Phase I Clinical Study to Evaluate the Safety and Tolerability of PRO-201 Ophthalmic Solution Applied on the Ocular Surface of Healthy Volunteers.
Phase: PHASE1
Conditions: Myopia
Sponsor: Laboratorios Sophia S.A de C.V.

 
Protocol: 
SOPH201-0521/I 
Title: Phase I clinical study, to evaluate the safety and tolerability of the PRO-
201 ophthalmic solution, prepared by Laboratorios Sophia, S.A. de C.V. on the 
ocular surface of ophthalmologically and clinically healthy subjects
Information about the molecule under study 
Generic name:    Atropine Sulfate 0.01%  
Distinctive name:    PRO-201  
Indication:     Delayed progression of myopia    Delayed progression of myopia  
Protocol Information 
Study Phase:    I  
Version: 1.0 
Release Date: 10-ago-21 
This protocol has been carried out in accordance with the principles 
of the Declaration of Helsinki and will be carried out in accordance 
with Good Clinical Practices and in compliance with the ICH 
guidelines and current local legislation. 
Sponsor: Laboratorios Sophia, S.A. de C.V. 

 

Content 
Content
 
Table and Figure Index
 
Responsible for the study
 
Signature Page
 
From the sponsor
 
Investigator Agreement
 
List of abbreviations
 
1. Protocol Summary
 
1.1 Synopsis
 
1.2 Study diagram
 
1.3 Subject Timeline
 
2. Introduction and background
 
2.1Theoretical framework
 
2.2 Background on the investigational product
 
2.2.1 Preclinical study to evaluate the feasibility of a 1.0% atropine sulfate-induced model of dry eye in 
albino rabbits New Zealand
 
2.2.2 Clinical study of safety, tolerance and mydriatic efficacy of a topical ophthalmic reformulation of 
1% atropine sulfate
 
2.3 Background on the investigation
 
2.3.1 From the research question
 
2.4 Risk-benefit assessment
 
2.4.1 Known potential risks
 

2.4.2 Known potential benefits
 
2.5 Problem statement
 
2.6 Rationale for the study
 
3. Objectives and hypotheses
 
3.1 Objectives
 
3.1.1 Main objective:
 
3.1.2 Specific objectives:
 
3.2 Hypothesis
 
4. Study design
 
4.1 Study Overview
 
4.2 Rationale for the study design
 
4.3 Expected duration
 
5. Study population
 
5.1 Eligibility Criteria
 
5.1.1 Inclusion criteria
 
5.1.2 Exclusion criteria:
 
5.2 Criteria for elimination and/or substitution of subjects
 
5.2.1 Elimination Criteria
 
5.2.2 Substitution of subjects
 
5.3 Counting failures
 
5.4 Recruitment and retention strategies
 
5.5 Procedure in case of early discontinuation
 

5.6 Identification of the subject
 
6. Investigational product and treatment
 
6.1 Managed Products
 
6.1.1 Investigational product
 
6.1.2 Investigational Product Dosage
 
6.1.3 Treatment with the investigational product
 
6.2 Storage and handling of the investigational product in the study center
 
6.3 Concomitant treatments and medications (permitted and prohibited)
 
6.4 Procedure for monitoring and measuring adherence
 
6.5 Strategies to improve adherence
 
7. Study methods and procedures
 
7.1 Research center
 
7.2 Clinical Study Registration
 
7.3 Assignment of treatment
 
7.4 Outcome variables
 
7.4.1 Primary outcome variables
 
7.4.2 Secondary outcome variables
 
7.4.3 Exploratory variables
 
7.4.4 Definition of variables, methods and scales for their measurement
 
7.4.5 Description of the variables, methods and scales for their measurement
 
7.5 Description of procedures or assessments during the study
 
7.5.1 Signing of informed consent
 

7.5.2 Preparation of medical history (includes ophthalmological and general medical history)
 
7.5.3 Eligibility Criteria
 
7.5.4 Assigning Subject Code
 
7.5.5 Adverse events
 
7.5.6 Vital Signs Measurement
 
7.5.7 Urine Pregnancy Test
 
7.5.8 Ophthalmological evaluation
 
7.5.9 Application of Medications During Visits
 
7.5.10 Delivery of material for the subject
 
7.5.11 Delivery of Study Medication
 
7.5.12 Evaluation of concomitant medicinal products
 
7.5.13 Assessment of adherence to treatment
 
7.5.14 Return of Study Medication
 
7.5.15 Subject Journal Removal
 
7.6 Study Diagram and Timeline
 
7.6.1 Study diagram
 
7.6.2 Study timeline
 
7.7 Procedures to be carried out per visit
 
7.7.1 Baseline visit
 
7.7.2 Visit 1
 
7.7.3 Final Visit
 
7.7.4 Security Call
 

7.7.5 Unscheduled follow-up visits
 
7.8 Data collection
 
7.8.1 Source documents
 
7.8.2 Electronic forms of data collection
 
7.8.3 Archiving
 
8. Evaluation and management of adverse events
 
8.1 Regulation and regulations on adverse events
 
8.2 Definition of Adverse Event
 
8.3 Use of adverse events as a study safety variable
 
8.4 Definitions relevant to the classification of adverse events
 
8.5 Responsibilities of the researcher
 
8.5.1 Recording of adverse events in the electronic case report form
 
8.5.2 Adverse Event Tracking
 
8.5.3 Procedures for a serious adverse event
 
8.5.4 Causation assessment
 
8.6 Unanticipated Issues
 
8.6.1 Reporting Unanticipated Issues
 
9. Study Monitoring
 
9.1 Monitoring the Study Site
 
9.2 Audit and quality control
 
9.2.1 Pre-study audit
 
9.2.2 Audit during the conduct of the study
 

10. Sample size calculation and statistical analysis
 
10.1 Sample Size Calculation
 
10.1.1 Calculation methodology
 
10.1.2 Size Calculation
 
10.2 Clinical Data Management
 
10.3 Statistical methodology
 
10.3.1 Population Analysis
 
10.3.2 Safety and tolerability analysis
 
10.3.3 Exploratory Analysis
 
10.4 Procedure for handling missing data
 
11. Ethical considerations
 
11.1 Approval of the committees
 
11.2 Amendments to the protocol
 
11.3 Early Study Termination
 
11.4 Informed Consent
 
11.4.1 Obtaining
 
11.4.2 Special considerations
 
11.4.3 Modifications to informed consent
 
11.5 Confidentiality
 
11.6 Conflict of interest
 
11.6.1 Declaration of Interests
 
11.7 Access to Information
 

11.8 Ancillary and post-study care
 
12. Biosecurity aspects
 
13. Posting Policy
 
13.1 Final Report
 
13.2 Communication of results
 
13.3 Publication of results
 
14. Financing and Insurance
 
14.1 Compensation to Study Participants
 
14.2 Insurance for Study Participants
 
15. Bibliography
 
16. Annexes
 
16.1 ETDRS Primer for Near Vision
 
16.2 SICCA Eye Staining Rating (modified from Whitcher et al, 2010)
 
16.3 Ocular Comfort Index (OCI)
 
16.4 Fraction to LogMAR conversion table.
 
16.5 Number of events/sample size for the exact method vs. the Wu method, for different values of the 
parameter β, cumulative time a, and tracking time f
 

Table and Figure Index 
 
 
 
 
 
 
 
 
 

Responsible for the study 
The administrative structure of the sponsoring party, corresponding to Laboratorios Sophia, S.A. de 
C.V., is shown in Table 1and Figure 1. Responsible for the study. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  

Signature Page 
From the sponsor  
  
  
  
 

Investigator Agreement    
I agree to conduct this clinical study according to the design and guidelines of this protocol, in 
accordance with its provisions. I declare that I will conduct the study in accordance with the 
standards of Good Clinical Practice, and I will report all information or data in accordance with the 
protocol, particularly any adverse event. I will also handle clinical supplies, provided by the sponsor, 
strictly in accordance with this protocol. I understand that the information that identifies me may 
be used by the sponsor. Because the information contained in this protocol and the Investigator's 
Manual is confidential, I understand that it is prohibited from sharing it with any third party, which 
is not involved in the approval, supervision, or conduct of the study. I will ensure that I take the 
necessary precautions to protect the information from loss, inadvertent disclosure, or access by 
unauthorized third parties.  
 
Name:  
[Write full name of the 
researcher 
Title:  
Principal Investigator  
Name of the center:  
[Insert Name of Study Site] 
Geographic location (city/state/country)  
[Enter the geographical data of the center]  
  
Signature  
  
Date  
  
  
  
  
 

List of abbreviations 
Ad 
Adherence 
AO 
Both eyes 
VA 
Visual acuity 
NVA 
Near visual acuity 
BCVA 
Best-corrected visual acuity 
NBCVA 
Near Best Corrected Visual Acuity 
GCP 
Good clinical practice 
CDM 
 Clinical Data Management 
REC 
Research Ethics Committee 
CR 
Research Committee 
COFEPRIS 
Federal Commission for the Protection against Sanitary Risks 
ESR 
Eye Staining Rating 
SD 
Standard deviation 
IUD 
Intrauterine device 
EA 
Adverse Event 
eCFR 
 Electronic case form report 
USA 
United States of America 
HR 
Heart rate 
ICF 
Informed consent form 
RF 
Respiratory rate 
H0 
Null hypothesis 
H1 
Alternate hypothesis 
ICH 
International Council for Harmonisation 
ICMJE 
International Committee of Medical Journal Editors 
ECI 
Eye Comfort Index 
PI 
Principal Investigator 
ITT 
By intention to treat 
BPM 
Beats per minute  
Mmhg 
Millimeters of mercury  

NOM 
Official Mexican Standard 
WHO 
World Health Organization 
BDP 
Diastolic blood pressure 
RP 
Research product 
PIO 
Intraocular pressure 
PNA 
Unanticipated problems 
PP 
Population by protocol 
ADR 
Adverse drug reaction 
NCTR 
National Clinical Trials Registry 
SDV 
Source document verification 
SICCA 
Sjögren's international collaborative clinical Alliance 
SPSS 
Statistical package for the social sciences   
SADR 
Suspected adverse drug reaction 
FS 
Fluorescein staining 
TRPL 
Tear film rupture time 
SLG 
Staining with Lysmine Green 
UFTLS 
Pharmacovigilance and Technovigilance Unit of Sophia Laboratories 
CV 
Current version 
x2 
Chi-square 
°C 
Celsius degrees  
 
 
 
 
 
 
 

1. Protocol Summary 
1.1 Synopsis 
Title of the study: 
Phase I clinical study, to evaluate the safety and tolerability of the PRO-201 ophthalmic solution, 
prepared by Laboratorios Sophia, S.A. de C.V. on the ocular surface of ophthalmologically and 
clinically healthy subjects 
Study Number: 
   SOPH201-0521/I   
Date of creation: 
28/May/2021 
Protocol version: 
   1.0  
Release Date:  
Aug/10/2021 
Therapeutic indication: 
Adjuvant in treatment to delay the progression 
of myopia. 
Use: 
Myopia 
Estimated duration of the study (from the first 
visit of the first patient to the preparation of the 
final report): 
6 months 
Clinical Development Phase: I 
Objectives: 
Main objective:  
• 
To evaluate the safety and tolerability of the PRO-201 formulation manufactured by 
Laboratorios Sophia S.A. de C.V. on the ocular surface of ophthalmologically and clinically 
healthy subjects. 
Specific objectives: 
• 
Primary 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular 
surface in healthy volunteers, through the incidence of unexpected adverse 
events (AEs) related to the investigational product (PI). 

o To assess the tolerability of PRO-201 ophthalmic solution applied to the ocular 
surface in healthy volunteers using the incidence of photophobia. 
• 
Secondary 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular 
surface in healthy volunteers, by evaluating the pupillary diameter by the OPD 
scan. 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular 
surface in healthy volunteers, using the incidence of expected PI-related AEs. 
o To evaluate the tolerability of PRO-201 ophthalmic solution applied to the ocular 
surface in healthy volunteers, by evaluating the best corrected near visual acuity 
(CVA) with the ETDRS primer. 
• 
Exploratory 
o To assess the safety of PRO-201 ophthalmic solution applied to the ocular surface 
in healthy volunteers, by assessing best-corrected visual acuity (BCVA). 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular 
surface in healthy volunteers, by assessing intraocular pressure (IOP). 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular 
surface in healthy volunteers, by evaluating corneal and conjunctival staining with 
fluorescein and lysmine green.  
o To assess the safety of PRO-201 ophthalmic solution applied to the ocular surface 
in healthy volunteers, by assessing vital signs (heart rate and blood pressure). 
o To evaluate the tolerability of PRO-201 ophthalmic solution by means of the 
Ocular Comfort Index (ILO) score. 
Hypothesis: 
H0= The PRO-201 ophthalmic solution is not safe in its ophthalmic application as it has an incidence 
of unexpectedly related adverse events (AEs) greater than 10% and is not tolerable as it presents 
an incidence of photophobia greater than 15% of the study population. 
𝐻0:𝑝−𝑝0 ≤−𝛿 
H1= PRO-201 ophthalmic solution is safe in ophthalmic application with an incidence of 
unexpected, related adverse events (AEs) of less than 10% and is tolerable with an incidence of 
photophobia of less than 15% of the study population. 
𝐻1:𝑝−𝑝0 > −𝛿 
Study Design: 
Phase I Clinical Trial, Controlled, Non-Comparative, Open-label, Single-Center 
Number of subjects (planned and analyzed): 

Number of planned subjects: 29 evaluable subjects (both eyes) 
Diagnosis and main inclusion criteria: 
- 
Ophthalmologically and clinically healthy subjects. 
Selection criteria: 
Inclusion criteria: 
− 
Be clinically healthy 
− 
Can voluntarily give their signed informed consent (CRF) 
− 
Be able and willing to comply with scheduled visits, treatment plan, and other study 
procedures 
− 
Be between 18 and 35 years old. 
− 
No history of contact lens use 
− 
Women of childbearing potential must ensure continued use (initiated ≥ 30 days prior to 
signing the CRF) of hormonal contraceptive or intrauterine device (IUD) use during the 
study period 
− 
Have a better far-corrected visual acuity of 20/30 (logMAR 0.2) or better in both eyes. 
− 
Have a better corrected visual acuity close to 20/25 (logMAR 0.1) or better in both eyes. 
− 
Have vital signs within normal parameters.  
− 
Have an IOP ≥10 and ≤ 21 mmHg. 
Exclusion criteria: 
− 
Be a user of topical ophthalmic products of any kind.  
− 
Allergy or known intolerance to ingredients in atropine eye drops and other derivatives of 
muscarinic antireceptor agents. 
− 
Being a user of medicines, or herbal products (plant extracts, infusions, naturopathic 
preparations, homeopathics, etc.), by any other means of administration. 
− 
For women: being pregnant, breastfeeding, or planning to become pregnant within the 
study period. 
− 
Have participated in any investigational clinical study 90 days prior to inclusion in the 
present study. 
− 
Have previously participated in this same study. 
− 
Who are unable to follow the lifestyle considerations described in Annex 1. 
− 
Have a history of any chronic-degenerative disease, including diabetes and high blood 
pressure. 
− 
Have an active inflammatory or infectious disease at the time of study entry. 
− 
Have unresolved injuries or trauma at the time of study entry. 
− 
Have a history of any type of eye surgery. 
− 
Having undergone surgical, non-ophthalmologic procedures within the past 3 months. 
− 
Be or have an immediate family member (e.g., spouse, parent/legal guardian, sibling, or 
child) who is employed by the research site or sponsor, and who is directly involved in this 
study. 

