Conditions: Mydriasis Dilation
Keywords: Nyxol Pharmacologically Induced Mydriasis Dilation
Allocation: RANDOMIZED | Intervention model: CROSSOVER
Enrollment: 32
Healthy volunteers: yes

Brief summary:
The objectives of this study are:

* To evaluate the efficacy of Nyxol (phentolamine mesylate ophthalmic solution 1%) to expedite the reversal of pharmacologic mydriasis
* To evaluate the safety of Nyxol
* To evaluate the effect of Lumify® to suppress conjunctival hyperemia (redness) potentially associated with administration of Nyxol

Detailed description:
Randomized, 2-arm cross-over, double-masked Phase 2b study in approximately 32 healthy subjects, evaluating safety and efficacy of Nyxol in subjects with pharmacologically induced mydriasis.

At the first visit subjects will be screened for study eligibility.

After screening, eligible subjects will be randomized 1:1 to one of the two treatment sequences:

Treatment sequence 1: Placebo (Visit 1), Nyxol (Visit 2).

Treatment sequence 2: Nyxol (Visit 1), Placebo (Visit 2).

Randomization will be stratified by mydriatic agent (2.5% phenylephrine or 1% tropicamide). Approximately one half of the randomized subjects will receive 2.5% phenylephrine and one half will receive 1% tropicamide. Subjects will receive their mydriatic agent 1 hour before treatment. Each subject will receive the same mydriatic agent throughout the study.

At each visit, pupil diameter (PD), accommodation, near and distance visual acuity (VA) and redness in each eye will be measured before (-1 hour/baseline) and 1 hour after (maximum/0 minutes) the mydriatic agent instillation in each eye (i.e., right before the study treatment is administered), and at 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours after treatment dosing.

As needed, two hours post treatment, subjects may request the administration of Lumify® in the non-study eye.

Arms:
arm 1: 1 drop in each eye, 1 hour post medically-induced mydriasis arm 2: 1 drop in each eye, 1 hour post medically-induced mydriasis

Interventions: intervention 1: Phentolamine Mesylate Ophthalmic Solution 1% intervention 2: Phentolamine Mesylate Ophthalmic Solution Vehicle (Placebo)
Intervention descriptions: intervention 1: 1% phentolamine mesylate ophthalmic solution (Nyxol), a non-selective alpha-1 and alpha-2 adrenergic antagonist intervention 2: Topical Sterile Ophthalmic Solution

Protocol document:
Prot_002.pdf: 
 
 
 
 
CONFIDENTIAL 
 
Clinical Study Phase 2 Protocol 
OPI-NYXRM-201 
MIRA-1 
 
Randomized, Cross-Over, Double-Masked, Placebo-Controlled Study of the 
Safety and Efficacy of Phentolamine Mesylate Ophthalmic Solution to Reverse 
Pharmacologically Induced Mydriasis in Normal Healthy Subjects 
 
 
Ocuphire Pharma, Inc. 
 
 
 
 
 
 
 
Version: 
02 
Original: 
June 30th, 2019 
Amendment 1: 
 
July 24, 2019 
 
 
CONFIDENTIAL 
This document contains Ocuphire Pharma, Inc. (Ocuphire) information that is confidential, a 
trade secret and/or proprietary in nature.  It is loaned to you for your confidential use on behalf 
of Ocuphire and is not to be photocopied, disclosed or transmitted to any other person or party 
who is not covered by a Confidential Disclosure Agreement with Ocuphire.  As the Principal 
Investigator you are responsible for the safekeeping and return of this document to Ocuphire 
upon request.  You will be sent updated information and/or amendments as they become 
available.




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INVESTIGATOR’S AGREEMENT 
 
OPI-NYXRM-201 
MIRA-1 
 
Randomized, Cross-Over, Double-Masked Placebo-Controlled Study of the Safety and 
Efficacy of Phentolamine Mesylate Ophthalmic Solution to Reverse Pharmacologically 
Induced Mydriasis in Normal Healthy Subjects 
 
Version: 
 
 
02 
 
 
Original: 
 
 
June 30, 2019 
Amendment 1: 
 
July 24, 2019 
 
Investigator Agreement: 
I, the undersigned, have reviewed this protocol and I agree to conduct this protocol in accordance 
with Good Clinical Practice, the ethical principles set forth in the Declaration of Helsinki and 
with the U.S. Code of Federal Regulations governing the protection of human subjects 
(21 CFR 50), Institutional Review Boards (21 CFR 56) and the obligations of clinical 
investigators (21 CFR 312). 
 
Signature: ____________________________ Date: _________________________ 
 
Printed Name: _________________________ 
 
 
 




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ABBREVIATIONS AND TERMS 
 
 
Abbreviation 
Full term 
AE 
Adverse Event 
ANCOVA 
Analysis of Covariance 
ARP 
All Randomized Population 
BCDVA 
Best Corrected Distance Visual Acuity  
BP 
Blood Pressure 
°C 
Degree Centigrade 
CFR 
Code of Federal Regulations 
CHF 
Congestive Heart Failure 
CI 
Confidence Interval 
CRF 
Case Report Form 
CRO 
Clinical Research Organization 
CSR 
Clinical Study Report 
°F 
Degree Fahrenheit 
DB 
Database 
DCNVA 
Distance Corrected Near Visual Acuity 
FAS 
Full Analysis Set 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practice 
HR 
Heart Rate 
IB 
Investigators’ Brochure 
ICH 
International Council for Harmonisation 
IND 
Investigational New Drug Application 
IOP 
Intra-Ocular Pressure 
IRB 
Institutional Review Board 


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ITT 
Intent-To-Treat 
IUD 
Intra-Uterine Device 
LSM 
Least Squares Mean 
max 
Maximum 
MedDRA 
Medical Dictionary for Regulatory Activities 
Nyxol 
Phentolamine Mesylate Ophthalmic Solution 1% (Nyxol®) 
OR 
Odds Ratio 
OTC 
Over The Counter 
OU 
Oculus Uterque (both eyes) 
PD 
Pupil Diameter 
SAE 
Serious Adverse Event 
SE 
Standard Error 
SOC 
System Organ Class 
SP 
Safety Population 
TEAE 
Treatment Emergent Adverse Event 
US 
United States 
VA 
Visual Acuity 
 
 
 
 
 


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1. STUDY SUMMARY 
Study Number 
OPI-NYXRM-201 
Clinical Phase 
Phase 2b 
Type of Study 
Randomized, cross-over, double-masked, placebo-controlled study 
of the safety and efficacy of phentolamine mesylate ophthalmic 
solution to reverse pharmacologically induced mydriasis in normal 
healthy subjects 
Name of Investigational 
Product 
Nyxol® Eye Drops - 1% Phentolamine Mesylate ophthalmic 
solution (Nyxol) 
Duration of Study  
Up to 10 days, including screening and treatment 
Rationale 
Phentolamine mesylate is a non-selective alpha-1 and alpha-2 
adrenergic antagonist that acts on the adrenergic receptors and 
inhibits contraction of the smooth muscle. Phentolamine 
mesylate inhibits contraction of the iris dilator muscle, 
resulting in a smaller pupil size. 
 
Pharmacologically induced mydriasis is achieved either by 
stimulating the iris dilator muscle with the use of alpha-1 agonists 
(e.g. phenylephrine), or by blocking the iris sphincter muscle with 
the use of muscarinic antagonists (e.g., tropicamide). 
 
