Clinical trial NCT04784091
Conditions: Blepharitis
Keywords: Demodex
Sponsor: Tarsus Pharmaceuticals, Inc.
Start date: 2021-04-29; Completion date: 2022-05-09
Enrollment: 412 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
The purpose of this study is to compare the safety and efficacy of TP-03, an eyedrop, to its vehicle control for the treatment of blepharitis due to Demodex, a microscopic mite frequently found in human hair follicles.

Detailed description:
This Phase 3 study is a randomized, controlled, multicenter, double-masked, parallel trial to compare the safety and efficacy of TP-03 to vehicle control for the treatment of Demodex blepharitis. The primary objective of the study is to assess the safety and efficacy of TP-03, 0.25% compared to its vehicle from Day 1 to Day 43 in adult participants with mild to severe Demodex blepharitis. The primary efficacy endpoint will be cure based upon collarettes. Safety will be determined by assessing adverse effects related to the treatment as well as evaluating any changes in visual acuity, intraocular pressure, slit lamp biomicroscopy, endothelial cell density, hematology, blood chemistry, and urinalysis.

Arms: arm 1: TP-03, lotilaner ophthalmic solution, 0.25%, administered topically twice a day for approximately 43 days arm 2: Vehicle of TP-03 ophthalmic solution, administered topically twice a day for approximately 43 days
Interventions: intervention 1: TP-03 intervention 2: TP-03 Vehicle
Intervention descriptions: intervention 1: TP-03, lotilaner ophthalmic solution, 0.25%, administered twice a day intervention 2: Vehicle of TP-03 ophthalmic solution, administered twice a day

Protocol documents:
SAP_004.pdf: 
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STATISTICAL ANALYSIS PLAN 
Randomized, Controlled, Multicenter, Double-Masked, Parallel, Phase 
3 Trial to Evaluate the Safety and Efficacy of TP-03 for the Treatment 
of Demodex Blepharitis (Saturn-2)
Sponsor: 
Tarsus Pharmaceuticals, Inc. 
Protocol Number: 
TRS-010 
 
 
 
 
Date: 
22MAR2022 
 
 
Registration No: NCT04784091


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Table of Contents 
1. 
Introduction............................................................................................................................... 7 
2. 
Study Objectives ...................................................................................................................... 7 
2.1 Primary Endpoint ...................................................................................................................... 7 
2.2 Secondary Endpoints ............................................................................................................... 7 
 
2.4 Safety and Other Assessments................................................................................................ 8 
2.5 Statistical Hypotheses .............................................................................................................. 8 
2.6 Estimands ................................................................................................................................. 9 
2.7 Study Milestones for Analyses ............................................................................................... 10 
3. 
Study Design and Procedures ............................................................................................... 11 
3.1 General Study Design ............................................................................................................ 11 
3.2 Schedule of Activities ............................................................................................................. 13 
4. 
Study Treatments ................................................................................................................... 14 
4.1 Method of Assigning Subjects to Treatment Arms ................................................................. 14 
4.2 Masking and Unmasking ........................................................................................................ 14 
5. 
Sample Size and Power Considerations ................................................................................ 14 
6. 
Data Preparation .................................................................................................................... 16 
6.1 Input Data ............................................................................................................................... 16 
6.2 Output Data ............................................................................................................................ 17 
7. 
Analysis Populations .............................................................................................................. 17 
7.1 Full Analysis Set ..................................................................................................................... 17 
7.2 COVID-19 Analysis Set .......................................................................................................... 17 
7.3 Per-Protocol Set ..................................................................................................................... 18 
7.4 Safety Set ............................................................................................................................... 18 
8. 
General Statistical Considerations ......................................................................................... 18 
8.1 Unit of Analysis....................................................................................................................... 18 
8.2 Missing or Inconclusive Data Handling .................................................................................. 18 
8.3 Definition of Baseline ............................................................................................................. 19 
8.4 Data Analysis Conventions .................................................................................................... 19 
8.5 Adjustments for Multiplicity ..................................................................................................... 20 
9. 
Disposition of Subjects and Protocol Deviations.................................................................... 20 
10. 
Demographic and Pretreatment Variables ............................................................................. 21 
10.1 Demographic Variables .......................................................................................................... 21 
11. 
Medical History and Concomitant Medications ...................................................................... 21 
11.1 Medical History ....................................................................................................................... 22 
11.2 Concomitant Medications ....................................................................................................... 22 