Elimination criteria: 
− 
Withdrawal of the letter from the FCI. 
− 
Presentation of a serious adverse event related or not to the investigational product, which 
at the discretion of the PI and/or sponsor could affect the patient's ability to continue with 
the study procedures safely. 
− 
Non-tolerability or hypersensitivity to any of the compounds used during the tests 
(fluorescein, lysmine green, tetracaine). 
− 
No tolerability or hypersensitivity to the investigational drug. 
− 
Adherence < 90% determined by the subject's diary 
Research Product (PI): 
- 
 PRO-201 . Atropine Sulfate 0.01%. Zapopan, Jalisco, Mexico.   Ophthalmic solution   
Laboratorios Sophia, S.A. de     C.V  
- 
Dosage: 1 drop once a day (QD) in the evening, in both eyes (AO). 
- 
Route of administration: Ophthalmic 
Duration of treatment: 
14 days 
Duration of the subject in the study: 
21 days 
Evaluation criteria: 
Primary outcome variables 
− 
Incidence of unexpected PI-related AEs (Time of Assessment [TE]: days 7, 16 and 21) 
− 
Incidence of IP-related photophobia. (TE: 7th, 16th and 21st) 
Secondary outcome variables: 
− 
Changes in pupillary diameter (ET: day 7 and 16). 
− 
Incidence of expected PI-related AEs (TE: days 7, 16, and 21).  
− 
Changes in visual acuity (TE: day 7 and 16). 
Exploratory variables: 
− 
Changes in the AVMC (TE: day 7 and 16). 
− 
Intraocular pressure (IOP) changes (ET: day 7 and 16). 
− 
Changes in corneal and conjunctival staining with fluorescein and lysmine green (ET: day 
7 and 16). 
− 
Changes in vital signs (heart rate and blood pressure) (ET: day 7 and 16). 
− 
Changes in the OCI (TE: day 16). 
Statistical methodology 
The analysis of the data collected in the study will be carried out using the SPSS version 19.0 
statistical package. For continuous and discrete quantitative variables, data will be expressed by 

measures of central tendency: mean, standard deviation, and ranges. Qualitative data will be 
presented as frequencies and percentages. The statistical analysis to rule out differences will be 
carried out by means of non-parametric statistics. The Kolmogorov-Smirnov statistic will be used 
for a sample and the Mann-Whitney statistic (when applicable). The analysis for qualitative 
variables will be performed by means of Pearson's X2 (Chi-square) test or Fisher's Exact test (when 
applicable). For all analyses, a p < 0.05 and an alpha of 0.05 (two-tailed) will be considered 
significant.  

1.2 Study diagram 
Obtaining informed consent. Compliance with 
all inclusion criteria and none of exclusion 
criteria. 
 Obtaining baseline values 
N= 29. 
Clinical Evaluations 
V1 (Day 7) 
VB 
D 1 
VF 
D 16 (±1) 
 
V1 
D 7 (± 1) 
BV= Basal visit  
 
V1 = Visit 1 
 
VF= Final visit. Evaluation of outcome variables. 
SC = Safety Call 
QD= Once a day 
AO= Both eyes 
 
 
 
In parentheses () is the allowed window period. 
 
Starting the PRO-201 Application 
1 drop QD AO 
6 days after baseline 
visit 
Clinical Evaluations 
VF (day 16) 
13 days after baseline 
 20 days after baseline 
visit 
Security Call (Day 21) 
End of application (day 14) 
 
D 14 
  
SC 
D 21 (± 1) 
  

1.3 Subject Timeline 
Procedures 
VB 
D 1 
V1 
D 7 ± 1 
VF 
D 16 ± 1 
SC 
D21 ± 1 
Informed Consent Form (FCI) Signature 
X 
 
 
 
Medical history 
X 
 
 
 
Eligibility Criteria 
X 
 
 
 
Concomitant Drug Evaluation 
X 
X 
X 
 
Urine pregnancy test 
X 
 
X 
 
Vitals signs 
X 
X 
X 
 
Best Corrected Visual Acuity (BCVA) 
X 
X 
X 
 
Best Corrected Near Visual Acuity (BCNVA) 
X 
X 
X 
 
Pupillary diameter measurement 
X 
X 
X 
 
Evaluation of fluorescein staining and 
lysmine green 
X 
X 
X 
 
PIO 
X 
X 
X 
 
Comprehensive ophthalmological 
evaluation 
X 
X 
X 
 
Adverse Event (AE) Assessment 
X 
X 
X 
X 
Subject Code Assignment and Research 
Product (PI) 
X 
 
 
 
Eye Comfort Index (ECI) 
X 
 
X 
 
Delivery of the PI and initiation of 
intervention 
X 
 
 
 
Daily subject delivery 
X 
 
 
 
Adherence assessment 
 
X 
X 
 
Return / evaluation of the subject's diary 
 
 
X 
 
Return of PI 
 
 
X 
 

2. Introduction and background 
2.1Theoretical framework 
Myopia is a refractive error of the eye in which light rays converge on a focal point in front of the 
retina, instead of converging on the retina itself, making it difficult to focus well on distant objects. In 
progressive myopia, this defect occurs due to an exaggerated growth of the eyeball that causes a 
progressive thinning of all layers of the eye, with the retina being the most affected. [1] 
Myopia is one of the five priorities for the World Health Organization (WHO) in the initiative "Vision 
2020: The Right to Sight" [2]. It is estimated that half of the world's population will be myopic by 
2050. [3] 
Although the exact etiology of myopia remains difficult to explain, it appears to have both genetic 
and environmental components, making prevention and treatment challenging and individualized. 
Stopping the progression of myopia has the potential to positively affect quality of life and eye 
health.[4] 
The initial problem with myopia is that patients who suffer from it do not see far away, but what is 
really important Limiting the progression of myopia is explained by the increased risk of blindness 
with ≤ −6 diopters and/or ≥ 26 mm axial length (macular neovascular membrane, macular 
chorioretinal atrophy, retinal detachments, glaucoma, etc.). [5] [6] 
Myopia usually appears between the ages of six and eight and progresses on average diopter per year 
until the age of 15 or 16, but there are patients who continue to increase for practically their entire 
lives. [1] 
We are facing an increase in myopia globally; in the United States of America, 42% of the population 
is myopic, a percentage that in 1970 was 25%. In some areas of Asia such as Taiwan, Singapore, and 
Hong Kong, young adults have myopia rates of 80 to 90%.  [7] 
There are different studies that show that the most effective way to slow down the progression of 
this type of myopia is the use of topical antimuscarinic drugs. Among the antimuscarinic drugs, 
atropine eye drops have been confirmed to be the safest and most effective treatment.  [8] 
Atropine is the most studied and used non-selective antimuscarinic agent to stop the progression of 
myopia. The first references, in German and English publications, to the use of atropine in the 
treatment of myopia date back to the 1970s in the 1970s. [9, 10] 
The ATOM studies (for its acronym in English Atropine for the Treatment of Myopia) they have used 
atropine in different concentrations (1%, 0.5%, 0.1% and 0.01%) for a period of up to 5 years, 
demonstrating a reduction in the progression of myopia; as well as in the increase of axial length. 
Finding a small percentage of adverse effects in the population studied. [11, 12, 13, 14] 

The effect of decreasing the progression of myopic refractive error using 0.01% atropine is currently 
recognized. However, today 0.01% atropine is only marketed in Asia and magisterial preparations are 
made in American countries such as Canada. In clinical trials, patients are provided with dilutions 
prepared with artificial tears. Although there are ophthalmologists who make this dilution in their 
clinics, this practice can present risks of dosage, stability and asepsis if it is not carried out under 
adequate conditions. [15] 
2.2 Background on the investigational product 
We do not have clinical studies of an ophthalmic solution containing 0.01% atropine sulfate so far. 
However, we have the experience in clinical studies with atropine sulfate at concentrations higher 
than PRO-201. 
2.2.1 Preclinical study to evaluate the feasibility of a 1.0% atropine sulfate-induced model 
of dry eye in albino rabbits New Zealand 
Fifteen healthy New Zealand albino rabbits were given a 1% atropine sulfate solution three times 
daily for 14 days in both eyes, and five other rabbits served as a control group, as part of a preclinical 
study to evaluate the feasibility of a dry eye model. In the results of the study regarding the 
histopathological analysis of eyeballs, all the results of the 40 eyes studied were within normal limits, 
finding only slight changes such as those related to tissue preservation/fixation and no evidence of 
any pathological alteration. This is relevant because, together with the IOP levels evaluated (found 
within normal parameters) and the absence of AEs reported in this study, the histopathological result 
confirms the safety profile of the currently marketed product. [16] 
2.2.2 Clinical study of safety, tolerance and mydriatic efficacy of a topical ophthalmic 
reformulation of 1% atropine sulfate 
Open-label, single-center, observational, prospective clinical study included thirty healthy volunteers. 
The volunteers administered a drop of 1% atropine sulfate in reformulation once a day in a single 
dose and in only one eye, and the effect was assessed one hour after application. Biomicroscopy was 
performed at baseline and one hour after application, evaluating the presence or not of conjunctival 
hyperemia, conjunctival secretion, ciliary injection, chemosis, and corneal surface, as well as Rose 
Bengal and Fluorescein stains, grading on a scale of 0= absent, 1= mild, 2= moderate, and 3= severe, 
according to the staining of the ocular surface and the pupillary diameter, both horizontally and 
vertically. Of the 30 patients included in this study, there was no presence of drug residues or 
significant alterations of the conjunctival surface. 
Regarding mydriatic efficacy, pupillary dilation of up to twice the basal size was observed, both in the 
vertical diameter and the horizontal diameter. Regarding tolerance and safety, only 9 patients (30%) 
reported mild burning, with an average of 0.3 on a scale of 0 to 3. In the same area, there was also 
no uptake of fluorescein or Rose Bengal on the ocular surface. Based on the results presented, it can 
be concluded that 1% atropine sulfate (Atro Ofteno®) in reformulation has a good mydriatic efficacy 

one hour after its application, as well as being safe and well tolerated when applied to the ocular 
surface of healthy volunteer patients. [17] 
2.3 Background on the investigation 
2.3.1 From the research question 
Is the application of PRO-201 ophthalmic solution safe and tolerable in clinically healthy subjects? 
The importance of the question that precedes this paragraph is that Laboratorios Sophia S.A. de C.V. 
does not have clinical studies with products identical to the PRO-201 solution, although there are 
with similar products (different concentration and excipients).  
2.4 Risk-benefit assessment 
2.4.1 Known potential risks 
Formulations containing atropine sulfate at the same concentrations as PRO-201 are products that 
are marketed in Asia, as well as in compounding formulations in other countries in the Americas and 
Europe. The risks of using this product in specific populations are described below, however, the use 
of this drug in the study will be in a population group without the alterations or characteristics 
described below. To Concentration at 0.01% The potential risks are: photophobia, mydriasis, difficulty 
reading, allergic eye reactions (atopic dermatitis, allergic conjunctivitis and other allergic reactions), 
eye irritation, burning eyes, eye pain, tearing, foreign body sensation, dry eye, hyperemia and/or red 
eye, blurred vision, eye itching; Systemic adverse events include: headache, drowsiness, dizziness, 
tachycardia, flushing, restlessness, irritability, dry skin, dry mouth, skin rash and decreased alertness.   
[8]  [6] [18] [19] [20] [21] [22] [23] [24] 
2.4.2 Known potential benefits 
The benefit in the target population is to serve as an adjuvant in the treatment to delay the 
progression of myopia.  But for the study population, as it is a phase I study, it will be a healthy 
population. Therefore, a direct benefit for each individual is not expected, and the benefit provided 
by the research will be the[11, 12, 13, 14, 25] to know the behavior of the formulation; Have more 
data on their safety to be able to prevent or warn of unwanted effects on users or problems with 
their tolerance.   
2.5 Problem statement 
The effect of decreasing the progression of myopic refractive error using 0.01% atropine is currently 
recognized. However, today 0.01% atropine is only marketed in Asia and magisterial preparations are 
made in American countries such as Canada. In clinical trials, patients are provided with dilutions 
prepared with artificial tears. Although there are ophthalmologists who make this dilution in their 
clinics, this practice can present risks of dosage, stability and asepsis if it is not carried out under 
adequate conditions. [15] 

Therefore, the development of a 0.01% atropine solution manufactured and marketed by 
Laboratorios Sophia, S.A. de C.V., would represent a great opportunity as a treatment to prevent 
progression in the face of the increase in myopia cases worldwide that has been recorded in recent 
years. 
The Laboratory has experience in preclinical and clinical studies with products with 1% atropine, as 
well as experience in commercialization. However, having no experience with the formulation of PRO-
201, it is important to develop the appropriate clinical research. 
2.6 Rationale for the study 
Laboratorios Sophia, S.A. de C.V., has carried out preclinical and phase I clinical studies of a similar 
formulation, but with a higher concentration of the component. The decrease in the concentration 
of the main component (atropine sulfate) in the PRO-201 solution to this formulation studied could 
mean a greater safety of PRO-201, not necessarily a greater tolerance, since the excipients may be 
involved with clinical manifestations unrelated to the main components.  
Therefore, the present study serves to test the safety and tolerability of the PRO-201 product, in a 
controlled environment with healthy subjects.  
 
 

3. Objectives and hypotheses 
3.1 Objectives 
3.1.1 Main objective:  
• 
To evaluate the safety and tolerability of the PRO-201 formulation manufactured by 
Laboratorios Sophia S.A. de C.V. on the ocular surface of ophthalmologically and clinically 
healthy subjects. 
3.1.2 Specific objectives: 
3.1.2.1 Primary 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular surface 
in healthy volunteers, through the incidence of unexpected adverse events (AEs) 
related to the investigational product (PI). 
o To assess the tolerability of PRO-201 ophthalmic solution applied to the ocular 
surface in healthy volunteers using the incidence of photophobia. 
3.1.2.2 Secondary 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular surface 
in healthy volunteers, by evaluating the pupillary diameter by the OPD scan. 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular surface 
in healthy volunteers, using the incidence of expected PI-related AEs. 
o To evaluate the tolerability of PRO-201 ophthalmic solution applied to the ocular 
surface in healthy volunteers, by evaluating the best corrected near visual acuity 
(CVA) with the ETDRS primer. 
3.1.2.3 Exploratory 
o To assess the safety of PRO-201 ophthalmic solution applied to the ocular surface in 
healthy volunteers, by assessing best-corrected visual acuity (BCVA). 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular surface 
in healthy volunteers, by assessing intraocular pressure (IOP). 
o To evaluate the safety of PRO-201 ophthalmic solution applied to the ocular surface 
in healthy volunteers, by evaluating corneal and conjunctival staining with 
fluorescein and lysmine green.  
o To assess the safety of PRO-201 ophthalmic solution applied to the ocular surface in 
healthy volunteers, by assessing vital signs (heart rate and blood pressure). 
o To evaluate the tolerability of PRO-201 ophthalmic solution by means of the Ocular 
Comfort Index (OCI) score. 

3.2 Hypothesis 
H0= The PRO-201 ophthalmic solution is not safe in its ophthalmic application as it has an incidence 
of unexpectedly related adverse events (AEs) greater than 10% and is not tolerable as it presents an 
incidence of photophobia greater than 15% of the study population. 
𝐻0:𝑝−𝑝0 ≤−𝛿 
H1= PRO-201 ophthalmic solution is safe in ophthalmic application with an incidence of unexpected, 
related adverse events (AEs) of less than 10% and is tolerable with an incidence of photophobia of 
less than 15% of the study population. 
𝐻1:𝑝−𝑝0 > −𝛿 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

4. Study design 
4.1 Study Overview 
Phase I Clinical Trial, Controlled, Non-Comparative, Open-label, Single-Center 
4.2 Rationale for the study design  
The design of the study (clinical trial) is considered the highest standard of data quality when seeking 
to explore the effect of an intervention. The drug development phase (phase I) corresponds to the 
objective of the study, which is to evaluate safety and tolerability, so the intervention time is short 
and the sample size required is smaller than that of an efficacy clinical trial. Blinding in the study was 
not considered because there will be only one treatment group.  
4.3 Expected duration  
The total duration of the study, from the first visit of the first patient to the preparation of the final 
report, is estimated to be 6 months.  
The planned recruitment period is 4 months. Considering that the proposed sample is 29 subjects, 
the total average recruitment rate during the study should be no less than 2 subjects included each 
week. 
The approximate duration of each subject in the study is up to 22 days. 
 