Nyxol, either by directly antagonizing the alpha-1 agonist or by 
indirectly antagonizing the pupil dilation effect of muscarinic 
blocking, can expedite the reversal of mydriasis prior to natural 
reversal. 
Study Objectives 
The objectives of this study are: 
• To evaluate the efficacy of Nyxol to expedite the reversal 
of pharmacologic mydriasis 
• To evaluate the safety of Nyxol 
• To evaluate the effect of Lumify® to suppress conjunctival 
hyperemia (redness) potentially associated with 
administration of Nyxol 
Design 
Placebo-controlled, randomized, double-masked, 2-arm cross-over, 
Phase 2b study in 32 randomized subjects, evaluating safety and 
efficacy of Nyxol in subjects with pharmacologically induced 
mydriasis. 


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TREATMENTS WILL BE ADMINISTERED TO BOTH EYES (OU). 
Following the successful completion of screening each subject will 
be randomized 1:1 to one of two treatment sequences.  
 
Treatment sequence 1: Placebo (Visit 1): Nyxol (Visit 2) 
Treatment sequence 2: Nyxol (Visit 1): Placebo (Visit 2) 
 
Randomization will be stratified by mydriatic agent 
(phenylephrine 2.5% or tropicamide 1%), with randomization into 
each stratum capped at 16 subjects; that is, approximately half of 
the randomized subjects will receive phenylephrine 1 hour before 
treatment and approximately half will receive tropicamide 1 hour 
before treatment. Each subject will receive the same mydriatic 
agent throughout the study. 
Photopic lighting conditions will remain the same  during the two 
treatment visits. 
Two hours post treatment, subjects may request the administration 
of Lumify® (to be provided) in the non-study eye as needed (site 
shall record such usage). 
Patient Population  
Thirty-two (32) normal healthy subjects 
Inclusion Criteria 
1. Males or females ≥ 18 and ≤ 45 years of age with brown irides 
only (See Appendix 2). 
2. Otherwise healthy and well controlled subjects. 
3. Ability to comply with all protocol-mandated procedures and 
to attend all scheduled office visits. 
4. Willing to give written informed consent to participate in this 
study. 
Exclusion Criteria 
Ophthalmic (in either eye): 
1. Clinically significant ocular disease as deemed by the 
Investigator (e.g., cataract, glaucoma, corneal edema, 
uveitis, severe keratoconjunctivitis sicca) that might 
interfere with the study. 
2. Unwilling or unable to discontinue use of contact lenses 
during treatment visits. 
3. Ocular trauma, ocular surgery or non-refractive laser 
treatment within the 6 months prior to screening. 




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childbearing potential unless she is 1 year postmenopausal 
or 3 months post-surgical sterilization. All females of 
childbearing potential must have a negative urine 
pregnancy test result at Visit 1/Screening and Visit 2 
examinations and must intend to not become pregnant 
during the study. 
6. Resting HR outside the normal range (50-110 beats per 
minute) at the Screening Visit. HR may be repeated only 
once if outside the normal range following at least a 5-
minute rest period in the sitting position. 
7. Hypertension with resting diastolic BP > 105 mmHg or 
systolic BP > 160 mmHg at the Screening Visit. BP may 
be repeated only once if outside the specified range 
following at least a 5-minute rest period in the sitting 
position. 
Visit Schedule 
At the first visit subjects will be screened for study eligibility.  
After screening, eligible subjects will be randomized 1:1 to one of 
the two treatment sequences. The first treatment sequence will 
entail subjects receiving placebo on the first treatment day (Visit 
1/Day 1) and receiving active drug on the second treatment day 
(Visit 2/Day 8+2 days).  The second treatment sequence will entail 
subjects receiving active drug on the first treatment day (Visit 
1/Day 1) and receiving placebo on the second treatment day (Visit 
2/Day 8+2 days).   
At Visit 1, subjects completing screening will receive one of two 
approved mydriatic agents, 1% tropicamide or 2.5% 
phenylephrine, approximately 1 hour prior to receiving study 
treatment.  Randomization will be stratified on mydriatic agent, 
with randomization into each stratum capped at approximately 16 
subjects (one half of the randomized subjects will receive 
phenylephrine, one half will receive tropicamide). 
Each subject will receive the same mydriatic agent throughout the 
study. Each subject will receive 1 drop of mydriatic agent in each 
eye. If the drop is missed, the Investigator should give the drop 
again. 
The study eye is defined as the eye with the largest pupil diameter 
at maximum (1 hour after instillation of the mydriatic agent) at 
Visit 1.  If both eyes have the same pupil diameter at maximum, 
then the study eye will be the right eye.  This is the study eye for 
both Visit 1 and Visit 2 assessments.  
 
ALL TREATMENTS WILL BE ADMINISTERED TO BOTH 
EYES (OU). 








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Duration of Subject 
Participation 
and Study 
The total length of subject participation is approximately 10 days 
with 2 clinic visits, as summarized below: 
• Screening/Treatment-study Visit 1 Day 1 (1 week) 
• Treatment-study Visit 2 Day 8+2 days (1 day) 
The execution of the entire study (first subject screen through last 
randomized subject completed) is expected to be approximately 1 
to 2 months. 


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TABLE OF CONTENTS 
SPONSOR SIGNATURE & CONTACTS ........................................................................................................... 2 
INVESTIGATOR’S AGREEMENT ..................................................................................................................... 3 
ABBREVIATIONS AND TERMS ........................................................................................................................ 5 
1. 
STUDY SUMMARY ..................................................................................................................................... 7 
2. 
INTRODUCTION ....................................................................................................................................... 17 
2.1. 
Investigational products .................................................................................................................... 17 
2.2. 
Findings from nonclinical and clinical studies ................................................................................ 17 
2.3. 
Route of administration, dosage regimen, and treatment period .................................................. 19 
2.4. 
Compliance ......................................................................................................................................... 19 
2.5. 
Study Population ................................................................................................................................ 19 
3. 
OBJECTIVES AND PURPOSE ................................................................................................................ 20 
4. 
STUDY DESIGN ......................................................................................................................................... 21 
4.1. 
Primary and secondary endpoints .................................................................................................... 21 
4.2. 
Description and schedule of visits and procedures ......................................................................... 22 
4.3. 
Measures taken to minimize/avoid bias ........................................................................................... 24 
4.4. 
Study medications .............................................................................................................................. 24 
4.4.1. Packaging and labeling ........................................................................................................................ 24 
4.4.2. Storage of study medication ................................................................................................................ 24 
4.4.3. Study medication accountability ......................................................................................................... 24 
4.4.3.1. Receipt and disposition of study medication ............................................................................... 24 
4.4.3.2. Return of study medication .......................................................................................................... 24 
4.5. 
Expected duration of subject participation ..................................................................................... 24 
4.6. 
Randomization and procedure for breaking the code .................................................................... 25 
4.7. 
Collection of data ............................................................................................................................... 25 
4.8. 
Completed subject .............................................................................................................................. 25 
4.9. 
Non-completing subject ..................................................................................................................... 25 
4.9.1. Study medication discontinuation ....................................................................................................... 25 
4.9.2. Reasons for Withdrawal from Study ................................................................................................... 26 
4.9.3. Entire study terminated ........................................................................................................................ 26 
4.9.4. Actions after discontinuation ............................................................................................................... 26 
4.10. 
Completed study ................................................................................................................................ 26 
4.11. 
Procedure after the completion of the study ................................................................................... 26 
5. 
SUBJECT INCLUSION AND EXCLUSION CRITERIA ...................................................................... 27 
5.1 
Subject inclusion criteria ................................................................................................................... 27 
5.2 
Subject exclusion criteria .................................................................................................................. 27 
6. 
TREATMENT OF SUBJECTS .................................................................................................................. 29 
6.1. 
Treatment adherence ......................................................................................................................... 29 
6.2. 
Concomitant medications .................................................................................................................. 29 
7. 
ASSESSMENT OF EFFICACY ................................................................................................................ 31 
7.1. 
Specification of the efficacy parameters .......................................................................................... 31 
7.2. 
Assessing, recording, and analyzing of efficacy parameters .......................................................... 31 
7.2.1. Screening/Day 1 .................................................................................................................................. 32 
7.2.2. Treatment Visit 1/Day 1 ...................................................................................................................... 32 
7.2.3. Treatment Visit 2/Day 8 + 2 Days ....................................................................................................... 33 
7.2.4. Unscheduled Visits .............................................................................................................................. 34 
7.2.5. Visit variation ...................................................................................................................................... 34 
8. 
ASSESSMENT OF SAFETY ..................................................................................................................... 35 
8.1. 
Specification of safety parameters .................................................................................................... 35 
8.2. 
Assessing, recording, and analyzing safety parameters ................................................................. 35 
8.3. 
Adverse events and Serious Adverse Events ................................................................................... 35 
8.3.1. Adverse event definitions .................................................................................................................... 36 


