Conditions: Ophthalmological Agent Toxicity Bacterial Conjunctivitis
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 37
Healthy volunteers: yes

Brief summary:
This is a phase I clinical study evaluating the safety and tolerability of PRO-231 ophthalmic solution through the incidence of unexpected adverse events, incidence of conjunctival hyperemia and chemosis, changes in Best Corrected Visual Acuity (BCVA), changes in ocular surface integrity, compared to VIGAMOXI®.

Detailed description:
This is a Phase I, controlled, comparative, parallel-group, single-blind, single-center, controlled clinical trial. The variables to be evaluated include:

Primary (safety):

* Incidence of unexpected adverse events related to the interventions
* Incidence of conjunctival hyperemia and chemosis
* Changes in Best Corrected Visual Acuity (BCVA) assessed with Snellen chart
* Changes in ocular surface integrity using fluorescein staining, using the standard Oxford scale.

Primary (tolerability):

\- Changes in the ocular comfort index (OCI) score

Secondary (safety):

\- Incidence of unexpected adverse events related to the interventions(excluding conjunctival hyperemia and/or chemosis).

Secondary (tolerability):

\- Presence of other ocular symptoms (burning, foreign body sensation, pruritus and lacrimation).

Arms:
arm 1: * PRO-231: Moxifloxacin 0.5% Ophthalmic solution.
* Dosage: 1 drop every 6 hours (3 daily applications \[TID\], in the right eye).
* Route of administration: Topical ophthalmic. arm 2: * VIGAMOXI® :Moxifloxacin 0.5% Ophthalmic Solution.
* Dosage: 1 drop every 6 hours (3 daily applications \[TID\], in the right eye).
* Route of administration: Topical ophthalmic.

Interventions: intervention 1: PRO-231 intervention 2: VIGAMOXI®
Intervention descriptions: intervention 1: Moxifloxacin 0.5% Ophthalmic solution. intervention 2: Moxifloxacin 0.5% Ophthalmic solution.

Protocol document:
Prot_SAP_000.pdf: 
 
 
 
 
 
Protocol: 
SOPH231-1221/I 
 
Title: Phase I clinical study, to evaluate the safety and tolerability of PRO-231 
ophthalmic solution, versus VIGAMOXI,® on the ocular surface of 
ophthalmologically and clinically healthy subjects. 
Information about the molecule under study 
Generic name:    Moxifloxacin     0.5%  
Distinctive name:    PRO-23    1  
Indication:     Conjuntivitis bacteriana.  
 
Protocol Information 
Study Phase:    I  
Version: 2.0 
Release Date: 27-jul.-22 
 
 
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. 


 
 
 
 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  I 
Change History 
The following changes were made to generate version 2.0 of the protocol, dated Jul 27-22 (replaces 
version 1.0 dated Feb 18-22). 
• 
Updated version number and date in the corresponding sections and in the header.  
• 
Added operational definition of hypotheses in the Summary, 3.2 Hypotheses sections. The 
wording was improved and information regarding the statistical methodology was updated 
in the sections Summary, 10.3 Statistical methodology and in section 10.3.1 Analysis of 
populations. In section 7.4 Outcome variables, in section 7.4.3 Definition of variables, the 
statistical test "Student's t-test of independent groups" is added for the variable incidence 
of expected AEs and related to the interventions (excluding conjunctival hyperemia and/or 
chemosis) (bis) included in Table 2. Operational definition of variables. 
• 
Information is added regarding the adverse reactions reported with moxifloxacin 
ophthalmic solution 5mg/mL, classified by apparatus and systems, as well as by their 
criterion of frequency of presentation. 
• 
Annex 16.5 LogMAR Fraction Conversion Table was amended in section 16. Annexes. 
• 
Spelling corrections were made, writing improvements were made, and bibliographic 
citations were updated that correspond to the modifications made throughout the 
document. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  1 of 90  
Content 
Responsible for the study .................................................................................................................... 8 
Signature Page ................................................................................................................................... 10 
From the sponsor ........................................................................................................................... 10 
Investigator Agreement ..................................................................................................................... 11 
List of abbreviations .......................................................................................................................... 12 
1. Protocol Summary ......................................................................................................................... 15 
1.1 Synopsis ................................................................................................................................... 15 
1.2 Study diagram .......................................................................................................................... 20 
1.3 Subject Timeline ....................................................................................................................... 21 
2. Introduction and background ........................................................................................................ 22 
2.1 Theoretical framework ............................................................................................................. 22 
2.2 Background on the investigation ............................................................................................. 25 
2.2.1 From the research question .............................................................................................. 25 
2.3 Problem statement .................................................................................................................. 25 
2.4 Rationale for the study ............................................................................................................. 25 
3. Objectives and hypotheses ............................................................................................................ 27 
3.1 Objectives ................................................................................................................................ 27 
3.1.1 Main objective: .................................................................................................................. 27 
3.1.2 Specific objectives: ............................................................................................................ 27 
3.2 
Study hypothesis................................................................................................................ 28 
4. Study design .................................................................................................................................. 29 
4.1 Study Overview ........................................................................................................................ 29 
4.2 Rationale for the study design ................................................................................................. 29 
4.3 Expected duration of the study ................................................................................................ 29 
4.4 Duration of the subject in the study/treatment ....................................................................... 29 
5. Study population............................................................................................................................ 30 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  2 of 90  
5.1 Eligibility Criteria ...................................................................................................................... 30 
5.1.1 Inclusion criteria ................................................................................................................ 30 
5.1.2 Exclusion criteria: .............................................................................................................. 30 
5.2 Criteria for elimination and/or substitution of subjects ........................................................... 31 
5.2.1 Elimination Criteria ............................................................................................................ 31 
5.2.2 Substitution of subjects ..................................................................................................... 31 
5.3 Counting failures ...................................................................................................................... 31 
5.4 Recruitment and retention strategies ...................................................................................... 32 
5.5 Procedure in case of loss of follow-up ..................................................................................... 32 
5.6 Identification of the subject ..................................................................................................... 33 
6. Investigational product and treatment .......................................................................................... 33 
6.1 Interventions ............................................................................................................................ 33 
6.1.1 Investigational product ...................................................................................................... 33 
6.1.2 Investigational Product Dosage ......................................................................................... 34 
6.1.3 Treatment with the investigational product ...................................................................... 34 
6.1.4 Comparison Product .......................................................................................................... 34 
6.1.5 Comparator Product Dosage ............................................................................................. 34 
6.1.6 Treatment with the comparator product .......................................................................... 34 
6.2 Storage and handling of the investigational product in the study center ................................ 34 
6.3 Concomitant treatments and medications (permitted and prohibited) .................................. 35 
6.3.1 Permitted Medications ...................................................................................................... 35 
6.3.2 Prohibited medicines: ........................................................................................................ 36 
6.4 Procedure for monitoring and measuring adherence .............................................................. 36 
6.5 Strategies to improve adherence ............................................................................................. 37 
7. Study methods and procedures ..................................................................................................... 38 
7.1 Research centre ....................................................................................................................... 38 
7.2 Clinical Study Registration ........................................................................................................ 38 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  3 of 90  
7.3 Assignment of treatment ......................................................................................................... 39 
7.4 Outcome variables ................................................................................................................... 39 
7.4.1 Primary outcome variables ................................................................................................ 39 
7.4.2 Secondary outcome variables ........................................................................................... 40 
7.4.3 Definition of variables, methods and scales for their measurement ................................. 40 
7.4.4 Description of the variables, methods and scales for their measurement ........................ 42 
7.5 Description of procedures or assessments during the study ................................................... 46 
7.5.1 Signing of informed consent .............................................................................................. 46 
7.5.2 Preparation of medical history (includes ophthalmological and general medical history) 46 
7.5.3 Eligibility Criteria ................................................................................................................ 47 
7.5.4 Assigning Subject Code ...................................................................................................... 47 
7.5.5 Adverse events .................................................................................................................. 47 
7.5.6 Vital Signs Measurement ................................................................................................... 47 
7.5.7 Urine Pregnancy Test ........................................................................................................ 47 
7.5.8 Ophthalmological evaluation ............................................................................................. 47 
7.5.9 Application of Medications During Visits ........................................................................... 49 
7.5.10 Delivery of material for the subject ................................................................................. 49 
7.5.11 Delivery of Study Medication .......................................................................................... 49 
7.5.12 Evaluation of concomitant medicinal products ............................................................... 49 
7.5.13 Assessment of adherence to treatment .......................................................................... 50 
7.5.14 Return of Study Medication ............................................................................................ 50 
7.5.15 Subject Journal Removal ................................................................................................. 50 
7.6 Study Diagram and Timeline .................................................................................................... 50 
7.6.1 Study diagram.................................................................................................................... 50 
7.6.2 Study timeline ................................................................................................................... 51 
7.7 Procedures to be carried out per visit ...................................................................................... 52 
7.7.1 Baseline visit ...................................................................................................................... 52 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  4 of 90  
7.7.2 Visit 1 ................................................................................................................................. 52 
7.7.3 Final Visit ........................................................................................................................... 53 
7.7.4 Security Call ....................................................................................................................... 53 
7.7.5 Unscheduled follow-up visits ............................................................................................. 53 
7.8 Data collection ......................................................................................................................... 53 
7.8.1 Source documents ............................................................................................................. 53 
7.8.2 Electronic forms of data collection .................................................................................... 54 
7.8.3 Archiving ............................................................................................................................ 54 
8. Evaluation and management of adverse events ............................................................................ 55 
8.1 Regulation and regulations on adverse events ........................................................................ 55 
8.2 Definition of Adverse Event ..................................................................................................... 55 
8.3 Use of adverse events as a study safety variable ..................................................................... 55 
8.4 Definitions relevant to the classification of adverse events ..................................................... 55 
8.5 Responsibilities of the researcher ............................................................................................ 56 
8.5.1 Recording of adverse events in the electronic case report form ....................................... 57 
8.5.2 Adverse Event Tracking ..................................................................................................... 58 
8.5.3 Procedures for a serious adverse event ............................................................................ 60 
8.5.4 Causation assessment ....................................................................................................... 62 
9. Study Monitoring ........................................................................................................................... 65 
9.1 Monitoring the Study Site ........................................................................................................ 65 
9.2 Audit and quality assurance ..................................................................................................... 65 
10. Sample size calculation and statistical analysis ............................................................................ 67 
10.1 Sample Size Calculation .......................................................................................................... 67 
10.1.1 Calculation methodology ................................................................................................. 67 
10.1.2 Size Calculation ................................................................................................................ 68 
10.2 Clinical Data Management ..................................................................................................... 68 
10.3 Statistical methodology .......................................................................................................... 69 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  5 of 90  
10.3.1 Population Analysis ......................................................................................................... 70 
10.3.2 Safety and tolerability analysis ........................................................................................ 71 
10.4 Other analyses ....................................................................................................................... 72 
10.4.1 Procedure for handling missing data ............................................................................... 72 
11. Ethical considerations .................................................................................................................. 73 
11.1 Approval of the committees .................................................................................................. 73 
11.2 Amendments to the protocol ................................................................................................. 73 
11.3 Early Study Termination ......................................................................................................... 74 
11.4 Informed Consent .................................................................................................................. 75 
11.4.1 Obtaining ......................................................................................................................... 75 
11.4.2 Special considerations ..................................................................................................... 76 
11.4.3 Modifications to informed consent ................................................................................. 76 
11.5 Confidentiality ........................................................................................................................ 76 
11.6 Conflict of interest ................................................................................................................. 77 
11.6.1 Declaration of Interests ................................................................................................... 77 
11.7 Access to Information ............................................................................................................ 77 
11.8 Ancillary and post-study care ................................................................................................. 78 
12. Biosecurity aspects ...................................................................................................................... 79 
13. Posting Policy ............................................................................................................................... 80 
13.1 Final Report ............................................................................................................................ 80 
13.2 Communication of results ...................................................................................................... 80 
13.3 Publication of results .............................................................................................................. 80 
14. Financing and Insurance .............................................................................................................. 81 
14.1 
Compensation to Study Participants ................................................................................. 81 
14.2 
Study Insurance ................................................................................................................. 81 
15. Bibliography ................................................................................................................................. 82 
16. Annexes ....................................................................................................................................... 86 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  6 of 90  
16.1 Cartilla de Snellen .................................................................................................................. 86 
16.2. Efron Scale for Conjunctival Hyperemia ................................................................................ 87 
16.3 Oxford scale ........................................................................................................................... 87 
16.4 Eye Comfort Index (JI) ............................................................................................................ 88 
16.5 Fraction to LogMAR conversion table. ................................................................................... 90 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  7 of 90  
Table and Figure Index 
Table 1. Responsible for the study ....................................................................................................... 8 
Table 2. Operational definition of variables ....................................................................................... 40 
Table 3. Shaffer's classification .......................................................................................................... 48 
Table 4. Study timeline ...................................................................................................................... 51 
Table 5. Triangulation of concepts..................................................................................................... 70 
 
Figure 1. Administrative structure. ...................................................................................................... 9 
Figure 2. Study diagram ..................................................................................................................... 50 
Figure 3. Adverse Event Care ............................................................................................................. 61 
Figure 4. Handling discrepancies. ...................................................................................................... 69 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  8 of 90  
Responsible for the study 
The administrative structure of the sponsoring party, corresponding to Laboratorios Sophia, S.A. de 
C.V., is shown in Table 1. Responsible for the study 
Function§ 
Name/ Contact 
Affiliation ¥ 
Medical Head of the Study 
Dr. Leopoldo Martín Baiza Durán 
leopoldo.baiza@sophia.com.mx 
Regional Director of Medical Affairs  
Study Director 
Dr. Oscar Olvera Montaño 
oscar.olvera@sophia.com.mx 
Regional Medical Affairs Manager 
Author of the Protocol 
Dra. Gisela García Sánchez 
gisela.garcia@sophia.com.mx 
Specialist in Therapeutic Area 
Biostatistics 
Dr. Patricia del Carmen Muñoz Villegas 
patricia.munoz@sophia.com.mx 
Head of Biostatistics 
Operational Manager 
M. en C. Miriam Isabel Serrano Andrade 
miriam.serrano@sophia.com.mx 
Regional Manager of Clinical Research 
¥ Employees of Laboratorios Sophia, S.A. de C.V Av. Paseo del Norte No.5255, Col. Guadalajara Technology Park, 
Carretera Guadalajara-Nogales Km13.5 C.P. 45010 Zapopan, Jalisco, Mexico Tel +52(33) 3000 4200 
§ In the event of staff turnover, the medical head of the study will be in charge of appointing the interim manager. 
 