5. Study population  
5.1 Eligibility Criteria 
5.1.1 Inclusion criteria 
− 
Be clinically healthy 
− 
Can voluntarily give their signed informed consent (CRF) 
− 
Be able and willing to comply with scheduled visits, treatment plan, and other study 
procedures 
− 
Be between 18 and 35 years old. 
− 
No history of contact lens use 
− 
Women of childbearing potential must ensure continued use (initiated ≥ 30 days prior to 
signing the CRF) of hormonal contraceptive or intrauterine device (IUD) use during the study 
period 
− 
Have a better far-corrected visual acuity of 20/30 (logMAR 0.2) or better in both eyes. 
− 
Have a better corrected visual acuity close to 20/25 (logMAR 0.1) or better in both eyes. 
− 
Have vital signs within normal parameters.  
− 
Have an IOP ≥10 and ≤ 21 mmHg. 
5.1.2 Exclusion criteria: 
− 
Be a user of topical ophthalmic products of any kind.  
− 
Allergy or known intolerance to ingredients in atropine eye drops and other derivatives of 
muscarinic antireceptor agents. 
− 
Being a user of medicines, or herbal products (plant extracts, infusions, naturopathic 
preparations, homeopathics, etc.), by any other means of administration. 
− 
For women: being pregnant, breastfeeding, or planning to become pregnant within the study 
period. 
− 
Have participated in any investigational clinical study 90 days prior to inclusion in the present 
study. 
− 
Have previously participated in this same study. 
− 
Who are unable to follow the lifestyle considerations described in Annex 1. 
− 
Have a history of any chronic-degenerative disease, including diabetes and high blood 
pressure. 
− 
Have an active inflammatory or infectious disease at the time of study entry. 
− 
Have unresolved injuries or trauma at the time of study entry. 
− 
Have a history of any type of eye surgery. 
− 
Having undergone surgical, non-ophthalmologic procedures within the past 3 months. 
− 
Be or have an immediate family member (e.g., spouse, parent/legal guardian, sibling, or child) 
who is employed by the research site or sponsor, and who is directly involved in this study. 
− 
 

5.2 Criteria for elimination and/or substitution of subjects 
5.2.1 Elimination Criteria 
▪ Withdrawal of the letter from the FCI. 
▪ Presentation of a serious adverse event related or not to the investigational product, which 
at the discretion of the PI and/or sponsor could affect the patient's ability to continue with 
the study procedures safely. 
▪ No tolerability or hypersensitivity to any of the compounds used during the tests (fluorescein, 
lysmine green or tetracaine). 
▪ No tolerability or hypersensitivity to any of the investigational drugs. 
▪ Adherence < 90% determined by the subject's diary 
5.2.2 Substitution of subjects 
The sponsor, with the prior authorization of the research ethics committees, may decide to replace 
the subjects who withdraw their FCI or those who present loss of follow-up, until the necessary 
sample size is completed. 
5.3 Counting failures 
A screening failure is defined as those participants who agree to participate in the study, giving their 
consent, but who are not assigned to the treatment group, i.e., do not enter the study. It is necessary 
to report at least the following information on counting failures:   
- 
Demographics.  
- 
Details of the failure to count (specify whether it is due to eligibility criteria, which one, or 
some other reason for the failure).  
- 
Presence of serious adverse events during the count.  
 
This is necessary to comply with the CONSORT (Consolidated Standards of Reporting Trials) guidelines 
for the publication of results or to respond to possible questions from regulatory authorities.  
5.4 Recruitment and retention strategies  
The duration of the subject's participation in the study is approximately 21 days, during which time 
they must attend three visits in total and a safety call will be made, which corresponds to the baseline 
visits, visit 1, final visit and safety call. Strategies to improve subject retention include, but are not 
limited to:  
• 
Clearly inform the objectives of the study. 
• 
Make calls or send text messages to remind you of appointments or activities to do.   
• 
Provide a printed calendar and an identification card to remember appointments and 
activities that will be carried out, in addition to the estimated time of their duration.  

• 
Systematic organization of the study procedures, so that the subject does not last longer 
than necessary in his visit.  
• 
Minimize subject wait times.  
All materials to be delivered to the subject or recruitment strategies implemented by the center will 
be submitted for approval by the corresponding committees.  
5.5 Procedure in case of early discontinuation  
For this protocol, early discontinuation is defined as those subjects who were assigned to the PI, who 
at some points were active subjects of the study, but their final evaluation could not be completed.  
If the subject does not terminate their participation due to withdrawal of consent or major deviation, 
the last visit, in which their withdrawal was determined, will be considered their final visit. Subjects 
who are removed due to the presence of AEs will continue with the follow-up that is defined until 
the closure of their AE.  
In cases where the participating subject does not attend their appointment, the research center must 
make a call to find out the reason and will try to make a new appointment within the established 
window period or an unscheduled appointment. If it is not possible to make an appointment, the 
subject will be considered as loss of follow-up and will be asked about the presence of adverse events 
and the reason for leaving the study, as minimum data.  
5.6 Identification of the subject  
Study subjects will be identified by a number and the initials of their name.  
The initials of the study subject will be obtained starting with the first letter of the name, followed 
by the first letter of the first surname and the first letter of the second surname, obtaining a 
maximum of three letters, in case the person has two names or compound surname, the first letter 
will always be used.  
Example:  
 
A) Arieh Daniel Carrizalez Market                                    B) Juan De la Torre Orozco  
 
to. Initials: AMC     
 
 
 
b. Initials: JDO  
In the counting stage, the participant number will be assigned consecutively, using 3 consecutive 
digits.   
Once the subject has been selected, they will be assigned a number with which they will be identified 
throughout the study. This code will be made up of eight numbers in the following order from left to 
right:   
 

• 
three digits of the molecule under study according to the name by the sponsor.    
• 
Two digits correspond to the research center number.   
• 
three digits of the number following their inclusion assigned in the research center.  
Example of assigned number:  
 
 
 
201 - 01 - 001   
Key to the  
molecule 
  
Number of  
inclusion   
Number of  
center   

6. Investigational product and treatment 
6.1 Managed Products 
6.1.1 Investigational product  
• 
 PRO-201 . Atropine Sulfate 0.01% Ophthalmic solution. Zapopan, Jalisco, Mexico.   
Laboratorios Sophia, S.A. de     C.V  
• 
Route of administration: Ophthalmic. 
6.1.2 Investigational Product Dosage 
- 
Dosage: 1 drop once a day (QD) in the evening, in both eyes (AO). 
6.1.3 Treatment with the investigational product 
The investigational product will be delivered at the conclusion of the baseline visit, and the 
investigational subject will administer the drops at night, applying a total of one drop at night to each 
eye during the 14 days of application.  
The baseline visits and visit 1 should be done in the morning shift, before 4:00 p.m., so that the patient 
can have enough time to apply PRO-201 at the indicated time.  
6.2 Storage and handling of the investigational product in the study center  
The delivery will be made by means of a courier service contracted by the sponsor, expressly selected 
for this purpose, to the address of the research center according to the study plan.  
The reception will be carried out by the assigned personnel of the research team. You will need to 
check that the primary packaging (box) is in good condition. If you show alterations or defects in its 
integrity that in your judgment could have damaged the content, you must report it to the sponsor. 
If the package does not show significant defects, it will proceed to open it.   
Inside the shipment you must locate the acknowledgment document and the temperature meter 
(data logger). You must check that the temperature recorded complies with what is specified for 
transport and safekeeping. It will verify the content (PI) with what is reported in the document. In 
case the document corresponds to the content, you will sign the receipt and send it to the sponsor. 
Otherwise, it will notify the sponsor.  
Storage and safeguarding are the responsibility of the research center. The medication must be 
stored in a secure area with restricted access.   
The storage temperature should be between 2° and 30°C.  
From its receipt at the center and until there is no stock of PI stored, the research center has the 
obligation to review the storage conditions of the PI daily and manually record, in the designated 

format, the temperature set by the data logger (current, minimum and maximum temperature). Such 
data will be reviewed by the clinical monitor during their monitoring visits according to the records 
stored in the data logger's memory. 
In the event of loss of material, it must be documented in the logbook of inputs and outputs along 
with a clear description of the mechanism by which the loss occurred.   
Upon completion of the protocol, all study material will be retrieved by the sponsor as part of the 
closing visit. The final delivery of materials will be made by the principal investigator or the person 
designated by him to deliver material at the end of the study.   
The sponsor reserves the right to initiate civil and criminal action against the principal investigator in 
the event of a lack of undocumented material at the conclusion of the study.  
6.3 Concomitant treatments and medications (permitted and prohibited)  
Any medication that is used, in addition to appearing in the clinical note, must be registered in the 
concomitant medications section of the eCRF. 
Allowed medications:  
- 
Ophthalmic:  
All permitted medications applied via ophthalmic medication during the study must wait a minimum 
period of 10 minutes from the last application of the treatments under study or reference. The above 
is to avoid the interaction of treatments in the tear film, based on the flow index and physiological 
tear volume.  [26] 
 
o Tetracaine 0.5%  
o Tropicamide 0.8% /Phenylephrine 5%  
o Hypromellose 2% 
o Fluorescein 
o Lysmine Green 
Prohibited Medications:  
- 
Any ophthalmic medications that are not on the list of allowed medications  
- 
Any systemic medication 
6.4 Procedure for monitoring and measuring adherence  
For more than four decades, there has been a lot of research on the appropriate way to measure and 
quantify medication adherence, but none has reached a consensus to stand as the gold standard, 
both in cross-sectional and longitudinal studies. [27, 28, 29, 30, 31, 32, 33, 34] 

There are different procedures to measure adherence to pharmacological interventions. The most 
common procedure includes self-reports, which include patient interviews, questionnaires and self-
monitoring diaries. Its strengths are speed, flexibility, low cost and ease of implementation; They have 
a high degree of specificity for non-adherence, however, the sensitivity and reliability for adherence 
is low. [34, 35] 
The biochemical measurement of the drug, or its metabolite, is one of the methods that best confirms 
the use of the drug. However, in addition to raising costs and being impractical, it is of little use in the 
context of ophthalmic applications, since concentrations at the peripheral level could be 
undetectable; and samples of other tissues involve more invasive methods that would not be 
advisable. [34] 
There is no standardized parameter to define adequate adherence, it must be defined and delineated 
by the objectives of the research.  [34] 
For this study, a minimum adherence of 90% (per subject's diary) will be necessary to meet the 
objectives of the research. Therefore, subjects with adherence of less than 90% will not be 
considered for the evaluation of exploratory variables but will be considered for the safety and 
tolerability variables in the intention-to-treat population group.  
6.5 Strategies to improve adherence  
1. The PI will sensitize the subject to the importance, to achieve the objectives of the study, 
of the correct application of the concomitant treatment.  
2. Direct questioning by the PI about the application of the concomitant treatment.  
3. Delivery of a printed calendar specifying the date of the visit and its activities.   
If deemed necessary, text messages may be sent as reminders. The content of these messages must 
be approved in advance by the CEI.  
 

7. Study methods and procedures  
7.1 Research center  
This study will be carried out in a research center previously evaluated by the sponsor. The center 
will be an institution or establishment where health research is carried out that complies with current 
regulations.  
The research center will be responsible for forming a multidisciplinary research team to execute the 
clinical study according to the protocol. It is their prerogative to design the organization and select 
the personnel who will perform the functions. However, it is necessary for the sponsor to sponsor 
that the PI and sub-investigator be ophthalmology specialists.  
Any person who is designated, under the responsibility of the PI, as a part of the study monitoring 
(sub-investigator, nurse, etc.) or a specific function of participation in the study (pharmacist, 
administrative assistant, study coordinator, etc.) must appear in the "Delegation of Responsibilities".   
The competence and training of any person who has direct participation in the activities of the study 
must be verified prior to the performance of any activity related to the protocol. The above must be 
recorded and the documents that constitute evidence of this competence and/or training must be 
kept in the master file of the study. The competence and training of the staff who have functions in 
the study, both at the central level and in the study center, is the responsibility of the sponsor.  
The sponsor must ensure that all study site personnel participating in the study are adequately 
trained on the study (research protocol, investigator's manual, amendments, standard operating 
procedures, etc.) and on ICH Good Clinical Practice, prior to the start of their participation in the 
study. Training must be recorded in writing, and those records must be filed in the master record of 
the study.  
7.2 Clinical Study Registration  
This clinical study will be registered by the sponsor in public clinical trial registries before its start 
(inclusion of the first subject): National Registry of Clinical Trials (RNEC) of the Federal Commission 
for the Protection against Health Risks (COFEPRIS) and in a WHO primary registry platform. WHO 
Primary Registries meet specific criteria for content, quality and validity, accessibility, unique 
identification, technical capacity and administration. WHO Primary Registries meet the requirements 
of the International Committee of Medical Journal Editors (ICMJE).  
 
 

7.3 Assignment of treatment 
- 
It is a non-comparative, open-label study. All patients will receive the same treatment. 
7.4 Outcome variables  
7.4.1 Primary outcome variables  
− 
Incidence of unexpected PI-related AEs. 
− 
Incidence of IP-related photophobia. 
7.4.2 Secondary outcome variables  
− 
Changes in pupillary diameter. 
− 
Incidence of expected PI-related AEs. 
− 
Changes in visual acuity. 
7.4.3 Exploratory variables 
− 
Changes in the AVMC. 
− 
Intraocular pressure (IOP) changes. 
− 
Corneal and conjunctival staining changes with fluorescein and lysmine green. 
− 
Changes in vital signs (heart rate and blood pressure). 
− 
Changes in OCI. 
7.4.4 Definition of variables, methods and scales for their measurement 
Variable 
Conceptual Definition 
Operational 
Definition 
Measurement 
Type 
Reference 
value 
Statistical 
test 
Incidence of 
unexpected-
IP-related AEs 
Any adverse medical event 
that occurs in the clinical 
research subject to whom 
the PI was administered 
that has a certain causal 
relationship with the PI 
and is not expected.  [36] 
All AEs (according to 
the protocol) in which 
the PI suspects a 
causal relationship 
with the PI will be 
reported. 
• Discrete 
quantitative 
• Nominal 
qualitative 
< 10% 
• U for Mann 
Whitney 
• Pearson X2 
or Fisher 
Exact* 
Photophobia 
incidence 
Expected-PI-related AE  
Present / Absent 
• Discrete 
quantitative 
• Nominal 
qualitative 
< 15% 
• U for Mann 
Whitney 
• Pearson X2 
or Fisher 
Exact* 
Pupillary 
diameter  
The natural pupil of the 
human eye is usually 
approximately circular, for 
a given subject, the 
pattern of aberrations, 
diffraction, depth of field 
and retinal illumination 
depends on the pupillary 
diameter, which in turn 
The pupillary diameter 
results from the 
balance between the 
pupil sphincter muscle 
and the radial fibers of 
the iris that have only 
autonomic 
innervation.  [37] 
It will be evaluated 
through the OPD scan. 
Continuous 
quantitative 
2 – 6 mm 
Kolmogorov-
Smirnov or 
Mann 
Whitney U* 

Variable 
Conceptual Definition 
Operational 
Definition 
Measurement 
Type 
Reference 
value 
Statistical 
test 
varies depending on the 
ambient lighting. 
Incidence of 
AD  
Any adverse medical event 
that occurs in the clinical 
research subject to whom 
the PI was administered.  
[36] 
All AEs (according to 
the protocol) will be 
reported and not just 
those where the PI 
suspects a causal 
relationship with the 
PI 
• Discrete 
quantitative 
• Nominal 
qualitative 
• Incidence 
• Seriousness 
(severity) 
• Causality 
 