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8.3.2. Follow-up of subjects after adverse events ......................................................................................... 39 
9. 
STATISTICS ............................................................................................................................................... 40 
9.1. 
Sample Size ......................................................................................................................................... 40 
9.2. 
Analysis Populations .......................................................................................................................... 40 
9.3. 
Statistical Methods ............................................................................................................................. 40 
9.3.1. General Considerations ....................................................................................................................... 40 
9.3.2. Demographic and Baseline Characteristics ......................................................................................... 40 
9.3.3. Subject Disposition .............................................................................................................................. 40 
9.3.4. Medical History and Prior/Concomitant Medications ......................................................................... 41 
9.3.5. Analysis of Efficacy ............................................................................................................................ 41 
9.3.6. Analysis of Safety ................................................................................................................................ 42 
9.4. 
Procedure for accounting for missing, unused, or spurious data .................................................. 42 
9.5. 
Procedure for reporting deviations from the statistical plan ........................................................ 43 
10. 
DIRECT ACCESS TO SOURCE DATA AND DOCUMENTS ............................................................. 44 
11. 
QUALITY CONTROL AND QUALITY ASSURANCE ........................................................................ 45 
12. 
ETHICAL CONSIDERATIONS AND GCP COMPLIANCE ............................................................... 46 
12.1. 
GCP compliance ................................................................................................................................. 46 
12.2. 
Institutional Review Board (IRB) .................................................................................................... 46 
12.3. 
Protocol Deviations/Violations .......................................................................................................... 46 
12.4. 
Informed consent requirements ....................................................................................................... 46 
13. 
DATA HANDLING AND RECORD KEEPING ..................................................................................... 48 
13.1. 
Data Entry .......................................................................................................................................... 48 
13.2. 
Data quality control and reporting .................................................................................................. 48 
13.3. 
Archiving of data ............................................................................................................................... 48 
13.4. 
Records retention ............................................................................................................................... 48 
13.5. 
Amendments to the protocol ............................................................................................................. 48 
APPENDIX 1: ADRENERGIC AND CHOLINERGIC DRUGS .................................................................... 49 
APPENDIX 2: IRIS COLOR CHART ............................................................................................................... 50 
APPENDIX 3: ORIGINAL SERIES SLOAN LETTER ETDRS CARD ........................................................ 51 
 
 








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3. OBJECTIVES AND PURPOSE 
The objectives of this study are: 
• To evaluate the efficacy of Nyxol to expedite the reversal of pharmacologic mydriasis 
• To evaluate the safety of Nyxol 
• To evaluate the effect of Lumify® to suppress conjunctival hyperemia (redness) potentially 
associated with administration of Nyxol 
 






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Table 1: 
Screening and Mydriatic/Treatment Schedule 




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• Treatment Visit 1 Day 1 (1 day; same day/same clinic visit as Screening Day) 
• Treatment Visit 2 Day 8+2 Days (1 day; 7+2 days after Treatment Visit 1) 
The execution of the entire study (first subject screened through last randomized subject 
completed) is expected to be approximately 1 to 2 months. 
4.6. Randomization and procedure for breaking the code 
A randomization code for allocating subjects to treatment sequence will be prepared by a 
masked biostatistician not connected with the study. Randomization will be stratified by 
mydriatic agent. 
At the initiation of study related procedures, every potential subject is assigned a 
Screening number in numerical order.  Once a subject is qualified for the study, the 
subject is assigned a randomization number in the order provided by the biostatistician. 
The study medications will be masked to both Investigator and study subjects, as well as 
Ocuphire.  Assignment to treatment sequence will be masked to the Investigator, 
Ocuphire, and the subjects.  Only in case of medical emergency or occurrence of SAEs 
will the randomization code be unmasked by the study pharmacist and made available to 
the Investigator, Ocuphire, and/or other personnel involved in the monitoring or conduct of 
this study. 
4.7. Collection of data 
The source documentation for all data collected in the study will maintained by the 
Investigator in the subject files at the study site.  All original data collected during this trial 
is to be recorded on paper case report forms (CRFs) and then electronically entered into 
the database following study completion. The paper CRF is considered to be the source 
documentation for this study. 
4.8. Completed subject 
A completed subject is defined as one who completes all planned dosing and procedures 
through the end of Visit 2. 
4.9. Non-completing subject 
A non-completing subject is defined as one who exits the study by their own volition or at 
the discretion of the Investigator and/or the Medical Monitor prior to completing all of the 
study procedures required in this protocol.  Any subject may decide to voluntarily 
withdraw from the study at any time without prejudice.   
4.9.1. Study medication discontinuation 
The study medication may be discontinued for the following reasons: 
• Adverse Events: AEs include clinically significant laboratory abnormalities and 
intercurrent diseases reported by the subject or observed by the Investigator with 
documentation on the CRF 
• Death: If a subject dies, the AE that caused the death should be documented on the 
CRF and be noted as serious and fatal. 
• Disallowed concurrent medication: Any medication not allowed by the protocol 
would be a protocol violation. 