Board 1. Responsible for the study 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  9 of 90  
 
Figure 1. Administrative structure. 
 
 
Clinical Study Director 
Medical Head of the 
Study 
Regional Strategic 
Alliances Leader 
Regional Clinical 
Operations Leader 
Specialist Home Est 
Clinics and Reg. 
Therapeutic Area 
Specialist/Head of 
Medical Affairs 
Operations 
Head of 
Biostatistics 
Regional Clinical 
Logistics Leader 
Clinical Logistics 
Associate 
Esp. 
Pharmacovigilance 
at Inv. Clínica 
Clinical Team 
Regional Clinical 
Trials Leader 
Monitor clínico  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  10 of 90  
Signature Page 
From the sponsor  
Name:  
Dr. Leopoldo Martín Baiza Durán  
Title:  
Medical Head of the Study  
 
  
 
 
Signature  
  
 
Date  
  
Name:  
Dr. Oscar Olvera Montaño 
Title:  
Study Director  
 
 
  
 
 
Signature  
  
 
Date  
  
Name:  
M. en C. Miriam Isabel Serrano Andrade  
Title:  
Operational Manager  
 
 
  
 
 
Signature  
  
 
Date  
  
Name:  
Dra. Gisela García Sánchez 
Title:  
Protocol Author  
 
 
  
 
 
Signature  
  
 
Date  
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  11 of 90  
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, in 
particular, 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:  
[Insert full name of 
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  
  
  
  
  
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  12 of 90  
List of abbreviations 
A 
Alpha 
°C 
Celsius degrees 
DNA 
Deoxyribonucleic acid 
AV 
Visual acuity 
AVMC 
Best-corrected visual acuity 
BPC 
Good clinical practice 
CBP 
How much is enough to 
CDM 
 Clinical Data Management 
CIS 
Research Ethics Committee 
CI 
Research Committee 
CIOMS 
Council for International Medical Sciences Organizations (ICE)  
en inglés, Council for International Organizations of Medical Sciences) 
COFEPRIS 
Federal Commission for the Protection against Sanitary Risks 
CONSORT 
Consolidated Standards of Reporting Trials 
IUD 
Intrauterine device 
EA 
Adverse Event/Adverse Events 
eCRF 
Electronic case report form 
FCI 
Informed consent form 
H0 
Null hypothesis 
H1 
Alternate hypothesis 
ICH 
International Council for Harmonisation 
ICMJE 
 International Committee of Medical Journal Editors 
ICO 
Eye Comfort Index 
IP 
Principal Investigator 
ITT 
By intention to treat 
LogMAR 
 Logarithm of the minimum angle of resolution 
Ml 
Milliliters 
Mmhg 
Millimeters of mercury  
NOM 
Official Mexican Standard 
OD 
Right eye (Latin for its acronym oculus dexter) 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  13 of 90  
WHO 
World Health Organization 
YOU 
Left eye (Latin for oculus sinister) 
PAE 
Statistical analysis plan 
PI 
Research Products 
PIO 
Intraocular pressure 
PP 
Population by protocol 
RAM 
Adverse drug reaction 
RNEC 
National Clinical Trials Registry 
S-W 
Saphiro-Wilk test 
SCE 
Foreign body sensation 
SDV 
Source document verification 
SPSS 
Statistical package for the social sciences 
SRAM 
Suspected adverse drug reaction 
TEA 
Evaluation time 
TF 
Fluorescein staining 
TID 
Three times a day (by its Latin acronym ter in die) 
UFTLS 
Pharmacovigilance and Technovigilance Unit of Sophia Laboratories 
USED 
United States of America 
x2 
Chi-square 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  14 of 90  
 
 
 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  15 of 90  
1. Protocol Summary 
1.1 Synopsis 
Title of the study: 
Phase I clinical study, to evaluate the safety and tolerability of PRO-231 ophthalmic solution, versus 
VIGAMOXI,® on the ocular surface of ophthalmologically and clinically healthy subjects. 
Study Number: 
   SOPH231-1    2    21/I   
Date of creation: 
Dec 1st-21 
Protocol version: 
   2.0  
Release Date:  
27-Jul-22 
Therapeutic indication: 
Ocular antibiotic. 
Use: 
Conjuntivitis bacteriana 
Estimated duration of the study (from the first 
patient's first visit to the preparation of the final report):  
5 to 6 months 
Clinical Development Phase:  
I 
Objectives: 
Main objective:  
• 
To evaluate the safety and tolerability of the PRO-231 formulation, versus VIGAMOXI,® on 
the ocular surface of ophthalmologically and clinically healthy subjects. 
 
Specific objectives: 
• 
Primary: 
Sponsor Name: 
Laboratorios Sophia, S.A. de C.V. 
Product Name: 
PRO-231 
Active ingredient name: 
Moxifloxacino hydrochloride 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  16 of 90  
o To assess the safety of PRO-231 ophthalmic solution applied to the ocular surface, 
in healthy volunteers, versus VIGAMOXI,® by: 
▪ 
To compare the incidence of adverse events (AEs), unexpected, related to 
interventions. 
▪ 
Incidence of conjunctival hyperemia and/or chemosis between 
interventions. 
▪ 
Changes in best-corrected visual acuity (MCVA) assessed with the Snellen 
chart between interventions. 
▪ 
Changes in ocular surface integrity using fluorescein staining, using the 
Oxford scale between interventions.  
o To evaluate the tolerability of PRO-231 ophthalmic solution applied to the ocular 
surface, in healthy volunteers, versus VIGAMOXI,® by:  
▪ 
Changes in Eye Comfort Index (JI) score between interventions. 
 
• 
Side: 
o To assess the safety of PRO-231 ophthalmic solution applied to the ocular surface, 
in healthy volunteers, versus VIGAMOXI,® by: 
▪ 
Incidence of expected and intervention-related AEs (excluding 
conjunctival hyperemia and/or chemosis). 
o To evaluate the tolerability of PRO-231 ophthalmic solution applied to the ocular 
surface, in healthy volunteers, versus VIGAMOXI,® by:  
▪ 
Presence of ocular symptoms (burning, foreign body sensation, pruritus 
and tearing) between interventions. 
 
Hypothesis: 
H0= PRO-231 ophthalmic solution has a safety and tolerability profile similar to that of VIGAMOXI® 
in clinically healthy subjects. 
𝐻0: 𝑆𝑇𝐴= 𝑆𝑇𝐵 𝑜 𝑆𝑇𝐴−𝑆𝑇𝐵= 0 
 
H1= PRO-231 ophthalmic solution does not have a similar safety and tolerability profile as 
VIGAMOXI® in clinically healthy subjects.  
𝐻1: 𝑆𝑇𝐴≠𝑆𝑇𝐵 𝑜 𝑆𝑇𝐴−𝑆𝑇𝐵≠0 
Operational definition of similarity: To reject H0, the p-value in the primary variables must be ≤ 
0.05 and the 95% CI of this difference must not include zero. Both conditions for hypothesis testing 
(p-value and 95% CI) must be met, otherwise the result will be interpreted as "absence of 
evidence", not "evidence of absence". 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  17 of 90  
Study Design: 
Phase I clinical study, controlled, comparative, parallel-group, single-blind, single-center. 
Number of subjects (planned and analyzed): 
Number of planned subjects: 38 (19 subjects per arm). 
Diagnosis and main inclusion criteria: 
Ophthalmologically and clinically healthy subjects. 
Selection criteria: 
Inclusion criteria: 
− 
Have the ability to voluntarily give their signed informed consent. 
− 
Ophthalmologically and clinically healthy subjects. 
− 
Be able and willing to comply with scheduled visits, treatment plan, and other study 
procedures. 
− 
Age 18 to 45 years. 
− 
Both sexes. 
− 
Women of childbearing potential must ensure the continuation (initiated ≥ 30 days prior 
to signing the informed consent form [ICF]) of the use of a hormonal contraceptive method 
or intrauterine device (IUD) during the study period. 
− 
AVMC of 20/30 or better in both eyes. 
− 
Corneal staining ≤ grade I on the Oxford Scale. 
− 
Have an intraocular pressure ≥ 10 and ≤ 21 mmHg. 
 
Exclusion criteria: 
− 
History of hypersensitivity to fluoroquinolones or any of the components of the PI. 
− 
Be a user of topical ophthalmic medications of any pharmacological group.  
− 
Be a user of medicines by any other means of administration. 
− 
History of eye surgery in the last 6 months. 
− 
Wearing contact lenses for less than two weeks prior to the start of the study, and during 
the study intervention period. 
− 
For women: being pregnant, breastfeeding, or planning to become pregnant within the 
study period. 
− 
Have participated in any investigational clinical study 30 days prior to inclusion in this 
study. 
− 
Have previously participated in this same study. 
− 
History of any chronic-degenerative disease, including Diabetes Mellitus or Systemic 
Arterial Hypertension. 
− 
Known diagnosis of liver or heart disease.  
− 
Have active inflammatory or infectious disease at the time of study entry. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  18 of 90  
− 
Have unresolved injuries or trauma at the time of study entry. 
− 
Having undergone surgical, non-ophthalmologic procedures within the past 3 months. 
− 
Have received doses of vaccine against the Influenza virus or Coronavirus, 7 days prior to 
the inclusion of this study, or plan to receive it during the study intervention. 
− 
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. 
− 
Positive smoking (specified as cigarette smoking regardless of quantity and frequency, 4 
weeks prior to study inclusion and during the study intervention period).  
− 
Positive alcoholism (specified as the consumption of alcoholic beverages, regardless of 
quantity and frequency, 72 hours prior to the inclusion of the study and during the study 
intervention period).  
 
Elimination criteria: 
− 
Withdrawal of the informed consent form. 
− 
Presentation of serious adverse event, whether or not related to the interventions, that, 
in the judgment of the Principal Investigator (PI) and/or sponsor, could affect the patient's 
ability to safely continue with study procedures. 
− 
Non-tolerability or hypersensitivity to any of the compounds used during the tests 
(fluorescein, tetracaine). 
− 
No tolerability or hypersensitivity to any of the investigational drugs. 
− 
Adherence < 80% determined by the subject's diary and corroborated by the final weight 
of the research products (PI) with respect to the initial weight. 
Duration of treatment: 
7 days 
Duration of the subject in the study: 
13 days 
Interventions: 
• 
Investigational product, dosage, and route of administration: 
- 
   PRO-    231 . Moxifloxacin 0.5% Ophthalmic Solution. Laboratorios Sophia, S.A. 
de C.V. Zapopan, Jalisco, Mexico. 
- 
Dosage: 1 drop every 6 hours (3 daily applications [TID], in the right eye [OD]). 
- 
Route of administration: Topical ophthalmic. 
 
• 
Comparator product, dosage and route of administration: 
- 
   VIGAMOXI    ®  Moxifloxacino 0.5% Solución oftálmica. Alcon Laboratories, Inc., 
Fort Worth, Texas, USA. 
- 
Dosage: 1 drop every 6 hours (TID) in OD. 
- 
Route of administration: Topical ophthalmic. 
 
Evaluation criteria: 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  19 of 90  
Primary outcome variables: 
Safety 
− 
Incidence of Intervention-Related Unexpected AEs (Time to Assessment [ET]: Days 3, 8, 
and 12) 
− 
Incidence of conjunctival hyperemia and chemosis. (TE: day 3 and 8) 
− 
Changes in the AVMC evaluated with the Snellen chart. (TE: day 3 and 8) 
− 
Changes in ocular surface integrity using fluorescein staining, using the Oxford scale. (TE: 
day 3 and 8). 
Tolerability 
− 
ICO score changes. (TEE: day 8) 
 
Secondary outcome variables: 
Safety: 
− 
Incidence of expected and intervention-related AEs (excluding conjunctival hyperemia 
and/or chemosis). (TE: day 3, 8 and 12) 
Tolerability: 
− 
Presence of eye symptoms (burning, foreign body sensation, pruritus and tearing). (TE: 
day 3 and 8) 
Statistical methodology 
The analysis of the data collected in the study will be carried out by means of specialized statistical 
software (SPSS available version or R software). The data will be expressed with measures of 
central tendency: mean, standard deviation and/or ranges for quantitative variables, and 
frequencies and/or percentages for qualitative variables. The Shapiro-Wilk test (S-W) will be used 
to determine if the distribution is normal in the data obtained in each study group (p>0.05). If the 
normality in the distribution of the data is verified, to rule out differences between groups, the 
Student's t-statistic will be used for independent groups, while intragroup differences will be 
analyzed with the Student's t-test for repeated measures. In case of p<0.05 in S-W, the statistical 
analysis to rule out differences between groups will be performed by means of the Mann-Whitney 
U test for quantitative variables, while intragroup differences will be performed by Wilcoxon's rank 
test. The difference between the qualitative variables will be analyzed by means of the X2 statistic 
(Chi-squared) or Fisher's exact (for expected values less than 5). Dichotomous variables will be 
analyzed by the McNemar test when applicable. An alpha (α) ≤0.05 will be considered as statistical 
significance, in addition to this, the 95% CI will be calculated for these differences. For primary 
outcome variables, H0 will be rejected, when p≤ 0.05 and the 95% CI does not include zero.  