• U for Mann 
Whitney 
• Pearson X2 
or Fisher 
Exact* 
 
Changes in 
the AVMCc 
Clinical assessment of near 
visual acuity allows visual 
ability to distinguish 
letters to be assessed.  
It will be evaluated by 
means of the ETDRS 
booklet for near vision 
(annex16.1 ETDRS 
Primer for Near 
Vision), using the font 
size in logMAR. 
Continuous 
quantitative 
0.1 or better 
Kolmogorov-
Smirnov or 
Mann 
Whitney U* 
Changes in 
the AVMC 
Visual spatial acuity is the 
ability to distinguish 
separate elements of an 
object and identify them. 
It is quantified as the 
minimum angle of 
separation (located at the 
nodal point of the eye) 
between two objects that 
allows them to be 
perceived as separate 
objects. 
Snellen Primer on 
logMAR 
Continuous 
quantitative 
0.2 or better  
Kolmogorov-
Smirnov or 
Mann 
Whitney U* 
Intraocular 
Pressure 
Tonometry is the objective 
measurement of IOP, 
based on the force 
required to flatten the 
cornea or the degree of 
corneal indentation 
produced by a fixed force.  
By means of 
Goldmann tonometry 
based on the Imberk-
Fick principle. 
Discrete 
quantitative 
10 – 21 mmHg 
Kolmogorov-
Smirnov or 
Mann 
Whitney U* 
Changes in 
ocular surface 
staining 
Detection of epithelial 
defects in the cornea and 
conjunctiva. 
Direct observation 
with a slit lamp will be 
graded according to 
SICCA (see annex 16.2 
SICCA Eye Staining 
Rating (modified from 
Whitcher et al, 2010) 
[38]. 
Discrete 
quantitative 
0 to 12 
Kolmogorov-
Smirnov or 
Mann 
Whitney U* 
Changes in 
vital signs 
 
It includes the 
measurement of HR, SBP 
and DBP. They are clinical 
parameters that reflect 
the physiological state of 
the organism. 
HR, SBP and DBP will 
be measured with an 
electronic blood 
pressure monitor or 
manually.  
Discrete 
quantitative 
HR: 60 – 100 
bpm 
SBP: 120 – 139 
mmHg 
DBT: 80 – 89 
mmHg 
Kolmogorov-
Smirnov or 
Mann 
Whitney U* 
OCI Score 
The OCI is a questionnaire 
designed to measure 
irritation of the ocular 
surface, it assesses 
symptoms focused on the 
comfort associated with 
ocular surface alterations. 
The evaluator will 
apply the 
questionnaire to the 
subject and will allow 
the subject to answer 
it calmly without any 
pressure and/or 
Discrete 
quantitative 
0 – 100 
Kolmogorov-
Smirnov or 
Mann 
Whitney U* 

Variable 
Conceptual Definition 
Operational 
Definition 
Measurement 
Type 
Reference 
value 
Statistical 
test 
Elevated values indicate 
more severe symptoms. 
coercion (see annex 
16.3 Ocular Comfort 
Index (OCI)  
Abbreviations: AVMC, best corrected visual acuity; CVA, near best corrected visual acuity; AE, adverse event; HR, heart rate; 
OCI, ocular comfort index; PI, principal investigator; bpm, beats per minute; DBP, diastolic blood pressure; SBP, systolic blood 
pressure; IOP, intraocular pressure; rpm, breaths per minute; TF, fluorescein staining; IP; principal investigator; X2, Chi-square. 
*When applicable. 
7.4.5 Description of the variables, methods and scales for their measurement 
For clinical purposes, ophthalmological evaluations will be carried out in both eyes, and their record 
will be recorded in the clinical file. However, to obtain results and comply with the assumption of 
independence within the variables, only examinations and qualifications of the right eye will be 
recorded in the eCRF when appropriate. Next, the description of the variables is made, which are not 
in order of execution.  
7.4.5.1 Intraocular pressure  
Tonometry is the objective measurement of IOP, based primarily on the force required to flatten the 
cornea or the degree of corneal indentation produced by a fixed force. Goldman tonometry is based 
on the Imbert-Fick principle. [39] 
The tonometry will be performed, after instillation of the topical anesthetic, with fluorescein and the 
use of the cobalt blue filter (after the evaluation of the surface staining). 2 samples will be taken, and 
the average will be calculated, which will be recorded in the clinical file. The average will be recorded 
in the eCRF.  
Management as AE: IOP peaks ≥24mmHg should be reported as AEs. 
7.4.5.2 Heart rate 
Heart rate is the number of times the heart ventricles contract per unit of time, usually per minute. 
The heart rate measurement will be done by direct auscultation in the chest with a stethoscope. In 
the case of measuring, it in women, it may be allowed to measure it on the pulse of the wrist. 
However, the method chosen for the patient will be the same to be used on all visits.  [40] 
For the measurement of the heart rate, it will be necessary for the patient to be calm and in a state 
of rest. You will be asked to rest for at least 20 minutes upon arrival at the clinic, before your 
measurement, and when you are due to be assessed after the application of the medication, you 
should also have a 20-minute rest period.  
For protocol issues, we refer to rest or rest as the state where the patient does not have agitation 
because of unnecessary physical effort. The patient should be seated for most of the rest of the 
period, being able to get up to walk to go to the bathroom. 

Management as AE: Patients with heart rates greater than 100 bpm or less than 60 bpm.  
7.4.5.3 Systemic Blood Pressure 
It is the force exerted by blood circulation on the walls of the arteries. Blood pressure is assessed 
with two measurements: systolic (measured when the heart beats and is when the pressure is at the 
highest) and diastolic (measured between beats, when the pressure is at the lowest point). When it 
is reported, the systolic is written first and then the diastolic. [41] 
A calibrated aerobic or mercury sphygmomanometer and stethoscope will be used to measure it. 
The left arm should be used for the measurement, and it should be done at rest.  
Management as AE: Patients with systolic blood pressure of 140 mmHg or higher and diastolic blood 
pressure of 90 mmHg or higher.  
7.4.5.4 Photophobia 
Photophobia is a sensory disturbance caused by light, it is considered a common symptom seen in 
neurological and ophthalmological conditions. According to Albilali (2018) it can be induced by glare; 
It was described as "uncomfortable vision, based on the diffusion of light through the ocular means 
or on a transient or permanent lack of adaptation." It involves an uncomfortable sensation of 
excessive brightness without pain, and occurs mainly with eye diseases, such as toxic, amblyopia, 
albinism, rod monochromatism, and corneal, lenticular, and vitreous opacities. In this study, it may 
occur secondary to pharmacological mydriasis generated using atropine sulfate. [42] 
Photophobia will be assessed within the adverse event interrogation according to the 8. Evaluation 
and management of adverse events, without questioning in a directed way within visit 1, final visit 
and security call. It will be reported as present or absent.  
7.4.5.5 Pupillary diameter 
It is the hole in the middle of the iris through which light penetrates the eyeball. Its diameter results 
from the balance between the pupil sphincter muscle and the radial fibers of the iris (pupil dilator). 
Normal values are 2-6 mm. [37] 
In higher concentrations this product is mydriatic, however, due to the nature of the research and 
the physicochemical characteristics this reaction is not expected to occur, we will consider as 
significant the change that is generated in the pupil of the research subjects > 2 mm (pharmacological 
mydriasis), whether this change in photopic or mesopic vision. The method to perform the 
measurement will be using the following tool: OPD-Scan III (NIDEK CO., LTD, Tokyo, Japan). It is 
requested that the tool used be the same for all patients of the research center and for all their visits. 
It is necessary that the measurement is always done in the same office, with the same artificial light 
conditions and that the patient has at least 5 minutes inside the place before the measurement.  

If there is a change greater than 2 mm between baseline and any of the other measurements, the 
product will be considered to have produced pharmacological mydriasis. This change will also be 
considered an adverse event.  
7.4.5.6 Best-corrected visual acuity 
Visual acuity (VA) is a test of visual function. Visual spatial acuity is the ability to distinguish separate 
elements of an object and identify them. It is quantified as the minimum angle of separation (located 
at the nodal point of the eye) between two objects that allow them to be perceived as separate 
objects.  
Snellen's notation is described as the distance at which the test is performed, divided by the distance 
at which the letter is vertically equivalent to 5 arc minutes. Thus, at 6 meters a letter 6/6 (20/20) is 
equivalent to 5 minutes of arc, a letter 6/12 (20/40) is equivalent to 10 minutes, and a letter 6/60 
(20/200) is equivalent to 50 minutes. The Snellen fraction can also be expressed as a decimal place 
(i.e., 20/20 = 1 and 20/40 = 0.5). [34]   
VA will be evaluated at baseline, without refractive correction with the Snellen chart. It will be in a 
place with adequate lighting (natural or artificial) and at 3m from the subject to be evaluated. Visual 
acuity of the right eye will be taken initially, asking the subject to hold both eyes open and using an 
occlude to cover the contralateral eye; the subject will read aloud the lines that the evaluator points 
out, the line of smaller letters that he can see will be noted by the evaluator in fraction as the AV of 
OD in the clinical record and the eCRF, then the same evaluation will be performed in the left eye 
and its record only in the clinical file. 
Next, the subject's best refractive correction (obtained by autokeratorefractometer and subjective 
tests) will be evaluated and the examination will be repeated using the refraction obtained. This 
result will be reported as best-corrected visual acuity, both data will be reported for statistical 
purposes in logarithmic figures (LogMAR) and in eCRF.  
A fraction conversion table is attached to LogMAR. (see annex 16.4 Fraction to LogMAR conversion 
table. ) 
Management as AE: A decrease in 2 or more lines of sight in BCVA compared to that obtained at 
baseline will be considered an adverse event. 
7.4.5.7 Best Corrected Visual Acuity Near 
Visual Acuity (VA) is the characteristic of vision that determines the ability to detect, discriminate, 
and recognize objects on a background at a close distance. It assesses macular function and provides 
information on the accuracy of retinal focus, as well as the integrity of the neurological elements of 
the eye and the interpretive ability of the brain.  
Near visual acuity (NVA) will be assessed, without refractive correction with the ETDRS chart. It will 
be in a place with adequate lighting (natural or artificial) and at 35 to 40 cm from the subject to be 

evaluated. Visual acuity of the right eye will be taken initially, asking the subject to hold both eyes 
open and using an occluder to cover the contralateral eye; the subject will read aloud the lines that 
the evaluator points out, the line of smaller letters that he can see will be noted by the evaluator in 
fraction as the AVc of OD in the clinical record and the eCRF, then the same evaluation will be 
performed on the left eye and its record only in the clinical file. 
Next, the subject's best refractive correction (obtained by autokeratorefractometer and subjective 
tests) will be evaluated and the examination will be repeated using the refraction obtained. This result 
will be reported as best corrected near visual acuity (CVA), both data will be reported for statistical 
purposes in logarithmic figures (LogMAR) and in the eCRF. It is necessary that the measurement is 
always done in the same office, with the same light conditions. 
A fraction conversion table is attached to LogMAR. (see annex 16.4 Fraction to LogMAR conversion 
table. ) 
Management as AE: A decrease in 2 or more lines of sight in the CVVA compared to that obtained 
at baseline will be considered an adverse event 
7.4.5.8 SICCA Ocular Surface Staining 
The sequence of staining of the ocular surface is important for its reproduction and accuracy. The 
application of fluorescein should be done before the application of lysmine green. 
Fluorescein (TF) staining: A drop of topical anesthetic will be instilled in the bottom of the conjunctival 
sac, then a second drop will be applied to the tip of the fluorescein strip, allowing it to sit on the strip 
for 5 seconds to elute the dye, shaking off the excess at the end. A small contact of the strip is made 
with the conjunctiva at the fundus of the temporal sac, while the patient looks upwards, without 
damaging the conjunctiva. After evaluating the TRPL, between 4 and 8 minutes after the instillation 
of the stain, the evaluation of the fluorescein stain in the cornea is performed, with the slit lamp with 
a cobalt blue filter. It will be graded according to the Sjögren International Clinical Collaboration 
Alliance (SICCA) Eye Staining Rating (CTO).  [43] 
According to the CTO, grade 0 corresponds to the absence of punctate epithelial erosions (PES); grade 
1 is defined as the presence of 1-5 EEP; grade 2 corresponds to 6-30 EEP; and >30 EEP will be classified 
as grade 3. Additionally, one rating point will be added if: 1) EEP is present in the central portion of 
the cornea with a diameter of 4mm; 2) filaments are observed and 3) confluent staining patches are 
observed, including linear stains. See Figure 2.  [43] 
The PI will record in the file and the eCFR the grade granted for the corneal staining of OD and OS, 
respectively. The maximum score per eye is 6. 
Lysmine green (TVL) staining: After revision with fluorescein, a drop of saline solution will be applied 
to the tip of the lysmine green strip allowing it to sit on the strip for 5 seconds to elute the dye. A 
drop of the strip is instilled at the bottom of the temporal sac, while the patient looks upwards, 

without damaging the conjunctiva. The patient may be asked to blink repeatedly to prevent 
accumulations in the conjunctival folds. The examination should be done between 1 and 4 minutes 
after installation through a neutral density filter or with the red-free filter. It will be graded according 
to the CTO of the SICCA. [43] 
In CTO, grade 0 is defined as the presence of 0 to 9 spots of lysmine green staining in the 
interpalpebral bulbar conjunctiva (grading the temporal and nasal portion separately); grade 1 is 
defined by the presence of 10 to 32 points; grade 2 by 33 to 100; and grade 3 for >100 points. Due 
to the difficulty in counting individual points in a moving eye, any area ≥ 4mm2 of confluent points is 
considered >100 points. See Figure 2. [43] 
The PI will record, in the file and in the eCFR, the sum of the grades given to the temporal and nasal 
portion for the OD, then the same evaluation will be performed in the OI and its record only in the 
clinical file. The maximum score per eye is 6. 
Green Lysmine 
(conjunctiva only)
Fluorescein 
(cornea only) 
Grade       Points 
Grade       Points 
0-9 
1 
10-32 
2 
33-100 
3 
>100 
0 
0 
1 
1-5 
2 
6-30 
3 
>30 
Score by staining pattern: 
Additional points for fluorescein 
staining only: 
+1 if there are confluent staining 
patches 
+1 if there is staining in the pupillary 
area 

Ocular surface staining by SICCA assessment is determined by the sum of corneal fluorescein staining 
and conjunctival staining with lysmine green. As seen in Figure 2. The values that can be obtained for 
each eye are from 0 to 12, and abnormal staining is considered when the value is 3 or greater. 
Management as an EA: It will be considered an adverse event when the grade is greater than or equal 
to 3 points.  
7.4.5.9 Adverse events 
As described in the 8. Evaluation and management of adverse events An adverse event is defined as 
any unfavorable medical occurrence in a subject to whom an investigational product is administered, 
regardless of causal attribution.  
Adverse event management shall be carried out as described in section 8. Evaluation and 
management of adverse events and incidents.  
The Principal Investigator will record in the corresponding section of the eCRF the adverse events 
that the study subjects may present in addition to referring them in the clinical record.  
For an adequate evaluation of adverse events, in addition to directed questioning, it is necessary to 
perform a comprehensive ophthalmological evaluation at each visit, which consists of 
ophthalmological examination of the eyelids and adnexa; anterior and posterior segment that is 
performed in a routine ophthalmological check-up, whose procedures are not specifically included 
in the study variables. Posterior pole evaluation can be with direct or indirect ophthalmoscopy, with 
or without pharmacological mydriasis, at the discretion of the PI. An assessment of the fundus will 
be carried out in search of abnormalities that alter the result of the study. IOP will be measured in 
this evaluation, with the PI choice instrument, it should be measured after the evaluation of stains. 
The result of the assessment will be recorded in the clinical file. In the eCRF, only findings that are 
considered so by the Principal Investigator will be reported as an adverse event.  
The adverse events that can occur with PRO-201 according to what is reported in the literature are: 
photophobia, mydriasis, difficulty reading, ocular allergic reactions (atopic dermatitis, allergic 
conjunctivitis and other allergic reactions), eye irritation, eye burning, eye pain, tearing, foreign body 
sensation, dry eye, hyperemia and/or red eye, blurred vision, eye itching; Systemic adverse events 
include: headache, drowsiness, dizziness, tachycardia, flushing, restlessness, irritability, dry skin, dry 
mouth, skin rash and decreased state of alert.   [8]  [6] [18] [19] [20] [21] [22] [23] [24] 
7.4.5.10 Respiratory rate 
It is the measurement of the breaths per minute that the subjects have per research. It is discreetly 
evaluated by observing, without the subject being aware of it, the movements of the rib cage, 
counting as one breath each increase/elevation of the subject's rib cage. 12 to 24 breaths per minute 
is considered normal.  