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• Lack of efficacy: A subject may elect to discontinue participation in the study for a 
perceived lack of efficacy. 
• Investigator decision: A subject may be discontinued for reasons other than those 
bulleted previously if the Investigator thinks it is not in the best interest of the 
subject to continue. 
• Other: If there is any other reason for subject discontinuation this should be noted 
on the CRF. 
The reason for premature study medication discontinuation should be entered onto the 
appropriate CRF.   
4.9.2. Reasons for Withdrawal from Study 
• Subject withdraws consent. 
• Subject is lost to follow-up. 
• Subject withdraws for other reason. 
4.9.3. Entire study terminated 
The entire study may be terminated by Investigators or Ocuphire.  Prompt, written notice 
of reasonable cause to the other party (Ocuphire or Investigators, respectively) is required.  
Prompt notice to the IRB and to regulatory authorities is also required. 
4.9.4. Actions after discontinuation 
All subjects who discontinue study medication due to a report of an AE must be followed-
up and provided appropriate medical care until their signs and symptoms have remitted or 
stabilized or until abnormal laboratory findings have returned to acceptable or pre-study 
limits. 
For any subject who chooses to withdraw consent or who is non-compliant, every possible 
effort should be made by the Investigator to assure there is a final telephone call that 
includes assessments for AEs, concomitant medications and conjunctival redness.   
4.10. Completed study 
The study is completed when all randomized subjects have completed the study, all CRFs 
have been completed, and all CRF data entered into the database.  Final DB lock will 
occur after the last randomized subject completes, all data has been entered and all queries 
resolved.   
4.11. Procedure after the completion of the study 
When the study is completed, the CRO (Oculos) will provide Ocuphire and the 
Investigator with a brief (i.e., one to three pages) report, containing a description of the 
study, the number of subjects enrolled, the number of subjects completed the number of 
subjects who dropped out and why, efficacy findings and AEs. 
 




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2. Clinically significant systemic disease (e.g., uncontrolled diabetes, myasthenia 
gravis, cancer, hepatic, renal, endocrine or cardiovascular disorders) that might 
interfere with the study 
3. Initiation of treatment with or any changes to the current dosage, drug or regimen of 
any topical or systemic adrenergic or cholinergic drugs up to 7 days prior to 
Screening, or during the study 
4. Participation in any investigational study within 30 days prior to screening 
5. Women of childbearing potential who are pregnant, nursing, planning a pregnancy, 
or not using a medically acceptable form of birth control. Acceptable methods 
include the use of at least one of the following: intrauterine device (IUD), hormonal 
(oral, injection, patch, implant, ring), barrier with spermicide (condom, diaphragm), 
or abstinence. An adult woman is considered to be of childbearing potential unless 
she is 1 year postmenopausal or 3 months post-surgical sterilization. All females of 
childbearing potential must have a negative urine pregnancy test result at Visit 1 
Screening and Visit 2 examinations and must intend to not become pregnant during 
the study 
6. Resting HR outside the normal range (50-110 beats per minute) at the Screening 
Visit. HR may be repeated only once if outside the normal range following at least 
a 5-minute rest period in the sitting position 
7. Hypertension with resting diastolic BP > 105 mmHg or systolic BP > 160 mmHg at 
the Screening Visit. BP may be repeated only once if outside the specified range 
following at least a 5-minute rest period in the sitting position 
 
 




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inclusion of this subject’s subsequent visits in the safety and efficacy analysis will be made 
by Ocuphire. 
All medications which the subject has taken within 30 days prior to the Screening Visit 
and during the study will be recorded in the CRF.  The name of the drug, dose, route of 
administration, duration of treatment and indication will be recorded for each medication.  
For combination products (e.g., Contac®), the brand name is required.  For 
non-combination products, the generic name is desired.  The use of routine ophthalmic 
diagnostic pharmaceutical agents (e.g., fluorescein and local anesthetic) will be allowed, 
and individual documentation not required.  Any change in dosing parameters should also 
be recorded in the CRF. 
 








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7.2.4. Unscheduled Visits 
An Unscheduled Visit may be any visit to the Investigator other than the specific visits 
requested in the protocol as possibly required for the subject’s ophthalmic condition.  The 
Investigator will perform all procedures necessary to evaluate the study participant at these 
visits and record any AEs in the CRF. 
As noted in Section 4.9.4, every possible effort should be made by Investigators to assure 
that subjects who discontinue early from the study have a final telephone follow-up that 
includes assessments of AEs, concomitant medications and conjunctival redness. 
7.2.5. Visit variation 
  
 
 




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Documentation of AEs/adverse reactions includes start date and end date, severity, 
relationship to study medications, action(s) taken, seriousness and outcome.   
8.3.1. Adverse event definitions 
The following definitions of terms apply to this section: 
Adverse event. An AE is any untoward medical occurrence in a patient or clinical study 
subject administered a study medication (pharmacological/biological product) that does 
not necessarily have a causal relationship to this medication. An AE can therefore be any 
unfavorable and unintended sign (including abnormal laboratory findings), symptom, or 
disease temporarily associated with the use of the study medication, whether or not related 
to the study medication.  Study medication includes the investigational drug under 
evaluation and the comparator product or vehicle placebo that is given or administered 
during any phase of the study.  
Medical conditions/diseases present before starting the investigational treatment are only 
considered AEs if they worsen after starting the investigational treatment. Abnormal test 
results constitute AEs only if they induce clinical signs or symptoms, are considered 
clinically significant, or require therapy. 
The occurrence of AEs should be sought by open-ended questioning of the subject at each 
visit during the study. At each clinic visit, study personnel should ask the following 
question: “Have you had any problems since your last visit?”. AEs also may be detected 
when they are volunteered by the subject during or between visits or through study 
assessments. 
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 Ocuphire, its occurrence places the subject or subject at immediate risk of 
death.  It does not include an AE or suspected adverse reaction that, had it occurred in a 
more severe form, might have caused death.  
Serious adverse event or serious suspected adverse reaction.  An AE or suspected adverse 
reaction is considered ‘‘serious’’ if, in the view of either the Investigator or Ocuphire, it 
results in any of the following outcomes at any dose:  
• Death 
• Life threatening 
• Initial or prolonged hospitalization 
• Disability or permanent damage 
• Congenital anomaly or birth defect 
• Needs intervention to prevent impairment 
• Other medically important serious event 
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 or subject may require medical or surgical 
intervention to prevent one of the outcomes listed in this definition.  
Treatment on an outpatient emergency basis that does not result in hospital admission, or a 
hospitalization that is elective or is a preplanned treatment for a pre-existing condition that 