Laboratorios Sophia, S.A. de C.V. 
 
 
 
Study No.    SOPH23    1    -1    2    
21/I  
 
 
 
 
 
 
Version 2.0 
 
   
 
 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  20 of 90  
1.2 Study diagram 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  21 of 90  
1.3 Subject Timeline 
Procedures 
Scrutiny / Basal 
Visit 
Day 0 
Visit 1 
 
Day 3 ± 1 
Final Visit 
 
Day 8 
Call Security 
Day 12 ± 1 
Signing the informed consent form 
X 
 
 
 
Comprehensive medical history 
X 
 
 
 
Ophthalmological medical history 
X 
 
 
 
Urine pregnancy test 
X 
 
X 
 
Eligibility Criteria 
X 
 
 
 
Eye symptoms 
X 
X 
X 
 
AVMC Measurement 
X 
X 
X 
 
Previous Segment Evaluation 
X 
X 
X 
 
Fluorescein staining evaluation 
X 
X 
X 
 
Intraocular pressure measurement 
X 
X 
X 
 
Posterior Segment Evaluation 
X 
 
X 
 
Subject and IP code assignment 
X 
 
 
 
Delivery of the PIs and indicate the start of 
application the next day (day 1) 
X 
 
 
 
Return of IP 
 
 
X 
 
Adherence assessment  
 
X 
X 
 
Evaluation of adverse events 
X 
X 
X 
X 
Eye Comfort Index 
X 
 
X 
 
Subject Diary Delivery 
X 
 
 
 
Return / Evaluation of the subject's Journal 
 
 
X 
 
Subject continuity assessment 
 
X 
 
 
Abbreviations: AVMC, best corrected visual acuity; PI, research products. 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  22 de 90 
2. Introduction and background 
2.1 Theoretical framework 
Humans are exposed to a variety of microorganisms, including bacteria, viruses, and fungi. In most 
cases, these microorganisms do not cause infection thanks to the external barriers (skin and mucous 
membranes) that protect us from their invasion, and the immune system. However, some 
microorganisms can directly invade these external barriers or be introduced into the body through 
surgical or traumatic wounds; Additionally, the patient's immune system may be compromised, 
allowing microorganisms, which do not normally cause any problems, to generate an infection. [1] 
Various compounds have been used to help the immune system fight pathogenic microorganisms. 
An important property of anti-infective drugs is their selective toxicity, eliminating microorganisms 
while causing minimal or no adverse reactions to the host. [1] 
These drugs are not usually active against all species of microorganisms in the same category; The 
species against which the drug shows intrinsic activity is called Spectrum of activity of the drug. 
Narrow-spectrum drugs are only effective against a few species of microorganisms, while broad-
spectrum drugs have activity against a wide variety of species. At the same time, microorganisms can 
develop resistance mechanisms against drugs to which they were originally susceptible, limiting the 
effectiveness of anti-infective drugs. [1] 
Bacteria are the largest contributor to eye infections in the world and can affect different eye 
structures causing conjunctivitis, keratitis, endophthalmitis, blepharitis, orbital cellulitis and 
dacryocystitis.  [2, 3] 
Bacterial conjunctivitis is one of the most common eye infections and is estimated to account for up 
to 1% of consultations at the first level of care. It is characterized by eye discomfort, conjunctival 
injection, and mucopurulent discharge, and has a significant economic and social impact, as a result 
of disruption of the patient's normal activities, as well as absenteeism from work and/or school. It is 
usually self-limiting (duration of 1 to 2 weeks); however, empirical treatment with broad-spectrum 
anti-infective drugs is recommended to shorten the course of the disease, improve patient comfort, 
decrease transmissibility, promote early return to work or school, and reduce the risk of serious 
complications that may compromise vision [1, 2, 3, 4, 5, 6, 7] 
The pathogens most commonly implicated in bacterial conjunctivitis include strains of Staphylococcus 
aureus, Streptococcus pneumonie, Streptococcus viridians, Haemophilus influenzae, Serratia 
marcescens and Pseudomona aeruginosa. [8, 9]
Although broad-spectrum topical antibiotics are usually used empirically, several factors need to be 
considered when selecting a treatment, such as how quickly it kills the pathogen causing the 
infection, how well it penetrates eye tissues, and the potential impact on the development of 
bacterial resistance. [9] 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  23 de 90 
Moxifloxacin is a broad-spectrum antimicrobial that belongs to the fourth generation 
fluoroquinolones and thanks to its 8-methoxy group, it has improved activity against strains of 
Streptococcus and Staphylococcus, as well as moderate to excellent activity against gram-negative 
organisms that cause ocular pathologies. [6] 
Moxifloxacin is a dose-dependent bactericide. Like the other fluoroquinolones, it interferes with the 
bacterial life cycle by binding to DNA gyrase (topoisomerase II) and topoisomerase IV. Both are 
essential enzymes for the replication, translation, repair, and recombination of bacterial 
deoxyribonucleic acid (DNA).  Human cells lack these enzymes, so they are not affected by 
fluoroquinolones.[6, 8, 9, 10] [1, 8] 
The antimicrobial activity or spectrum of fluoroquinolones will depend on their class, due to the 
variation in affinity and binding strength to topoisomerases, as well as to the bacterial species. [10] 
They are generally well-tolerated drugs with a low incidence of adverse reactions after systemic 
administration. When they occur, these reactions are usually mild and reversible, the most common 
being: gastrointestinal (nausea, vomiting and diarrhea), dermatological (rash, urticaria, pruritus) and 
central nervous system (headache). However, they have also been associated with serious adverse 
events, including Clostridium difficile, prolongation of the QT interval, tendinitis, dysglycemia, liver 
toxicity, phototoxicity, acute renal failure and seizures. [1, 8, 11, 12] 
The frequency of adverse events with topical fluoroquinolone administration is low; The most 
frequent ocular adverse reactions reported after the application of the 0.5% moxifloxacin ophthalmic 
solution are: decreased visual acuity, dry eye, keratitis, eye discomfort, ocular hyperemia, eye pain, 
pruritus, subconjunctival hemorrhage, tearing and burning. The most frequent non-ocular adverse 
events reported are: dysgeusia, fever, cough, otitis media, pharyngitis, rash and rhinitis. . [1][13, 14] 
The following adverse reactions have been reported during clinical studies with Moxifloxacin 5 mg/mL 
ophthalmic solution and are classified according to the following criteria: very common, occurring in 
1 or more in 10 patients (≥1/10); common, occurring in 1 or more in 100 patients (≥1/100 to <1/10); 
rare, occur in 1 or more in 1,000 patients (≥1/1,000 to <1/100); rare, occurring in 1 or more in 10,000 
patients (≥1/10,000 to <1/1,000); very rare, there is less than one case in every 10,000 patients 
(<1/10,000); or unknown frequency (a number cannot be estimated from the available data). Adverse 
reactions are ordered by organ or system affected, from the most severe to the mildest: [15, 16, 17] 
Classification by organs and 
systems 
Frequency 
Adverse Reactions [MedDRA Preferred Terms 
(v.15.1)] 
Blood and lymphatic system 
disorders 
Strange 
Decreased hemoglobin 
Nervous system disorders 
Rare 
Headache 
Strange 
Paresthesia (tingling or numbing sensation) 
Eye disorders 
Common 
Eye pain, eye irritation 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  24 de 90 
Rare 
Dry eye, conjunctival hemorrhage, red eye, 
itchy eyes, eyelid edema (swelling of the 
eyelids), eye discomfort. 
Strange 
Corneal alteration, conjunctivitis, blepharitis 
(inflammation of the edge of the eyelids), 
conjunctival edema, blurred vision, decreased 
visual acuity, asthenopia (visual fatigue), 
redness of the eyelids. 
Respiratory disorders 
Strange 
Nasal discomfort, pharyngolaryngeal pain 
(sore throat), foreign body sensation in the 
throat 
Gastrointestinal disorders 
Rare 
Dysgeusia (bad taste in the mouth) 
Strange 
Vomit 
 
Additional adverse reactions identified in post-marketing surveillance include the following: 
frequency cannot be estimated from available data. Within each classification by organ and system, 
adverse reactions are presented from the most serious to the mildest. [15, 16, 17] 
Classification by organs and systems 
Adverse reactions [MedDRA preferred terms 
(v.15.1)] 
Immune system disorders 
Hypersensitivity (allergic reactions) 
Nervous system disorders 
Seasickness 
Eye disorders 
Corneal ulcers, increased tearing, photophobia 
(sensitivity to light), eye discharge. 
Heart disorders 
Palpitations 
Respiratory disorders 
Dyspnea (shortness of breath) 
Gastrointestinal disorders 
Nausea 
Skin disorders 
Erythema (redness of the skin), purito (itching), 
rash (hives or blisters on the skin), hives 
(reddish hives that may cause itchy skin) 
 
Available ophthalmic fluoroquinolones are indicated for the treatment of bacterial conjunctivitis and 
keratitis, with a treatment regimen of one drop three times daily for 7 days (the total duration of 
therapy will depend on the severity of the clinical picture and the course of the infection).  In addition, 
they have been used off-label as prophylaxis in surgical procedures and ocular trauma to prevent 
endophthalmitis. However, because new molecules such as gatifloxacin and moxifloxacin have a 
higher spectrum and lower resistance, they should be reserved for the treatment of more severe eye 
infections.[8, 9, 15, 16] [17, 18] [17, 19] 
There is no antimicrobial that provides ideal coverage for all pathogens at any site of infection. 
However, moxifloxacin 0.5% ophthalmic solution is a unique, preservative-free antibiotic (thanks to 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  25 de 90 
its concentration it does not require the addition of preservatives to the formula), which offers broad-
spectrum activity, and a moderate to high clinical success rate against common ocular pathogens. In 
addition to this, thanks to its concentration and its high penetrance to the ocular tissues, it allows a 
dosing regimen at infrequent intervals, simplifying its administration and making it more convenient 
for patients. [6, 7, 9, 10]
 
2.2 Background on the investigation 
2.2.1 From the research question 
Is the application of PRO-231 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-231 solution, although there are 
with similar products (different fluoroquinolones and excipients).  
 
2.3 Problem statement 
Bacterial conjunctivitis is one of the most common eye infections and it is estimated that it represents 
up to 1% of consultations at the first level of care, generating a significant economic and social impact, 
as a result of the interruption of the patient's normal activities, as well as absenteeism from work 
and/or school.  Although it is usually a self-limiting condition, with an average duration of 1 to 2 
weeks, empirical treatment with broad-spectrum anti-infective drugs is recommended to shorten the 
course of the disease, improve patient comfort, decrease transmissibility, promote early return to 
work or school, and reduce the risk of serious complications that may compromise vision [9][1, 2, 3, 
4, 5, 6, 7] 
Fluoroquinolones are antibiotics whose usefulness in the area of ophthalmology has already been 
proven, as they have been used for decades as a treatment and/or prophylaxis for eye infections. Its 
effectiveness and safety have given this family of drugs a significant place within the therapeutic 
options of today's ophthalmologist; however, despite being generally well-tolerated drugs with low 
toxicity, it was decided to compare PRO-231 with VIGAMOXI,® the topical formulation of moxifloxacin 
0.5% currently marketed in our country, to verify that its safety and toxicity profile is equal to the 
latter. 
 
2.4 Rationale for the study 
Although eye infections can be considered minor infections, in some cases, they can become 
complicated and leave irreversible sequelae that can compromise the patient's vision. For this reason 
it is important to prescribe topical antibiotics for the treatment of eye infections, or as prophylaxis in 
surgical procedures and eye trauma. An important property of anti-infective drugs is their selective 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  26 de 90 
toxicity, eliminating microorganisms while causing minimal or no adverse reactions to the host. [17, 
18] [1] 
Moxifloxacin hydrochloride, the active ingredient in PRO-231 ophthalmic solution, is a fourth-
generation fluoroquinolone, which has already been approved for use in the treatment of bacterial 
conjunctivitis and keratitis caused by susceptible strains; and it is already marketed by international 
ophthalmic product laboratories. It has the advantage of being preservative-free, and in addition, 
offers broad-spectrum activity, and a moderate to high clinical success rate against common ocular 
pathogens. In addition to this, thanks to its concentration and its high penetrance to the ocular 
tissues, it allows a dosing regimen at infrequent intervals, simplifying its administration and making 
it more convenient for patients. [6, 7, 9, 10] 
In our country, this antibiotic formulation is marketed and approved under the name VIGAMOXI® 
(Alcon Laboratories, Inc. Fort Worth, Texas, USA) for the treatment of bacterial conjunctivitis. Clinical 
studies are required to evaluate that the safety and tolerability of PRO-231, prepared by Laboratorios 
Sophia S.A. de C.V., is comparable to that of VIGAMOXI,® and which are the basis of future clinical 
studies that prove its efficacy. The current commitment of Laboratorios Sophia, S.A. de C.V. is to 
generate its own information for its products that serves for the pertinent regulatory procedures, 
and as a scientific basis for their commercialization. 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  27 de 90 
3. Objectives and hypotheses 
3.1 Objectives 
3.1.1 Main objective:  
To evaluate the safety and tolerability of the formulation PRO-231 versus VIGAMOXI,® on the ocular 
surface of ophthalmologically and clinically healthy subjects. 
 