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11.3 Concomitant Procedures ........................................................................................................ 22 
12. 
Dosing Compliance ................................................................................................................ 23 
12.1 Dosing Compliance ................................................................................................................ 23 
13. 
Efficacy Analyses ................................................................................................................... 23 
13.1 Primary Analysis .................................................................................................................... 25 
13.1.1 
Sensitivity Analyses.................................................................................................... 26 
13.1.1.1 
Sensitivity Analysis I ........................................................................................... 27 
13.1.1.2 
Sensitivity Analysis II .......................................................................................... 27 
13.1.1.3 
Sensitivity Analysis III ......................................................................................... 27 
13.1.1.4 
Sensitivity Analysis IV ........................................................................................ 28 
13.1.1.5 
Sensitivity Analysis V ......................................................................................... 28 
13.2 Secondary Analyses .............................................................................................................. 28 
13.2.1 
Secondary Efficacy Endpoints ................................................................................... 29 
 
14. 
Safety Analyses ..................................................................................................................... 31 
14.1 Treatment Exposure ............................................................................................................... 31 
14.2 Adverse Events ...................................................................................................................... 31 
14.3 Corrected Distance Visual Acuity (CDVA) ............................................................................. 33 
14.4 Slit-Lamp Biomicroscopy Examination ................................................................................... 33 
14.5 Corneal Staining ..................................................................................................................... 34 
14.6 Intraocular Pressure (IOP) ..................................................................................................... 34 
14.7 Endothelial Cell Density ......................................................................................................... 35 
14.8 Blood Sample Collection for Hematology, Blood Chemistry, and Urinalysis ......................... 35 
15. 
Other Analyses ....................................................................................................................... 35 
15.1 Drug Concentration Analysis .................................................................................................. 35 
15.2 Urine Pregnancy Test ............................................................................................................ 36 
15.3 Eyelid Photos ......................................................................................................................... 36 
15.4 Drop Comfort .......................................................................................................................... 36 
16. 
Interim Analyses ..................................................................................................................... 37 
17. 
Changes from Protocol-Stated Analyses ............................................................................... 37 
 
 
 
 
 
 
 
 
 
 
 


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List of Abbreviations 
Abbreviation 
Definition 
ADaM 
Analysis Data Model 
AE 
Adverse Event 
ANCOVA 
Analysis of Covariance 
ATC 
Anatomical Therapeutic Chemical 
BID 
Bis in die (Two Times a Day) 
CDVA 
Corrected Distance Visual Acuity 
CI 
Confidence Interval 
COVID-19 
Coronavirus Disease 2019 
CS 
Clinically Significant 
eCRF 
Electronic Case Report Form 
EDC 
Electronic Data Capture 
ETDRS 
Early Treatment Diabetic Retinopathy Study 
FAS 
Full Analysis Set 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
IOP 
Intraocular Pressure 
IRB 
Institutional Review Board 
IRT 
Interactive Response Technology 
logMAR 
Logarithm of the Minimum Angle of Resolution 
LS 
Least Squares 
MCMC 
Markov Chain Monte Carlo 
MedDRA 
Medical Dictionary for Regulatory Activities 
MI 
Multiple Imputation 
NCS 
Not Clinically Significant 
OD 
Oculus dexter (Right Eye) 
OS 
Oculus sinister (Left Eye) 
PDF 
Portable Document Format 
PP 
Per Protocol 
PT 
Preferred Term 
RDC 
Remote Data Capture 
RTF 
Rich Text Format 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SD 
Standard Deviation 
SDC 
Statistics & Data Corporation 
SDTM 
Study Data Tabulation Model 
SE 
Standard Error 
SOC 
System Organ Class 
TEAE 
Treatment-Emergent Adverse Event 
TP-03 
Lotilaner Ophthalmic Solution, 0.25% 
WHODrug 
World Health Organization Drug Dictionary 
 
 