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is unrelated to the medication under study and has not worsened since the start of the 
study, is not considered an SAE. 
Suspected adverse reaction means any AE for which there is a reasonable possibility that 
the drug caused the AE.  For the purposes of IND safety reporting, ”reasonable possibility” 
means there is evidence to suggest a causal relationship between the drug and the AE.  
Suspected adverse reaction implies a lesser degree of certainty about causality than 
adverse reaction, which means any AE caused by a drug.  
Unexpected adverse event or unexpected suspected adverse reaction. An AE or suspected 
adverse reaction is considered ”unexpected” if it is not listed in the Investigators’ Brochure 
or is not listed at the specificity or severity that has been observed; or, if an Investigators’ 
Brochure is not required or available, is not consistent with the risk information described 
in the general investigational plan or elsewhere in the current application, as amended.  
“Unexpected,” as used in this definition, also refers to AEs or suspected adverse reactions 
that are mentioned in the Investigators’ Brochure as occurring with a class of drugs or as 
anticipated from the pharmacological properties of the drug but are not specifically 
mentioned as occurring with the particular drug under investigation. 
The study medication relationship for each AE/adverse reaction should be determined by 
the Investigator using these explanations: 
• Not related 
• Unlikely related 
• Possibly related 
• Probably related 
• Definitely related 
• Unknown 
Unless the relationship is considered to be “Not related” or “Unlikely related” and there is 
any valid reason, even if undetermined, for suspecting a possible cause-and-effect 
relationship between the study medication and the occurrence of the AE, then the AE 
should be considered “related”.  
If the relationship between the AE/SAE and the investigational product is determined by 
Ocuphire to be anything other than “Not related” or “Unlikely related” the event will be 
considered to be related to the investigational product for the purposes of expedited 
regulatory reporting. 
Severity of an AE is defined as a qualitative assessment of the level of discomfort of an 
AE as is determined by the Investigator or reported to him/her by the subject.  The 
assessment of severity is made irrespective of study medication relationship or seriousness 
of the event and should be evaluated according to the following scale: 
• 1 = Mild: present, but not distressing, and no disruption of normal daily activity 
• 2 = Moderate: discomfort sufficient to reduce or affect normal daily activity 
• 3 = Severe: incapacitating, with inability to work or perform normal daily activity 
A change in severity for a reported AE will require an end date for the previous severity 
and a new start and end date for the new severity.  For example, a change in severity may 
go from mild to severe or from severe to moderate.  In either case the start or end dates 
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Should a pregnancy occur, it must be reported and recorded on the pregnancy form. 
Pregnancy in itself is not regarded as an AE unless there is a suspicion that an 
investigational product may have interfered with the effectiveness of a contraceptive 
medication. The subject will be withdrawn from study medication and followed through 
conclusion of pregnancy. The outcome of all pregnancies (spontaneous miscarriage, 
elective termination, normal birth or congenital abnormality) must be followed up and 
documented. 
Expedited reporting of Serious and Unexpected Adverse Events: All SAEs (related and 
unrelated) will be recorded following subject signature of the informed consent and until 
the final study visit (Visit 3). Any SAEs “suspected” to be related to the study medication 
and discovered by the Investigator at any time after the study should be reported. 
Any SAE that occurs must be reported to Oculos within 24 hours of its occurrence or within 
24 hours of learning of its occurrence.  Recurrent episodes, complications, or progression of 
the initial SAE must be reported to Oculos as follow-up to the original episode within 24 
hours of the Investigator receiving the information.  Information about all SAEs will be 
collected and recorded on the SAE form.  All pertinent medical records and information 
collected during the treatment and follow-up of the subject should be maintained at the site 
with a copy emailed 
  The Investigator must assess the 
SAE relationship and complete the SAE form.  Oculos may request additional information. 
Follow-up information (e.g., discharge summary) will be retained in the subject’s chart and 
a copy will be emailed to
.  In addition, all SAEs should be 
recorded on the AE CRF page with the serious question marked “Yes”. 
It is the Investigator’s responsibility to notify the approving IRB of any SAEs on a timely 
basis as instructed by Ocuphire following Ocuphire’s determination of causality.  All 
subjects who experience an SAE should be followed clinically and undergo the appropriate 
diagnostic evaluations until stabilization or resolution of the event.  Ocuphire will report 
all SAEs to the US Food and Drug Administration (FDA) on the appropriate schedule 
depending if the event is drug related or not drug related, expected, unexpected (based on 
the available information in the Investigators’ Brochure). 
Any death occurring during the study and follow-up period must be reported as an SAE. 
For any death occurring through the end of the study, regardless of the degree of 
relationship to study medication, the SAE resulting in the death must be reported to 
Oculos.  A death occurring after completion of the study including the Safety Follow-up 
Visits, that is not reasonably associated with study medication administration, does not 
require completion of the SAE form. 
8.3.2. Follow-up of subjects after adverse events 
If an AE/adverse reaction occurs, the Investigator will institute support and/or treatment as 
deemed appropriate.  All SAEs ongoing at the time of the last visit, or discontinuation 
from the study, will be followed up until the AE/adverse reaction is resolved or stabilized, 
the subject is lost to follow-up, or there is other resolution to the event.  
 
 








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9.5. Procedure for reporting deviations from the statistical plan 
Any deviations from the statistical plan will be described and a justification given in the 
final Clinical Study Report (CSR).   
 
 


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10. DIRECT ACCESS TO SOURCE DATA AND DOCUMENTS 
The Investigator will permit study-related monitoring visits, audits, IRB review, and 
regulatory inspection(s) by providing direct access to source data and documents. 


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11.  QUALITY CONTROL AND QUALITY ASSURANCE 
The progress of the study will be monitored by on-site, written, and telephone 
communications between personnel at the Investigator’s site and the Medical Monitor.  
The Investigator will allow Ocuphire, the Study Monitor, and the Medical Monitor to 
inspect all CRFs, subject records (source documents), signed consent forms, records of 
study medication receipt, storage, preparation, and disposition, and regulatory files related 
to this study. 
 


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12.  ETHICAL CONSIDERATIONS AND GCP COMPLIANCE  
12.1. GCP compliance 
The proposed study is subject to all applicable governmental rules and regulations 
concerning the conduct of clinical trials on human subjects.  This includes, but is not 
necessarily limited to, the approval of Institutional Review Boards, the Helsinki 
Declaration, U.S. FDA Law, ICH GCP guidelines, obtaining prospective informed 
consent, monitoring of the conduct of the study and the completeness of the CRFs by 
Ocuphire or its designee(s), and appropriate record retention by the Investigator.   
12.2. Institutional Review Board (IRB) 
This protocol, materials used to recruit subjects, and materials used to document consent 
must be approved by the IRB prior to initiation of the study.  Written IRB approval must 
adequately identify the protocol and informed consent.  In addition to approving the 
protocol, the IRB must also approve the Subject Information and Consent Form, as well as 
any advertising tools that will be used for the study.  Written approval also must indicate 
whether approval was granted based on full committee review or expedited review.  
Copies of all approved materials, all correspondence with the IRB and written approval 
from the IRB must be made available to Ocuphire, prior to the start of subject enrollment 
into the study. 
12.3. Protocol Deviations/Violations 
If the Investigator desires to modify the procedures and/or design of the study, he or she 
must contact and obtain the consent and approval of Ocuphire and the IRB, regarding the 
proposed changes, prior to their implementation.  Changes implemented after obtaining 
this approval will be considered Protocol Deviations.  Changes implemented without prior 
approval will be considered Protocol Violations. 
12.4.  Informed consent requirements 
Written informed consent will be obtained from each subject.  A copy of the signed and 
dated consent document will be given to each subject.  The original signed and dated 
informed consent document must be maintained in the study files at the Investigator’s site. 
The Investigator is responsible for ensuring that no subject is subject to any study-related 
examination or activity before that subject has given informed consent.  The subject must 
give written consent after the receipt of detailed information.  The verbal explanation will 
cover all the elements specified in the written information provided for the subject. 
It should be emphasized that the subject is at liberty to withdraw consent to participate at 
any time, without penalty or loss of benefits to which the subject is otherwise entitled. 
Subjects who refuse to give, or withdraw, written informed consent may not be included or 
continued in this study, but this will not impact on their subsequent care.  
The Investigator will inform the subject of the aims, methods, anticipated benefits and 
potential hazards of the study, including any discomfort it may entail.  The subject must be 
given every opportunity to clarify any points he/she does not understand and if necessary, 
ask for more information.  At the end of the interview, the subject may be given time to 
reflect if this is required, or if the subject requests more time.  Subjects and/or legal 
guardian will be required to sign and date the informed consent form.  


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A copy of the signed and dated consent document will be given to each subject.  The 
original signed and dated informed consent document must be maintained in the study files 
at the Investigator’s site.  Signed informed consent must be obtained prior to the 
conductance of any study procedures. 
 