3.1.2 Specific objectives: 
3.1.2.1 Primary 
• 
To assess the safety of PRO-231 ophthalmic solution applied to the ocular surface, in healthy 
volunteers, versus VIGAMOXI,® by: 
o To compare the incidence of unexpected AEs related to interventions. 
o Incidence of conjunctival hyperemia and/or chemosis between interventions. 
o Changes in the AVMC, evaluated with the Snellen chart, between interventions. 
o Changes in ocular surface integrity using fluorescein staining, using the Oxford scale 
between interventions. 
• 
To evaluate the tolerability of PRO-231 ophthalmic solution applied to the ocular surface, in 
healthy volunteers, versus VIGAMOXI,® by: 
o Changes in Eye Comfort Index (JI) score between interventions. 
 
3.1.2.2 Secondary 
• 
To assess the safety of PRO-231 ophthalmic solution applied to the ocular surface, in healthy 
volunteers, versus VIGAMOXI,® by: 
o Incidence of expected and intervention-related AEs (excluding conjunctival 
hyperemia and/or chemosis). 
• 
To evaluate the tolerability of PRO-231 ophthalmic solution applied to the ocular surface, in 
healthy volunteers, versus VIGAMOXI,® by: 
o Presence of ocular symptoms (burning, foreign body sensation, pruritus and tearing) 
between interventions. 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  28 de 90 
3.2 Study hypothesis 
 
H0: PRO-231 ophthalmic solution has a similar safety and tolerability profile as VIGAMOXI® in clinically 
healthy subjects. 
 
𝐻0: 𝑆𝑇𝐴= 𝑆𝑇𝐵 𝑜 𝑆𝑇𝐴−𝑆𝑇𝐵= 0 
 
 
H1: PRO-231 ophthalmic solution does not have a similar safety and tolerability profile  as VIGAMOXI® 
in clinically healthy subjects.  
 
𝐻1:𝑆𝑇𝐴≠𝑆𝑇𝐵 𝑜 𝑆𝑇𝐴−𝑆𝑇𝐵≠0 
 
Operational definition of similarity: To reject H0, the p-value in the primary variables must be ≤ 
0.05 and the 95% CI of this difference must not include zero. Both conditions for hypothesis testing 
(p-value and 95% CI) must be met, otherwise the result will be interpreted as "absence of evidence", 
not as "evidence of absence".  
 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  29 de 90 
4. Study design 
4.1 Study Overview 
Phase I clinical study, controlled, comparative, parallel-group, single-blind, 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.  
Although this is a comparative study with two intervention groups, only the principal investigator and 
the person responsible for the statistical analysis will be blinded, because the primary packaging of 
the comparator cannot be modified to resemble the product under investigation. Despite this, the 
secondary packaging for the products will be the same and the researcher will not know the 
allocation. 
 
4.3 Expected duration of the study  
The total duration of the study, from the first subject's first visit to the preparation of the final report, 
is estimated to be 5 to 6 months. 
 
4.4 Duration of the subject in the study/treatment  
The approximate duration of each subject in the study is up to 13 days. 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  30 de 90 
5. Study population  
5.1 Eligibility Criteria 
5.1.1 Inclusion criteria 
• 
Have the ability to voluntarily give their signed informed consent. 
• 
Ophthalmologically and clinically healthy subjects. 
• 
Be able and willing to comply with scheduled visits, treatment plan, and other study 
procedures. 
• 
Age 18 to 45 years. 
• 
Both sexes. 
• 
Women of childbearing potential must ensure continued use (initiated ≥ 30 days prior to 
signing the CRF) of the use of a hormonal contraceptive method or IUD during the study 
period. 
• 
AVMC of 20/30 or better in both eyes. 
• 
Corneal staining ≤ grade I on the Oxford Scale. 
• 
Have an intraocular pressure ≥ 10 and ≤ 21 mmHg. 
 
5.1.2 Exclusion criteria: 
• 
History of hypersensitivity to fluoroquinolones or any of the components of the PI. 
• 
Be a user of topical ophthalmic medications of any pharmacological group.  
• 
Be a user of medicines by any other means of administration. 
• 
History of eye surgery in the last 6 months. 
• 
Wearing contact lenses for less than two weeks prior to the start of the study, and during the 
study intervention period. 
• 
For women: being pregnant, breastfeeding, or planning to become pregnant within the study 
period. 
• 
Have participated in any investigational clinical study 30 days prior to inclusion in this study. 
• 
Have previously participated in this same study. 
• 
History of any chronic-degenerative disease, including Diabetes Mellitus or Systemic Arterial 
Hypertension. 
• 
Known diagnosis of liver or heart disease.  
• 
Have active inflammatory or infectious disease at the time of study entry. 
• 
Have unresolved injuries or trauma at the time of study entry. 
• 
Having undergone surgical, non-ophthalmologic procedures within the past 3 months. 
• 
Have received doses of vaccine against the Influenza virus or Coronavirus, 7 days prior to the 
inclusion of this study, or plan to receive it during the intervention of the same. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  31 de 90 
• 
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. 
• 
Positive smoking (specified as cigarette smoking regardless of quantity and frequency, 4 
weeks prior to study inclusion and during the study intervention period).  
• 
Positive alcoholism (specified as the consumption of alcoholic beverages, regardless of 
quantity and frequency, 72 hours prior to the inclusion of the study and during the study 
intervention period).  
 
5.2 Criteria for elimination and/or substitution of subjects 
5.2.1 Elimination Criteria 
• 
Withdrawal of the consent form under information. 
• 
Presentation of a serious adverse event, whether or not related to the PIs, that, in the 
judgment of the PI and/or the sponsor, could affect the patient's ability to continue with 
study procedures safely. 
• 
Non-tolerability or hypersensitivity to any of the compounds used during the tests 
(fluorescein, tetracaine). 
• 
No tolerability or hypersensitivity to any of the investigational drugs. 
• 
Adherence < 80% determined by the subject's diary and corroborated by the final weight of 
the PI with respect to the initial weight. 
5.2.2 Substitution of subjects 
The sponsor may decide to replace subjects who withdraw their FCI or those who have 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.  
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  32 de 90 
5.4 Recruitment and retention strategies  
The duration of the subject's participation in the study is approximately 13 days, during which time 
the subject will be required to attend three visits in total and will be given a safety call, 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 in order 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 loss of follow-up  
For this protocol, loss of follow-up is defined as those subjects who were assigned the PI, who at 
some point were active subjects of the study, but their final evaluation could not be completed.  
In the event that 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. In the event that 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.  
A loss of follow-up <20% is considered not to be a problem for the validity of the results obtained. 
[20, 21] 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  33 de 90 
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:  
 
To. Arieh Daniel Mercado Carrizalez   
 
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 corresponding to the research centre number.   
• 
three digits of the number following their inclusion assigned in the research centre.  
Example of assigned number:  
 
6. Investigational product and treatment 
6.1 Interventions 
6.1.1 Investigational product  
• 
   PRO-    23    1 . Moxifloxacin 0.5%    Ophthalmic solution . . Zapopan, Jalisco, Mexico.   
Laboratorios Sophia, S.A. de     C. V  
• 
Route of administration: Ophthalmic. 
• 
Pharmaceutical form: Ophthalmic solution. 
• 
Presentation: multi-dose dropper bottle, 5 mL. 
• 
Prepared by: Laboratorios Sophia, S.A. de C.V. 
231 - 01 - 001   
Key to the  
molecule   
Number of  
inclusion   
Number of  
center   


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  34 de 90 
• 
Solution description: transparent, free of visible particles and yellow in color. 
• 
Packaging description: White bottle made of low-density polyethylene filled to 5 mL. 
6.1.2 Investigational Product Dosage 
Dosage: 1 drop three times a day (DID) for 7 days, in the right eye (OD). 
6.1.3 Treatment with the investigational product 
The product under investigation will be delivered at the conclusion of the baseline visit. The subject 
under investigation will begin the application the next day (day 1) by administering one drop every 6 
hours, applying a total of one drop three times a day to the right eye during the 7 days of application.  
6.1.4 Comparison Product 
• 
   VIGAMOXI    ®  Moxifloxacino 0.5%    Ophthalmic solution .    Alcon Laboratories, Inc.     Fort 
Worth, Texas, USA.   
• 
Route of administration: Ophthalmic. 
• 
Pharmaceutical form: Ophthalmic solution. 
• 
Presentation: multi-dose dropper bottle, 5 mL. 
• 
Prepared by: Alcon Laboratories, Inc. 
• 
Description of the solution: transparent, greenish-yellow solution. 
• 
Packaging description: Natural low-density polyethylene bottle and a dosing cap with 
polypropylene closure (Drop-trainer® dispensing system)  filled to 5 mL. 
6.1.5 Comparator Product Dosage 
Dosage: 1 drop TID for 7 days, in OD. 
6.1.6 Treatment with the comparator product 
The product under investigation will be delivered at the conclusion of the baseline visit. The subject 
under investigation will begin the application the next day (day 1) by administering one drop every 6 
hours, applying a total of one drop three times a day to the right eye during the 7 days of application.  
 
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. In the event that 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. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  35 de 90 
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 is the responsibility of the research center. The medication must be stored 
in a secure area with restricted access. 
The storage temperature should be 2° to 25°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 on a daily basis 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 material 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 medication section of the eCRF ( electronic Case Report Form). 
6.3.1 Permitted 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 
in order to avoid the interaction of treatments in the tear film, based on the flow index and 
physiological tear volume.  [22] 
o Tetracaína 0.5%  
o Tropicamide 0.8% /Phenylephrine 5%  
o Hipromelosa 2% 
o Fluorescein 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  36 de 90 
- 
Systemic: 
In case participants of childbearing potential decide to use a hormonal contraceptive method, 
the following will be allowed:  
o Oral contraceptives. 
o Subdermal implant. 
o Parenteral injection. 
o HORMONAL DIU. 
 
6.3.2 Prohibited medicines:  
- 
Any ophthalmic or systemic medications that are not on the list of allowed medications. 
 
6.4 Procedure for monitoring and measuring adherence  
For more than four decades, numerous investigations have been conducted on the appropriate way 
to measure and quantify medication adherence, however, none has reached a consensus to stand as 
the gold standard, both in cross-sectional and longitudinal studies. [23, 24, 25, 26, 27, 28, 29, 30] 
There are different procedures to measure adherence to pharmacological interventions. The most 
common procedure includes self-reports, which include: interviews with the subject, 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. [30, 31] 
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. [30] 
There is no standardized parameter to define adequate adherence, it must be defined and delineated 
by the objectives of the particular research.  [30] 
For this study, a minimum adherence of 80% (per subject's diary) will be considered to be necessary 
to meet the objectives of the research; In turn, the information in the diary will be corroborated with 
the final weight of the research product with respect to its initial weight. Therefore, subjects with 
adherence of less than 80% will not be considered for the study. 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  37 de 90 
6.5 Strategies to improve adherence  
1. The PI will sensitize the subject to the importance, in order 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 previously approved by the Research Ethics Committee (ERC). 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  38 de 90 
7. Study methods and procedures  
7.1 Research centre  
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 that the PI 
and sub-investigator be ophthalmology specialists.  
Any person who is designated, under the responsibility of the PI, 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 personnel who have 
functions in the study, both at the central level and in the study center, is the responsibility of the 
principal investigator.  [32] 
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 the Good Clinical Practices of the ICH (International Council on 
Harmonization) International Council for Harmonisation), before the start of their participation in it. 
Training must be recorded in writing and those records must be filed in the master record of the 
study. [32] 
 