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1. 
Introduction 
The purpose of this statistical analysis plan (SAP) is to describe the planned analyses and reporting for 
protocol TRS-010, Version 4.0 dated 09DEC2021. 
This SAP is being written with due consideration of the recommendations outlined in the most recent 
International Conference on Harmonisation (ICH) E9 Guideline entitled Guidance for Industry: Statistical 
Principles for Clinical Trials and the most recent ICH E3 Guideline entitled Guidance for Industry: Structure 
and Content of Clinical Study Reports. 
This SAP describes the data that will be analyzed and the subject characteristics, efficacy, and safety 
assessments that will be evaluated. This SAP provides details of the specific statistical methods that will 
be used. The statistical analysis methods presented in this document will supersede the statistical analysis 
methods described in the clinical protocol. If additional analyses are required to supplement the planned 
analyses described in this SAP, they may be completed and will be identified in the clinical study report. 
2. 
Study Objectives 
The overarching objective of this study is to demonstrate the safety and efficacy of TP-03, 0.25%, for the 
treatment of Demodex blepharitis. 
The primary objective of this study is to demonstrate the safety and efficacy of lotilaner ophthalmic solution 
(TP-03), 0.25%, compared to its vehicle as a cure of mild to severe Demodex blepharitis. 
The secondary objectives of this study are: 
• 
To demonstrate the efficacy of TP-03, 0.25%, to eradicate Demodex mites from the eyelid margin;  
• 
To demonstrate the efficacy of TP-03, 0.25%, to eliminate collarettes and erythema from the eyelid 
margin; and 
• 
To demonstrate the efficacy of TP-03, 0.25%, to eliminate erythema from the eyelid margin. 
 
 
 
 
2.1 
Primary Endpoint  
The primary efficacy endpoint is cure based on a collarette score of 0 for the upper eyelid of the analysis 
eye at Day 43. The primary safety endpoint is treatment-related adverse effects. 
2.2 
Secondary Endpoints 
The secondary efficacy endpoints are: 
• 
Demodex mite eradication (mite density of 0 mites/lash) from the analysis eye, at Day 43 


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• 
Composite cure based on collarette score (0) and erythema score (0) for the upper eyelid of the 
analysis eye at Day 43 
• 
Erythema cure based on erythema score (0) for the upper eyelid of the analysis eye at Day 43 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
2.4 
Safety and Other Assessments 
The safety and other assessments to be performed include the following: 
• 
Adverse events (AEs) 
• 
Corrected distance visual acuity (CDVA) 
• 
Slit lamp biomicroscopy assessment 
• 
Corneal staining 
• 
Intraocular pressure (IOP) measurement 
 
 
 
 
 
 
 
 
 
 
2.5 
Statistical Hypotheses 
The null and alternative hypotheses, based on the primary efficacy endpoint, are: 
H01: The proportion of subjects cured at Day 43 based on collarette score of 0 for the upper 
eyelid of the analysis eye, by treatment with TP-03, 0.25%, is ≤ the proportion cured 
by treatment with vehicle. 
H11: The proportion of subjects cured at Day 43 based on collarette score of 0 for the upper 
eyelid of the analysis eye, by treatment with TP-03, 0.25%, is > the proportion cured 
by treatment with vehicle. 


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The secondary efficacy hypotheses for Demodex eradication are: 
H02: The proportion of subjects with Demodex eradicated at Day 43 (mite density of 0 
mites/lash) in the analysis eye by treatment with TP-03, 0.25%, is ≤ the proportion 
with Demodex eradicated by treatment with vehicle. 
H12: The proportion of subjects with Demodex eradicated at Day 43 (mite density of 0 
mites/lash) in the analysis eye by treatment with TP-03, 0.25%, is > the proportion 
with Demodex eradicated by treatment with vehicle. 
The secondary efficacy hypotheses for composite of collarette and erythema cure are: 
H03: The proportion of subjects with a composite cure, collarette score of 0 and erythema 
score of 0, for the upper eyelid of the analysis eye at Day 43 by treatment with TP-03, 
0.25%, is ≤ the proportion with a composite cure by treatment with vehicle. 
H13: The proportion of subjects with a composite cure, collarette score of 0 and erythema 
score of 0, for the upper eyelid of the analysis eye at Day 43 by treatment with TP-03, 
0.25%, is > the proportion with a composite cure by treatment with vehicle. 
The secondary efficacy hypotheses for erythema cure are: 
H04: The proportion of subjects with an erythema cure, score of 0, for the upper eyelid of 
the analysis eye at Day 43 by treatment with TP-03, 0.25%, is ≤ the proportion with an 
erythema cure by treatment with vehicle.  
H14: The proportion of subjects with an erythema cure, score of 0, for the upper eyelid of 
the analysis eye at Day 43 by treatment with TP-03, 0.25%, is > the proportion with an 
erythema cure by treatment with vehicle. 
 