Management as AE: Any value outside of the normal will be considered an adverse event. It is 
possible that in some cases the research subject should be considered as a possible case of a patient 
with COVID-19, and the investigator should refer the patient to medical attention and withdraw the 
subject from the investigation.  
7.4.5.11 Body temperature 
It is the degree or thermal level of a body. Normal values range from 35°C to 37.4°C.  It can be taken 
by a contact thermometer or not, in different parts of the body. In the study, any thermometer 
calibrated at the research center may be used, if the same one is used for all patients and for all 
their visits.  
Management as AE: Any value outside of the normal will be considered an adverse event. It is 
possible that in some cases the research subject should be considered as a possible case of a patient 
with COVID-19, and the investigator should refer the patient to medical attention and withdraw the 
subject from the investigation. 
7.4.5.12 Ocular comfort index 
It is a questionnaire designed to measure ocular surface irritation with Rasch analysis to produce 
estimates on a linear interval scale (ratings: 0-100). Like the index for ocular surface diseases, the 
ocular comfort index (OCI) assesses symptoms. The OCI contains 8 items (one positive and eight 
negative) that focus on discomfort associated with ocular surface alterations. Each of these questions 
has two parts, which separately inquire about the frequency and severity of symptoms.  [44]See 
annex 16.3 Ocular Comfort Index (OCI) 
At the baseline visit and final visit, the evaluator will deliver the questionnaire to the subject and allow 
the subject to answer it calmly without any pressure and/or coercion, only assisting him if he has 
difficulty understanding any of the questions. 
7.5 Description of procedures or assessments during the study 
The different procedures that will be done during the study are described below. The list may not be 
in order, and it may be arranged in the most optimal way according to the needs of the research 
center. 
7.5.1 Signing of informed consent 
Procedure by which it is guaranteed that the subject under investigation has voluntarily expressed 
his intention to participate in this research, after having understood the information that has been 
given to him about the objectives of the research, benefits, discomforts, and possible risks. 

7.5.2 Preparation of medical history (includes ophthalmological and general medical history) 
It must be carried out in accordance with the provisions of NOM-004-SSA3-2012 of the clinical file. 
Considering that the clinical record is the unique set of information and personal data of a patient, 
which is integrated into any type of establishment for medical care, whether public, social or private, 
which consists of written, graphic, imaging, electronic, magnetic, electromagnetic, optical, magneto-
optical and any other type of documents, in which,  Health personnel must make the records, 
annotations, where appropriate, certificates and certifications corresponding to their intervention in 
the patient's medical care, in accordance with the applicable legal provisions. [45] 
And it will consist of: 
• 
Medical history: It must be prepared by medical personnel and other professionals in the 
health area, in accordance with the specific information needs of each of them, it must have, 
in the order indicated, the following sections: 
- 
Interrogation. - It must have at least: identification form, if applicable, ethnic group, 
hereditary-family history, pathological and non-pathological personal history, current 
condition (inquire about previous conventional, alternative and traditional treatments) 
and interrogation by devices and systems (especially ophthalmological). 
- 
Physical exam. - You must have at least: vital signs (temperature, blood pressure, heart 
and respiratory rate), weight and height (when applicable). 
- 
Previous and current results of laboratory, cabinet and other studies (when applicable). 
- 
Diagnoses or clinical problems (when applicable). 
- 
Prognosis (when applicable). 
- 
Therapeutic indication (when applicable). 
7.5.3 Eligibility Criteria 
It is the assessment that the patient meets all the inclusion criteria and none of the exclusion or 
elimination criteria. 
7.5.4 Assigning Subject Code 
It is the granting of a code that represents the subject in the study. This code is assigned when the 
research subject is included in the study. 
7.5.5 Adverse events 
Described in the 8. Evaluation and management of adverse events 
7.5.6 Vital Signs Measurement 
It is the measurement of heart rate, respiratory rate, blood pressure, and body temperature. These 
measurements can be made with a stethoscope, sphygmomanometer and mercury or digital 
thermometer. 

Vital signs are described in the following sections: 7.4.5.2 Heart rate,7.4.5.3 Systemic Blood Pressure, 
7.4.5.10 Respiratory rate and 7.4.5.11 Body temperature 
7.5.7 Urine Pregnancy Test 
The pregnancy test will be performed at the baseline visit and at the final visit. The Researcher must 
provide the research subject with a female, of reproductive age (there is no natural or induced 
menopause, defined as 12 consecutive months of amenorrhea). To perform the test, the patient will 
be allowed to go to the bathroom, have privacy for the performance of the test, and after the test is 
performed, the Investigator must corroborate the result by observing the medical device.  [46] 
7.5.8 Ophthalmological evaluation 
Evaluation of your eyeball, eyelids, eyelashes, and other structures of your eyes by inspection, slit 
lamp (biomicroscopy), as well as palpation (touch). As part of this evaluation, visual acuity is taken 
with better corrected visual acuity, the integrity of the ocular surface is assessed, the anterior 
segment, intraocular pressure is taken, gonioscopy is assessed, and posterior segment (fundoscopy) 
is assessed. 
7.5.8.1 Best-corrected visual acuity 
Described in the 7.4.5.6 Best-corrected visual acuity 
7.5.8.2 Best Corrected Visual Acuity Near 
Described in the 7.4.5.7 Best Corrected Visual Acuity Near 
7.5.8.3 Ocular Surface Integrity 
This will be done by means of biomicroscopy using the slit lamp of the research center. An inspection 
of the cornea, conjunctiva, tear film, and eye stains with lysmine green and fluorescein will be carried 
out (in the section 7.4.5.8 SICCA Ocular Surface Staining stains are described.) 
7.5.8.4 Previous Segment 
It is the evaluation of the structures of the anterior segment [cornea, conjunctiva, anterior chamber, 
iris, pupil (includes measurement of pupillary diameter), lens, aqueous humor]. The evaluation will 
be made by means of the slit lamp. 
7.5.8.5 Intraocular pressure 
Described in the 7.4.5.1 Intraocular pressure  

7.5.8.6 Gonioscopy 
It is an assessment of the iridotrabecular angle by attaching a gonioscopy lens to the patient's cornea. 
In some cases, the lens will be filled with a high viscosity solution (2% hypromellose) to improve the 
coupling of it with your cornea.  
For the classification of the angle, the Shaffer system will be used. 
Board 2. Shaffer's classification 
DEGREE 
ANGULAR OPENING 
DESCRIPTION 
RISK OF OCCLUDING 
4 
45°-35° 
Open 
Impossible 
3 
35°-20° 
Open 
Impossible 
2 
20° 
Narrow 
Possible 
1 
≤ 10° 
Extremely narrow 
Likely 
0 
0° 
Closed 
Occluded 
 
Angular closure or suspected angular closure shall be Grade 2 or less in more than 180° of the angular 
circumferences.  
7.5.8.7 Posterior Segment (Fundoscopy) 
Also called ophthalmoscopy. This is the test done with a light and a magnifying glass to look at the 
fundus (optic nerve and retina) through the pupil. Sometimes it will be necessary to dilate the pupil 
so that the fundus of the eye can be evaluated much better.  
7.5.9 Application of Medications During Visits 
During patient visits, different medications or medical devices may be applied to the ocular surface 
for ophthalmological examination.  
Examples:  
• 
Tetracaine 0.5% ophthalmic solution, used to anesthetize the patient and facilitate the 
measurement of intraocular pressure, application of dyes for staining and placement of the 
lens for gonioscopy. 
• 
Ophthalmic solution of tropicamide 0.8% and phenylephrine 5%, used to dilate the pupil and 
better assess the posterior segment.  
• 
Hypromellose ophthalmic solution 2%, may be used by the Investigator for the assessment 
of gonioscopy.  

7.5.10 Delivery of material for the subject 
It refers to the delivery of the subject's identification card and the subject's diary. The ID card will 
serve as an ID with the treatment assignment number; this card may serve as an appointment card. 
The Subject's Diary serves to keep track of the times the treatment is applied. 
7.5.11 Delivery of Study Medication 
Once the patient is in the study, the investigational product will be given, and the dosage of the 
product will be explained (one drop once a day at night in both eyes). 
7.5.12 Evaluation of concomitant medicinal products 
It is the question about the use of medications that they are taking regularly or that they have used 
in the last month. If you have required injected eye drug therapy, you should ask about the injected 
drugs in the last 6 months.   
7.5.13 Assessment of adherence to treatment 
It refers to indirectly evaluating the number of applications during the period between visits. To 
assess the approximate number of drops, the Subject's Journal can be reviewed to find out the 
applications registered. 
7.5.14 Return of Study Medication 
It refers to the return made by the research subject of the product under investigation.  
7.5.15 Subject Journal Removal 
It refers to the delivery made by the research subject of the subject's diary.  
 
 
 
 
 
 
 
 
 
 

7.6 Study Diagram and Timeline 
7.6.1 Study diagram 
 
 
Figure 2. Study diagram 
BV= Basal visit  
 
SC = Safety Call  
VF= Final visit. Evaluation of outcome variables                           QD= Once a day                  
V1 = Visit 1 
                                                                             AO= Both eyes  
Obtaining informed 
consent. Compliance with 
all inclusion criteria and 
none of exclusion criteria. 
 Obtaining baseline 
values
Clinical Evaluations 
V1 (Day 7)
VB 
VF 
D 16 (±1) 
V1 
(
)
Starting the PRO-
201 Application 
1 drop QD AO 
6 days after baseline
Clinical Evaluations 
VF (day 16)
13 days after baseline 
 20 days after 
baseline visit 
Security Call (Day 21) 
End of application (day 
  
D 14 
SC 
D 21 (± 1) 

7.6.2 Study timeline 
Board 3. Study timeline 
Procedures 
VB 
D 1 
V1 
D 7 ± 1 
VF 
D 16 ± 1 
LLS 
D 21 ± 1 
FCI Signature 
X 
 
 
 
Medical history 
X 
 
 
 
Eligibility Criteria 
X 
 
 
 
Concomitant Drug Evaluation 
X 
X 
X 
 
Urine pregnancy test 
X 
 
X 
 
Vitals 
X 
X 
X 
 
Best-corrected visual acuity  
X 
X 
X 
 
Near Best Corrected Visual Acuity 
X 
X 
X 
 
Pupillary diameter measurement 
X 
X 
X 
 
Evaluation of fluorescein staining and 
lysmine green 
X 
X 
X 
 
PIO 
X 
X 
X 
 
Comprehensive ophthalmological 
evaluation 
X 
X 
X 
 
Pharmacologic mydriasis 
(phenylephrine/tropicamide) 
X 
 
 
 
Anterior chamber angle assessment 
(gonioscopy) 
X 
 
 
 
Evaluating AEs 
X 
X 
X 
X 
Subject and IP code assignment 
X 
 
 
 
Eye comfort index  
X 
 
X 
 
Delivery of the PI and initiation of 
intervention 
X 
 
 
 
Daily subject delivery 
X 
 
 
 
Adherence assessment 
 
X 
X 
 
Return / evaluation of the subject's diary 
 
 
X 
 
Return of PI 
 
 
X 
 
Abbreviations: D, day; AEs, adverse events; FCI, informed consent form; LlS, security call; PI, research 
product; IOP, intraocular pressure; V, visit.  

7.7 Procedures to be carried out per visit 
Listed below are the procedures that will be performed at each visit, which may not show the optimal 
order for the Research Center. The investigator should order it according to his or her needs, and the 
needs of the study and sponsor. 
The measurement of the pupillary diameter should always be done prior to pharmacological 
mydriasis in the case of the baseline visit and prior to the ophthalmological check-up at all visits. In 
the evaluation of adherence to treatment, it can be reviewed in the subject's diary. 
7.7.1 Baseline visit  
- 
Signing of informed consent 
- 
Preparation of general and ophthalmological medical history 
- 
Measurement of vital signs (temperature, blood pressure, heart rate, respiratory rate) 
- 
Concomitant Drug Evaluation 
- 
Pregnancy test (where applicable) 
- 
Best-corrected visual acuity measurement 
- 
Best Corrected Visual Acuity Measurement Near 
- 
Pupillary Diameter Assessment 
- 
Ophthalmological evaluation 
- 
Fundus evaluation under pharmacologic mydriasis 
- 
Evaluating the angle in the anterior chamber 
- 
Fluorescein and Lysmine Green Eye Staining 
- 
Intraocular pressure measurement 
- 
Evaluation of adverse events 
- 
Ocular Comfort Index (OCI) 
- 
Subject Code Assignment and Research Product Delivery 
- 
Submission of the subject's diary 
7.7.2 Visit 1 
- 
Measurement of vital signs (temperature, blood pressure, heart rate, respiratory rate) 
- 
Concomitant Drug Evaluation 
- 
Best-corrected visual acuity measurement 
- 
Best Corrected Visual Acuity Measurement Near 
- 
Ophthalmological evaluation 
- 
Pupillary Diameter Assessment 
- 
Fluorescein and Lysmine Green Eye Staining 
- 
Intraocular pressure measurement 
- 
Evaluation of adverse events 
- 
Adherence Assessment 

7.7.3 Final Visit 
- 
Measurement of vital signs (temperature, blood pressure, heart rate, respiratory rate) 
- 
Concomitant Drug Evaluation 
- 
Pregnancy test (where applicable) 
- 
Best-corrected visual acuity measurement 
- 
Best Corrected Visual Acuity Measurement Near 
- 
Ophthalmological evaluation 
- 
Pupillary Diameter Assessment 
- 
Fluorescein and Lysmine Green Eye Staining 
- 
Intraocular pressure measurement 
- 
Evaluation of adverse events 
- 
Ocular Comfort Index (OCI) 
- 
Adherence Assessment 
- 
Return of the research product 
- 
Return of subject's diary 
7.7.4 Security Call 
- 
Evaluation of adverse events 
7.7.5 Unscheduled follow-up visits 
At the request of the subject or personnel related to the study, unscheduled follow-up visits may be 
carried out for the reporting of adverse events or any situation that warrants it. During these visits, 
all pertinent data on the adverse events reported should be collected and, where appropriate, an 
appropriate management plan should be established.  
7.8 Data collection 
7.8.1 Source documents 
Source documents are all written or printed records derived from automated processes (e.g., 
printouts of laboratory results issued by automated analysis equipment) where the information is 
recorded for the first time, and which is part of the permanent records of the subject's history. 
Examples of source documents are medical history, clinical evolution notes, laboratory reports, 
cabinet study reports, nursing notes, follow-up notes, surgery records, etc.  
The PI is obliged to accept the monitoring of the information related to the study, audits, review by 
ethics and research committees, and inspections by the health authority, this obligation implies 
direct access to the source documents.  

7.8.2 Electronic forms of data collection 
All data related to the protocol will be captured through an electronic case report form (eCRF) by the 
investigation team staff. The data related to the protocol should NOT be captured directly in the eCRF 
but should be transcribed from the corresponding source document. This procedure allows 
monitoring to verify the information captured in the eCRFs. It is the responsibility of the researcher 
to ensure that the information is transcribed into the eCRFs in a correct, complete, and timely 
manner. It is understood that all data captured and sent by the eCRF to data analysis is approved by 
the Researcher.  
7.8.3 Archiving 
The data collected in this database is anonymous (it only stores the subject number together with 
other information of interest). The program used for data capture and storage covers the traceability 
requirements necessary for the execution of clinical studies. The data collected will be stored by the 
sponsor or the clinical research organization designated for this purpose and its storage will have a 
duration of 10 years. The records of assignments of subject number will remain in the participating 
institutions in charge of the PI or his work team and must be kept for at least 5 years.  
 