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 Sanitary Risks (COFEPRIS) and in a primary registry platform of the World 
Health Organization (WHO). 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).  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  39 de 90 
7.3 Assignment of treatment 
The assignment/randomization of the treatment will be carried out centrally using a validated 
electronic system from an external provider, previously evaluated and authorized by Laboratorios 
Sophia S.A. de C.V. There are two treatment groups. Participants will be randomized in a 1:1 ratio to 
treatment with PRO-231 or VIGAMOXI.® Having the same probability of being assigned to one or the 
other treatment. After signing the FCI, the subject will receive a participant number with which all 
their information will be encrypted pseudonymity during collection and completely anonymized 
during analysis. The generation will be carried out by a third party, authorized by Laboratorios Sophia, 
S.A. de C.V., through its electronic system, through a list of random numbers generated by software 
(SAS Institute Inc, Cary, NC, USA). The information pertaining to this third party will be on file. Only 
the principal investigator and the person responsible for the statistical analysis will be blinded, 
because the primary packaging of the comparator cannot be modified to resemble the product under 
investigation. Despite this, the secondary packaging for the products will be the same and the 
researcher will not know the allocation. IP will be identified by means of labels in accordance with 
current and applicable regulations, which must contain at least: 
- 
Sponsor's name, address, and phone number. 
- 
Dosage form and route of administration. 
- 
Number of doses. 
- 
Batch number. 
- 
Legend "Exclusively for clinical studies". 
- 
Expiration date. 
7.4 Outcome variables  
7.4.1 Primary outcome variables  
7.4.1.1 Security 
- 
Incidence of unexpected AEs related to interventions. 
- 
Incidence of conjunctival hyperemia and chemosis. 
- 
Changes in the AVMC evaluated with the Snellen chart. 
- 
Changes in ocular surface integrity using fluorescein staining, using the Oxford scale. 
7.4.1.2 Tolerability 
- 
ICO score changes. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  40 de 90 
7.4.2 Secondary outcome variables  
7.4.2.1 Security 
- 
Incidence of expected and intervention-related AEs (excluding conjunctival hyperemia 
and/or chemosis). 
7.4.2.2 Tolerability 
- 
Presence of eye symptoms (burning, foreign body sensation, pruritus and tearing). 
7.4.3 Definition of variables, methods and scales for their measurement 
Board 2. Operational definition of variables 
Variable 
Conceptual 
Definition 
Operational 
Definition 
Measurement 
Type 
Reference value 
Statistical test 
Incidence of 
Intervention-
Related 
Unexpected AEs 
Any adverse 
medical event 
that occurs in 
the clinical 
research subject 
to whom any of 
the 
interventions 
were 
administered 
and that has a 
certain 
coincidental 
relationship to 
the 
interventions 
[33] 
All AEs 
(according to 
protocol) in 
which the PI 
suspects a 
causal 
relationship 
with the 
interventions 
will be reported 
• Discrete 
quantitative 
• Nominal 
qualitative 
• Relationshi
p with IP 
• Severity 
• Causality 
• Student's t-test 
of independent 
groups 
• U de Mann 
Whitney* 
• Pearson X2 or 
Fisher Exact* 
Incidence of 
conjunctival 
hyperemia 
It is the reaction 
of the 
conjunctiva to a 
stimulus, a red 
appearance 
secondary to 
the vasodilation 
of the 
conjunctiva 
vessels of 
variable 
intensity is seen 
Direct 
observation. 
Classification by 
Efron scale. See 
attachment 
16.2. Efron 
Scale for 
Conjunctival 
Hyperemia 
• Ordinal 
qualitative 
Degrees: 
0 = Normal, 
I = Muy live, 
II = Mild, 
III = Moderate and 
IV = Severe 
• Pearson X2 or 
Fisher Exact* 
• McNemar Test* 
Incidence of 
Chemosis 
It is defined as 
conjunctival 
edema, the 
result of an 
inflammatory 
reaction. It is 
classified as 
present or 
absent 
The evaluator 
will use a 
narrow beam of 
light at 60° and 
will measure 
whether the 
conjunctiva 
separates ≥ 1/3 
of the total 
• Nominal 
qualitative 
Present / Absent 
• Pearson X2 or 
Fisher Exact* 
• McNemar Test* 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  41 de 90 
Variable 
Conceptual 
Definition 
Operational 
Definition 
Measurement 
Type 
Reference value 
Statistical test 
eyelid opening 
or if it exceeds 
the gray line 
Changes in the 
AVMC 
Visual spatial 
acuity is the 
ability to 
distinguish 
separate 
elements of an 
object and 
identify them as 
a whole. 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 
Cartilla de 
Snellen. Ver 
anexo 16.5 
Fraction to 
LogMAR 
conversion 
table.  
• Continuous 
quantitative 
20/30 (0.2 
LogMAR) or 
better 
• Student's t-test 
of independent 
groups 
• U de Mann 
Whitney* 
Changes in the 
integrity of the 
ocular surface 
Detection of 
corneal and 
conjunctival 
epithelial 
defects by 
means of the CT 
using the Oxford 
scale 
Direct 
observation 
with slit lamp, 
graduation on 
Oxford scale. 
See attachment 
16.3 Oxford 
scale 
• Ordinal 
qualitative 
Degrees: 
The TF is 
presented in a 
series of panels (A 
– E). The staining 
points vary from 0 
– 5 for each panel 
and from 0 – 15 
for the total 
exposed area of 
the conjunctiva 
and cornea 
• Pearson X2 or 
Fisher Exact* 
• McNemar Test* 
ICO score 
changes 
The ICO is a 
questionnaire 
designed to 
measure 
irritation of the 
ocular surface, 
it assesses 
symptoms 
focused on the 
comfort 
associated with 
ocular surface 
alterations. 
Elevated values 
indicate more 
severe 
symptoms 
The evaluator 
will apply the 
questionnaire to 
the subject and 
allow the 
subject to 
answer it calmly 
without any 
pressure and/or 
coercion. See 
annex 16.4 Eye 
Comfort Index 
(JI) 
• Continuous 
quantitative 
0 – 100 
• Student's t-test 
of independent 
groups 
• U de Mann 
Whitney* 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  42 de 90 
Variable 
Conceptual 
Definition 
Operational 
Definition 
Measurement 
Type 
Reference value 
Statistical test 
Incidence of 
expected and 
intervention-
related AEs 
(excluding 
conjunctival 
hyperemia 
and/or 
chemosis). 
(bis) 
Any adverse 
medical event 
that occurs in 
the clinical 
research subject 
who was 
administered 
any of the 
interventions 
and that has a 
casual 
relationship to 
them [33] 
All intervention-
related AEs will 
be reported 
according to the 
protocol. 
• Discrete 
quantitative 
• Nominal 
qualitative 
• Incidence 
• Severity 
• Causality 
 
• Independent 
group student's 
t-test 
• U de Mann 
Whitney* 
• Pearson X2 or 
Fisher Exact* 
 
Incidence of eye 
symptoms 
(burning, SCE, 
pruritus, and 
tearing) 
Burning: refers 
to the sensation 
of stinging or 
eye irritation. 
SCE: refers to 
the sensation of 
"sand" inside 
the eye. 
Pruritus: This 
refers to the 
itching or 
tickling 
sensation 
caused by the 
need to rub or 
scratch. 
Tearing: It refers 
to excessive 
tear production 
[34, 35, 36, 37] 
The evaluator 
will perform the 
interrogation to 
rule out or 
confirm the 
general 
presence of 
symptoms 
• Nominal 
qualitative 
Present / Absent 
• Pearson X2 or 
Fisher Exact* 
• McNemar Test* 
Abbreviations: AVMC, best corrected visual acuity; AE, adverse event; ICO, eye comfort index; IP; principal investigator; 
SCE, foreign body sensation; TF, fluorescein staining; X2, Chi-square. *When applicable. 
7.4.4 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, in order to obtain results and comply with the 
assumption of independence within the variables, only the 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.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  43 de 90 
7.4.4.1 Conjunctival hyperemia  
It is defined as the simplest reaction of the conjunctiva to a stimulus, with a red appearance 
secondary to vasodilation of the conjunctival vessels of variable intensity. It will be graduated using 
the Efron scale. See annex  [38]16.2. Efron Scale for Conjunctival Hyperemia 
The PI will record in the file and the eCRF the grade, in grades (Grade 0 = normal, grade 1 = traces, 
grade 2 = mild, grade 3 = moderate, grade 4 = severe), granted for conjunctival hyperemia OD and 
OS respectively. 
Management as AE: An adverse event will be considered when the grade is greater than or equal to 
grade 3 (moderate).  
7.4.4.2 Chemosis 
It is defined as conjunctival edema, the result of an inflammatory reaction. It is qualified as present 
or absent. The evaluator will use a narrow beam of light at 60° and will measure whether the 
conjunctiva separates from the sclera by ≥ 1/3 of the total eyelid opening or if it exceeds the gray 
line.  [39] 
The PI will record in the file and the eCRF if the chemosis is absent or present. 
Management as AE: An adverse event will be considered when present. 
7.4.4.3 Ocular symptomatology 
The subject will be questioned directly about the presence in general (since the last visit) of the 
following symptoms: burning, foreign body sensation, pruritus, tearing. 
7.4.4.3.1 Ardor 
Feeling of irritation or stinging in the eyes. [37] 
7.4.4.3.2 Foreign Body Sensation 
Feeling of sand inside the eye. [36] 
7.4.4.3.3 Pruritus 
It is an itching or tickling sensation that causes the need to rub or scratch the eye. [34] 
7.4.4.3.4 Lagrimeo  
Excessive tear production or flow. [35, 36] 
The subject will respond only if the symptom is absent or present  
These symptoms will be analyzed as one of the variables of the study and will not be considered an 
adverse event. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  44 de 90 
7.4.4.4 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 as a whole. 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'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). [40] 
VA will be evaluated at baseline, without refractive correction with the Snellen chart (see annex 16.1 
Cartilla de Snellen). It will be located in a place with adequate lighting (natural or artificial) and at a 
distance of 3m from the subject to be evaluated. Visual acuity of the right eye (OD) 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 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 autokeratorrefractometer 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 (AV and AVMC) will be reported for statistical purposes in logarithmic figures (LogMAR) in 
the eCRF.  
A fraction conversion table is attached to LogMAR. (see annex 16.5 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. 
7.4.4.5 Ocular surface staining  
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 subject looks upwards, without 
damaging the conjunctiva. Between 4 and 8 minutes after the instillation of the staining, the 
assessment of the fluorescein stain in the cornea is carried out, with the slit lamp with a cobalt blue 
filter. It will be graded according to the Oxford Scale.  See attachment [41, 42]16.3 Oxford scale 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  45 de 90 
The PI will record in the file and the eCRF the grade, in grades (Grade 0 = Absent, grade I = Minimal, 
grade II = Mild, grade III = Moderate, grade IV = Marked, grade V = Severe), granted for the corneal 
staining of OD and OS respectively. 
Management as AE: An adverse event will be considered when the rating is greater than or equal to 
grade III (moderate). 
7.4.4.6 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 Goldmann tonometer, 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 ocular adverse events that can occur with PRO-231, according to what is reported in the 
literature, are: ocular burning, dysgeusia after instillation, foreign body sensation, ocular pruritus, 
conjunctival hyperemia, chemosis, photophobia and whitish corneal precipitates.  The frequency of 
systemic adverse events with ophthalmic fluoroquinolone administration is low.[1, 13] [1] 
7.4.4.7 Eye 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). Similar to the index for ocular surface diseases, 
the ocular comfort index (ICO) assesses symptoms. The ICO 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.  [43]See 
attachment 16.4 Eye Comfort Index (JI) 
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. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  46 de 90 
As it is a subjective measurement scale according to the subject's symptomatology, variations in the 
ICO cannot be managed as AEs.  
 
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 subject, 
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 subject's medical care, in accordance with the applicable legal provisions. [44] 
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 in particular, 
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). 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  47 de 90 
- 
Therapeutic indication (when applicable). 
 
7.5.3 Eligibility Criteria 
It is the assessment that the subject 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 respiratory rate, heart rate and blood pressure. These measurements can 
be made with a stethoscope and mercury or digital sphygmomanometer. 
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). Women of childbearing potential 
with contraceptive methods including bilateral tubal obstruction should take a pregnancy test. To 
perform the test, the subject 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.  [45] 
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 7.4.4.4 Best-corrected visual acuity 
7.5.8.2 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 fluorescein eye staining will be done (in section 7.4.4.5 
Ocular surface staining ). 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  48 de 90 
7.5.8.3 Previous segment 
It is the evaluation of the structures of the anterior segment (cornea, conjunctiva, anterior chamber, 
iris, pupil, lens, aqueous humor). The evaluation will be made by means of the slit lamp. 
7.5.8.4 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. Goldmann tonometry is based 
on the Imbert-Fick principle. [46] 
The tonometry will be performed with a Goldmann tonometer, 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. 
7.5.8.5 Gonioscopy 
It is the assessment of the iridocorneal angle by attaching a gonioscopy lens to the subject's cornea. 
In some cases the lens will be filled with a high viscosity solution (2% hypromellose) to improve the 
coupling of it with the subject's cornea.  
This evaluation will be carried out only at the baseline visit as part of the comprehensive 
ophthalmological check-up and will be useful to corroborate the subject's ophthalmological health 
status. 
For the classification of the angle, the Shaffer system will be used. 
Board 3. Shaffer's classification 
DEGREE 
ANGLED APERTURE 
DESCRIPTION 
RISK OF OCCLUDING 
4 
45°-35° 
Open 
Impossible 
3 
35°-20° 
Open 
Impossible 
2 
20° 
Narrow 
Possible 
1 
≤ 10° 
Extremely narrow 
Probable 
0 
0° 
Closed 
Occluded 
 
Angular closure or suspected angular closure shall be considered to be Grade 2 or less in more than 
180° of the angular circumference.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  49 de 90 
7.5.8.6 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 the pupil will need to be dilated so that 
the fundus of the eye can be fully evaluated.  
This evaluation will be carried out only at the baseline visit as part of the comprehensive 
ophthalmological check-up under pharmacological mydriasis and will be useful to corroborate the 
subject's ophthalmological health status. 
 
