 
regulations of the country or countries in which the research is to be performed as well as 
applicable international norms and standards but these must not be allowed to reduce or eliminate 


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any of the protections for research subjects set forth in this Declaration. The committee must have 
the right to monitor ongoing studies. The researcher must provide monitoring information to the 
committee, especially information about any serious AEs. No change to the protocol may be made 
without consideration and approval by the committee. 
 
16. Medical research involving human subjects must be conducted only by individuals with the 
appropriate scientific training and qualifications. Research on patients or healthy volunteers 
requires the supervision of a competent and appropriately qualified physician or other health care 
professional. The responsibility for the protection of research subjects must always rest with the 
physician or other health care professional and never the research subjects, even though they have 
given consent. 
 
17. Medical research involving a disadvantaged or vulnerable population or community is only justified 
if the research is responsive to the health needs and priorities of this population or community and 
if there is a reasonable likelihood that this population or community stands to benefit from the 
results of the research. 
 
18. Every medical research study involving human subjects must be preceded by careful assessment of 
predictable risks and burdens to the individuals and communities involved in the research in 
comparison with foreseeable benefits to them and to other individuals or communities affected by 
the condition under investigation. 
 
19.  Every clinical trial must be registered in a publicly accessible database before recruitment of the 
first subject. 
 
20. Physicians may not participate in a research study involving human subjects unless they are 
confident that the risks involved have been adequately assessed and can be satisfactorily managed. 
Physicians must immediately stop a study when the risks are found to outweigh the potential 
benefits or when there is conclusive proof of positive and beneficial results. 
 
21. Medical research involving human subjects may only be conducted if the importance of the 
objective outweighs the inherent risks and burdens to the research subjects. 
 
22. Participation by competent individuals as subjects in medical research must be voluntary. Although 
it may be appropriate to consult family members or community leaders, no competent individual 
may be enrolled in a research study unless he or she freely agrees. 
 
23. Every precaution must be taken to protect the privacy of research subjects and the confidentiality of 
their personal information and to minimize the impact of the study on their physical, mental and 
social integrity. 
 
24. In medical research involving competent human subjects, each potential subject must be adequately 
informed of the aims, methods, sources of funding, any possible conflicts of interest, institutional 
affiliations of the researcher, the anticipated benefits and potential risks of the study and the 


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discomfort it may entail, and any other relevant aspects of the study. The potential subject must be 
informed of the right to refuse to participate in the study or to withdraw consent to participate at 
any time without reprisal. Special attention should be given to the specific information needs of 
individual potential subjects as well as to the methods used to deliver the information. After 
ensuring that the potential subject has understood the information, the physician or another 
appropriately qualified individual must then seek the potential subject’s freely-given informed 
consent, preferably in writing. If the consent cannot be expressed in writing, the non-written 
consent must be formally documented and witnessed. 
 
25. For medical research using identifiable human material or data, physicians must normally seek 
consent for the collection, analysis, storage and/or reuse. There may be situations where consent 
would be impossible or impractical to obtain for such research or would pose a threat to the validity 
of the research. In such situations the research may be done only after consideration and approval of 
a research ethics committee. 
 
26. When seeking informed consent for participation in a research study the physician should be 
particularly cautious if the potential subject is in a dependent relationship with the physician or may 
consent under duress. In such situations the informed consent should be sought by an appropriately 
qualified individual who is completely independent of this relationship. 
 
27. For a potential research subject who is incompetent, the physician must seek informed consent from 
the legally authorized representative. These individuals must not be included in a research study 
that has no likelihood of benefit for them unless it is intended to promote the health of the 
population represented by the potential subject, the research cannot instead be performed with 
competent persons, and the research entails only minimal risk and minimal burden. 
 
28. When a potential research subject who is deemed incompetent is able to give assent to decisions 
about participation in research, the physician must seek that assent in addition to the consent of the 
legally authorized representative. The potential subject’s dissent should be respected. 
 
29. Research involving subjects who are physically or mentally incapable of giving consent, for 
example, unconscious patients, may be done only if the physical or mental condition that prevents 
giving informed consent is a necessary characteristic of the research population. In such 
circumstances the physician should seek informed consent from the legally authorized 
representative. If no such representative is available and if the research cannot be delayed, the study 
may proceed without informed consent provided that the specific reasons for involving subjects 
with a condition that renders them unable to give informed consent have been stated in the research 
protocol and the study has been approved by a research ethics committee. Consent to remain in the 
research should be obtained as soon as possible from the subject or a legally authorized 
representative. 
 
30. Authors, editors and publishers all have ethical obligations with regard to the publication of the 
results of research. Authors have a duty to make publicly available the results of their research on 
human subjects and are accountable for the completeness and accuracy of their reports. They should 


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adhere to accepted guidelines for ethical reporting. Negative and inconclusive as well as positive 
results should be published or otherwise made publicly available. Sources of funding, institutional 
affiliations and conflicts of interest should be declared in the publication. Reports of research not in 
accordance with the principles of this Declaration should not be accepted for publication. 
 
C.   ADDITIONAL PRINCIPLES FOR MEDICAL RESEARCH COMBINED WITH 
MEDICAL CARE 
 
31. The physician may combine medical research with medical care only to the extent that the research 
is justified by its potential preventive, diagnostic or therapeutic value and if the physician has good 
reason to believe that participation in the research study will not adversely affect the health of the 
patients who serve as research subjects. 
 
32. The benefits, risks, burdens and effectiveness of a new intervention must be tested against those of 
the best current proven intervention, except in the following circumstances: 
• 
The use of placebo, or no treatment, is acceptable in studies where no current proven 
intervention exists; or 
• 
Where for compelling and scientifically sound methodological reasons the use of placebo is 
necessary to determine the efficacy or safety of an intervention and the patients who receive 
placebo or no treatment will not be subject to any risk of serious or irreversible harm. Extreme 
care must be taken to avoid abuse of this option. 
 
33. At the conclusion of the study, patients entered into the study are entitled to be informed about the 
outcome of the study and to share any benefits that result from it, for example, access to 
interventions identified as beneficial in the study or to other appropriate care or benefits. 
 
34. The physician must fully inform the patient which aspects of the care are related to the research. 
The refusal of a patient to participate in a study or the patient’s decision to withdraw from the study 
must never interfere with the patient-physician relationship. 
 
35. In the treatment of a patient, where proven interventions do not exist or have been ineffective, the 
physician, after seeking expert advice, with informed consent from the patient or a legally 
authorized representative, may use an unproven intervention if in the physician's judgment it offers 
hope of saving life, re-establishing health or alleviating suffering. Where possible, this intervention 
should be made the object of research, designed to evaluate its safety and efficacy. In all cases, new 
information should be recorded and, where appropriate, made publicly available. 
 


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PROTOCOL EYN-MYD-TP-32 
VERSION C 
 
 
A MULTI-CENTER, DOUBLE-MASKED, PLACEBO-CONTROLLED, PHASE 3 STUDY OF 
THE SAFETY AND EFFICACY OF FIXED COMBINATION PHENYLEPHRINE 2.5%- 
TROPICAMIDE 1%OPHTHALMIC SOLUTION ADMINISTERED WITH A MICRODOSE 
DISPENSER FOR DILATION OF THE PUPIL 
(THE MIST-2 STUDY) 
 
 
SPONSOR: 
 
EYENOVIA, INC. 
295 MADISON AVENUE, SUITE 2400 
NEW YORK NY  10017 
USA 
PHONE:  917-289-1117 
 
 
I have read this protocol in its entirety.  I agree to: 
 
• 
Implement and conduct this study in compliance with this study protocol; conditions of 
approval imposed by FDA and my reviewing Institutional Review Board (IRB); Good Clinical 
Practice (GCP); and any other applicable laws and regulations.   
• 
Maintain all study-related information supplied by Eyenovia in a confidential manner.  
 
Protocol Amendment 
 
Modification of the study protocol is prohibited without prior written agreement in the form of a 
protocol amendment. All amendments will be created by the Sponsor and must be approved 
by the reviewing IRB prior to implementation at your site, except when required to mitigate immediate 
safety risks or when the changes involve only logistical or administrative revisions.  
 
 
 
 
___________________________        ___________________________          ______________                       
Print Name of Investigator 
 
       
Signature 
 
 
      Date 

SAP_001.pdf: 
 
 
STATISTICAL ANALYSIS PLAN  
A MULTI-CENTER, DOUBLE-MASKED, PLACEBO-CONTROLLED, 
PHASE 3 STUDY OF THE SAFETY AND EFFICACY OF FIXED 
COMBINATION 
PHENYLEPHRINE 
2.5% 
- 
TROPICAMIDE 
1% 
OPHTHALMIC SOLUTION ADMINISTERED WITH A MICRODOSE 
DISPENSER FOR DILATION OF THE PUPIL (THE MIST-2 STUDY) 
 
 
 
Sponsor:  
 
EYENOVIA, INC. 
 