2.6 
Estimands 
The primary comparisons in this trial will be between active drug versus vehicle at Day 43 in the Full 
Analysis Set (FAS, as outlined in Section 7.1) using the following estimands.  
• 
Population: subjects with Demodex Blepharitis defined through enrollment criteria 
• 
Endpoint:  
o 
Proportion of subjects cured, based on a collarette score of 0, for the upper eyelid of the 
analysis eye at Day 43 
o 
Proportion of subjects with their Demodex mites eradicated, mite density of 0 mites/lash, from 
the analysis eye at Day 43. 
o 
Proportion of subjects that achieved a composite cure, based on collarette and erythema 
scores of 0, for the upper eyelid of the analysis eye at Day 43. 


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o 
Proportion of subjects that achieved an erythema cure, based on erythema score 0, for the 
upper eyelid of the analysis eye at Day 43 
• 
Intercurrent event:  
o 
Discontinuation of study medication and non-optimal compliance is ignored [treatment policy 
strategy].  
o 
Withdrawal due to lack of efficacy or AEs: missing data will be imputed as failure [hypothetical 
strategy]. 
o 
Missing data without withdrawal or withdrawal due to reasons other than lack of efficacy or 
AEs: missing data will be imputed employing Multiple Imputation (MI) using randomized 
treatment-based Markov Chain Monte Carlo (MCMC) methodology [hypothetical strategy]. 
• 
Population-level summary:  
o 
Difference in proportion of subjects cured, based on a collarette score of 0 for the upper eyelid 
of the analysis eye, at Day 43 
o 
Difference in proportion of subjects with their Demodex mites eradicated, mite density of 0 
mites/lash, from the analysis eye at Day 43. 
o 
Difference in proportion of subjects that achieved a composite cure, based on collarette and 
erythema scores of 0 for the upper eyelid of the analysis eye, at Day 43. 
o 
Difference in proportion of subjects that achieved an erythema cure, based on erythema score 
0 for the upper eyelid of the analysis eye, at Day 43 
 
If the percentage of data missing at Day 43 due to discontinuation of study medication and non-optimal 
compliance exceeds 2%, these missing data will be imputed as failures and the primary efficacy analysis 
will be repeated in a separate analysis.  
Sensitivity analyses on the primary efficacy endpoint and secondary efficacy endpoints are described in the 
Efficacy Analysis section (Section 13).  
2.7 
Study Milestones for Analyses 
Two study milestones for analysis purposes are defined based on subject completion of key visits:  
Milestone 1 will be met after all subjects have completed efficacy assessment visits: The last 
efficacy measures are scheduled at Day 57 for Cohort 1 subjects and Day 43 for Cohort 2 subjects. 
Milestone 1 will be used to assess efficacy and safety during the on-treatment interval, which is 
defined as screening through Day43/57. After this milestone is met, the database will be cleaned, 
locked, unmasked and all tables, listings, and figures will be produced with data for the on-treatment 
interval (excluding Day 90). However, all available data for adverse events and concomitant 
medications and procedures with start dates within the on-treatment interval will be included in the 
tables and listings, and thus AEs or concomitant medications or procedures with start dates during 