7.5.9 Application of Medications During Visits 
During the visits to the subjects, 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 subject's ocular surface and 
facilitate the measurement of intraocular pressure, application of dyes for stains 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 subject is in the study, the investigational product will be given, and the dosage of the 
product will be explained (one drop three times a day in the right eye). 
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. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  50 de 90 
7.5.13 Assessment of adherence to treatment 
It refers to indirectly evaluating the number of applications during the period between visits. In order 
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 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  51 de 90 
7.6.2 Study timeline 
Procedures 
Scrutiny / Basal 
Visit 
Day 0 
Visit 1 
 
Day 3 ± 1 
Final Visit 
 
Day 8 
Call Security 
Day 12 ± 1 
Signing the informed consent form 
X 
 
 
 
Medical history 
X 
 
 
 
Ophthalmological medical history 
X 
 
 
 
Urine pregnancy test 
X 
 
X 
 
Eligibility Criteria 
X 
 
 
 
Eye symptoms 
X 
X 
X 
 
AVMC Measurement 
X 
X 
X 
 
Previous Segment Evaluation 
X 
X 
X 
 
Fluorescein staining evaluation 
X 
X 
X 
 
Intraocular pressure measurement 
X 
X 
X 
 
Posterior Segment Evaluation 
X 
 
X 
 
Subject and IP code assignment 
X 
 
 
 
Delivery of the PIs and indicate the start of 
application the next day (day 1) 
X 
 
 
 
Return of IP 
 
 
X 
 
Adherence assessment  
 
X 
X 
 
Evaluation of adverse events 
X 
X 
X 
X 
Eye Comfort Index 
X 
 
X 
 
Subject Diary Delivery 
X 
 
 
 
Return / Evaluation of the subject's Journal 
 
 
X 
 
Subject continuity assessment 
 
X 
 
 
Abbreviations: AVMC, best corrected visual acuity; PI, research products. 
Board 4. Study timeline 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  52 de 90 
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. 
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 
- 
Pregnancy test (where applicable) 
- 
Best-corrected visual acuity measurement 
- 
Previous Segment Evaluation 
- 
Fundus evaluation under pharmacologic mydriasis 
- 
Evaluation of the angle in the anterior chamber (gonioscopy) 
- 
Fluorescein Eye Staining 
- 
Intraocular pressure measurement 
- 
Evaluation of adverse events 
- 
Eye symptoms 
- 
Eye Comfort Index (ILO) 
- 
Subject Code Assignment and Research Product Delivery 
- 
Submission of the subject's diary 
7.7.2 Visit 1 
- 
Best-corrected visual acuity measurement 
- 
Previous Segment Evaluation 
- 
Fluorescein Eye Staining 
- 
Intraocular pressure measurement 
- 
Evaluation of adverse events 
- 
Adherence Assessment 
- 
Eye symptoms 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  53 de 90 
7.7.3 Final Visit 
- 
Pregnancy test (where applicable) 
- 
Best-corrected visual acuity measurement 
- 
Previous Segment Evaluation 
- 
Posterior Segment Evaluation 
- 
Fluorescein Eye Staining 
- 
Intraocular pressure measurement 
- 
Evaluation of adverse events 
- 
Eye Comfort Index (ILO) 
- 
Adherence Assessment 
- 
Eye symptoms 
- 
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 the medical history, clinical evolution notes, laboratory reports, 
cabinet study reports, nursing notes, follow-up notes, surgery records, etc.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  54 de 90 
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 master file will remain in the participating institutions in charge of the PI or 
its work team and must be safeguarded for at least 5 years.  
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  55 de 90 
8. Evaluation and management of adverse events 
8.1 Regulation and regulations on adverse events 
The registration and reporting of adverse events will be carried out in accordance with the guidelines 
established in NOM-220-SSA1-2016 and the international guidelines ICH E6.   [33][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-231 is paramount to the study, therefore, it is considered 
important to report any unwanted manifestations or diseases that occur during the course of 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 (Medically important event 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  56 de 90 
or reaction: That clinical manifestation or adverse event that in the opinion of the doctor may not be 
immediately life-threatening,  result in death or hospitalization, but which could endanger the subject 
or require medical intervention to prevent the occurrence of any of the criteria listed in the definition 
of serious adverse reaction).  [50] 
Severity (mild, moderate, or severe). Mild are 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.   [33][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.     [33] [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. [33] [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 3. Adverse 
Event Care).  
The sponsor's final report will include the report of adverse events in compliance with current 
national and international regulations. [33] [47] 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  57 de 90 
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 their participation will be in 
accordance with the stipulations of NOM-012-SSA3-2012. [33] 
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  
o 
Recovered/resolved with sequelae  
o 
Recovering/Resolving 
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 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  58 de 90 
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, the dose, 
the route of administration, start and end dates of its 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 authority.  
▪ 
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: 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  59 de 90 
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 points of assessment of the subject. 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 the 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 AS. 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 of 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. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  60 de 90 
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 as long as 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 subject should continue until the outcome of pregnancy. Pregnancy is not 
by 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:  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  61 de 90 
 
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 authority. However, it may be suspected that 
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 
Document 
Attention and  
treatment  
relevant 
Categorization 
Preparation of a medical note 
eCRF Registration and Reporting 
 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 
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 
 
Notify to: 
+ Pharmacovigilance Specialist 
in Clinical Research in 
maximum 24 hours 
+Corresponding Health 
Authority 
The 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  62 de 90 
or Summary of the Comparator Drug, 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, in the event that 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 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 virtue of the fact that, in cases of serious adverse events, the clinical monitor 
of the study must be notified within 24 hours after learning of it, so that in turn it informs 
the Clinical Team and the Pharmacovigilance and Technovigilance Unit of Sophia 
Laboratories (UFTLS), 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 drug will be reported to the appropriate regulatory authority 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, the World Health Organization and the Uppsala Monitoring Center.  
[33][51] 
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] 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  63 de 90 
For all adverse events, the Principal Investigator is responsible for examining and evaluating the 
subject to determine the association of the event with the clinical study and the intervention, 
whether 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 subject considers that a relationship exists or not. 
[49] 
UFTLS may employ the causation categories 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: [33] [51] 
• 
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. [33] [51] 
• 
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 [33] [51] 
• 
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. [33] [51] 
• 
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. [33] [51] 
• 
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. [33] [51] 
• 
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. [33] [51] 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  64 de 90 
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 take 
into account 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. [52] 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  65 de 90 
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 centre, at least one start 
visit 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 assurance  
To ensure compliance with Good Clinical Practice (GCP) 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.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  66 de 90 
Details about the audit process are set out separately in the Audit Plan.  
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. In the event that the audit has 
not been scheduled by the sponsor, the facility must notify Laboratorios Sophia, S.A. de C.V. 
immediately.  
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  67 de 90 
10. Sample size calculation and statistical analysis 
10.1 Sample Size Calculation 
Bacterial conjunctivitis is commonly self-limiting, resolving after about 10 days, and can rarely last 
more than 3 weeks. Its most common treatment is the use of antibiotics with a broad spectrum of 
action on the variety of ocular pathogens. Proper management of bacterial conjunctivitis reduces the 
time to resolution of the disease, minimizes the risk of sequelae, helps prevent the spread of 
infection, and decreases the time the patient can spend away from work or school. Moxifloxacin 
hydrochloride is a fourth-generation fluoroquinolone, its use in topical ophthalmic presentation for 
the treatment of bacterial conjunctivitis has been approved since 2003 in the United States. 
Moxifloxacin is one of the most potent fluoroquinolones evaluated for ophthalmic use for bacterial 
eradication. There is sufficient evidence on the safety and tolerability of moxifloxacin. Commonly 
reported AEs are eye pain, eye burning/irritation, hyperemia, and tearing; most of them of mild 
intensity and transient.[53] [54, 55] [54] [53, 55, 56, 57, 58] 
Considering this information and the primary objective of the study; To evaluate the safety and 
tolerability of PRO-231 ophthalmic solution (moxifloxacin 0.5%) on the ocular surface of clinically 
healthy subjects, the following proposal was made for the sample size. 
10.1.1 Calculation methodology 
Although sample estimation in phase 1 studies is common in treatments whose toxicity/risk is high, 
as it is in clinical studies for the management/survival of cancer; It was considered pertinent to 
estimate the sample size according to the safety profile reported in the literature after the use of 
moxifloxacin hydrochloride. Considering the primary safety variables of the protocol: [59] 
• 
Incidence of AD 
• 
Changes in ophthalmologic signs  
• 
Changes in the AVMC 
• 
Changes in fluorescein staining of the ocular surface 
 
The equation was used to compare two proportions: two samples, two sides of equality. This 
calculation is useful when you want to test whether the proportions of two groups are different 
(Incidence of AD in VIGAMOXI® is different from the incidence of AD in PRO-231). The following 
formulas are used for the calculation to compute the sample size and power respectively:  [60] 
 
𝑛𝐴= 𝑘𝑛𝐵 𝑦 𝑛𝐵= (𝑝𝐴(1 −𝑝𝐴)
𝑘
) (
𝑧1−𝛼2
⁄ + 𝑧1−𝛽
𝑝𝐴−𝑝𝐵
)
2
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  68 de 90 
1 −𝛽= Φ (𝑧−𝑧1−𝛼2
⁄ ) + Φ (−𝑧−𝑧1−𝛼2
⁄ ) , 𝑧=
𝑝𝐴−𝑝𝐵
√𝑝𝐴(1 −𝑝𝐴)
𝑛𝐴
+ 𝑝𝐵(1 −𝑝𝐵)
𝑛𝐵
 
 
Where:  
k = nA/nB: ratio between the sample sizes of the two groups 
Φ: Function of Standard Normal Distribution 
Φ -1: The standard normal quantile function  
α: error tipo I 
β: error tipo II 
The proportion of subjects who had at least one AE (ocular and non-ocular) was considered in the 
study by Garg et al. (2015), where they evaluated the clinical and antibacterial efficacy, as well as the 
safety of besifloxacin ophthalmic suspension compared to moxifloxacin 0.5% (besifloxacin; n=61, 
moxifloxacin; n=62). A total of 6 ocular AEs were present in subjects treated with moxifloxacin and at 
least one AE occurred in 3 patients. The most common AE was conjunctivitis (3.2%). For the expected 
proportion of subjects of PRO-231, 50% was used, which is a conservative value and gives larger 
samples. The calculation was made using the free software R (R Core Team, 2021), using a code 
available online for this purpose. [56] [61] [62] 
10.1.2 Size Calculation 
PRO-231 ophthalmic solution is expected to be safe in its ophthalmic application as it presents an 
incidence of AE similar to that presented by VIGAMOXI.® 
On this basis, a sample of 17 ophthalmologically and clinically healthy subjects was estimated. This 
value increased by 10% considering possible losses. Based on this consideration, 19 subjects per arm 
(38 subjects in total) are required to provide one eye (OD) to respond to the primary objective of the 
study. 
 
10.2 Clinical Data Management 
The clinical data management (or clinical data management, CDM), allows the generation of high-
quality, reliable data with statistical value. CDM is the process of collecting, cleaning, and managing 
the information of the subjects of a study in accordance with regulatory standards (guidelines 21 CRF 
Part 11, ICH and GCP). It covers the design of the electronic case report form (eCRF), comments on 
the eCRF, 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) .[63] 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  69 de 90 
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 
the [63, 64]Figure 4. 
The following roles will be included in the CDM team:  
• 
Data Manager 
• 
Database Designer/Programmer 
• 
Medical Coder 
• 
Clinical Data Coordinator 
• 
Quality Control 
• 
Capturist (data entry associate) 
 
 
Figure 4. Handling discrepancies. 
(DCF, medical note; CRA, clinical monitor; PI, principal investigator; SEC, self-evident correction) [63] 
10.3 Statistical methodology 
The statistical analysis plan (PAE) was developed considering the evaluation criteria described in the 
study protocol. 
Data Validation 
Discrepancy? 
DCF 
IP 
CRA 
SDV 
Data 
manager 
SEC 
Resolution 
Database 
NO 
YES 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  70 de 90 
The statistical analysis will be carried out by personnel of Laboratorios Sophia, S.A. de C.V. Specialized 
statistical software (SPSS version available [IBM Corporation, Armonk, NY, USA] or R [The R 
Foundation for Statistical Computing; http://www.R-project.org]), for Windows. 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 platform of the selected statistical package. 
The variables will be categorized according to their nature (see Board 2). The statistical tests used will 
be following the corresponding assumptions for parametric statistics as long as the assumption of 
normality is met (Shapiro-Wilk, p>0.05). 
10.3.1 Population Analysis 
The statistical analysis will be presented to give a general summary of the subjects entered into 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 and/or ranges. Changes in the AVMC and ICO will be expressed as continuous 
variables. Whereas, the incidence (number) of AEs will be expressed as a discrete variable. 
The result of the nominal and ordinal qualitative variables will be presented in frequencies, 
proportions and/or percentages. For these, tables of frequencies p x q will be constructed. All 
percentages will be presented with a decimal. 
The level of difference to consider significance will be an alpha (α) of 0.05 or less, the 95% CI will be 
calculated for these differences. The triangulation between the type of variable and the 
measurements is shown in Table 5. [63] 
The operational definition of the study for hypothesis testing will be as follows:  
 
Hypothesis 
testing 
value 
Result 
Interpretation 
P value 
≤ 0.05 
H0 
rejected 
Both treatments are different in terms of safety and 
tolerability 
95% CI 
Does not 
include zero 
P value 
≥ 0.05 
H0 
accepted 
Both treatments are similar in terms of safety and 
tolerability (they are not different) 
95% CI 
Includes zero 
 
Both conditions for hypothesis testing (p-value and 95% CI) must be met, otherwise the result 
will be interpreted as "absence of evidence", not as "evidence of absence".  
 