 
 
295 MADISON AVENUE, SUITE 2400 
NEW YORK NY 10017 
USA 
 
 
Protocol Number: 
EYN-MYD-TP-32 VERSION C 
 
 
Author:  
 
Brian Kilgallen, MS 
 
 
 
Senior Principal Biostatistician 
Statistics & Data Corporation 
 
Date:   
 
18-Jan-2019 
Version:  
 
Final 
 


 
 
 
Page 2 of 33 
A Multi-Center, Double-Masked, Placebo-Controlled, Phase 3 Study 
of the Safety and Efficacy of Fixed CombinationPhenylephrine 2.5% 
- Tropicamide 1% Ophthalmic Solution Administered with a 
Microdose Dispenser for Dilation of the Pupil (The MIST-2 Study) 
 
 
 
Protocol Number: 
EYN-MYD-TP-32 Version C 
Version:  
 
Final  
Date:   
 
18-Jan-2019 
 
Statistical Analysis Plan Approval 
 
 
Prepared by:  
 
 
 
 
 
 
 
 
 
 
Brian Kilgallen, MS 
 
 
 
Date 
 
 
Senior Principal Biostatistician 
 
 
Statistics & Data Corporation 
 
 
 
Reviewed by:  
 
 
 
 
     
___________ 
 
 
Kirk Bateman, MS 
 
 
 
Date 
 
 
Senior Director, Biostatistics 
 
 
Statistics & Data Corporation 
 
 
 
Approved by:  
 
 
 
 
 
 
 
 
 
 
Ginger Clasby, MS 
 
  
 
Date 
Vice President, Clinical Operations 
 
 
Eyenovia, Inc. 
 


Protocol EYN-MYD-TP-32 Version C SAP, Final Version 
 
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Confidential & Proprietary 
Page 3 of 33 
 
 
Table of Contents 
1. 
Introduction ......................................................................................................................... 7 
2. 
Study Objectives ................................................................................................................. 7 
3. 
Study Variables ................................................................................................................... 7 
3.1 Primary Variable ................................................................................................................. 7 
3.2 Exploratory Variables .......................................................................................................... 7 
3.3 Safety Variables .................................................................................................................. 8 
3.4 Statistical Hypotheses ......................................................................................................... 8 
4. 
Study Design and Procedures ............................................................................................. 8 
4.1 General Study Design ......................................................................................................... 8 
4.2 Schedule of Visits and Assessments ................................................................................... 9 
5. 
Study Treatments .............................................................................................................. 12 
5.1 Method of Assigning Subjects to Treatment Groups .......................................................... 12 
5.2 Masking and Unmasking ................................................................................................... 12 
6. 
Sample Size and Power Considerations ............................................................................ 13 
7. 
Data Preparation ............................................................................................................... 14 
8. 
Analysis Populations ......................................................................................................... 14 
8.1 All Enrolled Subjects ......................................................................................................... 14 
8.2 Randomized Subjects ....................................................................................................... 14 
8.3 Intent to Treat ................................................................................................................... 15 
8.4 Per Protocol ...................................................................................................................... 15 
8.5 Modified Per Protocol ........................................................................................................ 15 
8.6 Safety ............................................................................................................................... 15 
9. 
General Statistical Considerations ..................................................................................... 15 
9.1 Unit of Analysis ................................................................................................................. 15 
9.2 Missing or Inconclusive Data Handling .............................................................................. 15 
9.3 Definition of Baseline ........................................................................................................ 16 
9.4 Data Analysis Conventions................................................................................................ 16 
9.5 Adjustments for Multiplicity ................................................................................................ 17 
10. 
Disposition of Subjects ...................................................................................................... 17 
11. 
Demographic and Pretreatment Variables ......................................................................... 18 
11.1 Demographic Variables ..................................................................................................... 18 
11.2 Pretreatment Variables...................................................................................................... 19 
12. 
Medical History and Concomitant Medications ................................................................... 19 
12.1 Medical History ................................................................................................................. 19 


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Page 4 of 33 
 
12.2 Concomitant Medications .................................................................................................. 20 
13. 
Dosing Compliance and Treatment Exposure .................................................................... 20 
13.1 Dosing Compliance ........................................................................................................... 20 
13.2 Treatment Exposure .......................................................................................................... 20 
14. 
Efficacy Analyses .............................................................................................................. 21 
14.1 Primary Analysis ............................................................................................................... 21 
14.2 Secondary Analyses ......................................................................................................... 23 
14.2.1 
Pupil Diameter at 35 Minutes Responder Analysis ................................................. 23 
14.2.2 
Pupil Diameter at All Time points ........................................................................... 24 
14.2.3 
Maximal Pupil Dilation ........................................................................................... 24 
14.2.4 
Pupillary Light Reflex ............................................................................................. 24 
15. 
Safety Analyses ................................................................................................................ 25 
15.1 Adverse Events ................................................................................................................. 25 
15.2 Visual Acuity ..................................................................................................................... 28 
15.3 Slit Lamp Biomicroscopy Examination ............................................................................... 29 
15.4 Intraocular Pressure .......................................................................................................... 29 
16. 
Pharmacokinetic Analyses ................................................................................................ 30 
17. 
Pharmacodynamic Analyses ............................................................................................. 30 
18. 
Quality of Life Analyses ..................................................................................................... 30 
19. 
Interim Analyses ............................................................................................................... 30 
20. 
Changes from Protocol-Stated Analyses ........................................................................... 30 
21. 
References ....................................................................................................................... 30 
22. 
Revision History ................................................................................................................ 31 
23. 
Tables ............................................................................................................................... 31 
24. 
Listings ............................................................................................................................. 32 
25. 
Figures ............................................................................................................................. 33 
 
 
 


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Confidential & Proprietary 
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List of Abbreviations 
AE 
Adverse Event 
ATC 
Anatomical Therapeutic Chemical Classification 
BCDVA 
Best-Corrected Distance Visual Acuity 
CF 
Count Fingers 
CI 
Confidence Interval 
CPRA 
Cumulative Proportion of Responders Analysis 
CRO 
Contract Research Organization 
eCRF 
Eelectronic Case Report Form 
ETDRS 
Early Treatment of Diabetic Retinopathy Study 
GAT 
Goldmann Applanation Tonometer 
IB 
Investigator Brochure 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
IOL 
Intraocular Lens 
IOP 
Intraocular Pressure 
ITT 
Intent to Treat 
logMAR 
Logarithm of the Minimum Angle of Resolution 
LP 
Light Perception 
MAR 
Missing at Random 
MedDRA 
Medical Dictionary for Regulatory Activities 
MiDD 
Eyenovia MicroDose Dispenser 
MNAR 
Missing Not at Random 
MPP 
Modified Per Protocol 
OD 
Oculus Dexter (right eye) 
OS 
Oculus Sinister (left eye) 
OU 
Oculus Uterque (both eyes - in this protocol, OU does not connote simultaneous 
treatment/evaluation) 
PDF 
Portable Document Format 
PLR 
Pupillary Light Reflex 
PP 
Per Protocol 
PT 
Preferred Term 
RDC 
Remote Data Capture 
RTF 
Rich Text Format 


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SAP 
Statistical Analysis Plan 
SD 
Standard Deviation 
SDC 
Statistics and Data Corporation, Incorporated 
SLE 
Slit-Lamp Examination 
SOC 
System Organ Class 
TEAE 
Treatment-Emergent Adverse Event 
TLR 
Total Letters Read 
UCDVA 
Uncorrected Distance Visual Acuity 
US 
United States 
VA 
Visual Acuity 
WHO DD 
World Health Organization Drug Dictionary 
µD 
Microdose 
µL 
Microliter 
 
 


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1. Introduction 
The purpose of this statistical analysis plan (SAP) is to describe the planned analyses and reporting for 
Eyenovia protocol MYD-TP-32 version C, dated 20-November-2018. 
This SAP is being written with due consideration of the recommendations outlined in the most recent 
International Conference on Harmonisation (ICH) E9 Guideline entitled Guidance for Industry: Statistical 
Principles for Clinical Trials and the most recent ICH E3 Guideline, entitled Guidance for Industry: 
Structure and Content of Clinical Study Reports. 
This SAP describes the data that will be analyzed and the subject characteristics, efficacy, and safety 
assessments that will be evaluated. This SAP provides details of the specific statistical methods that will 
be used. The statistical analysis methods presented in this document will supersede the statistical 
analysis methods described in the clinical protocol. If additional analyses are required to supplement 
the planned analyses described in this SAP they may be completed and will be identified in the CSR. 
2. Study Objectives 
The primary objective of this study is to evaluate the safety and efficacy of Eyenovia’s fixed combination 
of combination of phenylephrine 2.5 %-tropicamide 1% microdose ophthalmic solution administered with 
Eyenovia’s microdose dispenser (MiDD) for dilation of the pupil as compared to placebo. 
3. Study Variables 
3.1 
Primary Variable 
The primary performance endpoint is mean change in pupil diameter at 35 minutes from the time of first 
treatment dose (T0) versus baseline, as measured by digital pupillometry in highly photopic conditions. 
The highly photopic condition will be established using a fully-charged transilluminator (muscle light) at 
the brightest setting. Baseline in the setting of the primary variable refers to the period-specific baseline 
on the day of treatment. 
3.2 
Exploratory Variables 
Additional exploratory outcomes include: 
• 
Proportion of eyes achieving pupil diameter of 6.0 mm or greater at 35 minutes 
• 
Proportion of eyes achieving pupil diameter of 7.0 mm or greater at 35 minutes 
• 
Mean change in pupil diameter at other timepoints (20, 50, 65, 80 120, and 180 minutes) 
• 
Distribution of pupil diameters at 20, 35, 50, 65, 80, 120, and 180 minutes 
• 
Time from baseline to maximal pupil dilation 
These exploratory outcomes will also be measured using digital pupillometry in highly photopic 
conditions. 