8. Evaluation and management of adverse events 
8.1 Regulation and regulations on adverse events 
The registration and reporting of adverse events and incidents will be carried out in accordance with 
the guidelines established in NOM-220-SSA1-2016 and the international guidelines ICH E6.   
[36][47][48][49] 
8.2 Definition of Adverse Event 
According to the International Council on Harmonization (ICH), an adverse event (AE) is any 
unfavorable medical appearance in a clinical investigational subject who is administered a 
pharmaceutical product, regardless of causal attribution. [47] [48] [49] 
Therefore, an AE can be any of the following: any unfavorable and unintentional disease, symptom, 
or sign (including an abnormal laboratory finding) that is temporally related to the use of a medical 
product, whether or not it is considered related to such a product; any new illness or exacerbation 
of an existing disease (worsening of the nature, frequency, or severity of a known condition); relapse 
of an intermittent medical condition (e.g., headache) not present at baseline; any deterioration in a 
laboratory value or other clinical test (e.g., electrocardiogram [ECG], x-ray) that is related to 
symptoms or that results in a change in study treatment or concomitant treatment or discontinuation 
of study medication. [47] [48] [49] 
As defined in the previous paragraph, an adverse event is defined as any event that occurs during 
treatment with a drug or device. However, the definition can also be applied as any unwanted event 
that occurs during a clinical trial, including behavioral disorders. [49] 
8.3 Use of adverse events as a study safety variable 
Measuring the safety of the use of PRO-201 is paramount to the study, therefore, it is considered 
important to report any unwanted manifestations or diseases that occur during the study, regardless 
of whether or not the manifestation is considered to be related to the treatment under investigation. 
[49] 
8.4 Definitions relevant to the classification of adverse events  
Severity (serious/non-serious), also called seriousness (serious/non-serious). Serious or serious is 
defined as any event that: results in death, threatens life, requires hospitalization or prolongs 
hospitalization, is a cause of permanent or significant disability or disability, is the cause of alterations 
or malformations in the newborn, other medically important conditions.  
Severity (mild, moderate, or severe). Mild is those that present with minimal symptoms, do not 
require treatment or suspension of the medication; moderate, when they interfere with usual 

activities, without threatening the subject's life, require treatment and may or may not require 
discontinuation of the medication; severe, those that interfere with usual activities and require 
pharmacological treatment and discontinuation of the medication.   [36][47][48] 
Causality. It is the relationship that is assigned between the pharmaceutical product and the adverse 
event: certainly caused by the pharmaceutical product, there is clear evidence of causality, i.e. the 
adverse event reappears with the administration of the pharmaceutical product; probably caused by 
the pharmaceutical product, there is a high suspicion of causality but no direct evidence is available 
or it is considered unnecessary or dangerous, i.e. the reaction disappears when the pharmaceutical 
product is discontinued; possibly caused by the pharmaceutical, there is additional information to 
suggest that the cause may be due to another pharmaceuticals or disease; unlikely to be caused by 
the pharmaceutical product, there is a clear explanation of the origin due to the underlying disease 
or the use of another pharmaceutical product; conditional, there is a lack of data to issue a clear 
causality; non-classifiable, those for which once all possible information has been obtained about the 
adverse event, it remains unclassifiable.     [36] [47] [48] [49] 
8.5 Responsibilities of the researcher  
It is the responsibility of the Investigator to verify AEs through questioning, pertinent physical 
examination, assessment of evolution, as well as appropriate medical and pharmacological 
management; as well as to follow up until the resolution or outcome and definitive discharge of the 
AE, following the definitions determined in national and international regulations. [36] [47] [48] 
In the event of an AE or any event that puts the health and well-being of the subjects at risk, pertinent 
medical care will be provided, either at the research center or will be referred to the Hospital Center 
with the highest resolution power with which the research center has a medical care agreement. The 
PI will notify the sponsor's clinical monitor, according to the times established in national and 
international regulations. In the case of serious adverse events, it will notify the sponsor and record 
the corresponding information in the eCRF and in turn, inform the IRB and IC.   
The attention of the AEs will be carried out according to the event care diagram (see Figure 4. Adverse 
Event Care).  
The sponsor's final report will include the report of adverse events in compliance with current 
national and international regulations. [36] [47] 
If the research subject debuts during their participation in the study with any chronic adverse event, 
such as diabetes or systemic arterial hypertension, they will be referred to the competent health 
professional for chronic treatment. The follow-up and termination of your participation will be in 
accordance with the stipulations of the ICH.  
8.5.1 Recording of adverse events in the electronic case report form  
The adverse event registry considers:  

▪ 
Subject identification information such as: subject number, age, gender, and if applicable 
specify the eye.   
▪ 
Information about the causality of the adverse event, its relationship to investigational 
products, or to another drug related to the study, as appropriate.  
▪ 
Important date information:  
o 
Date on which the adverse event occurs.  
o 
Date on which the Principal Investigator becomes aware of it.  
o 
Date of resolution or outcome, as applicable.   
▪ 
Information on diagnosis and clinical management.   
▪ 
Record the outcome or resolution of the event:   
o 
Recovered/resolved without sequelae 
o 
Recovered/resolved with sequelae  
o 
Not Recovered/Unresolved  
o 
Subject who presented death due to the adverse event  
o 
Subject who presented death and it is judged that the research product may have 
contributed  
o 
Subject who presented death and this is not related to the product under 
investigation 
o 
Unknown  
▪ 
Information about the investigational product or product associated with the Adverse Event, 
Incident, Adverse Incident, AMR or SRAM must be recorded. The information essential for 
registration is the generic name, distinctive name or code of the investigational product or of 
the product associated with the undesirable clinical manifestation; it will also be necessary to 
enter the data concerning the batch number, manufacturing laboratory, expiration date, dose, 
route of administration, start and end dates of administration and/or consumption, reason for 
prescription; according to whether it is an investigational product or drug (protocol in which 
the subject currently participates) or whether it is a drug that the research subject consumes 
for the treatment of underlying concomitant diseases or uses for the management of some 
transitory sign or symptom that does not correspond to the Natural History of the pathology 
that motivated its entry into the research protocol.  
▪ 
Indicate whether the removal of the suspected product (of causing the event) eliminates the 
adverse event. Also indicate if a dose adjustment is made, if the event changes in terms of 
intensity or seriousness, persistence of the reaction. It is important to indicate whether in 
those subjects who are exposed again to the product, which had previously been suspended, 
the AD reappears.   
▪ 
Information regarding concomitant pharmacotherapy. Indicate the generic name, dose, route 
of administration, start and end dates of use, as well as the reason for the prescription, 
regardless of whether it is in accordance with the prescribing information or technical data 

sheet or is used outside the regulations or what has been authorized by the local, national or 
international regulatory entity.  
▪ 
Relevant medical history information. The analysis of the AE considers the information 
previously narrated, despite the clinical context in which this harmful phenomenon occurs in 
the participants of the clinical research protocol, is of special interest, so the information about 
previous conditions, hypersensitivity or allergy phenomena, previous surgical procedures, 
laboratory analyses or cabinet examinations that have been performed on the participant,  
etc., that the researcher deems it appropriate to mention may do so.  
8.5.2 Adverse Event Tracking  
The Principal Investigator will provide care and follow-up of the adverse event presented by the 
participant until its outcome, according to what is referred to in the following section.  
8.5.2.1 Diagnosis against signs or symptoms 
Whenever possible, an AE should be assessed/reported as a diagnosis and not as a sign or 
symptom (e.g., liver failure or hepatitis rather than jaundice, asterixis, and elevated 
transaminases). If a diagnosis cannot be made from the signs or symptoms, then each sign or 
symptom should be recorded as an AE. If a diagnosis is made later, all AEs reported as signs 
or symptoms should be nullified and replaced by a single adverse event based on the 
diagnosis, with an onset date that corresponds to the onset date of the first symptom or sign 
of the eventual diagnosis. 
8.5.2.2 Adverse events secondary to other adverse events 
In general, AEs secondary to other AEs (cascading events or sequelae) should be identified 
according to the primary cause, with the exception of serious events. A medically important 
AE secondary to an AE that is separated in time from the initial event should be reported as 
a separate event in the EA section of the eCRF. For example: 
If vomiting results in mild dehydration without additional treatment in a normal adult, 
vomiting should only be reported on the eCRF. 
If vomiting results in severe dehydration, both events should be reported separately on the 
eCRF. 
If severe gastrointestinal bleeding leads to renal failure, both events should be reported 
separately on the eCRF. 
If a dizziness event leads to a fall and subsequent fracture, then all three events should be 
reported separately in the eCRF. 
If neutropenia is accompanied by infection, both events should be captured independently in 
the eCRF. 
All EAs must register separately if there are concerns regarding the association of events. 

8.5.2.3 Persistent or recurrent adverse events 
A persistent adverse event is one that spreads continuously without resolution between 
different patient assessment points. Such events only need to be registered once in the eCRF. 
The initial severity (intensity or degree) of the event will be recorded at the time of the first AD 
record. If a persistent AE becomes acute, the maximum severity should be recorded in the 
appropriate section of the eCRF. If the event becomes a serious event, it must be reported no 
later than 24 hours after the knowledge of the change in the status of the event. The eCRF 
section should be updated to reflect serious status and will record the date on which the event 
became serious, thus completing all relevant serious EA reporting data. 
A recurrent AE is one that resolves between assessment points and subsequently reappears. 
Each recurrence of the EA must be recorded separately in the eCRF. 
8.5.2.4 Abnormal vital signs 
Not all abnormal vital signs qualify as AE. For an abnormal vital sign to be reported as an AD, 
it must meet any of the following criteria:  
Be accompanied by clinical symptoms. It results in a change in treatment (dose modification, 
treatment interruption, etc.). 
The report on abnormal values should focus on obtaining a diagnosis and not just a 
description of the abnormality. 
8.5.2.5 Death 
Death should be considered an outcome and not an event. The event or condition that caused 
or contributed to the fatal outcome should be recorded as the EA in the eCRF section. 
8.5.2.6 Pre-existing medical conditions 
A pre-existing medical condition should be recorded as AE only if the frequency, severity, or 
characteristics of the condition worsen during the study. 
8.5.2.7 Hospitalization or prolonged hospitalization 
Any AE that results in hospitalization or prolongation of hospitalization must be documented 
or reported as a serious AE with the following exceptions: 
Hospitalization for pre-existing conditions if the following are met: 
Hospitalization was planned before the study. 
8.5.2.8 Pregnancies, miscarriages, and birth defects 
Fertile women should contact their doctor to report any suspected pregnancy during the 
study. A pregnancy report must be issued, and the sponsor must be informed immediately. 

Monitoring of the patient should continue until the outcome of the pregnancy. Pregnancy is 
not in itself an AE. 
Any abortion should be classified as a serious AE. 
Any birth defect or birth defect in a product from a woman who received the study drug should 
be classified as a serious AE. 
8.5.3 Procedures for a serious adverse event  
The process of attention to the adverse event considers the following stages:  
 
Figure 3. Adverse Event Care 
During the development and conduct of this study, undesirable harmful events or adverse 
reactions/incidents, of medical implication, may occur in the research subject, which do not 
necessarily have a causal relationship with the investigational products. These harmful phenomena 
can occur during the use of investigational pharmaceutical products at doses authorized for use in 
humans by a local, national or international regulatory entity. However, it may be suspected that the 
Document 
Attention and  
treatment  
relevant 
Categorization 
Preparation of a medical note 
Registration and reporting in eCRF 
 On-site investigation 
In hospital 1st-3rd level (if 
required) 
  
By outcome: 
Serious/Not Serious 
By severity: 
Mild, moderate, or severe 
Physical examination  
and evaluation of the  
EA 
EA not serious 
EA seriously
Follow up with the 
EA until its outcome 
The 
+ Inform, within 24 hours of 
their knowledge to the sponsor 
+ Follow up on the EA until its 
outcome 
+ Reporting to the IEC 
Notify to: 
+ Study Director 
+ Protocol author 
+ Pharmacovigilance Unit 
The 

investigational product causes some unwanted clinical manifestation. AEs, Incidents, Adverse 
Incidents, ADRs or SRAMs to one or more pharmaceutical products can occur during the systematic 
evaluation of the participants (on the days when the clinical review is scheduled, according to the 
schedule of activities) or suddenly, in such a way that:  
1. The investigator must be the first person to whom the subject notifies that he or she 
has developed or presented any harmful phenomenon of a clinical nature during his or 
her participation in this study.   
2. According to his clinical judgment, Based on the pertinent physical examination, 
interrogation, etc., as well as the analysis of the information available in the medical 
literature and what is referred to in the Investigator's Manual, Information to prescribe 
or Summary of the Comparator Drug Label, the principal investigator determines the 
pertinent care of the harmful event/reaction.  
3. Such care can be in the research center or in the hospital with the greatest resolution 
power. In such a way that, if the subject is sent by the PI to a hospital, he or she attends 
through a referral system. The reference can be with a card that identifies the subject as 
a study participant and links him or her to the pre-established agreement with the 
institution, or through a medical reference note issued by the Principal Investigator. 
Laboratorios Sophia, S.A. de C.V., will pay the expenses for the medical care of the 
participating subject, when the adverse event is associated with or is related to the 
product under investigation.  
4. Taking the clinical information collected, either during the care provided at the 
research center or that provided by the treating physician(s) in the hospital, the PI will 
record the AE in its clinical note, stating the seriousness, intensity (mild, moderate or 
severe) and relationship with the investigational product.   
5. The PI must migrate the relevant data to the eCRF and its respective adverse event 
section. By, in cases of serious adverse events, the clinical monitor of the study must be 
notified within 24 hours after learning it, so that in turn it informs the Clinical Team and 
the UTFLS, and that the CEI/CI is subsequently notified. Non-serious adverse events will 
be recorded and treated appropriately, and the safety profile of the investigational drug 
or PI will be reported to the appropriate regulatory entity in the final report of the clinical 
trial.  
The recording of the outcome of the AE depends substantially on the follow-up that the Principal 
Investigator performs on the subject, since it is expected that most of the harmful phenomena (see 
subsection 2.5 Risk-benefit assessment and consult the Investigator's Manual) are ophthalmic in 
nature, however, there may be systemic alterations. Therefore, in the opinion of the researcher, the 
withdrawal of the participant or their permanence will be considered.  