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the on-treatment interval and end dates after Day 43/57 will be shown in Milestone 1 tables and 
listings.  
Milestone 2 will be met after all eligible subjects have completed safety and other assessments at 
Day 90. Milestone 2 will be used to assess safety during the post-treatment interval, which is 
defined as after Day 43/57 through Day 90. After this milestone is met listings and tables for subject 
disposition, protocol deviations, analysis population inclusion, subjects effected by COVID-19 and 
the assessments performed at Day 90 (slit-lamp biomicroscopy, endothelial cell density, CDVA) 
will be re-run to include the Day 90 assessments. Adverse event and concomitant medication and 
procedure listings will be re-run to include the Day 90/post-treatment interval data, as will listings 
for any safety or efficacy endpoints collected during an unscheduled visit during the post-treatment 
interval. The tables for the AEs and concomitant medications and procedures with start dates 
during on-treatment interval will also be re-run and updated to include the Day 90/post-treatment 
interval data. Separate tables for AEs and concomitant medications and procedures with start dates 
after milestone 1, during the post-treatment period, will be generated for Milestone 2. 
3. 
Study Design and Procedures 
3.1 
General Study Design 
This Phase 3 study is a randomized, controlled, multicenter, double-masked, parallel trial to evaluate the 
safety and efficacy of TP-03, 0.25%, for the treatment of Demodex blepharitis.  
A subject’s participation is to start with screening at Day -14 to 1. At the Screening visit, potential subjects 
will be evaluated for eligibility. Prior to performing any study specific procedures, potential subjects must 
provide informed consent using the current Institutional Review Board (IRB) approved informed consent 
form (ICF). Potential subjects who meet all eligibility criteria can proceed directly to the Day 1 procedures 
or the Day 1 visit can be scheduled within the next 14 days. If a subject does not come in for the Day 1 visit 
within 14 days, the Screening visit procedures must be repeated. If the subject is considered eligible for the 
study at the end of the Screening visit, the subject will be randomized at Day 1 to receive one of the following 
treatments administered bilaterally two times a day (BID) for approximately 43 days: TP-03, 0.25%, or the 
TP-03 vehicle. Subjects in Cohort 1 (signed informed consent on or before 21 October 2021) will complete 
an observational study assessment visit (Day 57) while Cohort 2 subjects (signed informed consent on 22 
October 2021 or later) will not complete a Day 57 visit. Subjects from both cohorts at sites using specular 
microscopy will also attend a safety assessment including endothelial cell counts and end of study visit 
(Day 90). 
For subjects reporting for an unscheduled visit, assessments may be performed at the discretion of the 
investigator. At a minimum, CDVA and the occurrence of any AEs should be assessed. 
In this study, subjects will be instructed to administer a single drop of the study product BID (morning and 
evening) in each eye. The treatment period will be for approximately 43 days. 






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4. 
Study Treatments 
4.1 
Method of Assigning Subjects to Treatment Arms 
All subjects, investigators, study personnel involved with the conduct of the study and Sponsor will be 
masked with regard to treatment assignments. 
A balanced (1:1) randomization will be used for this study. A computer-generated blocked randomization 
schedule will be generated by 
. 
 will use the randomization schedule to 
package the study drug. The labelled study drug will be shipped to the US distribution center for distribution 
to the clinical sites. 
While undergoing Screening, potential study subjects will be identified by their initials and screening number 
in the screening and enrollment log. All subjects screened for the study who sign an ICF will be assigned a 
3-digit screening number. Screening numbers will be assigned in a sequential order beginning with 301. A 
dash (-) will be used in place of the middle initial for potential subjects who have no middle name. 
If a potential study subject has provided written informed consent and met all eligibility criteria, the subject 
is considered eligible to participate. If the potential subject elects to participate in the study, they can 
continue with the Day 1 procedure or be scheduled to return for Day 1. At Day 1, they will be dispensed the 
next available study drug bottle from the current site inventory by the Interactive Response Technology 
(IRT) system (
), which uses block stratification by site. The 
subject will be randomized and receive their assigned study drug at the completion of the Day 1 visit and 
the subject will be considered enrolled in the study. The subject will be assigned a unique identification 
number consisting of their site number followed by a hyphen and then the screening number. 
4.2 
Masking and Unmasking 
Throughout the study, the subject, investigator, site personnel performing study assessments and Sponsor 
will remain masked to the study medication. The active and vehicle control ophthalmic solutions will be 
indistinguishable in appearance and will be distributed in bottles identical in appearance. 
Treatment assignments should only be unmasked if needed for proper care of the subject. Sites will be 
provided instructions on the procedure to be followed to unmask the treatment assignment for a subject. 
Inadvertent unmasking will be reported to the sponsor and IRB as required. 
The IRT system will also be used to dispense any replacement bottles needed by a subject if they lose their 
current bottle or it becomes damaged. 
5. 
Sample Size and Power Considerations 
A total sample size of 300 subjects (150 subjects per arm) yields approximately 99% power to establish 
superiority of TP-03 to vehicle in the subjects meeting the primary efficacy endpoint assuming a response 
rate of 80.0% in TP-03 and 15.8% for vehicle treatments, respectively, using Pearson chi-squared to test a 
one-sided hypothesis with a significance level of 0.025. Response rates for sample size calculations are 