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Although not explicitly listed as an efficacy variable within the protocol, pupillary light reflex (PLR) will 
be collected at each treatment visit and also summarized within the analysis. 
3.3 
Safety Variables 
The safety variables include the following: 
• 
Slit-lamp examination (SLE) findings 
• 
Occurrence of adverse events (AE) 
• 
Intraocular pressure (IOP) measured at 65 minutes post-administration 
• 
Visual acuity (VA) changes 
3.4 
Statistical Hypotheses 
The null and alternative hypotheses for comparison of Eyenovia’s fixed combination phenylephrine 
2.5%-tropicamide 1% ophthalmic solution with placebo, are defined as follows: 
H0: The mean change per visit in pupil diameter at 35 minutes versus baseline, as measured by digital 
pupillometry in highly photopic conditions, is not different between Eyenovia’s fixed combination 
phenylephrine 2.5%-tropicamide 1% ophthalmic solution and placebo  
HA: The mean change per visit in pupil diameter at 35 minutes versus baseline, as measured by digital 
pupillometry in highly photopic conditions, is different between Eyenovia’s fixed combination 
phenylephrine 2.5%-tropicamide 1% ophthalmic solution and placebo 
4. Study Design and Procedures 
4.1 
General Study Design 
This trial is a double-masked, placebo-controlled, cross-over superiority study evaluating Eyenovia’s 
fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution versus placebo. 
Volunteer participants from 2-3 United States (US)-based investigational sites will be screened for study 
eligibility during a Screening Visit and enrolled after signing the study-specific informed consent form 
(ICF). Subjects meeting all inclusion/exclusion criteria will be scheduled for 3 treatment visits, which 
must occur at least 2 days, but no more than 7 days apart. At each treatment visit, baseline 
measurements will be taken, then either study drug or placebo will be administered in both eyes in two 
instances approximately 5 minutes apart, after which efficacy and safety assessments will be performed 
at specific time intervals. The study drug administration schedule for subjects will be equally randomized 
across 2 sequences of 2 study treatments and 3 periods.  
This study will be double-masked. There will be no differences in the presentation of study drug or 
placebo administered and all study personnel conducting ophthalmic assessments will be masked to 
treatment assignment. Study drug administration will be performed by at least 3 different trained 
personnel during the study. 


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4.2 
Schedule of Visits and Assessments 
Study visits include a Screening Visit, which must occur between 1 and 14 days prior to treatment; 
followed by Treatment Day 1, Treatment Day 2, and Treatment Day 3. Treatment visits must occur at 
least 2 days and not more than 7 days apart. The study visit schedule is presented in flow chart form in 
Figure 1. 
Figure 1: Study Visit Flow Chart 
  
The schedule of visits and assessments is provided below in Table 1. 


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Table 1: Schedule of Medication Administration and Examinations 
 
Assessment/Procedure 
Screening 
Visit 
(Day -14 to 
Day -1) 
Treatment  
Visit 1 (Day 1)1, Visit 21 and Visit 31 
Baseline0 
Time 02 
Time 1 
T0+20 min 
(± 2 min) 
Time 2 
T0+35 min 
(± 5 min) 
Time 3 
T0+50 min 
(± 5 min) 
Time 4 
T0+65 min 
(± 5 min) 
Time 5 
T0+80 min 
(± 10 min) 
Time 6 
T0+120 min 
(± 10 min) 
Time 7 
T0+180 min 
(± 10 min) 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
Medical history 
X 
 
 
 
 
 
 
 
 
 
Ocular history 
X 
 
 
 
 
 
 
 
 
 
Prior/concomitant medication use 
X 
 
 
 
 
 
 
 
 
 
Urine pregnancy test3 
X 
X3 
Manifest refraction (OU) 
X 
 
 
 
 
 
 
 
 
 
BCDVA (OU)4 
X 
X 
 
 
 
 
 
 
 
X 
Study drug administration sequence 
determination (OU) 
 
X 
 
 
 
 
 
 
 
 
Study drug administration (OU) 
 
 
X 
 
 
 
 
 
 
 
Slit lamp biomicroscopy (OU)5 
X 
X 
 
 
 
 
 
 
 
X 
Van Herrick Angle Assessment (OU)6 
X 
 
 
 
 
 
 
 
 
 
IOP (OU)7 
X 
X 
 
 
 
 
X 
 
 
 
Pupil diameter assessment (OU)8 
X 
X 
 
X 
X 
X 
X 
X 
X 
X 
Pupillary light reflex (OU) 
 
X 
 
 
X 
 
X 
 
 
X 
Study eligibility determination 
X 
 
 
 
 
 
 
 
 
 
Dilated fundus exam (OU) 
X 
 
 
 
 
 
 
 
 
 
AE assessment 
 
X 
 
 
X 
 
 
 
 
X 
0 Baseline refers to evaluations made at each Treatment Day prior to study medication administration. 
1 Treatment Visit 1, 2 and 3 must be separated by at least 2 days; but may be up to 7 days apart to allow for scheduling flexibility. 
2 Time 0 starts at the point the first of two study medication doses is administered in second eye of the study subject 
3 A urine pregnancy test will be conducted in females of childbearing potential at the Screening Visit and at the last visit (Treatment Visit 3). The test may be administered at any time 
during the Screening and Treatment Day 3 Visits.  If the subject terminates study participation prior to Treatment Visit 3, this test must be administered promptly at the time of 
termination. 
4 BCDVA to be measured using ETDRS methods.  For younger pediatric subjects, UCDVA may be measured using age-appropriate methods per investigator’s usual practice. 
5 For younger pediatric subjects who cannot cooperate with a traditional SLE, a portable slit lamp model may be used, if necessary. 
6 Performed as part of slit lamp examination. 
7 IOP to be measured using Goldmann Applanation tonometry. For younger pediatric subjects, IOP may be measured using age-appropriate methods per investigator’s usual practice. 
8 Performed using Neuroptics pupillometer – VIP 300.  For younger pediatric subjects for whom the pupillometer cannot be successfully used, a ruler or pupil gauge may be used.  At 
Treatment Visit 3 - Time 7, subjects whose pupil diameter is larger than baseline may be followed at hourly intervals up to 6 hours after medication administration to gather additional 
data regarding the duration of study treatment effect.  


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Study visits will be referred to in all tables and listings. The following table shows the scheduled study 
visits and their planned timing for each study visit: 
Scheduled Visit 
Visit Timing 
Screening 
Day -14 to Day -1 
 Treatment Visit 1 
Day 1 
Treatment Visit 2 
Day +3 to Day +8 
Treatment Visit 3 
Day +5 to Day +15 
Subjects will receive Eyenovia fixed combination or placebo control at more than one treatment visit, 
and based on planned seqences of ABB/BAA, subjects will receive the same treatment at both 
Treatment Visits 2 and 3. For summaries by visit, a derived “all visits combined” will also be defined for 
analysis (as necessary) as follows: 
• 
For continuous outcomes, the value for analysis will be defined as the single value if subjects 
have a given treatment once (eg, the value of treatment A at Visit 1 for subjects are randomized 
to sequence ABB), or based on the mean of the two values if the subject has received the 
treatment twice (eg the mean of values from Visits 2 and 3 for subjects are randomized to 
sequence BAA).  
• 
For categorical and time to event outcomes, the value for analysis will be derived from the values 
at Visits 1 and 2, values form Visit 3 will not be considered. 
In addition, time point number and planned study time will be included in tables and listings to enable 
reviewers to understand the assessment timing without referring to the protocol schedule for time points 
within a treatment visit. The following table shows the scheduled study time points, their planned study 
time, and the acceptable time window for each study visit: 
Scheduled Time 
Planned Study Time 
Time Window 
Time 0 
Minute 0 (pre-treatment) 
N/A 
Time 1 
20 Minutes Post-Dose 
±2 minutes 
Time 2 
35 Minutes Post-Dose 
±5 minutes 
Time 3 
50 Minutes Post-Dose 
±5 minutes 
Time 4 
65 Minutes Post-Dose 
±5 minutes 
Time 5 
80 Minutes Post-Dose 
±10 minutes 
Time 6 
120 Minutes Post-Dose 
±10minutes 
Time 7 
180 Minutes Post-Dose 
±10 minutes 
Data from unscheduled visits, unscheduled time points or unplanned repeat assessments will be 
included in the data listings. In general, these data will be excluded from the summary tables unless 
otherwise specified. 
 


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5. Study Treatments 
Two study treatments administered by Eyenovia’s MiDD will be evaluated in this study. The treatments 
evaluated are: 
• 
Eyenovia’s fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution 
• 
Placebo 
5.1 
Method of Assigning Subjects to Treatment Groups 
At the Screening Visit, subjects who provide verbal and written informed consent will be assigned a 
unique subject ID number. This subject ID will be used to identify subjects in all datasets and listings for 
the study. 
Each eligible subject will be randomly assigned to receive one of 2 sequences of drug administration 
(ABB and BAA, where “A” is one of the two study treatments and “B” is the other) where 1 of the 2 study 
medications will be administered in both eyes at each of the 3 treatment visits. After Treatment Visit 3, 
each study subject will have been treated with both study medications, and one of the study treatments 
will have been received twice. The study drug administration schedule is equally distributed across 2 
randomization sequences, balanced for carryover. This is the optimal three-period, two-group design 
(Jones and Kenward, 2015, section 3.5).  
Subject treatment assignments will be prepared using a computer-generated randomization scheme by 
an independent statistician who is not involved in the day-to-day conduct of the study. Each individual 
subject’s treatment assignment sequence will be provided in paper format contained in a sealed 
envelope. The study randomization scheme will be stratified by iris color. Note that if the study includes 
more than one site, the randomization will effectively be stratified by site in order to supply randomization 
envelopes to each site and maintain treatment sequence balance within each site. 
5.2 
Masking and Unmasking 
The study drug administered will be masked to the study subject, the Investigator, and study staff 
administering the drug and/or performing clinical assessments. The Sponsor (or designee) involved in 
day-to-day study management will also be masked to treatment assignments. 
This study will be double-masked so that there are no differences in presentation (Eyenovia’s fixed 
combination mydriasis agent is formulated as a multi-dose microdroplet spray and placebo is also 
administrated as a microdroplet spray). To maintain masking of the study drug assignments for staff 
administering investigational product and/or performing clinical evaluations, study inventory storage and 
management will be performed by a pharmacy associated with the study site. The Sponsor, or designee, 
will ship study drug kits to each site pharmacy. Each kit will be labeled with the study protocol number, 
study drug code, product storage requirements, and the Caution – New Drug – Limited by Federal (US) 
Law to Investigational Use label. 