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13.  DATA HANDLING AND RECORD KEEPING 
All procedures for the handling and analysis of data will be conducted using good 
computing practices meeting ICH and U.S. FDA guidelines for the handling and analysis 
of data for clinical trials. 
13.1.  Data Entry 
All study data will be entered into a paper CRF. CRFs are considered to be source 
documents for this study. Data will be entered from the CRF into the study database. 
13.2. Data quality control and reporting 
After data have been entered into the study database, a system of computerized data 
validation checks will be implemented and applied to the database.  Query reports 
pertaining to data omissions and discrepancies will be forwarded to the Clinical 
Investigator and Ocuphire for resolution.  The study database will be updated in 
accordance with the resolved query reports.  All changes to the study database will be 
documented. 
13.3.  Archiving of data 
Archived versions of the database will be saved by Ocuphire consistent with ICH Good 
Clinical Practices (GCP) Guidelines, complying with whichever of the requirements is 
longer. Ocuphire will notify the Investigator when documents should be returned. 
13.4.  Records retention 
The Investigator’s site and clinical laboratory will retain all records related to the study in 
compliance with ICH GCP Guidelines. 
13.5.  Amendments to the protocol 
Modifications of the signed protocol are only possible by approved protocol amendments 
and with the agreement of all responsible persons.  The procedure for approval of a 
protocol amendment is identical to that for approval of the protocol.  The 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 by the committee.  This 
should be fully documented. 
The Investigator must not implement any deviation from or change to the protocol, without 
discussion with, and agreement by Ocuphire and prior review and documented 
approval/favorable opinion of the amendment from the relevant ethics committee, except 
where it is necessary to eliminate an immediate hazard to study subjects, or where the 
change(s) involves only logistical or administrative aspects of the study [e.g., change in 
monitor(s), change of telephone number(s)]. 
Protocol amendments will be submitted to the appropriate authority(ies) as required by the 
applicable regulatory requirement(s). 
 
 
 


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APPENDIX 1: ADRENERGIC AND CHOLINERGIC DRUGS 


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APPENDIX 2: IRIS COLOR CHART 
 


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APPENDIX 3: ORIGINAL SERIES 
 CARD 



SAP_003.pdf: 




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1. 
TABLE OF CONTENTS 
1. 
TABLE OF CONTENTS ............................................................................................................................... 2 
2. 
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS .................................................................. 5 
3. 
INTRODUCTION ......................................................................................................................................... 7 
3.1. 
Preface .............................................................................................................................................. 7 
3.2. 
Purpose of Analyses .......................................................................................................................... 7 
3.3. 
Summary of Statistical Analysis Changes to the Protocol ................................................................ 7 
4. 
STUDY OBJECTIVES AND ENDPOINTS ................................................................................................. 8 
4.1. 
Study Objectives ............................................................................................................................... 8 
4.2. 
Study Endpoints ................................................................................................................................ 8 
4.2.1. 
Primary Endpoints ........................................................................................................... 8 
4.2.2. 
Secondary Endpoints ....................................................................................................... 8 
5. 
STUDY METHODS .................................................................................................................................... 10 
5.1. 
General Study Design and Plan ....................................................................................................... 10 
5.2. 
Inclusion – Exclusion Criteria and General Study Population ........................................................ 13 
5.3. 
Randomization and Blinding ........................................................................................................... 13 
5.4. 
Analysis Variables .......................................................................................................................... 13 
6. 
SAMPLE SIZE ............................................................................................................................................ 14 
7. 
GENERAL CONSIDERATIONS ............................................................................................................... 15 
7.1. 
Analysis Populations ....................................................................................................................... 15 
7.1.1. 
Full Analysis Set Population (FAS) .............................................................................. 15 
7.1.2. 
Per Protocol Population (PP) ......................................................................................... 15 
7.1.3. 
All Randomized Population (ARP) ............................................................................... 15 
7.1.4. 
Safety Population (SP) .................................................................................................. 15 
7.2. 
Covariates and Subgroups ............................................................................................................... 15 
7.2.1. 
Planned Covariates ........................................................................................................ 15 
7.2.2. 
Planned Subgroups ........................................................................................................ 15 
7.3. 
Management of Analysis Data ........................................................................................................ 16 
7.3.1. 
Data Handling ............................................................................................................... 16 
7.3.2. 
Missing Data ................................................................................................................. 16 
7.3.2.1. 
Handling of Missing Date Values ............................................................ 16 
7.3.2.2. 
Missing Baseline Data ............................................................................. 17 
7.3.2.3. 
Imputation Methods ................................................................................. 17 
7.3.3. 
Handling of Early Termination Visit Information ......................................................... 18 
7.3.4. 
Pooling of Investigational Sites ..................................................................................... 18 
7.3.5. 
Coding Conventions for Events and Medications ......................................................... 19 
7.3.6. 
Analysis Software ......................................................................................................... 19 
7.3.7. 
Study Data ..................................................................................................................... 19 
7.4. 
Planned Study Analyses .................................................................................................................. 20 
7.4.1. 
Statistical Summaries: Descriptive and Inferential ....................................................... 20 
7.4.2. 
Interim Analyses and Data Monitoring ......................................................................... 21 
7.4.3. 
Final Analysis and Publication of Study Results .......................................................... 21 
7.5. 
Multiple Testing Procedures ........................................................................................................... 21 
7.6. 
Baseline Values ............................................................................................................................... 21 




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LIST OF TABLES 
 
Table 1 
Screening and Mydriatic/Treatment Schedule .............................................................. 11 
 
 
LIST OF FIGURES 
 
Figure 1 
SDTM, ADaM, and TFL Development and Validation ................................................ 20 
 
 
 






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3. 
INTRODUCTION 
3.1. 
Preface 
This document presents a statistical analysis plan (SAP) for Ocuphire Pharma, Inc. Protocol 
OPI-NYXRM-201 (MIRA-1) (Randomized, cross-over, double-masked, placebo-controlled 
study of the safety and efficacy of phentolamine mesylate ophthalmic solution to reverse 
pharmacologically induced mydriasis in normal healthy subjects). 
Reference materials for this statistical plan include the protocol OPI-NYXRM-201 
Amendment 1 (24JUL2019) and Case Report Forms (CRFs; Final Version 18JUL2019).  
The SAP described hereafter is an a priori plan.  The SAP will be finalized and approved 
prior to unmasking of any study data.     
For the reasons stated here, the conduct of the study in the field is considered to be 
independent of any study outcome that might materialize upon enactment of the currently 
proposed statistical plan.  
3.2. 
Purpose of Analyses 
The purposes of the planned analyses described in this SAP are to evaluate the efficacy of 
Nyxol to expedite the reversal of pharmacologic mydriasis, to evaluate the safety of Nyxol, 
and to evaluate the effect of Lumify® to suppress conjunctival hyperemia (redness) 
potentially associated with administration of Nyxol. Results from the analyses completed will 
be included in the final clinical study report (CSR) for OPI-NYXRM-201, and may also be 
utilized for regulatory submissions, manuscripts, or other clinical development activities. 
Post-hoc exploratory analyses not identified in this SAP may be performed to further 
examine the study data.  These analyses will be clearly identified, where appropriate, in the 
final CSR. Additional analyses not prospectively identified in this SAP may also be 
completed for publications, or regulatory or funding inquiries. These analyses, if performed, 
may not be reported in the CSR, but will be fully detailed in the document containing the 
additional analyses. 
3.3. 
Summary of Statistical Analysis Changes to the Protocol 
The analyses described in this analysis plan are consistent with the analyses described in the 
study protocol. 