8.5.4 Causation assessment  
Causality assessment is the methodology used to estimate the probability of attributing the observed 
adverse event to a pharmaceutical product. It considers probabilistic categories according to the 
available evidence and the quality of the information, based on the national pharmacovigilance and 
technovigilance regulations.  [36][50] 
An adverse event may or may not be related to the clinical trial. A causal relationship means that the 
intervention caused (or is reasonably likely to have caused) the adverse event. This usually implies a 
relationship between the time of the intervention and the adverse event (e.g., the adverse event 
occurred shortly after the subject under investigation received the intervention). [49] 
For all adverse events, the Principal Investigator is responsible for examining and evaluating the 
patient to determine the association of the event with the clinical study and the intervention, 
whether it is related to the experimental treatment, concomitant, surgical procedure, or diagnostic 
procedures performed during the study. [49] 
Accepting that the adverse event is related to the clinical study requires a plausible mechanism of 
action—that is, that there is a logical sequence between the event that occurred and the intervention 
that caused it. In some cases, it is helpful to know the opinion of other doctors directly or indirectly 
involved in the research; as well as whether the patient considers that there is a relationship. [49] 
The Pharmacovigilance and Technovigilance Unit of Sophia Laboratories (UFTLS) may use the 
categories of causality described by the Uppsala Monitoring Centre, to categorize the likelihood of 
adverse event to the investigational product or concomitant treatments or treatments used during 
visits: [36] [50] 
• 
Definitive (certain): a clinical event, including alterations in laboratory tests, that manifests 
itself with a plausible temporal sequence in relation to the administration of the drug, and 
that cannot be explained by concurrent disease, or by other drugs or substances. The 
response to drug withdrawal (withdrawal) must be clinically plausible. The event must be 
definitive from a pharmacological or phenomenological point of view, using, if necessary, a 
conclusive re-exposure (challenge) procedure. [36] [50] 
• 
Probable: A clinical event, including alterations in laboratory tests, that manifests itself with 
a reasonable time sequence in relation to the administration of the drug, that is unlikely to 
be attributed to concurrent disease, or to other drugs or substances, and that a clinically 
reasonable response is presented upon withdrawal. It is not necessary to have re-exposure 
(challenge) information to assign this definition [36] [50] 
• 
Possible: A clinical event, including alterations in laboratory tests, that manifests itself with a 
reasonable time sequence in relation to the administration of the medicinal product, but 
which can also be explained by concurrent disease, or by other drugs or substances. 
Information regarding the withdrawal of the drug may be missing or unclear. [36] [50] 

• 
Unlikely: A clinical event, including alterations in laboratory tests, that manifests itself with 
an unlikely temporal sequence in relation to the administration of the drug, and that can be 
more plausibly explained by concurrent disease, or by other drugs or substances. [36] [50] 
• 
Conditional/Unclassified: A clinical event, including alterations in laboratory tests, reported 
as an adverse reaction, for which further data are essential for appropriate evaluation, or 
additional data are under review. [36] [50] 
• 
Non-assessable/Unclassifiable: A notification that suggests an adverse reaction but cannot 
be judged because the information is insufficient or contradictory, and that cannot be 
verified or completed in its data. [36] [50] 
Thus, the degree of certainty to establish the investigational product as the causal agent of the 
harmful phenomenon that occurs to the subject of the clinical study, can be indicated directly by the 
Principal Investigator based on his clinical experience or by applying the categories of causality 
described by the Uppsala Monitoring Centre. It is important that the researcher and the UFTLS 
consider the following arguments in favor of the causal relationship:   
a) 
Strength of association, which refers to the number of cases in relation to those 
exposed.   
b) 
The consistency of the data, i.e. the presence of a common characteristic or pattern.   
c) 
The exposure-effect pattern, which determines the relationship with the site of 
appearance, time, dose and reversibility after deletion.   
d) 
Biological plausibility, which refers to the possible pharmacological or 
pathophysiological mechanisms involved in the development or presentation of the adverse 
event.   
e) 
Experimental findings, e.g., the appearance of abnormal metabolites or high levels 
of the drug or its biotransformation product.   
f) 
Analogy, which refers to the experience gained with other related drugs, adverse 
reactions frequently produced by the same family of pharmacological agents.   
g) 
Nature and characteristics of the data, i.e. objectivity, accuracy and validity of the 
relevant documentation. [51] 
8.6 Unanticipated Issues  
Unanticipated problems (PNA) consider those situations that pose risks to the participating subjects, 
in general, any incident, experience or result that meets all the following criteria:  
• 
Unexpected in terms of its nature, severity, or frequency in relation to: 1) study-related 
documents such as investigator's manual, study protocol, and informed consent form; and 2) the 
characteristics of the population being studied.  
• 
Related or possibly related to your participation in the study (possibly related means that 
there is a reasonable possibility that the incident, or results, was caused by study procedures).  

• 
Indicative that the research places participants at a higher risk of harm (including physical, 
psychological, economic, or social) than previously recognized.  
8.6.1 Reporting Unanticipated Issues 
The PI will be responsible for reporting NAPs to the sponsor, IC and IEC. The report must contain the 
following information:  
• 
Identification of the study: protocol title and number, name of the PI and, if applicable, of 
the center.  
• 
Detailed description of the event, incident, experience, or outcome.  
• 
Explanation, justification of the reasons why the incident represents a NAP.  
• 
Description of changes to the protocol or corrective actions taken or proposed in response 
to the NAP.  
NAPs that are SEA must be reported to the IEC/IC and the sponsor within the first 24 hours of the PI 
becoming aware of it.   
Any other NAPs will be reported to the IEC/IC and the sponsor within the first 5 business days after 
the PI becomes aware of it.  
 

9. Study Monitoring  
The study sponsor is responsible for monitoring the study. Monitoring activities include, but are not 
limited to general safety monitoring, general study quality monitoring, monitoring by study site, 
detection monitoring, reporting and tracking of adverse events, monitoring for resolution of 
discrepancies in data capture, etc.  
Responsibility for monitoring activities and ultimate responsibility for monitoring rests with the 
sponsor.  
The details of the monitoring activities are specified in a separate document from this protocol in a 
Monitoring Plan.  
9.1 Monitoring the Study Site  
The research center participating in the study will be monitored. For each center, at least one start 
visits and one closing visit must be carried out, which does not exclude the carrying out of one or 
more follow-up visits between these two mandatory visits.  
The initial visit must be carried out before the inclusion of the first participant in that center; In it, 
the monitor will verify that the material to be used during the study has been received and that the 
personnel who will participate in the study activities have been trained on the study, as well as verify 
that the regulatory requirements and applicable standard operating procedures are met.  
At the follow-up visit(s), the monitor will conduct a review of the study documents to confirm that 
the applicable research protocol and standard operating procedures are being followed, data 
completion is complete and timely, and that adverse event reports are being conducted 
appropriately. At each visit, the monitor will discuss the findings with the researcher and define the 
actions to be taken.  
The closure visit will take place at the end of the study, once the last site participant has been 
discharged from follow-up. On this visit the monitor will verify that the site has all the necessary 
documents for archiving, that all biological samples have been analyzed, that all IP (used and unused) 
has been returned to the sponsor, and that all unused material has been recovered.  
Details on monitoring are set out in the corresponding plan.  
9.2 Audit and quality control  
To ensure compliance with GCPs and all applicable regulatory requirements, Laboratorios Sophia, 
S.A. de C.V. may conduct quality assurance audits. Regulatory agencies could also conduct a 
regulatory inspection of this study.  
Details about the audit process are set out separately in the Audit Plan.  

9.2.1 Pre-study audit  
The research center included in the study will be subject to a feasibility visit prior to the selection of 
the center, where it will be verified that they meet the minimum requirements indicated by the 
sponsor.  
9.2.2 Audit during the conduct of the study  
They may take place at any time before, during, or after the conclusion of the study. If any audit or 
inspection is conducted, the investigator and the institution shall agree to allow the auditor/inspector 
direct access to all relevant documents and shall allocate their time and that of their staff to the 
auditor/inspector to discuss the findings and any pertinent problems. If the audit has not been 
scheduled by the sponsor, the facility must notify Laboratorios Sophia, S.A. de C.V. immediately.  

10. Sample size calculation and statistical analysis 
10.1 Sample Size Calculation 
The sample size calculation was performed based on the primary objective of the study, to evaluate 
the safety and tolerability of the PRO-201 ophthalmic solution manufactured by Laboratorios Sophia, 
S.A. de C.V. on the ocular surface of ophthalmologically and clinically healthy subjects. To meet this 
objective, it is estimated that 29 subjects will contribute one eye (right eye [OD]) to the study. 
10.1.1 Calculation methodology 
Atropine sulfate 0.01% has sufficient scientific evidence that confirms its efficacy and safety as an 
adjuvant in the treatment to delay the progression of myopia in children. In the meta-analysis carried 
out by Zhao et al (2019), in a sample of 1079 subjects from 7 clinical studies (505 in the 0.01% atropine 
group and 574 in the control), the incidence of AEs in 0.01% atropine sulfate was statistically lower 
compared to the control group [OR=0.26, 95% CI (0.11, 0.61)] . While, in the review by Li et al (2019), 
children who received 0.01% atropine had a better balance between the use of the treatment-
incidence of AD, being well tolerated at this concentration (only 2% had photophobia). On the other 
hand, in the meta-analysis by Wang et al (2020), no study reported severe/serious AEs after 
administration of 0.01% atropine. Finally, the meta-analysis by Huang et al. (2016), in agreement with 
these results, mentions that low-dose atropine (0.01%) is one of the most effective interventions with 
a minimal prevalence of clinical symptoms. Considering this information and the primary objective of 
the study, the following proposal was made for the sample size.   [52, 53, 54, 55, 25] [52] [25][53][54] 
For one-arm clinical studies (single-arm trials) where the primary endpoint depends on the duration 
of the study (e.g., survival rate, AD incidence, etc.), there are options available in the literature for 
sample size calculation. The most common options are the log-rank and assuming exponential 
distribution. Another option for calculation is to use a parametric exact test, following a Chi-square 
distribution.  [56] 
The probability that a subject will experience no improvement during the study is given by: 
 
𝑑= ∫𝐺(𝑡) ∙𝑓1(𝑡)
∞
0
𝑑𝑡 
Where F1(t) is f(t) with θ= θ1 (θ is a scale parameter, and β is the form of the parameter that 
determines the shape of the risk function [if β>1 the risk increases over time, and if β<1 the risk 
decreases over time, when β=1 represents a special case of exponential distribution with constant 
risk]). Dividing the number of events by d given the sample size adjusted for possible losses. [56] 

Considering the OR calculated in the meta-analysis by Zhao et al, and the time of exposure to the 
product (14 days = 0.47 months) and follow-up time (21 days = 0.7 months), we can expect according 
to the simulation performed by Phadnis, a rate of 0.09 AE/subject in the study. Considering this 
incidence rate, in a sample of 24 subjects, 9% could present AE during one month of follow-up, see  
[52] [56]annex 16.5 Number of events/sample size for the exact method vs. the Wu method, for 
different values of the parameter β, cumulative time a, and tracking time f 
 10.1.2 Size Calculation 
The PRO-201 ophthalmic solution is expected to be safe in its ophthalmic application when it has an 
incidence of unexpected, related AEs of less than 10%, and is tolerable when it has an incidence of 
photophobia that is less than 15% of the study population. 
On this basis, a sample of 24 ophthalmologically and clinically healthy subjects was estimated. This 
value was increased by 20% considering possible losses. Based on this consideration, 29 subjects (who 
will contribute one eye, OD) are required to respond to the primary objective of the study. 
10.2 Clinical Data Management 
Clinical Data Management (CDM) Clinical data management), enables the generation of high-quality, 
reliable and statistically valuable data. CDM is the process of collecting, cleaning, and managing the 
information of the subjects of a study in accordance with regulatory standards (guidelines 21 CFR 
Part 11, ICH and GCP). It covers eCFR design, comments on the eCFR, database design, capture (Data 
entry), validation (source document verification, SDV), discrepancy management (You want), medical 
coding (Medical Coding), extraction (Soft lock) and database shutdown (Hard Lock). [57] 
In accordance with roles and responsibilities, multiple users can be created, whose types of access to 
the eCRF can be limited to capture (principal investigator, IP), medical coding, database design, or 
quality control (Quality Check). The handling of discrepancies will be carried out based on the flow in 
Figure 5.[57, 58] 
The following roles will be included in the CDM team:  
• 
Data Manager, 
• 
Database designer/programmer, 
• 
Medical coder, 
• 
Clinical Data Coordinator, 
• 
Quality control, and 
• 
Capturist (data entry associate). [57] 
 
 

 
Figure 4. Management of discrepancies (MN, medical note; CRA, clinical monitor; SEC self-evident 
correction)  
10.3 Statistical methodology 
The statistical analysis plan was developed considering the evaluation criteria described in the study 
protocol. 
The statistical analysis will be carried out by personnel of Laboratorios Sophia, S.A. de C.V. The SPSS 
statistical package version 19.0 for Windows (IBM Corporation, Armonk, NY, USA) will be used. The 
coding will be done using consecutive numbers. The data will be collected and sorted in an Excel 
spreadsheet (Microsoft® Office). The data will then be exported to the SPSS package platform. The 
variables will be categorized according to their nature (see Board 4 Triangulation of concepts 
The statistical tests used will be following the corresponding assumptions for non-parametric 
statistics.  
10.3.1 Population Analysis 
The statistical analysis will be presented to give a general summary of the subjects entered the study 
and a view of the safety and tolerability of its results. The data provided by the research site will be 
summarized for this purpose, according to its nature. The Shapiro-Wilk test will be performed to find 
out if the quantitative data are distributed in a normal way (p>0.05).  

The results of the quantitative variables will be presented in measures of central tendency: mean, 
standard deviation, maximum and minimum. Changes in pupillary diameter, near visual acuity, BCVA 
and OCI will be expressed as continuous variables. Changes in IOP, HR, systemic blood pressure, and 
TF will be expressed as discrete variables. 
The result of the nominal and ordinal qualitative variables will be presented in frequencies, 
proportions and/or percentages. For these, 2 x 2 frequency tables will be built. All percentages will 
be presented with decimal. 
The level of difference to consider significance will be an alpha (α) of 0.05 or less. The triangulation 
between the variable type and the measurements is shown in the Board 4 Triangulation of concepts. 
 
Variable 
Type 
Variable 
A1 
A2 
B1 
C1 
C2 
D1 
D2 
D3 
E1 
E2 
E3 
E4 
E5 
Selection 
A1 
Demographic 
D 
 
 
 
 
 
 
 
 
 
 
 
 
A2 
Medical History/Selection 
Criteria 
 
D 
 
 
 
 
 
 
 
 
 
 
 
Basal 
B1 
Comprehensive 
ophthalmological 
evaluation 
 
DT 
D 
TB 
 
 
TB 
 
 
 
 
 
 
Safety & Tolerability 
C1 
Incidence of AE* 
 
 
 
TB 
 
 
TB 
 
 
 
 
 
 
C2 
Incidence of photophobia 
 
 
 
TB 
TB 
 
TB 
 
 
 
T 
 
 
Secondary Security Outcome 
D1 
Pupillary diameter 
 
 
 
 
 
B 
TB 
 
 
 
 
 
 
D2 
Incidence of AD 
 
 
 
TB 
TB 
 
TB 
TB 
TB 
TB 
TB 
TB 
 
D3 
Near visual acuity 
 
D 
 
 
 
 
TB 
B 
 
 
 
 
 
Exploratory 
E1 
BCAV 
 
D 
 
 
 
 
TB 
 
B 
 
 
 
 
E2 
PIO 
 
D 
 
 
 
 
TB 
 
 
B 
 
 
 
E3 
Corneal and conjunctival 
stains with fluorescein and 
lysmine green 
 
D 
 
 
 
 
TB 
 
 
 
B 
 
 
E4 
Vitals signs 
 
D 
  
 
 
 
TB 
 
 
 
 
B 
 
E5 
OCI 
 
D 
 
 
 
 
 
 
 
 
 
 
B 
Abbreviations: BCVA, best corrected visual acuity; B, bivariate analysis; D, descriptive statistics; AE, adverse event; OCI, 
ocular comfort index; M, multivariate analysis; IOP, intraocular pressure; T, 2x2 contingency table. *Unexpected-related 
AEs. 

10.3.2 Safety and tolerability analysis 
10.3.2.1 Analysis for Primary Variables 
An analysis of safety and tolerability outcomes will be carried out in the safety population, defined as 
any subject who has received at least one dose of the investigational product (PRO-201) regardless 
of adherence to it or adherence to the protocol (intention-to-treat population, ITT). 
The statistical analysis for the primary variables of a quantitative type will be estimated by means of 
the Kolmogorov-Smirnov statistic for the difference in measurements, once the values have been 
adjusted with respect to their baseline within everyone. In cases where applicable, the nonparametric 
Mann-Whitney U test will be used. 
For conjunctival hyperemia, Pearson's X2 or Fisher's exact test will be used at expected values less 
than 5. 
10.3.2.2 Analysis for secondary variables 
For the analysis of these variables, the same primary analysis will be carried out as long as they have 
the necessary measurements to do so.  
10.3.3 Exploratory Analysis 
The analysis of exploratory variables will be carried out in those subjects who conclude their 
participation without deviations from the study protocol (population by protocol, PP). The same 
primary analysis will be carried out if they have the necessary measurements to do so. 
10.4 Procedure for handling missing data 
There is no imputation procedure for missing data. 
 