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For each treatment day, one or more new MiDD base units and cartridges containing the study drug(s) 
to be administered will be provided by the pharmacy for use. Each MiDD will appear the same except 
for a label indicating the study drug code. New MiDDs must be prepared by the pharmacy for use by 
site personnel on each treatment day. To maintain masking of site personnel, the MiDD containing each 
of the 3 study drugs will be labeled with the appropriate study randomization code, pharmacy preparation 
date and “use by” date, then forwarded to study drug administrator for use. The label containing the 
study drug code should not be revealed to individual subjects or otherwise disclosed in any way to 
personnel at the site who are performing post-drug administration ophthalmic assessments during the 
study. 
Unmasking of treatment assignments can only occur in the event of a medical emergency or occurrence 
of an AE that, in the opinion of the Investigator, warrants such action. In the absence of medical need, 
the treatment assignment code will not be available to the Investigator, site staff, or Sponsor (or 
designee) representative involved in day-to-day study management until study completion and database 
lock. 
In the event an Investigator considers unmasking of a subject’s treatment assignment to be necessary, 
he/she must first contact the study Medical Monitor (or designee) to discuss the case. Only after 
consultation with the study Medical Monitor will the decision be made regarding unmasking. Treatment 
assignments will be revealed only for the subjects approved for unmasking, thus leaving masking on 
remaining subjects intact. 
6. Sample Size and Power Considerations 
Up to 90 subjects who have provided informed consent for study participation will be enrolled in the 
study. It is estimated that 65 subjects will be randomized in order to achieve the goal of having 54 
subjects evaluable for the primary efficacy endpoint. 
A sample size of 27 subjects per sequence, 54 total (the study has two sequences, ABB and BAA where 
A is the Eyenovia fixed combination, and B is the placebo control), will have 95% power to detect a 
difference in means of 0.5 mm, assuming a standard deviation of differences of 1.0 mm, using a paired 
t-test with a 0.05 two-sided significance level. 
Power calculations are provided for additional exploratory outcomes below on an informational basis.  
When the sample size is 54, a two-sided 95% exact confidence interval for a single proportion will have 
a half-width of 13.9% or less.  
• 
For evaluation of the proportion of eyes achieving pupil diameter of 6.0 mm or greater at 35 
minutes: if the observed proportion in the fixed combination treatment is 70.4% (38/54), then 


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the exact 95% confidence interval for the proportion will be [56.4%, 82.0%], with a half-width of 
12.8%. 
• 
For evaluation of the proportion of eyes achieving pupil diameter of 7.0 mm or greater at 35 
minutes: if the observed proportion in the fixed combination treatment is 40.7% (22/54), then 
the exact 95% confidence interval for the proportion will be [27.6% 55.0%], with a half-width of 
13.7%. 
7. Data Preparation 
All reported study data will be recorded on the electronic case report forms (eCRF) supplied by Statistics 
and Data Corporation, Incorporated (SDC). Only the Principal Investigator and authorized study staff 
according to the Delegation of Responsibilities log are entitled to make entries in the eCRF.  
After data are entered into the clinical study database, electronic edit checks and data review will be 
performed. All data validation specifications and procedures are detailed in the Data Validation Manual 
as a separate document. When the database has been declared to be complete and accurate, the 
database will be locked. Any changes to the database after data have been locked can only be made 
with the approval of the Sponsor and the clinical contract research organization ([CRO], if applicable), 
in consultation with SDC.  
All analyses outlined in this document will be carried out after the following have occurred: 
• 
All data management requirements are met according to SDC standard operating procedures, 
including data entry, performance of edit and validation checks, documentation and resolution 
of data queries, and database lock with written authorization provided by appropriate SDC and 
Sponsor personnel; 
• 
Protocol deviations have been identified and status defined (major/minor deviations);  
• 
Analysis populations have been determined; and 
• 
Randomized treatment codes have been unmasked. 
8. Analysis Populations 
8.1 
All Enrolled Subjects 
The all enrolled subjects population will include all subjects who have provided informed consent and 
entered the study, including screen failures. 
8.2 
Randomized Subjects 
The randomized subjects population will include all subjects who were randomized to a treatment 
sequence. 


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8.3 
Intent to Treat 
The intent to treat (ITT) population will consist of all randomized subjects who received a dose of study 
medication. Subjects will be analyzed according to the planned sequence of treatments. 
8.4 
Per Protocol 
The per protocol (PP) population will consist of all ITT subjects who completed all planned assessments 
(relevant to conducting the analysis of the primary performance endpoint) without major protocol 
violations. Subjects who have not received treatment at all three treatment visits will be excluded from 
the PP population. 
Subjects to be excluded from the PP population will be selected prior to unmasking treatment 
assignment. Subjects will be analyzed according to the sequence of treatments received. 
8.5 
Modified Per Protocol 
The modified per protocol (MPP) population is defined similarly to the PP population, except that 
classification will be made for each subject at the individual treatment visit level, rather than at the subject 
level. Unlike the PP population, subjects who only complete 1 or 2 of the 3 treatment visits may be 
included.  
The MPP will consist of all ITT subjects who completed all planned assessments without major protocol 
violations for the given treatment visit. Subjects to be excluded from the MPP population will be selected 
prior to unmasking treatment assignment. Subjects will be analyzed according to the sequence of 
treatments received. 
8.6 
Safety 
The safety population will consist of all ITT subjects. Subjects will be analyzed according to the sequence 
of treatments received. 
9. General Statistical Considerations 
9.1 
Unit of Analysis 
For measurements taken at the subject level, the unit of analysis will be the individual subject and for 
measurements taken at the eye level, the unit of analysis will be the individual eye unless otherwise 
indicated.  
9.2 
Missing or Inconclusive Data Handling 
By definition, the PP population is limited to subjects who complete the study and completed all planned 
assessments, therefore there should not be missing data for the primary efficacy analysis as data from 
individual subjects are wholly included or excluded. In the event of missing data for sensitivity analyses 
of the primary efficacy variable, the SAS PROC MIXED procedure uses all available data to produce 


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maximum likelihood parameter estimates. In such a setting the missing observations are ignorable if 
data are missing at random (MAR). 
Adverse events with unknown severity will be summarized as being severe, and adverse events with 
relationship to study drug not specified will be summarized as being related to study drug. 
For all other variables, missing data will not be imputed, observed values will be presented.  
9.3 
Definition of Baseline 
For all variables, the “pre-randomization” baseline value is defined as the last pre-treatment 
measurement taken prior to administration of study drug at Visit 1. Normally this will be the value 
obtained from the Visit 1 Baseline time point value, however for assessments collected at the Screening 
visit only (or if the value from Visit 1 Baseline is unavailable) the Screening visit value will be used. 
For variables collected after dosing at each Treatment Visit (eg, pupil diameter assessment), a “period-
dependent” baseline value is also defined as the last pre-treatment measurement taken prior to 
administration of study drug at that Visit (eg, Visit X Baseline time point, where X corresponds to 
Treatment Visit 1, 2, or 3). If Treatment Visits 2 or 3 are done but the baseline value is unavailable, the 
Baseline value will be imputed from previous treatment visit for analysis (eg, missing period-specific 
baseline value for Treatment Visit 2 would be imputed using the period-specific baseline value from 
Treatment Visit 1; missing period-specific baseline value for Treatment Visit 3 would be imputed using 
the period-specific baseline value from Treatment Visit 2). 
Change from baseline will be calculated as follow-up measure minus pre-randomization baseline/period-
dependent baseline value measure. 
Analyses of changes from baseline will include the applicable baseline value (pre-randomziation or 
period-dependent) as a covariate in the statistical model, or will adjust for baseline in the case of 
categorical analyses, unless otherwise specified.  
9.4 
Data Analysis Conventions 
All data analysis will be performed by SDC after the study is completed and the database has been 
locked and released for unmasking. Statistical programming and analyses will be performed using SAS® 
Version 9.4 or higher. Output will be provided in rich text format (RTF) for tables and portable document 
format (PDF) for tables, listings, and figures using landscape orientation. All study data will be listed by 
subject, treatment, and visit (as applicable) based on all randomized subjects unless otherwise 
specified. 
Summaries for continuous and ordinal variables will include the number of observations (n), arithmetic 
mean, standard deviation (SD), median, minimum, and maximum). Minima and maxima will be reported 
with the same precision as the raw values; means and medians will be presented to one additional 