Board 5. Triangulation of concepts. 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  71 de 90 
Variable 
Type 
Variable 
A1 
A2 
B1 
C1 
C2 
C3 
C4 
D1 
E1 
F1 
F2 
F3 
F4 
Selection 
A1 
Demographic 
D 
 
 
 
 
 
 
 
 
 
 
 
 
A2 
Medical History/Selection 
Criteria 
 
D 
 
 
 
 
 
 
 
 
 
 
 
Basal 
B1 
Comprehensive 
ophthalmological 
evaluation 
 
T 
D 
TB 
TB 
B 
T 
 
TB 
 
 
 
 
Primary Security 
C1 
Incidence of unexpected-
related AEs 
 
 
 
TB 
 
 
 
 
TB 
 
 
 
 
C2 
Incidence of conjunctival 
hyperemia and chemosis 
 
 
T 
 
TB 
 
 
 
TB 
 
 
 
 
C3 
AVMC 
 
 
B 
TB 
 
B 
 
 
TB 
 
 
 
 
C4 
Corneal and conjunctival 
stains with fluorescein 
 
 
B 
TB 
 
 
B 
 
TB 
 
 
 
 
Primary Tolerability 
D1 
ICO 
 
 
 
 
 
 
 
B 
 
 
 
 
 
Secondary Security 
E1 
Incidence of AE* 
 
 
T 
 
 
 
TB 
 
 
 
 
 
 
Secondary Tolerability 
F1 
Presence of burning 
 
 
 
 
 
 
 
 
B 
TB 
 
 
 
F2 
ETS Presence 
 
 
 
 
 
 
 
 
B 
 
TB 
 
 
F3 
Presence of pruritus 
 
 
 
 
 
 
 
 
B 
 
 
TB 
 
F4 
Presence of tearing 
 
 
  
 
 
 
 
 
B 
 
 
 
TB 
Abbreviations: AVMC, best corrected visual acuity; B, bivariate analysis; D, descriptive statistics; AE, adverse event; ICO, 
eye comfort index; SCE, foreign body sensation; T, contingency table p x q. *EAs related or not, with the research product. 
 
10.3.2 Safety and tolerability analysis 
10.3.2.1 Analysis for Primary Variables 
The analysis of the primary outcome variables of safety and tolerability will be carried out in the per-
protocol population (PP), defined as any subject with an adherence ≥ 80% determined by the 
subject's diary and corroborated by the final weight of the research product to which it was assigned 
(PRO-231 or VIGAMOXI)® with respect to its initial weight.  and that they conclude their participation 
without deviations from the study protocol. 
The Shapiro-Wilk test (S-W) will be used to determine if the distribution is normal in the data obtained 
in each study group (p>0.05). If the normality in the distribution of the data is verified, to rule out 
differences between groups, the Student's t-statistic will be used for independent groups, while 
intragroup differences will be analyzed with the Student's t-test for repeated measures.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  72 de 90 
In case of p<0.05 in S-W, the statistical analysis to rule out differences between groups will be 
performed by means of the Mann-Whitney U test for quantitative variables, while intragroup 
differences will be performed by Wilcoxon's rank test. 
For qualitative variables, Pearson's X2 (Chi-square) or Fisher's exact test will be used in expected 
values less than 5. Dichotomous variables will be analyzed by the McNemar test when applicable. 
10.3.2.2 Analysis for secondary variables 
The analysis of the secondary outcome variables of safety and tolerability will be carried out in the 
safety population, defined as any subject who has received at least one dose of the investigational 
product (PRO-231 or VIGAMOXI)® regardless of their adherence to it or adherence to the protocol 
(intention-to-treat population, ITT). 
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. 
 
10.4 Other analyses 
The final report of the results will be shown in tables or graphs, as appropriate. 
10.4.1 Procedure for handling missing data 
There is no imputation procedure for missing data. 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  73 de 90 
11. Ethical considerations 
11.1 Approval of the committees 
The present study will be conducted in accordance with the principles 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,  applicable by the regulatory authorities.  
The study will not begin at the research center if there are no confidentiality agreements and 
economic proposal 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 considered to be 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 that may be amended will be: protocol, form of informed consent, 
researcher's manual, documents for the subject, measurement scales and schedule of activities.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  74 de 90 
Any amendment must be approved by the sponsor and/or the principal investigator, the amended 
document(s), once reviewed and approved by the CEI and the IC 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 aforementioned Committees. It is the investigator's 
responsibility to take action 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 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 temporarily suspended or terminated prematurely if there is a sufficiently 
reasonable cause. Written notice, documenting the reason for the suspension or early termination, 
must be delivered by the party executing 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) considers it for security alerts.  
3. The Sponsor determines 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 are not assessable or are not sufficiently complete.  
8. The determination that the primary objective has been achieved.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  75 de 90 
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; as long as 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 form 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, the informed consent form 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 be in compliance with 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 and every 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 informed 
consent form 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 guaranteeing 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.  
In the event that a subject is illiterate, acceptance will be with his or her fingerprint, and in the event 
that the subject is unable to give adequate written informed consent, a representative of the "legally 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  76 de 90 
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, Research Ethics and COFEPRIS Committees.  
Such amendments may be implemented only after obtaining the written approval of the Research 
Ethics Committee and authorization from COFEPRIS (as applicable), except for an amendment that is 
required to eliminate an immediate danger to study subjects.  
A process of re-consent of each subject affected by the amendment must be carried out under the 
same conditions as those described above, in order 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 as long as 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 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  77 de 90 
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 of 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 
the 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.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  78 de 90 
 
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.  
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  79 de 90 
12. Biosecurity aspects 
NO BIOSECURITY IMPLICATIONS  
This protocol, entitled: "Phase I clinical study, to evaluate the safety and tolerability of the ophthalmic 
solution PRO-231, versus VIGAMOXI,® on the ocular surface of ophthalmologically and clinically 
healthy subjects", and number: SOPH231-1221/I has no biosafety implications, since infectious-
contagious biological material; pathogenic strains of bacteria or parasites; viruses of any type; 
radioactive material of any type; animals and/or cells will NOT be used and/or genetically modified 
plants; toxic, dangerous or explosive substances; any other material that endangers the health or 
physical integrity of the research centre'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 laboratory, farm or wildlife animals be used. 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  80 de 90 
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 Regional Management Department of Medical Affairs 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 IC and the IEC. It will be the responsibility of the PI to communicate it to 
the research subjects.   
Laboratorios Sophia, S.A. de C.V. will maintain at all times 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, in the event that 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.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  81 de 90 
14. Financing and Insurance 
14.1 Compensation to Study Participants 
Subjects who participate in the study will not receive financial compensation for their participation 
in the study. However, the subjects will receive financial support for travel expenses in each 
scheduled visit to which they attend punctually. Such support, as well as the amount, may be 
specified in the form of informed consent. 
 
14.2 Study Insurance 
In accordance with current local regulations, Laboratorios Sophia S.A. de C.V. has contracted a civil 
liability policy, in order to comply with the responsibility of providing medical treatment and 
compensation to which a subject would be legally entitled, in the case of damages directly caused by 
this research. 
In the event of a medical emergency, the research center must have personnel, material, equipment 
and procedures for its immediate management. 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  82 de 90 
15. Bibliography 
 