11. Ethical considerations 
11.1 Approval of the committees 
The present study will be conducted in accordance with the standards of the Declaration of Helsinki, 
World Medical Association 2013. Nuremberg Code; Nuremberg Trial by the International Tribunal of 
Nuremberg, 1947. Belmont Report, National Commission for the Protection of Biomedical and 
Behavioral Research Subjects, 1979.  It will be conducted in accordance with the scientific and 
technical requirements necessary for the registration of medicines for human use by the 
International Council for Harmonisation (ICH) Guide to Good Clinical Practice. Council for 
International Organizations of Medical Sciences (CIOMS, 2002) International Ethical Guidelines for 
Biomedical Research in Human Subjects. Council for International Organizations of Medical Sciences 
(CIOMS, 2008) International Ethical Guidelines for Epidemiological Studies. The Research Ethics 
Committee and the Research Committee will evaluate the protocol before conducting the study and 
will issue their approval or possible modifications for its performance, these Committees must be 
notified of any significant changes to the protocol. In addition to the above, the current regulations 
of the regulatory authority will also be complied with.  
The personnel authorized by the sponsor will submit to evaluation by the Research Ethics 
Committees, Research Committees, and when applicable to the Biosafety Committee the essential 
documentation of the research project: research protocol, informed consent form, researcher's 
manual, subject material, as well as other documents additionally requested, according to local 
requirements, national or international applicable by the regulatory entities.   
The study will not begin at the research center if there are no confidentiality agreements and 
economic proposals from each of the principal investigators, duly signed and without having 
previously obtained the favorable opinion and/or approval of the Research Ethics Committees, 
Research Committees, and when applicable by the Biosafety Committee.  Corresponding.   
The study will not begin without having complied with the relevant local, national or international 
regulatory requirements and without having the corresponding health authorization.  
The study is a study with a greater than minimum risk, in accordance with the Regulations of the 
General Health Law on Health Research, Title Two, Chapter I, Article 17, Section III, published in the 
Official Gazette of January 6, 1987.  
11.2 Amendments to the protocol  
The amendment procedure will be pertinent when there is a need to make any change to a document 
that is part of the research project or protocol, derived from variations in the methodological 
structure, substitution of the principal investigator or in the face of the identification of risks in the 
research subjects. The documents subject to amendment will be protocol, letter of informed 

consent, researcher's manual, documents for the subject, measurement scales and schedule of 
activities.  
Any amendment must be approved by the sponsor and/or the principal investigator, the amended 
document(s), once reviewed and approved by the Research Ethics Committee and the Research 
Committee or, when applicable, by the Biosafety Committee, (entities that issued the initial favorable 
opinion for the conduct of the research) will be sent for authorization by COFEPRIS.   
Amendments that substantially modify the protocol confer an additional or different risk on the 
research subjects must be approved by the Committees. It is the investigator's responsibility to act 
in situations that require immediate action to avoid unnecessary harm to study participants.  
The principal investigator is responsible for communicating to the Research Ethics Committee any 
amendments to the protocol that may eventually affect the rights, safety, or welfare of research 
participants. Likewise, it must inform us of any situation or new knowledge that will show a greater 
risk for the participants, the premature termination or suspension of the study, the reasons and the 
results obtained so far. It must also report on the conclusion of the study upon completion of the 
research protocol.  
11.3 Early Study Termination  
The study may be suspended or terminated prematurely if there is a sufficiently reasonable cause. 
Written notice, documenting the reason for the suspension or early termination, shall be delivered 
by the party enforcing the suspension. The PI should promptly inform the study participants, the IC, 
and the IRC providing the reasons.   
Situations in which study suspension or early termination will be contemplated include, but are not 
limited to:  
1. The presence of serious adverse events in more than 10% of participants in a study 
group.  
2. The regulatory authority (COFEPRIS) considered it for security alerts.  
3. The Sponsor determined it for its convenience or eventualities such as financial 
support, manufacturing errors, etc.  
4. The identification of unexpected risks to the participants, which are significant or 
unacceptable.  
5. Obtaining new relevant safety information.  
6. Insufficient adherence to the requirements of the protocol.  
7. The data obtained is not assessable or is not sufficiently complete.  
8. The determination that the primary objective has been achieved.  
9. The determination of futility.  

In the event of suspension, the study may be resumed once the situations that led to the suspension 
have been corrected; if this justification is sufficient for the sponsor, CI, CEI and regulatory 
authorities.  
11.4 Informed Consent  
The FCI contains complete and understandable information about the study and the investigational 
product, in accordance with the applicable regulations in force and Good Clinical Practice.   
The FCI will be considered as a source document and will be filed as such. The site's principal 
investigator is responsible for ensuring that all new versions of the informed consent are submitted 
to the appropriate approvals (the same as those to which the original informed consent letter was 
submitted) and that the most current approved version is the one presented to the study subjects.  
11.4.1 Obtaining  
Informed consent must be obtained before the subject undergoes any procedure indicated in the 
protocol. For this purpose, a letter of informed consent must be signed.  
Written consent documents will incorporate the elements of informed consent described in the 
Declaration of Helsinki and the ICH Guide to Good Clinical Practice and will follow all applicable laws 
and regulations.  
The PI, or the study staff delegated by him/her, will provide the prospective participant with all the 
information regarding the characteristics of the study, its potential benefits, risks, objectives, and 
procedures.  
This information will be in a language understandable to the subject, it will be explained to the subject 
that he or she has the right to interrupt his or her participation in the study at any stage, without this 
affecting the relationship with the researcher and/or his or her future assistance. Informed consent 
will be put to the consideration of the potential participant; He must have enough time to analyze 
each one of the aspects mentioned above and in case he has any doubts, it will be clarified by the 
person in charge of obtaining the informed consent.   
Once the participant agrees to participate in the study, he/she must sign and date the letter of 
informed consent in the presence of two witnesses who are or are not related to the study subject, 
who will participate during the informed consent process and will sign guarantee that the process 
was carried out prior to any study procedure.  that the information of the study was clearly explained, 
and doubts were clarified if any.  
If a subject is illiterate, acceptance will be with his or her fingerprint, and if the subject is unable to 
give adequate written informed consent, a representative of the "legally authorized" subject may 
provide such consent for the subject in compliance with applicable laws and regulations.  
In the same way, the PI, or the study staff delegated by him, must sign and date this consent.  

The FCI must be signed in duplicate by all those involved, one copy will be filed in the researcher's 
folder, and the other will be given to the participant. The PI or delegated personnel must document 
the process of obtaining the Informed Consent by means of a detailed, precise and contemporary 
medical note, specifying the signed version, the date on which the document was signed and how 
the process was carried out.  
11.4.2 Special considerations  
The procedures that will be performed during the conduct of the study do not pose an additional risk 
that should be considered apart from the procedures listed for informed consent.  
11.4.3 Modifications to informed consent  
Any changes to the FCI constitute an amendment to this document and must be submitted for 
approval to the Research Ethics Committees and COFEPRIS.  
Such amendments may be implemented only after obtaining the written approval of the Research 
Ethics Committee and the Regulatory Entity (as applicable), except for an amendment that is required 
to eliminate an immediate danger to the study subjects.  
A process of re-consent for each subject affected by the amendment must be carried out under the 
same conditions as those described above, to communicate the new information contained in the 
document to them in a timely manner. The subject will be given a signed original of the amendment, 
and the researcher will keep the second original.  
11.5 Confidentiality  
All documents and information provided to the research center by the sponsor are strictly 
confidential. PI expressly agrees that data about your professional and clinical experience, provided 
to the sponsor on paper and stored in electronic form, is solely for use related to your activities with 
the clinical trial sponsor, in accordance with Good Clinical Practice.   
The PI agrees that he and his team members will use the information only within the framework of 
this study, to carry out the protocol. This agreement is binding if the confidential information has not 
been disclosed to the public by the sponsor.   
The clinical study protocol provided to the PI may be used by the PI and his team to obtain the 
informed consent of the subjects for the study. The clinical trial protocol, as well as any information 
taken from it, should not be disclosed to other parties without the written permission of the sponsor.   
The PI will not disclose any information without the prior written consent of Laboratorios Sophia, S.A. 
de C.V., except to the representatives of the Competent Authorities, and only at their request. In the 
latter case, the investigator undertakes to inform Laboratorios Sophia, S.A. de C.V. before disclosing 
the information to these authorities.   
 

The PI will fill out and maintain a record of the selection of the subjects, as well as the identification 
and enrollment list of each of the subjects participating in the study. The investigator agrees to give 
on-site access to the auditor and/or representatives of the Competent Authorities. The information 
will be processed in compliance with professional secrecy.  
In the eCRF and all communications related to study subjects, they will identify them only by the 
study subject's identification number, either by the scrutiny number or the assignment number. The 
information collected in this study will be exchanged between the sponsor and the research center 
and must be treated confidentially. The Health Authority, the CEI, the IC, the sponsor, the 
monitors/auditors and third-party auditors will be the only bodies authorized to review the study 
documentation. If publications arise from this research project, in no case will they contain 
information on the identification of the study subjects. If the results of the study are published, no 
personal information on the study subjects will be revealed.  
The protection of personal data will be carried out in accordance with the corresponding regulations 
in force.  
11.6 Conflict of interest  
The independence of the conduct of the study and its results from any current or perceived external 
influences is critical. For this reason, any current conflict of interest of any person who has a role in 
design, conduct, analysis, publication or any aspect of this study will be declared. Furthermore, those 
who have a perceived conflict of interest will be asked to handle it in a manner appropriate to their 
participation in the study.  
11.6.1 Declaration of Interests  
The PI undertakes to make a declaration of financial interests as well as conflict of interest prior to 
the start of the study.  
11.7 Access to Information  
The final database of the study will be the property of Laboratorios Sophia, S.A. de C.V. and its access 
will be restricted. The PI will not have access to it, unless it has prior written authorization from the 
sponsor.  
The information obtained, which is relevant to the safety of the subjects participating in the study, 
must be immediately shared with the research center, so that it can be notified to the study subjects.  
11.8 Ancillary and post-study care  
Upon completion of the study and closure of adverse events in accordance with section: 8, the 
sponsor shall not extend care to the research subject.  
 

12. Biosecurity aspects 
NO BIOSECURITY IMPLICATIONS  
This protocol, entitled: "Phase I clinical study, to evaluate the safety and tolerability of the PRO-201 
ophthalmic solution, prepared by Laboratorios Sophia, S.A. de C.V. on the ocular surface of 
ophthalmologically and clinically healthy subjects", and number: SOPH201-0521/I has no biosafety 
implications, since infectious and contagious biological material will NOT be used; pathogenic strains 
of bacteria or parasites; viruses of any kind; radioactive material of any kind; genetically modified 
animals and/or cells and/or plants; toxic, dangerous or explosive substances; any other material that 
endangers the health or physical integrity of the research center’s staff or research subjects or affects 
the environment. It is also declared that this project will not carry out cell, tissue or organ transplant 
procedures, or cell therapy, nor will laboratories, farm or wildlife animals be used.  
 

13. Posting Policy 
13.1 Final Report  
Once the statistical analysis is completed, the final report will be written with the results obtained, 
by the Team of the Medical Management Department of Laboratorios Sophia, S.A. de C.V. This report 
will be prepared following the recommendations of the E3 Step 4 Guide of the ICH.  
13.2 Communication of results  
Regardless of the results of the study, Laboratorios Sophia, S.A. de C.V., is committed to 
communicating the final report of the study to the principal investigators and COFEPRIS. These results 
will also be shared with the research committee and the IRC. It will be the responsibility of the PI to 
communicate it to the research subjects.   
Laboratorios Sophia, S.A. de C.V. will always maintain the rights over the publication and disclosure 
of the information contained.  
13.3 Publication of results  
Laboratorios Sophia, S.A. de C.V., acting as the sponsor of the study, assumes full responsibility for 
its function and retains exclusive ownership rights over the results of the study, which it may use in 
any way it deems appropriate.  
The PI undertakes not to publish or communicate data collected from the study, unless there is the 
prior written agreement of Laboratorios Sophia, S.A. de C.V. Any manuscript derived from the data 
obtained with this protocol must be submitted to review by the sponsor before any attempt to 
submit it for publication in any scientific journal or congress. 
However, if the sponsor is in the process of filing a patent application on the results of the study, the 
sponsor may delay its publication or communication of the results of the study until the date of 
registration or whenever it deems appropriate.  
The assignment of authorship of publications, which is the responsibility of the sponsor, will be the 
prerogative of the latter. However, the express authorization of the people who are invited to 
participate as authors must be obtained. Authors have the right to review the manuscript prior to its 
publication, as well as to issue comments and suggestions in this regard, such comments must be 
delivered within the first 15 calendar days from the date on which the project is received.  
 

14. Financing and Insurance 
14.1 Compensation to Study Participants  
Subjects who participate in the study will receive financial compensation for their participation in the 
study.  
14.2 Insurance for Study Participants  
The subjects participating in the study will sign the informed consent form, which specifies that 
Laboratorios Sophia, S.A. de C.V. agrees to pay for immediate treatment resulting from injuries or 
illnesses caused by the investigational products until their resolution, according to medical criteria.  
All study participants will be entitled to the coverage of a liability policy, contracted by Laboratorios 
Sophia, S.A de C.V. The information of the policy contracted will be available at the research center. 
In the event of a medical emergency, the research center must have personnel, material, equipment 
and procedures for its immediate management.  
 

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16. Annexes 
16.1 ETDRS Primer for Near Vision 

16.2 SICCA Eye Staining Rating (modified from Whitcher et al, 2010) 
Green Lysmine 
(conjunctiva only)
Fluorescein 
(cornea only) 
Grade Points 
Grade Points 
0-9 
1 
10-32 
2 
33-100 
3 
>100 
0 
0 
1 
1-5 
2 
6-30 
3 
>30 
Score by staining pattern: 
Additional points for fluorescein 
staining only: 
+1 if there are confluent staining 
patches 
+1 if there is staining in the pupillary 
area 

16.3 Ocular Comfort Index (OCI) 
Study No   SOPH201-0521/I                                                         Date        
Subject initials                                                        Participant number   201-       -   
 
Instructions:  
This questionnaire was designed to grade the comfort of your eyes. 
For each question, please circle your answer. 
 
Example: In the last week, how often were your eyes red? 
 
(Never     0      1      2     3      4      5      6     (Always) 
 
There are no right or wrong answers. Do not spend too long on any one question. 
 
1. In the last week, how often did your eyes feel dry? 
(Never     0      1      2     3      4      5      6    (Always) 
When your eyes felt dry, typically, how intense was the dryness? 
 
(Never     0      1      2     3      4      5      6    (Always) 
2. In the last week, how often did your eyes feel gritty? 
(Never     0      1      2     3      4      5      6     (Always) 
When your eyes felt gritty, typically, how intense was the grittiness? 
(Never     0      1      2     3      4      5      6     (Always) 
 
3. In the last week, how often did your eyes feel gritty? 
(Never     0      1      2     3      4      5      6     (Always) 
When your eyes stung, typically, how intense was the stinging? 
(Never     0      1      2     3      4      5      6     (Always) 
 
4. In the last week, how often did your eyes feel tired? 
(Never     0      1      2     3      4      5      6     (Always) 
When your eyes felt tired, typically, how intense was the tiredness? 
(Never     0      1      2     3      4      5      6     (Always) 

5. In the last week, how often did your eyes feel painful? 
(Never     0      1      2     3      4      5      6     (Always) 
When your eyes felt painful, typically, how intense was the pain? 
(Never     0      1      2     3      4      5      6     (Always) 
 
6. In the last week, how often did your eyes itch? 
(Never     0      1      2     3      4      5      6     (Always) 
When your eyes itched, typically, how intense was the itching? 
(Never     0      1      2     3      4      5      6     (Always) 

16.4 Fraction to LogMAR conversion table.  
LogMAR 
Snellen (feet) 
Snellen (meters) 
Decimal 
1.0 
20/200 
6/60 
0.10 
0.9 
20/150 
- 
- 
0.8 
20/120 
6/36 
0.15 
0.7 
20/100 
- 
0.20 
0.6 
20/80 
6/24 
- 
0.5 
20/60 
6/18 
0.30 
0.4 
20/50 
- 
0.40 
0.3 
20/40 
6/12 
0.50 
0.2 
20/30 
6/9 
- 
0.1 
20/25 
- 
0.75 
0.0 
20/20 
6/6 
1.00 
-0.1 
20/15 
6/5 
- 
-0.2 
- 
6/4 
1.50 
16.5 Number of events/sample size for the exact method vs. the Wu method, 
for different values of the parameter β, cumulative time a, and tracking time f 
a= Cumulative time in months, f= follow-up time in months. 
The term "exact" refers to the exact calculation using a Chi-square distribution. 
*Table taken and partially adapted from Phadnis MA, 2019. 
Annex taken from Phanis, 2019  [56]