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decimal place than reported in the raw values. Standard deviations will be presented to two additional 
decimal places than reported in the raw values. Summaries for discrete variables will include frequency 
counts and percentages. All percentages will be rounded to one decimal place (i.e., XX.X%). Change 
from baseline will be calculated as follow-up visit (or follow-up time point) minus baseline.  
All statistical tests will be two-sided with a significance level of 0.05 (α = 0.05) unless otherwise specified. 
Confidence intervals (CI) for differences between treatment groups will be two-sided at 95% confidence. 
All p-values will be rounded to four decimal places; p-values less than 0.0001 will be presented as 
“<0.0001”; p-values greater than 0.9999 will be presented as “>0.9999”. 
Unless otherwise specified, summaries will be presented by treatment sequence or treatment group 
and, where appropriate, visit and time point. 
For analyses where a ‘seed’ is specified within SAS® in order to replicate results, the seed for final 
analysis will be based on the database lock date (eg, 20190101 if the study had locked on 01-JAN-
2019). 
9.5 
Adjustments for Multiplicity 
All time points for pupil diameter will be analyzed separately; however, only the 35-minute time point will 
support statistical inference.  
As this study contains a single hypothesis being tested on a single endpoint with only two treatment 
groups, no adjustment for multiplicity is necessary. 
10. Disposition of Subjects 
Disposition of subjects will be summarized for all enrolled subjects. The total number of enrolled subjects 
with the number and percentage of not randomized subjects will be summarized. The reasons for screen 
failure will be displayed with the percentages calculated using total number of enrolled subjects as the 
denominator.  
Disposition of subjects will also be summarized for all randomized subjects. The number of subjects in 
each of the analysis populations (ITT, PP, MPP, and Safety) will be displayed by treatment sequence. 
Study completion will be assessed in terms of completion of treatment visits and overall study 
completion. The number and percentage of subjects completing each treatment visit, completing the 
study, prematurely discontinuing from the study and the reasons for study discontinuation will be 
summarized by treatment sequence for all randomized subjects. A subject listing will be provided which 
includes the date and reason for premature study discontinuation.  
Subject disposition will be presented in terms of the numbers and percentages of subjects who were 
randomized, completed the study, and discontinued from the study. Subjects who are not discontinued 


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from the study will be considered study completers. Disposition will be summarized by treatment 
sequence and for all subjects. 
The number and percentage of subjects with major protocol deviations will be summarized by category 
(eg, Informed Consent, Inclusion/Exclusion, etc.), treatment sequence and overall for all randomized 
subjects. The eCRF contains a pre-defined set of options for deviation category that is assigned by 
study site personnel, however, for consistency in reporting, sponsor-defined deviation categories will be 
determined by Eyenovia for use within the analysis during a review of all protocol deviations prior to 
database lock and unblinding. Protocol deviations will be classified as major or minor prior to the closure 
of the database during a masked review of each protocol deviation. Major deviations will be defined as 
those deviations that potentially impact the primary outcome of the study. A subject listing will be 
provided which includes the date of the deviation, the sponsor-defined deviation category, the deviation 
description and the classification of whether the deviation was judged to be major or minor. 
In addition, subject listings will be provided that include informed consent date, inclusion and exclusion 
criteria violations, and exclusions from the PP population. 
11. Demographic and Pretreatment Variables 
11.1 
Demographic Variables 
The demographic variables collected in this study include age, sex, race, ethnicity and iris color. 
Subjects who record more than one race will be grouped into a single category denoted as multi-racial. 
Demographic variables will be summarized for the ITT and PP population. 
Age (years) will be summarized, overall and by treatment sequence, using continuous descriptive 
statistics. Age will also be categorized using categories defined as < 17 years and ≥17 years, as well as 
based on observed quartiles in the actual data. Age will be reported in years and calculated using the 
following formula: 
Age = (informed consent date – date of birth) / 365.25 truncated as an integer 
Iris color will be combined into categories defined as: 
• 
Light: blue, green, hazel, gray 
• 
Dark: brown, black 
Any iris color values of “other” category will be allocated to the light or dark category in accordance with 
the stratum used for the study randomization. 
The number and percentage of subjects will be presented, overall and by treatment sequence, for age 
category, sex, race, ethnicity,iris color, and dichotomoized iris color .  


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A subject listing that includes all demographic variables will be provided. 
11.2 
Pretreatment Variables 
Pretreatment variables will be summarized for the ITT population. 
Baseline best-corrected distance visual acuity (BCDVA) at the Screening visit, expressed in logarithm 
of the minimum angle of resolution (logMar) units, will be summarized for each eye, for all subjects (refer 
to Section 15.2 for details) and by treatment sequence using continuous descriptive statistics. 
Manifest refraction is performed as a pre-BCDVA assessment at the Screening visit and will be 
summarized for each eye, for all subjects and by treatment sequence using continuous descriptive 
statistics. Manifest reaction results will be summarized within the baseline characteristics table and also 
listed.  
Van Herrick angle assessment will be performed as part of slit lamp examination at the Screening Visit. 
The distribution of grades will be tabulated for each eye, for all subjects and by treatment sequence. 
Using ophthalmoscopy, the fundus will be examined and evaluated by the Investigator to be either 
normal or abnormal at the Screening Visit. Results will be tabulated for each eye, for all subjects and by 
treatment sequence. 
Urine pregnancy test results from the Screening Visit will be listed only. 
A subject listing that includes all pretreatment variables will be provided. 
12. Medical History and Concomitant Medications 
12.1 
Medical History 
Medical history will be coded using Medical Dictionary for Regulatory Activities (MedDRA), version 21.0 
(or higher). Medical History will be summarized for the ITT Population. 
Ocular medical history is a subset of the overall medical history. Ocular medical history consists of those 
medical history records where the location field on the medical history eCRF is marked as location being 
oculus dexter (OD – right eye), oculus sinister (OS – left eye) or oculus uterque (OU – both eyes). 
Non-ocular medical history will be summarized using discrete summary statistics and presented across 
all subjects at the subject and event level by System Organ Class (SOC) and Preferred Term (PT) using 
the ITT population. Ocular medical history will be similarly summarized at the subject level. If a subject 
reports the same PT multiple times within the same SOC, that PT will only be reported once within that 
SOC. As with the PT, if a subject reports multiple conditions within the same SOC, that SOC will only 
be reported once.  
Listings of medical history will be generated separately for ocular and non-ocular data. 


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12.2 
Concomitant Medications 
Concomitant medications will be coded using World Health Organization Drug Dictionary (WHO DD), 
Enhanced B3, September 2018 (or higher), and summarized to the therapeutic drug class (Anatomical 
Therapeutic Chemical [ATC] Level 4 classification) and preferred name. If the ATC Level 4 classification 
is not provided, the next lowest classification that is provided in the coding dictionary will be used. The 
preferred name will be defined as the active ingredient; if the active ingredient is not provided or includes 
more than two ingredients (eg, multivitamins) then the drug name will be summarized as the preferred 
name. Any uncoded terms will be summarized under the ATC classification and preferred name of 
“Uncoded.” 
Prior medications are defined as those medications those with an end date before the first treatment 
(Visit 1) in the sequence. Concomitant medications for each treatment are defined as those that are in 
use at the start of study medication administration or started after dosing but initiation was prior to the 
next treatment in the sequence or study completion/discontinuation. With this definition, it should be 
noted it is possible for a medication to be concomitant for more than one treatment group. 
Concomitant medications will be summarized for the Safety Population, and will be summarized 
separately for ocular and non-ocular medications. Ocular medications are a subset of the overall 
medications, and consist of those medication records where the location field on the eCRF is marked 
as location being OD, OS, or OU. 
Medications will be tabulated for each treatment group using frequencies and percentages. Subjects 
may have more than 1 medication per ATC text. At each level of subject summarization, a subject will 
be counted once if he/she reports 1 or more medications. Percentages will be based on the number of 
subjects in each treatment group.  
Listings of prior and concomitant medications will be generated separately for ocular and non-ocular 
data. 
13. Dosing Compliance and Treatment Exposure 
13.1 
Dosing Compliance 
As this study involves in-office administrations of each study drug, administered to both eyes (OU) 
across two separate instances approximately 5 minutes apart, dosing compliance will not be 
summarized. A subject listing of exposure will contain dosing times for each eye. 
13.2 
Treatment Exposure 
Treatment exposure will be summarized using the Safety Population. 
The number and percent of subjects receiving their administration at each of the 3 treatment visits in 
this study will be tabulated along with the number of individual doses received per treatment visit. The 


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number and percent of subjects receiving at least one individual dose at all 3 treatment visits, and the 
number and percent of subjects receiving both individual doses of all 3 treatment visits will also be 
summarized.  
A subject listing of treatment exposure will also be produced. 
14. Efficacy Analyses 
14.1 
Primary Analysis 
The primary performance endpoint is mean change per visit in pupil diameter at 35 minutes versus 
period-dependent baseline, as measured by digital pupillometry in highly photopic conditions. The highly 
photopic condition will be established using a fully-charged transilluminator (muscle light) at the brightest 
setting.  
Pupil diameter in adults will be measured using Neuroptic pupillometer – VIP 300. For younger subjects 
for whom the pupillometer cannot be successfully used, a ruler or pupil gauge may be used, with each 
measurement repeated 2-3 times for optimal accuracy. The pupil diameter value used for analysis will 
be either the mean (if 2 measurements are taken) or the median (if 3 measurements are taken) of the 
repeated measurements in each eye. 
Primary efficacy analysis will be conducted on the PP population. 
The ABB/BAA crossover design used in this study allows for within-subject (i.e. within-eye) estimates of 
both treatment effects (A and B) and carryover effects (AA, AB, BA, BB). Thus, there is no need to 
include random effects for subject or eye in the analysis model, we will use fixed effects ANOVA to 
analyze these data (Jones and Kenward, 2015, section 5.2) and tests will be based on within-eye 
variances. A SAS REPEATED statement will be used to adjust for correlation between fellow eyes. 
Hypothesis tests will be based on confidence intervals produced by the SAS LSMEANS option for the 
pairwise differences between treatment. 
This study has both a “pre-randomization” covariate (iris color) and a “period-dependent” covariate 
(baseline pupil measurement) as discussed in Jones and Kenward (2015, section 5.4.4). We are 
assuming that the time between visits is sufficiently long that the baseline pupil measurement at each 
visit is unaffected by carryover. 
Comparisons among treatments on mean pupil diameter will be by a fixed-effects analysis of variance 
model. The model will contain an effect due to subject, eye, period (1, 2, or 3), direct effect of treatment, 
first order carryover, period-dependent baseline pupil diameter, and iris color (dark vs light). Proc MIXED 
coding in SAS would be: 
proc mixed; 
class subject eye period treatment carry iris; 