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4. 
STUDY OBJECTIVES AND ENDPOINTS 
Study objectives and endpoints defined in the protocol include safety and efficacy endpoints. 
Objectives and pre-specified endpoints are as follows: 
4.1. 
Study Objectives 
The objectives of this study are as follows: 
 To evaluate the efficacy of Nyxol to expedite the reversal of pharmacologic 
mydriasis. 
 To evaluate the safety of Nyxol. 
 To evaluate the effect of Lumify® to suppress conjunctival hyperemia (redness) 
potentially associated with administration of Nyxol  
4.2. 
Study Endpoints 
4.2.1. 
Primary Endpoints 
The primary efficacy endpoint from the protocol is the change (in mm) in pharmacologically 
induced mydriatic (max) pupil diameter (0 minutes) at 2 hours post treatment in the study 
eye. 
The primary safety measures are: 
 
 
 
 
 Adverse events (AEs) 
4.2.2. 
Secondary Endpoints 
Secondary endpoints for efficacy and safety assessments include the following: 
Efficacy: 
Secondary efficacy endpoints will be analyzed by study eye, non-study eye and both eyes 
(the pooled data from the study eye and non-study eye) unless otherwise indicated, and will 
include: 
 Change (in mm) from max pupil diameter (0 minutes) at each remaining time point 
(30 min, 1 hour, 4 hours, 6 hours). 






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Table 1 
Screening and Mydriatic/Treatment Schedule 


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7.3. 
Management of Analysis Data 
7.3.1. 
Data Handling 
Data from unscheduled visits will not be included in the analysis of efficacy or safety, but 
will be listed.   
7.3.2. 
Missing Data 
The primary efficacy endpoint is the change in pharmacologically induced mydriatic (max) 
pupil diameter (0 minutes) at 2 hours post treatment in the study eye. For the analysis of the 
primary efficacy endpoint, observed case data will be used (no imputation will be performed 
for missing efficacy data) for the analysis using the FAS.  However, confirmatory analyses 
will be performed using the ARP, with imputation performed for missing data as specified in 
Section 7.3.2.3.   
Otherwise there will be no substitutions made to accommodate missing data points for 
efficacy data. All data recorded on the CRF will be included in data listings that will 
accompany the CSR. 
Safety data will be imputed in limited situations. If the severity of an AE is missing, then the 
severity will remain missing.  If relationship of the AE to study drug is missing, the 
relationship will remain missing.  Missing or partial dates for AEs or concomitant 
medications will be imputed as described in Section 7.3.2.1. Otherwise, all summaries of 
safety endpoints will be completed using observed cases in the SP; no imputation will be 
completed. 
7.3.2.1. 
Handling of Missing Date Values 
Partial or Missing Dates 
The following conventions will be used to impute missing portions of dates for adverse 
events and concomitant medications, if warranted.  Note that the imputed values outlined 
here may not always provide the most conservative date.  In those circumstances, the 
imputed value may be replaced by a date that will lead to a more conservative analysis. 
A. 
Start Dates 
1) 
If the year is unknown, then the date will not be imputed and 
will be assigned a missing value.  
2) 
If the month is unknown, then: 
i) If the year matches the first dose date year, then impute 
the month and day of the first dose date. 
ii) Otherwise, assign ‘January.’  
3) 
 If the day is unknown, then: 


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i) If the month and year match the first dose date month 
and year, then impute the day of the first dose date. 
ii) Otherwise, assign the first day of the month. 
B. 
Stop Dates 
1) 
If the year is unknown, then the date will not be imputed and 
will be assigned a missing value. 
2) 
If the month is unknown, then assign ‘December.’ 
3) 
If the day is unknown, then assign the last day of the month. 
 
7.3.2.2. 
Missing Baseline Data 
Every effort will be made to ensure that accurate baseline information on the subjects is 
collected. In the event that a subject is missing baseline information, the subject will be 
included in the SP for assessment of safety, and excluded from the primary analyses. Each 
case of missing baseline data will be evaluated for potential inclusion in the exploratory 
endpoints. All baseline data will be observed cases, without imputation. 
7.3.2.3. 
Imputation Methods 
Imputation for efficacy data will only be performed for the confirmatory analysis of the 
primary efficacy endpoint, using the ARP. If 5% or fewer data are missing in all treatment 
groups, an analysis with last observation carried forward (LOCF) for missing data will be 
applied within a treatment group. If more than 5% of data in any treatment group are missing, 
multiple imputation will be employed to analyze incomplete data sets under the assumption 
that the mechanism responsible for the missing data is at worst characterized as missing at 
random (MAR).    
Multiple imputation is a simulation-based approach where missing values are replaced by 
multiple Bayesian draws from the conditional distribution of missing data given the observed 
data and covariates, creating multiple completed data sets.  These completed datasets can 
then be analyzed using standard analysis methods.  Rubin (1987) presented rules for how to 
combine the multiple sets of estimates to produce overall estimates, confidence intervals, and 
tests that adequately incorporate missing data uncertainty. 
Missing values for pupil diameter will be imputed simultaneously based on an underlying 
joint normal distribution using a Markov Chain Monte Carlo (MCMC) method.   
The imputations will be done separately for each treatment group and will include the 
following variables in the imputation model: pupil diameter at Day 1 (30 min, 1 hour, 2 
hours, 4 hours, 6 hours) and Day 8 (30 min, 1 hour, 2 hours, 4 hours, 6 hours). No imputation 
will be applied to the max pupil diameter (0 minutes) time points at Day 1 and Day 8. 
The number of imputations will be set to 500.  The outcomes of interest (change from 
baseline) will be calculated from these imputed datasets.  The treatment difference for each 
imputed dataset will be evaluated using mixed models.  See Section 11 for details on these 
models.  The estimates and standard errors of the differences in LS means based on the 500 








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sequence or group.  Expansion of descriptive table categories within each treatment may 
occur if such elaborations are thought to be useful. 
All study related data collected will be presented in listings.  Study related data not subject to 
analysis according to this plan will not appear in any tables or graphs but will be included in 
the data listings. 
7.4.2. 
Interim Analyses and Data Monitoring 
No formal interim analysis or safety monitoring committee is planned for this study.  
7.4.3. 
Final Analysis and Publication of Study Results 
The final analysis will be completed after all subjects have completed the study.   
7.5. 
Multiple Testing Procedures 
There will be no adjustments for multiplicity and no formal multiple testing procedures are to 
be implemented with this analysis plan. 
7.6. 
Baseline Values 
For all efficacy endpoints except pupil diameter, Baseline for a specific day (Visit 1 / Day 1, 
Visit 2 / Day 8) is defined as -1 hour prior to treatment. That is, change from baseline is 
based on -1 hour data.  For pupil diameter, change analyses for each visit will be based on 
max time point (0 minutes), during which maximum pupil diameter is expected.  That is, 
change from max is based on 0 minute data. 
 