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based upon a prior outside United States clinical study of TP-03, 0.25%, and vehicle for the treatment of 
Demodex blepharitis. 
Based on the results of a more recent Phase 2b/3 study, TRS-009 (Saturn-1), with a response rate of 44.0% 
for the TP-03 treatment group and 7.4% for the vehicle treatment group, a total sample size of 300 subjects 
still provides greater than 99% power. 
Similarly, 300 subjects (150 subjects per arm) yields 99% power to establish superiority of TP-03 to vehicle 
treatment in the proportion of subjects with Demodex eradicated and 99% power for the proportion of 
subjects with a composite of collarette and erythema cure at Day 43 assuming a response rate of 73.3% in 
TP-03 and 21.1% in vehicle for Demodex eradication and 73.3% in TP-03 and 10.5% in vehicle for 
composite cure using Pearson chi-squared to test a one-sided hypothesis with a significance level of 0.025. 
Based on the results from TRS-009 (Saturn-1) with response rates of 67.9% in the TP-03 treatment group 
and 17.6% in the vehicle treatment group for Demodex eradication and 13.9% in the TP-03 treatment group 
and 1.0% in the vehicle treatment group for composite cure, a sample size of 300 subjects still provides 
greater than 99% power to demonstrate superiority of TP-03 to vehicle for each endpoint. 
For erythema cure, a sample size of 300 subjects will provide 88% power to demonstrate superiority of TP-
03 to vehicle at a one-sided significance level of 0.025 based on the results from TRS-009 (Saturn-1), 
response rate of 19.1% for TP-03 and 7.4% for vehicle. 
The study will be considered a success and TP-03, 0.25% will be considered superior to vehicle in clinical 
cure if H01 is rejected in favor of H11. With this sample size, the study will have > 95% power to claim success 
for clinical cure. A closed hierarchical testing structure will be used such that the analysis will first be 
performed for the primary efficacy endpoint. Only if the null hypothesis for the primary efficacy endpoint is 
rejected will analyses be performed for the three secondary efficacy endpoints using the Hochberg testing 
strategy. Additional details can be found in Section 8.5. If in addition to success for clinical cure, the largest 
p-value among the secondary endpoints allows for rejection of the null hypothesis in favor of the alternative 
hypothesis at a significance level of 0.025 then the study will also claim superiority of TP-03, 0.25%, to 
vehicle for all secondary endpoints. If the study cannot reject the null hypothesis for the secondary endpoint 
with the largest p-value, the next largest p-value among the remaining two endpoints will be evaluated at a 
significance level of 0.0125.  The study will claim superiority of TP-03, 0.25%, to vehicle for both remaining 
endpoints if the null hypothesis is rejected in favor of the alternative hypothesis. If the study cannot reject 
the null hypothesis, the remaining secondary endpoint will be evaluated at a significance level of 0.0083. If 
the null hypothesis is rejected in favor of the alternative, the study will be able to claim success for the 
remaining secondary endpoint. 
Accounting for a 30% discontinuation, approximately 209 subjects per arm (approximately 418 total) will be 
randomized. The planned enrollment is much greater than typically planned to achieve 300 completed 


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subjects due to concerns regarding the impact of COVID-19 and the possibility that up to two sites may be 
unable to complete the study. 
6. 
Data Preparation 
6.1 
Input Data 
Electronic Case Report Forms (eCRF) will be developed by Statistics & Data Corporation (SDC). Data from 
source documents will be entered into the eCRF by site personnel. All users will complete role-based 
system and study-specific eCRF training prior to receiving access to the study database. User access will 
be granted based on a user’s role in the study and will be controlled through individual login credentials 
including a unique User ID and password. 
The clinical study database will be developed and tested in iMedNet™ v1.204.0 or higher. iMedNet™ is 
delivered as a single-instance multi-tenant Software-as-a-Service (SaaS) electronic data capture (EDC) 
system and is developed, maintained, and hosted by MedNet Solutions located in Minnetonka, Minnesota. 
Over the duration of the study, MedNet Solutions may apply system updates to the EDC system as part of 
their continuous improvement efforts. 
After data are entered into the clinical study database at each milestone, described in Section 2.7, electronic 
edit checks and data review will be performed. All data validation specifications and procedures are detailed 
in the Data Validation Manual as a separate document. When the database has been declared to be 
complete and accurate, the database will be locked. Any changes to the database after data have been 
locked can only be made with the approval of Tarsus Pharmaceuticals, Inc. in consultation with SDC and 
per SDC standard operating procedures (SOPs).  
Analyses outlined in this document will be carried out after the following have occurred for the first 
milestone: 
• 
All data management requirements are met according to SDC SOPs, including data entry, 
performance of edit and validation checks, documentation and resolution of data queries, 
generation of coding reports, SAE reconciliation, and database lock with written authorization 
provided by appropriate SDC and Tarsus Pharmaceuticals personnel. 
• 
Protocol deviations have been identified and status determined as major or minor as well as the 
potential impact on the primary efficacy endpoint. 
• 
Analysis populations have been determined. 
• 
Randomized treatment codes have been unmasked 
At the second milestone the following will occur: 
• 
All data management requirements are met according to SDCSOPs, including data entry, 
performance of edit and validation checks, documentation and resolution of data queries, 