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model deltaPD = subject*eye period treatment carry baselinePD iris / s; 
repeated eye / subject = subject type=cs; 
lsmeans treatment carry/pdiff cl; 
run; 
where  
• 
deltaPD=change from period-dependent baseline in pupil diameter at Time 2 (T0 + 35 minutes) 
• 
eye=OD, OS 
• 
period=1, 2, or 3 
• 
treatment: 
o 
Eyenovia’s fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution 
o 
Placebo 
• 
carry=first order carryover. An indicator variable will be constructed containing the treatment the 
subject received in the previous period (set to A or B for Periods 2 and 3, and arbitrary value for 
Period 1, three levels total). 
• 
baselinePD=period-dependent baseline pupil diameter 
• 
iris=iris color (dark, light) 
A compound symmetry covariance structure will be used for the model. If the model fails to converge, 
covariates will be removed one by one until the model converges. Iris color, carryover effect and 
period-dependent baseline pupil diameter are the candidate variables to be removed, in that order. 
Carryover effect included within the model will be tested to assess the carryover assumption. 
All time points for pupil diameter will be analyzed separately; however, only the 35-minute time point will 
support statistical inference. Success is defined as the fixed combination (provided by the LSMEANS 
statement) being statistically significantly better than placebo.  
The SAS® PROC MIXED procedure uses all available data to produce maximum likelihood parameter 
estimates, in such a setting the missing observations are ignorable if data are MAR. 
The primary analysis described above will be rerun on the MPP population as a sensitivity analysis.  
As a further sensitivity analysis, multiple imputation based on reference-based multiple imputation will 
be used to impute missing data at the 35-minute time point based on a missing not at random (MNAR) 
assumption. Since the PP population contains only subjects with non-missing values, this analysis will 
be conducted using the ITT population, which may contain subjects who have only partially available 
information. Placebo treatment will serve as reference group for this approach. Each subject will be 
included for all three treatment visits for this approach, regardless of how many treatment visits were 
actually completed. 
SAS code for the multiple imputation is as follows: 
proc mi out=outmi nimpute=20;  
   class trt period eye iris; 
   monotone Reg(PD_35 = period eye iris baselinePD); 
   mnar model(PD_35/ modelobs=(trt=’Placebo’)); 


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   var period eye iris baselinePD PD_35; 
  run; 
The resulting dataset, OUTMI, will then rerun using the model previously described for primary analysis, 
with an additional ‘by _imputation’ statement included, and the least squares means differences from 
this will be combined using Proc MIANALYZE.  
14.2 
Secondary Analyses 
The PP population will be used for the secondary analyses. Missing data will not be imputed. The 
secondary analysis variables will be presented in subject listings by visit. 
14.2.1 Pupil Diameter at 35 Minutes Responder Analysis 
The proportion of eyes achieving pupil diameter of 6.0 mm or greater at 35 minutes post-dose will be 
summarized by eye, by treatment group and visit (including across all visits) with descriptive statistics. 
Comparison of the fixed combination of phenylephrine 2.5%-tropicamide 1% microdose ophthalmic 
solution administered against placebo will be conducted with a generalized estimating equation (GEE) 
model for binomial outcome, implemented in SAS as follows: 
proc genmod descending; 
  class subject eye period treatment carry iris; 
  model resp = period treatment carry baselinePD iris / link=logit dist=binomial; 
  repeated subject = subject(eye)/ type=cs; 
  lsmeans treatment / oddsratio diff cl; 
run; 
where 
• 
resp=response; pupil diameter 6.0mm or greater at 35 minutes post-dose  
• 
eye=OD, OS 
• 
period=1, 2, or 3 
• 
treatment: 
o 
Eyenovia’s fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution 
o 
Placebo 
• 
carry=first order carryover. An indicator variable will be constructed containing the treatment the 
subject received in the previous period (set to A or B for Periods 2 and 3, and arbitrary value for 
Period 1, three levels total ). 
• 
baselinePD=baseline pupil diameter 
• 
iris=iris color (dark, light) 
The previously described analysis will be repeated using an alternate cutoff of 7.0mm. 
In addition, a cumulative proportion of responders analysis (CPRA) will be created to graphically display 
the proportions of subjects with a pupil diameter at 35 minutes post-dose over the entire range of 
possible pupil diameter cut-offs (Farrar 2006), with separate lines for each treatment, combined visit,eye 
combination.  


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14.2.2 Pupil Diameter at All Time points 
Pupil diameter, including actual value as well as change from period-dependent Baseline, will be 
summarized for each time point of treatment visits (20, 35, 50, 65, 80, 120, and 180 minutes) with 
descriptive statistics for continuous endpoint by treatment, by visit (including across all visits), and eye, 
and displayed graphically using mean and standard error. Missing values will not be imputed for these 
summaries. As pupil diameter at 35 minutes post-dose is the primary endpoint in the study, additional 
analyses of that time point are described in Section 14.1.  
The distribution of pupil diameters at each time point of treatment visits will be summarized by treatment 
group in a summary table, as well as graphically. 
14.2.3 Maximal Pupil Dilation 
Maximal pupil dilation is defined as the maximum pupil value after study drug is administered to both 
eyes, and time to maximal pupil dilation is the time point corresponding to first occurrence of this maximal 
value. For this analysis, the actual clock time will be used to determine number of minutes from study 
drug administration rather than the nominal time point value. In order to exclude increases in pupil 
diameter which might reasonably be attributed to measurement variability, increases less than 1.0mm 
will be excluded for consideration. In other words, maximal pupil dilation will be based on the subset of 
measurements  which are 1.0mm or greater. 
Time to maximal pupil dilation will be analyzed based on Kaplan-Meier approach by treatment, and visit 
(including across all visits). Each subject will be considered to have an event at the time where maximal 
(period-dependent) post-Baseline pupil dilation with 1.0mm or greater increase from BL was first 
observed. Subjects who do not have any (period-dependent) post-Baseline increases in pupil diameter 
in either eye will be censored at the time of the last non-missing pupil measurement. Subjects who 
discontinue prior to completing all post-dose assessments will be evaluated based on the assessments 
which are available.  
If all subjects experience an increase from period-dependent Baseline (ie, there are no censored 
observations), this analysis may be replaced by a summary of descriptive statistics for categorical 
outcome. 
14.2.4 PUPILLARY LIGHT REFLEX 
Pupillary light reflex (PLR) will be evaluated using a fully-charged transillumator, or muscle light. The 
response will be recorded on a scale ranging from 0 (non-responsive) to 3 (brisk). 
The responses at each time point will be summarized by eye using counts and percentages in each 
response category for each treatment group at each visit (including across all visits) and time point. 


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Shifts from period-dependent baseline value to worst (period-dependent) post-Baseline response will 
also be tabulated by treatment group. 
Pairwise comparisons of the responses at the 35 minute post-dose time point for the fixed combination 
of phenylephrine 2.5%-tropicamide 1% microdose ophthalmic solution administered against its 
individual components will be conducted with a generalized estimating equation (GEE) model for ordinal 
multinomial outcome, implemented in SAS as follows: 
proc genmod descending; 
  class subject eye period treatment carry iris; 
  model resp = period treatment carry baselinePLR iris / dist=multinomial aggregate=treatment; 
  repeated subject = subject(eye); 
  lsmeans treatment / oddsratio diff cl; 
run; 
where 
• 
resp=pupillary light response category (ordered categories as: non-responsive; minimally 
responsive; moderately responsive; and brisk) at 35 minutes post-dose  
• 
eye=OD, OS 
• 
period=1, 2, or 3 
• 
treatment: 
o 
Eyenovia’s fixed combination phenylephrine 2.5%-tropicamide 1% ophthalmic solution 
o 
Placebo 
• 
carry=first order carryover. An indicator variable will be constructed containing the treatment the 
subject received in the previous period (set to A or B for Periods 2 and 3, and arbitrary value for 
Period 1, three levels total). 
• 
baselinePLR=baseline pupillary light response category 
• 
iris=iris color (dark, light) 
A subject listing of PLR will also be produced. 
15. Safety Analyses 
All safety analyses will be conducted using the Safety Population. 
15.1 
Adverse Events 
An AE is defined as any untoward medical occurrence associated with the use of a drug in humans, 
whether or not the event is considered drug-related. An AE can therefore be any unfavorable and 
unintended sign (e.g., an abnormal laboratory finding), symptom, or disease occurring after the first dose 
of study drug, without any judgment about causality. Any pre-existing medical condition that worsens 
after first administration of study drug will also be considered a new AE. The AE reporting period ends 
upon study exit. Study drug includes refers to Eyenovia’s fixed combination or placebo which are 
administered by Eyenovia’s MiDD (refer to Section 4.1) given during any stage of the study. All AEs will 
be coded using MedDRA, version 21.0 (or higher) Treatment group for the AE will be determined as the 
most recent treatment received by the subject. For example, an AE reported after dosing at Treatment 