[1]  J. Bartlett y S. Jaanus, Clinical Ocular Pharmacology, 5 ed., Woburn, MA: Butterworth-Heinemann, 2008, 
pp. 151-154. 
[2]  J. Bartlett, «Ophthalmic Drug Facts,» 25 ed., St. Louis, MI, Wolters Kluwer Health, 2014, pp. 45-46. 
[3]  M. Teweldemedhin, H. Gebreyesus, A. H. Atsbaha, S. W. Asgedom y M. Saravanan, «Bacterial profile of 
ocular infections: a systematic review,» BMC Ophthalmology, vol. 17, nº 212, pp. 1 - 9, 2017.  
[4]  T. L. Comstock, M. R. Paterno, H. H. DeCory y D. W. Usner, «Safety and Tolerability of Besifloxacin 
Ophthalmic Suspension 0.6% in the Treatment of Bacterial Conjunctivitis. Data from Six Clinical and 
Phase I Safety Studies.,» Clin Drug Investig, vol. 30, nº 10, pp. 675 - 685, 2010.  
[5]  A. A. Azari y N. P. Barney, «Conjunctivitis A Systematic Review of Diagnosis and Treatment,» JAMA, vol. 
310, nº 16, pp. 1721 - 1729, 2013.  
[6]  B. A. Schlech y E. Alfonso, «Overview of the Potency of Moxifloxacin Ophthalmic Solution 0.5% 
(VIGAMOX®),» Survey of Ophthalmology, vol. 50, nº 1, pp. S7 - S15, 2005.  
[7]  C. Hutnik, H. Mohammad y Mohammad-Shahi, «Bacterial conjunctivitis,» Clinical Ophthalmology, vol. 4, 
pp. 1451 - 1457, 2010.  
[8]  Thomson Micromedex, Drug Information for the Health Care Professional, Taunton, Massachusetts: 
Quebecor World, 2007.  
[9]  G. M. Keating, «Moxifloxacin 0.5% Ophthalmic Solution In Bacterial Conjunctivitis,» Drugs, vol. 71, nº 1, 
pp. 89 - 99, 2011.  
[10] D. Miller, «Review of moxifloxacin hydrochloride ophthalmic solution in the tretment of bacterial eye 
infections,» Clinical Ophthalmology, vol. 2, nº 1, pp. 77 - 91, 2008.  
[11] L. S. M. Kuula, K. M. Vilijemaa, J. T. Backman y M. Blom, «Fluoroquinolone-related adverse events 
resulting in health service use and costs: A systematic review,» PLoS ONE, vol. 14, nº 4, pp. 1 - 17, 2019.  
[12] D. Baggio y M. R. Ananda-Rajah, «Fluoroquinolone antibiotics and adverse events,» Australian 
Prescriber, vol. 44, nº 5, pp. 161 - 164, 2021.  
[13] A. M. Thompson, «Ocular toxicity of fluoroquinolones,» Clinical and Experimental Ophthalmology, vol. 
35, pp. 566 - 577, 2007.  
[14] I. Alcon Laboratories, «Food and Drug Administration,» Jul 2011. [En línea]. Available: 
https://www.accessdata.fda.gov/drugsatfda_docs/label/2011/021598s017lbl.pdf. [Último acceso: 07 
Dic 2021]. 
[15] J. Benitez-del-Castillo, Y. Verboven, D. Stroman y L. Kodjikian, «The Role of Topical Moxifloxacin, a New 
Antibacterial in Europe, in the Treatment of Bacterial Conjunctivitis,» Clin Drug Investig, vol. 31, nº 8, 
pp. 543 - 557, 2011.  
[16] M. B. McDonald, E. E. Prtozko, L. S. Brunner, T. W. Morris, W. Haas, M. R. Paterno, T. L. Comstock y D. 
W. Usner, «Efficacy and Safety of Besifloxacin Ophthalmic Suspension 0.6% Compared with Moxifloxacin 
Ophthalmic Solution 0.5% for Treating Bacterial Conjunctivitis,» American Academy of Ophthalmology, 
vol. 116, nº 9, pp. 1615 - 1623, 2009.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  83 de 90 
[17] S. V. Scoper, «Review of Third- and Fourth-Generation Fluoroquinolones in Ophthalmology: In-Vitro and 
I-Vivo Efficacy,» Advances in Therapy, vol. 25, nº 10, pp. 979 - 994, 2008.  
[18] S. Pradhan y V. N. Prajna, «Topical Fluoroquinolones: Current Perspectives,» Delhi Journal of 
Ophthalmology, vol. 25, nº 4, pp. 267 - 271, 2015.  
[19] G. Høvding, «Acute bacterial conjunctivitis,» Acta Ophthalmologica, vol. 86, pp. 5 - 17, 2008.  
[20] D. Sacket, W. Richardson y W. Rosenberg, Evidence-Based Medicine: How to Practice and Teach, New 
York: Churchill Livingstone, 1997. .  
[21] J. Dettori, «Loss to follow-up,» Evid Based Spine Care J, vol. 2, nº 1, pp. 7-10, 2011.  
[22] R. Fiscella, «Ophthalmic Drug Formulations,» de Clinical Ocular Pharmacology, St Louis, MO, 
Butterworth Heinemann Elsevier, 2008, pp. 17-37. 
[23] L. Gordis, «General concepts for use of markers in clinical trials,» Control Clin Trials, vol. 5, pp. 481-487, 
1984.  
[24] M. Mattson y L. Friedman, «Issues in medication adherence assessment in clinical trials of the National 
Heart, Lung, and Blood Institute,» Control Clin Trials, vol. 5, pp. 488-496, 1984.  
[25] S. Norell, «Methods in assessing drug compliance,» Acta Med Scand, vol. S 683, pp. 34-50, 1984.  
[26] P. Rudd, R. Byyny, V. Zachary y e. al, «Pill count measures of compliance in a drug trial: variability and 
suitability,» Am J Hypertens, vol. 1, pp. 309-312, 1988.  
[27] K. Farmer, «Methods for measuring and monitoring medication regimen adherence in clinical trials and 
clinical practice,» Clin Ther, vol. 21, pp. 1074-1090, 1999.  
[28] H. Liu, C. Golin, L. Miller y e. al, «A comparison study of multiple measures of adherence to HIV protease 
inhibitors,» Ann Intern Med, vol. 134, pp. 968-977, 2001.  
[29] W. Lam y P. Fresco, «Medication Adherence Measures: An Overview,» BioMed Research International, 
vol. 2015, 2015.  
[30] M. Vitolins, C. Rand, S. Rapp, P. Ribisl y M. Sevick, «Measuring Adherence to Behavioral and Medical 
Interventions,» Controll Clin Trial, vol. 21, pp. 188S-194S, 2000.  
[31] J. Lee, K. Grace, T. Foster, M. Crawley y e. al, «How should we measure medication adherence in clinical 
trials and practice?,» Ther and Clin Risk Manag, vol. 3, nº 4, pp. 685-690, 2007.  
[32] I. C. f. Harmonisation, «ICH Harmonised Guideline,» 09 Nov 2016. [En línea]. Available: 
https://database.ich.org/sites/default/files/E6_R2_Addendum.pdf. [Último acceso: 14 Feb 2022]. 
[33] N. O. Mexicana, NOM-220-SSA1-2016, Facilities and Operations of pharmacovigilance., CDMX: Official 
Gazette of the Federation. , 2017.  
[34] D. Turbert, "American Academy of Ophthalmology," 03 Feb 2021. [Online]. Available: 
https://www.aao.org/eye-health/symptoms/itchiness. [Accessed 2022 Jan 20]. 
[35] D. Turbert, "American Academy of Ophthalmology," 04 Feb 2021. [Online]. Available: 
https://www.aao.org/eye-health/symptoms/tearing. [Accessed 20 Jan 2022]. 
[36] D. A. Palay y J. H. Krachmer, «Ophthalmic Differential Diagnosis,» de Primary Care Ophthalmology, 
Philadephia, PA., Elsevier Mosby, 2005, p. 34. 
[37] "American Academy of Ophthalmology," 25 Jan 2021. [Online]. Available: https://www.aao.org/eye-
health/symptoms/burning-eyes. [Accessed 20 Jan 2022]. 
[38] N. Efron, «Grading scales for contact lens complications,» Ophtalmic Physiol Opt, vol. 18, pp. 182-186, 
1998.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  84 de 90 
[39] European Group of Graves Orbitopathy, "Eugogo," ETA, [Online]. Available: 
http://www.eugogo.eu/_downloads/clincial_evaluation/CHEMOSIS-GO.pdf. [Accessed 11 April 2016]. 
[40] J. Kanski, Oftalmología Clínica, Barcelona: Elsevier, 2009.  
[41] International Dry Eye WorkShop 2007, «Methodologies to diagnose and monitor dry eye disease,» Ocul 
Surf, vol. 5, nº 2, pp. 108-152, 2007.  
[42] A. J. Bron, V. E. Evans y J. A. Smith, «Grading of corneal and Conjunctival Staining in the Context of Other 
Dry Eye Tests,» Cornea, vol. 22, nº 7, pp. 640 - 650, 2003.  
[43] M. Michel, W. Sickenberg y H. Pult, «The effectiveness of questionnaires in the determination of contact 
lens induced dry eye,» Ophthal Physiol Opt, vol. 29, pp. 479-486, 2009.  
[44] Ministry of Health, "Official Mexican Standard NOM-004-SSA3-2012 of the Clinical Record," Mexico City, 
2012. 
[45] A. P. Torres Jiménez and J. M. Torres Rincón, «Climacteric and menopause,» Revisa de la Facultad de 
Medicina UNAM, vol. 61, nº 2, pp. 51-58, 2018.  
[46] J. Kanski, Oftalmología Clínica, Barcelona: Elsevier, 2009.  
[47] International Conference on Harmanisation of Technical Requirements for Registration of 
Pharmaceuticals for Human Use, «Clinical Safety Data Management: Definitions and Standards for 
Expedited Reporting E2A,» ICH Harmonised Tripartite Guideline, 1994. 
[48] International Conference on Harmonisation of Technical Requierements for Registration of 
Pharmaceuticals for Human Use, «General Considerations for Clinical Trials,» ICH Topic E8, 1998. 
[49] Good Clinical Practice, «Participant Safety & Adverse Events,» gcp.nidatraining.org. 
[50] Ministry of Health Mexico, "NORMA Oficial Mexicana NOM-220-SSA1-2016, Instalación y operación de 
la farmacovigilance.," Official Gazette, 2016.  
[51] The Uppsala Monitoring Centre, "Monitoring the Safety of Medicines," World Health Organization, 
2001. 
[52] R. H. Meyboom, A. C. Egberts, I. R. Edwards, Y. A. hekster, F. H. de Koning y F. W. Gribnau, «Principles of 
signal detection in pharmacovigilance,» Drug Safety, vol. 16, pp. 355-365, 2997.  
[53] L. Baiza-Durán, O. Olvera-Montaño, A. Mercado-Sesma, A. Oregon-Miranda, A. Lizárraga-Corona, J. 
Ochoa-Tabares y e. al., «Efficacy and safety of 0.6% pazufloxacin ophthalmic solution versus 
moxifloxacin 0.5% and gatifloxacin 0.5% in subjects with bacterial conjuntivitis: a randomized clinical 
trial.,» J Ocul Pharmacol Ther, vol. 34, nº 3, pp. 250-255, 2018.  
[54] S. Tauber, C. Gale, R. Garber, J. Bartell, F. Vohra y D. Stroman, «Microbiological efficacy of a new 
ophthalmic formulation of moxifloxacin dosed twice-daily for bacterial conjunctivitis,» Adv Ther, vol. 28, 
nº 7, pp. 566-74, 2010.  
[55] G. Keating, «Moxifloxacin 0.5% ophthalmic solution: in bacterial conjunctivitis.,» Drugs, vol. 71, nº 1, pp. 
89-99, 2011.  
[56] P. Garg, U. Mathur, P. Sony, R. Tandon, T. Morris y T. Comstock, «Clinical and antibacterial efficacy and 
safety of besifloxacin ophthalmic suspension compared with moxifloxacin ophthalmic solution.,» Asia 
Pac J Ophthalmol (Phila)., vol. 4, nº 3, pp. 140-5, 2015.  
[57] T. Comstock, M. Paterno, H. Decory y D. Usner, «Safety and tolerability of besifloxacin ophthalmic 
suspension 0.6% in the treatment of bacterial conjunctivitis: data from six clinical and phase I safety 
studies.,» Clin Drug Investig., vol. 30, nº 10, pp. 675-85, 2010.  


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  85 de 90 
[58] M. McDonald, E. Protzko, L. Brunner, T. Morris, W. Haas, M. Paterno, T. Comstock y D. Usner, «Efficacy 
and safety of besifloxacin ophthalmic suspension 0.6% compared with moxifloxacin ophthalmic solution 
0.5% for treating bacterial conjunctivitis.,» Ophthalmology, vol. 116, nº 9, pp. 1615-1623.e1, 2009.  
[59] A. Mokdad, X. Xie, H. Zhu, D. Gerber and D. Heitjan, "Statistical justification of expansion cohorts in 
phase 1 cancer trials," Cancer, vol. 12416, pp. 3339 - 45 , 2018.  
[60] S. Chow, J. Shao and H. Wang, Sample size calculations in Clinical Trials, 2nd ed., Boca Raton, Fl: 
Chapman and Hall/CRC Biostatistics series, 2008, p. 89. 
[61] R Foundation for Statistical Computing, «R Core Team,» 2021. [En línea]. Available: www.R-project.org. 
[62] HyLown Consulting LLC, «Power and Sample Sizes,» HyLown Consulting LLC, 2021. [En línea]. Available: 
http://powerandsamplesize.com. [Último acceso: 2021]. 
[63] B. Krishnankutty, S. Bellary, N. Kumar and L. .. Mooddahadu, "Data management in clinical research: an 
overview.," Indian J Pharmacol, vol. 44, no. 2, pp. 168-72, 2012.  
[64] L. Su, "Clinical data management: current status, challenges, and future directions from industry 
prespectives.," OA J Clin Trials, vol. 2, pp. 93-105, 2010.  
[65] D. Azzam y Y. Ronquillo, «NCBI - StatPearls,» National Center for Biotechnology Information, 9 May 
2021. [En línea]. Available: https://www.ncbi.nlm.nih.gov/books/NBK558961/. [Último acceso: 28 Ene 
2022]. 
 
 
 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  86 de 90 
16. Annexes 
16.1 Cartilla de Snellen 
 
Cartilla de Snellen. Taken from Azzam and Ronquillo, 2021 [65] 
The above annex is a representation of the Snellen booklet, it is possible that the booklet designated for the study has variations in 
format. 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  87 de 90 
16.2. Efron Scale for Conjunctival Hyperemia 
 
 
 
 
 
 
16.3 Oxford scale 
Panel 
Staining pattern 
Degree 
Criterion 
Verbal Description 
A 
 
0 
Equal to or less than 
panel A 
Absent 
B 
 
I 
Equal to or less than 
panel B, greater than 
panel A 
Minimum 
C 
 
II 
Equal to or less than 
panel C, greater than 
panel B 
Lightweight 
D 
 
III 
Equal to or less than 
panel D, greater than C 
Moderate 
And 
 
IV 
Equal to or less than 
panel E, greater than 
panel D 
Marked 
>E 
 
 
V 
Larger than panel E 
Severe 
Oxford scale. Staining is represented by the dotted pattern (adapted from Bron AJ et al, 2003) [42] 
 


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  88 de 90 
16.4 Eye Comfort Index (JI) 
Fecha: 
Iniciales del sujeto:
No. de sujeto:
Indicaciones:
Este cuestionario fue diseñado para calificar el confort de sus ojos.
Para cada pregunta circule su respuesta
Ejemplo:
En la semana pasada, ¿qué tan seguido sus ojos estuvieron rojos?
No existen respuestas correctas o incorrectas. No tome demasiado tiempo en cada pregunta.
1
En la semana pasada, ¿qué tan seguido sus ojos se sintieron secos?
Cuando sus ojos se sentían secos, por lo general, ¿qué tan intensa era la sensación?
2
En la semana pasada, ¿qué tan seguido sus ojos se sintieron arenosos ?
Cuando sus ojos se sentían arenosos , por lo general, ¿qué tan intensa era la sensación?
3
En la semana pasada, ¿qué tan seguido sus ojos sintieron punzadas ?
Cuando sus ojos sentían punzadas , por lo general, ¿qué tan intensa era la sensación?
4
En la semana pasada, ¿qué tan seguido sus ojos se sintieron cansados ?
Cuando sus ojos se sentían cansados , por lo general, ¿qué tan intensa era la sensación?
5
6
Nunca
Siempre
Índice de Confort Ocular
Nunca
0
1
2
3
4
Ficha de Identificación
No. de estudio:
SOPH231-1221/I
6
Siempre
Severo
0
1
2
3
4
5
0
1
2
3
4
5
6
Nunca
Siempre
0
1
2
3
4
5
6
Severo
0
1
2
3
4
5
6
No he sentido
Nunca
Siempre
0
1
2
3
4
5
6
Severo
0
1
2
3
4
5
6
No he sentido
Nunca
Siempre
0
1
2
3
4
5
6
Severo
0
1
2
3
4
5
6
No he sentido
Hoja 1 de 2
No he sentido


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  89 de 90 
 
 
5
En la semana pasada, ¿qué tan seguido sus ojos se sintieron adoloridos ?
Cuando sus ojos se sentían adoloridos , por lo general, ¿qué tan intensa era la sensación?
6
En la semana pasada, ¿qué tan seguido sus ojos sintieron comezón ?
Cuando sus ojos sentían comezón , por lo general, ¿qué tan intensa era la sensación?
Nunca
Siempre
0
1
2
3
4
5
6
Severo
0
1
2
3
4
5
6
No he sentido
No he sentido
Índice de confort ocular
Hoja 2 de 2
Índice de confort ocular, traducido del Ocular Comfort Index disponible en: http://iovs.arvohournals.org
Severo
0
1
2
3
4
5
6
Nunca
Siempre
0
1
2
3
4
5
6


Laboratorios Sophia, S.A. de C.V.  
 
Study No: SOPH231-1221/I 
 
 
Version 2.0 
 
 
27-Jul-22 
C O N F I D E N C I A L 
   Page  90 de 90 
16.5 Fraction to LogMAR conversion table.  
LogMAR Value 
Snellen (pies) 
Snellen (metros) 
1.6 
20/800 
20/240 
1.5 
20/640 
6/190 
1.4 
20/500 
6/150 
1.3 
20/400 
6/120 
1.2 
20/320 
6/96 
1.1 
20/250 
6/75 
1.0 
20/200 
6/60 
0.9 
20/160 
6/48 
0.8 
20/125 
6/38 
0.7 
20/100 
6/30 
0.6 
20/80 
6/24 
0.5 
20/63 
6/19 
0.4 
20/50 
6/15 
0.3 
20/40 
6/12 
0.2 
20/32 
6/9.5 
0.1 
20/25 
6/7.5 
0.0 
20/20 
6/6 
-0.1 
20/15 
6/4.5 
-0.2 
20/12 
6/3.6 
Visual acuity equivalence table (adapted from Ophthalmic & Physiological Optics, 2016)  [69]