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8. 
SUMMARY OF STUDY DATA 
8.1. 
Subject Disposition 
A summary of the analysis sets includes the number and percentage of subjects by treatment 
sequence and overall for the following categories: subjects in the ARP, subjects in the SP, 
and subjects in the FAS. All percentages will be based on the number of subjects in the ARP.  
End of trial information will also be summarized in this table, including the number of 
subjects completing the study, the number of subjects who prematurely discontinued the 
study with reasons for withdrawal, the number of subjects completing the study medication 
dosing, and the number of subjects who prematurely discontinued the study medication with 
reasons for study medication discontinuation.   
A by-subject data listing of study completion information including the reason for premature 
study withdrawal, if applicable, will be presented. 
8.2. 
Protocol Deviations 
Major protocol deviations, as determined by a Sponsor blinded review of the data prior to 
database lock and unblinding of the study, may result in the removal of a subject’s data from 
the PP Population. The Sponsor or designee will be responsible for producing the final 
deviation file; this file will include a description of the protocol deviation and clearly identify 
whether this violation warrants exclusion from the PP Population. This file will be finalized 
prior to database lock. 
All protocol deviations will be presented in a by-subject data listing, with a flag to indicate if 
a deviation was considered major.  
8.3. 
Demographics and Baseline Characteristics 
Subject demographic data and baseline characteristics will be tabulated and summarized 
descriptively by treatment sequence and overall.  The demographic data and baseline 
characteristics will be summarized for the SP and FAS. 
The demographics and baseline characteristics consist of age (year), sex, race, ethnicity, and 
study eye (OD [right eye] or OS [left eye]), iris color (brown, non-brown), and mydriatic 
agent (phenylephrine or tropicamide). A subject’s age in years is calculated using the date of 
the informed consent and date of birth. Age will be summarized using descriptive statistics. 
The number and percentage of subjects by sex, race, ethnicity, study eye, iris color, and 
mydriatic agent will also be reported. Percentages will be based on the total number of 
subjects in the study population presentation. 
The following baseline characteristics will be summarized for OD and OS, and for study eye, 
fellow eye, and all eyes, at Visit 1 / Day 1 and Visit 2 / Day 8 using descriptive statistics: 


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 Pupil diameter (-1 hour) 
 Max Pupil diameter (0 minutes)  
 
 
 
 
 
 
All demographic and baseline information will be presented in by-subject listings. 
8.4. 
Medical History  
The number and percent of subjects with individual medical histories will be summarized for 
all subjects by treatment sequence and overall. Non-ocular and ocular medical history will be 
summarized separately.  
Medical history will be coded using the MedDRA Version 22.0. The number and percentage 
of subjects with any medical history will be summarized overall and for each system organ 
class (SOC) and preferred term (PT). Percentages will be calculated based on number of 
subjects in the SP.   
Subject medical history data including specific details will be presented in by-subject 
listings. 
8.5. 
Prior and Concurrent Medications 
The number and percentages of all concomitant medications will be summarized by 
treatment group, Anatomical Therapeutic Chemical (ATC) level 4, and PT.  The total number 
of concomitant medications and the number and percentages of subjects with at least 1 
concomitant medication will be summarized by treatment group. All summaries will be 
performed using the SP.   
A concomitant medication is defined as any medication taken on or after the day of first 
exposure to study drug. 
Prior medications are defined as any medication that has a start and stop date prior to the day 
of first exposure to any study drug, collected from up to 30 days prior to Screening. The total 
number of prior medications and the number and percentages of subjects with at least 1 prior 
medication will be summarized by treatment sequence. 
8.6. 
Treatment Administration 
The number and percentages of subjects with assigned treatment administered will be 
presented by treatment group and visit. The number and percentages of subjects with 
Lumify® administered in the non-study eye will also be presented. 


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9. 
EFFICACY ANALYSES 
Unless otherwise noted, all efficacy analyses will be completed using the FAS.  
Confirmatory analysis of the primary efficacy endpoint will be performed using the ARP and 
PP Population. The FAS will be the primary analysis population for efficacy. All efficacy 
analyses will be completed using the planned treatment.  
9.1. 
Clinical Efficacy   
The evaluations of clinical efficacy will be performed using the FAS (and ARP and PP 
Population, for primary efficacy), as specified below.  Baseline values are generally taken at 
the -1 Hour time point at each visit (Visit 1 / Day 1 and Visit 2 / Day 8). For pupil diameter, 
change analyses will be based on max time point (0 minutes) at each visit, during which 
maximum pupil diameter is expected. 
Primary Efficacy Endpoint: 
The primary efficacy endpoint is the change in pharmacologically induced mydriatic (max) 
pupil diameter (0 minutes) at 2 hours post-treatment in the study eye.  The primary efficacy 
endpoint will be analyzed using a mixed model with change from max pupil diameter (0 
minutes) to 2 hours in mean pupil diameter (mm) as the dependent variable, treatment 
sequence, period, treatment and mydriatic agent as fixed effects, subject within treatment 
sequence as a random effect, and max pupil diameter (0 minutes) as the covariate. Example 
SAS code is as follows: 
  
  
 
 
 
 
 
The analysis will be performed using the FAS, with subjects included in their randomized 
treatment sequence regardless of the treatment they actually received.  Observed case data 
only will be used; that is, missing values will not be imputed. The least-squares mean (LSM) 
and standard error (SE) will be provided for both treatment groups, along with the placebo-
corrected LSM, its 95% confidence interval (CI) and associated p-value.   
In addition, the primary efficacy endpoint will be analyzed by mydriatic agent using the same 
model as above but without mydriatic agent as a factor.   
A confirmatory analysis of the primary efficacy endpoint will be performed, using the ARP 
with missing Visit 1 / Day 1, Hour 2 and Visit 2 / Day 8, Hour 2 values imputed. See Section 
7.3.2 for imputation methods. 




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In addition, each secondary efficacy endpoint will be analyzed by mydriatic agent using the 
same model indicated above but without mydriatic agent as a factor.   
10. 
SAFETY ANALYSES 
All Safety analyses will be conducted using the SP.  All safety analyses will be completed 
using the actual treatment a subject received. Observed case data will be used; no imputation 
will be performed for missing safety data except for the limited situations described in 
Section 7.3.2. 
For HR and BP, Baseline for a specific day (Visit 1 / Day 1, Visit 2 / Day 8) is defined as the 
-1 hour baseline value.  HR and BP values will be summarized by treatment group and time 
point (-1 hour, 2 hours, 6 hours; at Visit 1 and at Visit 2).  Change from Baseline in the 
values will be summarized at the same timepoints, except for -1 hour. 
For IOP, the only pre-dose assessment is at the screening visit, so the screening IOP will be 
employed as the baseline value for all post-baseline change from baseline assessments. 
10.1. 
Adverse Events 
Adverse events will be coded using MedDRA, Version 22.0.   
Treatment-emergent adverse events (TEAEs) are defined as any adverse event that begins or 
worsens after initiation of the investigational product and through the subject’s last study 
visit (study completion or early termination) or follow-up telephone call.  Serious adverse 
events will be recorded from the date of informed consent, throughout the clinical trial.   
If the onset of an AE is on or after the date of first dose of study medication, or increasing in 
severity after first dose of study medication, then the AE will be considered treatment-
emergent. 
TEAEs will be assigned to the treatment group most recently administered to the subject as 
of the TEAE start date. 
Only TEAEs will be summarized; all AEs (TEAE, non-TEAE) will be included in a by-
subject listing.  
The number and percent of subjects with any TEAEs will be summarized by system organ 
class and preferred term by treatment group and overall.  At each level of tabulation (e.g., at 
the preferred term level) subjects will be counted only once if they had more than one such 
event reported during the AE collection period. 
 
Note that in MedDRA, ocular events are coded to the SOC of “special senses”. Thus, using 
SOC in the summaries will provide a separation of ocular and non-ocular adverse events. 








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12.2. 
List of Planned Listings 


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12.3. 
List of Planned Figures