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Visit 1 but prior to first dose at Treatment Visit 2 would be allocated to the treatment group given at 
Treatment Visit 1.  
Summaries of AEs will be generally presented by treatment, where AE data from periods 2 and 3 of this 
crossover design will be combined since they will correspond to the same treatment (due to planned 
sequences of ABB and BAA). 
Treatment-emergent adverse events (TEAE) are defined as any event that occurs or worsens on or after 
the day and time of first treatment is initiated. Only TEAEs are captured as AEs in this study, AEs with 
onset prior to first administration of study medication are recorded as Medical History for the subject. In 
the unexpected case that there are a dverse events recorded in the eCRF which began prior to 
treatment, these events will not be included in the summary tables but will be included in the AE data 
listings. 
Ocular AEs are a subset of AEs, and are identified as those AEs where the location field on the AE 
eCRF is marked as OD, OS, or OU. Since both eyes are treated in this study with study drug, summaries 
of ocular events will include AEs with onset in either eye. 
An overall summary will be presented that includes the number of TEAEs and the number and 
percentage of subjects who experienced at least one TEAE, by treatment group. This summary will also 
include breakdowns of TEAEs further categorized as ocular or non-ocular, serious TEAEs, TEAEs by 
maximum severity, TEAEs by maximum relationship, and TEAEs leading to subject withdrawal. 
Additional summaries of TEAEs will be provided showing the number and percentage of subjects who 
experienced at least one TEAE. These summaries will be presented by SOC and PT. Non-ocular TEAEs 
will be summarized using discrete summary statistics and presented by treatment group at the subject 
and event level by SOC and PT. Due to the fact that both eyes are treated with study drug, ocular TEAEs 
will be summarized in a similar manner (eg, separate summaries per eye are not planned). If a subject 
reports the same PT multiple times within the same SOC, that PT will only be reported once within that 
SOC. As with the PT, if a subject reports multiple conditions within the same SOC, that SOC will only 
be reported once. In the summary, SOC will be listed in ascending alphabetical order; PTs will be listed 
in order of descending frequency for all subjects within each SOC. The occurrence of non-ocular and 
ocular TEAEs will also be tabulated by SOC and PT for the following: maximal severity and suspected 
relationship to study drug. 
Separate summaries will be provided for the following categories of AEs:  
• 
Ocular TEAEs  
• 
Non-ocular TEAEs  
• 
Treatment-related ocular TEAEs 


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• 
Treatment-related non-ocular TEAEs 
• 
Expected TEAEs 
• 
Unexpected TEAEs 
• 
Serious TEAEs 
• 
Serious ocular TEAEs 
Severity of an AE is defined as a qualitative assessment of the degree of intensity of an AE as 
determined by the Investigator or reported to him/her by the subject. The assessment of severity is made 
irrespective of relationship to study drug or seriousness of the event and should be evaluated according 
to the following scale: 
• Mild: Subject is aware of sign or symptom, but it is easily tolerated 
• Moderate: Subject experiences discomfort enough to cause interference with usual activity. 
• Severe: AE is incapacitating to subject, causing inability to work or do usual activity. 
Summaries of TEAEs by maximal severity will be presented for ocular AEs and non-ocular AEs 
separately. The number of subjects with any TEAEs (along with percentages) will be tabulated by SOC 
and PT within each SOC by treatment group. To count the number of subjects with any TEAEs, if a 
subject has multiple TEAEs coded to the same PT within the same SOC, the subject will be counted 
once under the maximal severity. 
The relationship of each AE to the study drug should be determined by the Investigator using these 
explanations: 
• Not Related: Evidence exists that the AE has a cause other than the study drug (e.g. pre-existing 
condition or underlying disease, intercurrent illness, or concomitant medication) and does not 
meet any other criteria listed. 
• Possibly Related: A temporal relationship exists between event onset and administration of study 
drug. Although the AE may appear unlikely to be related to the study drug, it cannot be ruled out 
with certainty; and/or the event cannot be readily explained by the subject’s clinical state or 
concomitant therapies. 
• Probably Related: A temporal relationship exists between the event onset and administration of 
study drug; it appears with some degree of certainty to be related based on known therapeutic 
and pharmacologic actions of the study drug. It cannot be readily explained by the subject’s clinical 
state or concomitant therapies. 
• Definitely Related: Strong evidence exists that the study drug caused the AE. There is a temporal 
relationship between the event onset and administration of the study drug. There is strong 
therapeutic and pharmacologic evidence that the event was caused by the study drug. The 
subject’s clinical state and concomitant therapies have been ruled out as a cause 
For analysis purposes, AEs considered related to study drug (per Investigator) will include AEs with 
relationship equal to possibly related, probably related, or definitely related. Not related AEs will include 
AEs with relationship equal to “not related”. In the event relationship is not specified, the event will be 


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assumed to be related. Summaries of related TEAEs will be presented for ocular AEs and non-ocular 
AEs separately. 
Serious ocular AEs include, but are not limited to: 
• 
A decrease in BCDVA of ≥ 30 total letters read (TLR) or ≥ 0.6 logMAR from the most recent 
previous measurement of BCDVA 
• 
A decrease in BCDVA to light perception (LP) or worse that lasts > 1 hour 
• 
An ocular event that, in the opinion of the investigator, requires medical or surgical intervention 
to prevent permanent loss of sight 
• 
Severe intraocular inflammation; e.g., 4+ anterior chamber cell/flare or 4+ vitritis 
• 
Corneal decompensation 
• 
Retinal tear or detachment 
• 
Central or branch retinal vein occlusion 
An AE or suspected adverse reaction is considered "unexpected" if it is not listed in the Investigator 
Brochure (IB) or is not listed at the specificity or severity that has been observed; or, if an IB 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. "Unexpected," as used in this definition, also 
refers to AEs or suspected adverse reactions that are mentioned in the IB as occurring with a class of 
drugs or as anticipated from the pharmacological properties of the drug but are not specifically 
mentioned as occurring with the particular drug under investigation. The Investigator will initially classify 
the expectedness of the AE on the eCRF, but the final classification is subject to the Medical Monitor’s 
determination. 
Separate listings of AEs will be produced to list all AEs, AEs leading to study discontinuation, and serious 
AEs. 
15.2 
Visual Acuity 
Visual acuity measurements are done per eye, and assessed at Screening and at treatment visits at 
period-dependent Baseline and Time 7 (T0 + 180 minutes) time points. The pre-treatment evaluation of 
VA should be performed before the subject’s pupils have been dilated. Distance VA measurements 
should be obtained at a testing distance of 4 meters by a physician or trained technician using Early 
Treatment of Diabetic Retinopathy Study (ETDRS) lighted charts. 
Adult subjects’ BCDVA will be determined using subjective manifest refraction. The subject should be 
seated so that the distance from the subject's eyes to the ETDRS chart (Chart 1 for right eye and Chart 
2 for left eye) is 4 meters (157.5 inches) at a distance of 4 meters. Visual acuity in adults will be 
expressed in logMAR units, however, logMAR units may not be available for all pediatric methods. For 


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younger pediatric subjects who are unable to complete ETDRS testing, however, uncorrected distance 
visual acuity (UCDVA) should be measured using age-appropriate methods per the Investigator’s usual 
practice. 
The observed and change from period-dependent baseline in BCDVA will be summarized for adults by 
eye and by treatment and visit (including across all visits) using continuous descriptive summary 
statistics.  
The number and proportion of subjects who are able to perform ETDRS testing with a decrease from 
period-dependent baseline value in BCDVA of ≥ 30 total letters read (TLR) or ≥ 0.6 logMAR, and 
decreases in BCDVA to light perception (LP) will be summarized by treatment, visit and time point.  
Subject listings of visual acuity will also be produced, separately for adults and pediatric subjects who 
are unable to perform ETDRS testing. The listing for adults will include a variable that indicates if a 
subject had a visual acuity change from baseline of ≥0.6 on the logMAR scale. 
For younger pediatric subjects who are unable to complete ETDRS testing, visual acuity will only be 
listed because of the variable testing methods appropriate for this subgroup. 
15.3 
Slit Lamp Biomicroscopy Examination 
A slit lamp biomicroscopy examination of the anterior vitreous, cornea, conjunctiva, anterior chamber, 
iris, lens, lid, and sclera will be conducted on both eyes at the Screening Visit, and during treatment 
visits at the period-dependent Baseline and Time 7 (T0+180 minutes) time points. For pediatric subjects 
who cannot cooperate with a traditional SLE, a portable slit lamp model may be used according to the 
Investigator’s usual practice. The individual assessments will be graded as either normal/abnormal, or 
as none/mild/moderate/severe. Grading of each assessment is described in Section 10 of the protocol.  
The results (grade, or normal/abnormal classification, as applicable) of the assessment of each structure 
will be summarized using counts and percentages for each treatment group at each visit (including 
across all visits) and time point for each eye. Shift tables for the slit lamp biomicroscopy parameters will 
also be provided comparing the Time 7 (T0+180 minutes) result (grade, or normal/abnormal 
classification, as applicable) to baseline.  
A subject listing of the slit lamp biomicroscopy parameters will also be produced, results obtained from 
portable slit lamp model will be listed separately. 
15.4 
Intraocular Pressure 
After completion of slit lamp biomicroscopy, local anesthetic will be applied to facilitate intraocular 
pressure (IOP) measurements with the Goldmann Applanation Tonometer (GAT).  


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For adults who are able to undergo GAT, two consecutive IOP measurements of each eye must be 
obtained. If the 2 measurements differ by more than 2 mmHg, a third measurement must be obtained. 
IOP will be analyzed as the mean of these 2 measurements, or as the median of the 3 measurements. 
For pediatric subjects, IOP should be measured using age-appropriate methods per the Investigator’s 
usual practice. 
IOP will be collected at the Screening Visit, and during treatment visits at the period-dependent Baseline 
and Time 4 (T0+65 minutes) time points. 
The IOP values and changes from period-dependent baseline for each eye will be summarized for adults 
using continuous descriptive statistics for each eye by visit (including across all visits))and time point, 
for each treatment group. A subject listing of IOP in adults will also be produced. This listing will include 
a variable that indicates if a subject had an IOP increase from period-dependent baseline of ≥6 or ≥10 
mmHg. 
For younger pediatric subjects who are unable to undergo GAT, intraocular pressure will only be listed 
because of the variable testing methods appropriate for this subgroup. 
16. Pharmacokinetic Analyses 
Not applicable. 
17. Pharmacodynamic Analyses 
Not applicable. 
18. Quality of Life Analyses 
Not applicable. 
19. Interim Analyses 
No interim analyses are planned for this study. 
20. Changes from Protocol-Stated Analyses 
There are no changes from the protocol-stated analyses. 
21. References 
Farrar J.T., Dworkin R.H., Max M.B. Use of the Cumulative Proportion of Responders Analysis Graph 
to Present Pain Data Over a Range of Cut-Off Points: Making Clinical Trial Data More Understandable. 
Journal of Pain and Symptom Management. 2006; 31(4): 369-377. 
US Federal Register. (1998) International Conference on Harmonization; Guidance for Industry: 
Statistical Principles for Clinical Trials. Department of Health and Human Services: Food and Drug 
Administration. Federal Register, Vol. 63, No. 179, September 16, 1998, page 49583. (E9)  


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US Federal Register. (1996) International Conference on Harmonization; Guidance for Industry: 
Structure and Content of Clinical Study Reports. Department of Health and Human Services: Food and 
Drug Administration. Federal Register Vol. 61, July 17, 1996, page 37320. (E3) 
22. Revision History 
Documentation of revision to the SAP will commence after approval of the Final version 1.0. 
23. Tables 
Tables that will be included in the topline delivery are shown in boldface font.  
Table Number 
Title 
Population 
14.1.1.1 
Subject Disposition 
Enrolled 
Subjects 
14.1.1.2 
Subject DispositionReason for Screen Failure 
Enrolled 
Randomized 
Subjects 
14.1.2 
Major Protocol Deviations 
Randomized 
Subjects 
14.1.3.1 
Demographics 
ITT 
14.1.3.2 
Demographics 
Per Protocol 
14.1.4 
Baseline Characteristics 
ITT 
14.1.5.1 
Ocular Medical History 
ITT 
14.1.5.2 
Non-Ocular Medical History 
ITT 
14.1.6.1 
Ocular Concomitant Medications 
ITT 
14.1.6.2 
Non-Ocular Concomitant Medications 
ITT 
14.2.1.1 
Change from Baseline in Pupil Diameter at 35 Minutes – Fixed 
Effects ANOVA 
Per Protocol 
14.2.1.2 
Change from Baseline in Pupil Diameter at 35 Minutes – Fixed 
Effects ANOVA 
Modified Per 
Protocol 
14.2.1.3 
Change from Baseline in Pupil Diameter at 35 Minutes – 
Reference-Based Multiple Imputation 
ITT 
14.2.2.1 
Proportion of Eyes Achieving Pupil Diameter of 6.0mm or 
Greater at 35 minutes Post-Dose 
Per Protocol 
14.2.2.2 
Proportion of Eyes Achieving Pupil Diameter of 7.0mm or 
Greater at 35 minutes Post-Dose 
Per Protocol 
14.2.3 
Change from Baseline in Pupil Diameter by Time Point – Fixed 
Effects ANOVA 
Per Protocol 
14.2.4 
Distribution of Pupil Diameters by Time Point 
Per Protocol 
14.2.5 
Time to Maximal Pupil Dilation with ≥1.0mm Increase from 
Baseline 
Per Protocol 
14.2.6.1 
Pupillary Light Reflex Responses by Time Point 
Per Protocol 
14.2.6.2 
Pupillary Light Reflex Shift from Pre-Dose to Post-Dose 
Per Protocol 
14.3.1 
Overall Summary of TEAEs 
Safety 
Population 
14.3.2.1 
Ocular TEAEs by System Organ Class and Preferred Term 
Safety Population 
14.3.2.2 
Non-Ocular TEAEs by System Organ Class and Preferred Term 
Safety Population 
14.3.3.1 
Treatment Related Ocular TEAEs by System Organ Class and 
Preferred Term 
Safety Population 


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Table Number 
Title 
Population 
14.3.3.2 
Treatment Related Non-Ocular TEAEs by System Organ Class and 
Preferred Term 
Safety Population 
14.3.4.1 
Expected TEAEs by System Organ Class and Preferred Term 
Safety Population 
14.3.4.2 
Unexpected TEAEs by System Organ Class and Preferred Term 
Safety Population 
14.3.5.1 
Serious Ocular TEAEs by System Organ Class and Preferred Term 
Safety Population 
14.3.5.2 
Serious Non-Ocular TEAEs by System Organ Class and Preferred 
Term 
Safety Population 
14.3.6.1 
Ocular TEAEs by System Organ Class and Preferred Term and 
Maximum Severity 
Safety Population 
14.3.6.2 
Non-Ocular TEAEs by System Organ Class and Preferred Term and 
Maximum Severity 
Safety Population 
14.3.7 
Best-Corrected Distance Visual Acuity (logMAR) at 4 Meters  
Safety Population 
–Subjects Able to 
Undergo ETDRS 
Evaluation 
14.3.8.1 
Slit Lamp Biomicroscopy 
Safety Population 
14.3.8.2 
Slit Lamp Biomicroscopy Shifts from Pre- to Post-Dose 
Safety Population 
14.3.9 
Intraocular Pressure  
Safety Population 
–Subjects Who 
Are Able to 
Complete GAT 
14.3.10 
Treatment Exposure Across All Visits 
Safety Population 
24. Listings 
Listing Number 
Title 
Population 
16.1.7 
Randomization Schedule 
Randomized 
Subjects 
16.2.1 
Subject Disposition 
Enrolled Subjects 
16.2.2 
Protocol Deviations 
Randomized 
Subjects 
16.2.3.1 
Inclusion/Exclusion Criteria Subjects Excluded from Analysis 
Populations 
Enrolled Subjects 
Randomized 
Subjects 
16.2.3.2 
Subjects Excluded from Analysis Populations 
Enrolled Subjects 
16.2.4.1 
Demographics 
Randomized 
Subjects 
16.2.4.2 
Baseline Characteristics 
Randomized 
Subjects 
16.2.4.3 
Ocular Medical History 
Randomized 
Subjects 
16.2.4.4 
Non-Ocular Medical History 
Randomized 
Subjects 
16.2.4.5 
Prior and Concomitant Ocular Medications 
Randomized 
Subjects 
16.2.4.6 
Prior and Concomitant Non-Ocular Medications 
Randomized 
Subjects 
16.2.5 
Study Drug Administration 
Safety 


Protocol EYN-MYD-TP-32 Version C SAP, Final Version 
 
18-Jan-2019 
Confidential & Proprietary 
Page 33 of 33 
 
Listing Number 
Title 
Population 
16.2.6.1 
Pupillometry (mm) 
Randomized 
Subjects 
16.2.7.1 
All Adverse Events 
Randomized 
Subjects 
16.2.7.2 
Ocular Adverse Events 
Randomized 
Subjects 
16.2.7.3 
Non-Ocular Adverse Events 
Randomized 
Subjects 
16.2.7.4 
Adverse Events Leading to Study Discontinuation 
Randomized 
Subjects 
16.2.7.5 
Serious Adverse Events 
Randomized 
Subjects 
16.2.8.1 
Best-Corrected-Distance Visual Acuity (BCDVA) at 4 Meters 
using ETDRS Chart (logMAR) 
Randomized 
Subjects Able to 
Undergo ETDRS 
Evaluation 
16.2.8.2 
Uncorrected Distance Visual Acuity  
Randomized 
Subjects Unable to 
Perform ETDRS 
Evaluation 
16.2.8.3 
Slit Lamp Biomicroscopy – Traditional SLE 
Randomized 
Subjects 
16.2.8.4 
Slit Lamp Biomicroscopy – Portable Model 
Randomized 
Subjects 
16.2.8.5 
Intraocular Pressure (IOP) Using Goldmann Applanation 
Tonometer (GAT) 
Randomized 
Subjects Able to 
Complete GAT 
16.2.8.6 
Intraocular Pressure (IOP)  
Randomized 
Subjects Unable to 
Complete GAT 
16.2.8.7 
Pupillary Light Reflex (PLR) 
Randomized 
Subjects 
25. Figures 
Figure Number 
Title 
Population 
14.2.1.1 
Change from Baseline in Pupil Diameter by Eye, Treatment and Time 
Point 
Per Protocol 
14.2.1.2 
Change from Baseline in Pupil Diameter by Eye, Treatment and Time 
Point 
Modified Per 
Protocol 
14.2.1.1 
Pupil Diameter by Eye, Treatment and Time Point 
Per Protocol 
14.2.1.3 
Distribution of Pupil Diameters by Eye, Treatment and Time Points 
Per Protocol 
14.2.2 
Cumulative Proportion of Responders Analysis of Pupil Diameter at 35 
Minutes Post-Dose 
Per Protocol 
14.2.3 
Time to Maximal Pupil Dilation with ≥1.0mm Increase from Baseline 
Per Protocol