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generation of coding reports, SAE reconciliation, and database lock with written authorization 
provided by appropriate SDC and Tarsus Pharmaceuticals personnel. 
• 
Protocol deviations have been identified and status determined as major or minor. 
6.2 
Output Data 
Data from electronic data capture (EDC) and external data will be transferred to Biostatistics and 
incorporated into standard formats following the Study Data Tabulation Model (SDTM). Data will then be 
mapped to analysis datasets using the Analysis Data Model (ADaM). Both SDTM- and ADaM-formatted 
data will be used to create the subject listings, while all tables and figures will be based on the ADaM-
formatted data. 
SDTM will follow the SDTM version 1.7 model and will be implemented using the SDTM Implementation 
Guide version 3.3 and the latest version of the SDTM Controlled Terminology at the time of study start. 
ADaM data will follow the ADaM version 2.1 model and will be implemented using the ADaM 
Implementation Guide version 1.1. Both SDTM and ADaM will be validated using Pinnacle 21 version 3.1.2 
or later. Any discrepancies in the validation will be noted in reviewer’s guides accompanying the final data 
transfers. 
Define.xml will be created for SDTM and ADaM using the Define-XML version 2.0 model. 
7. 
Analysis Populations 
7.1 
Full Analysis Set 
The Full Analysis Set (FAS) will include all randomized subjects. This population will be the primary 
population for efficacy analyses and will be used to summarize all efficacy variables. Subjects in the FAS 
will be analyzed as randomized. 
7.2 
COVID-19 Analysis Set 
The COVID-19 pandemic is continuing during the treatment phase of this study and has impacted the 
conduct of this clinical study. Challenges from quarantines and potential site closures may lead to difficulties 
in meeting protocol-specified procedures. Protocol deviations related to COVID-19 will be documented at 
the subject level. Prior to unmasking, COVID-19-related protocol deviations will be assessed for their 
significance based on their impact to primary endpoint data. 
The COVID-19 analysis set includes all randomized subjects but excludes subjects that discontinued due 
to COVID-19 complications, subjects that would have discontinued due to COVID-19 had they remained 
on study, and subjects that had significant COVID-19-related protocol deviations. Subjects that would have 
discontinued due to COVID-19 had they remained on study will be determined and documented prior to 
unmasking. Protocol deviations will be assessed prior to unmasking for their significance based on their 
impact to primary endpoint data. Sensitivity analyses of the primary efficacy and key secondary analyses 
will be performed on the COVID-19 population and subjects will be analyzed as randomized if at least 5% 
of subjects met these criteria. 


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7.3 
Per-Protocol Set  
The per-protocol (PP) set will include subjects in the FAS who do not have significant protocol deviations 
that potentially could effect the primary endpoint analyses. Protocol deviations will be assessed prior to 
unmasking. Subjects in the PP set will be analyzed as treated. 
This population will be the secondary population for efficacy analyses and will be used to summarize a 
subset of efficacy variables. If the PP and FAS populations are exactly the same, then additional efficacy 
analyses on the PP population will not be performed.  
7.4 
Safety Set 
The Safety set will include all randomized subjects who have received at least one dose of study medication. 
This population will be used to summarize safety variables and will summarize subjects as treated. 
8. 
General Statistical Considerations 
8.1 
Unit of Analysis 
Safety endpoints will be analyzed for both eyes. For subject-level efficacy endpoints, the unit of analysis 
will be the subject. For efficacy endpoints assessed on each eye individually, the unit of analysis will be the 
analysis eye. The analysis eye is defined as the eye that met all of the inclusion criteria. If both eyes met 
all of the inclusion criteria, then the analysis eye will be the eye with the highest Demodex density at the 
Screening visit or, if both eyes have equal Demodex density, the right eye. 
8.2 
Missing or Inconclusive Data Handling 
Missing data due to intercurrent events will be handled as per Section 2.6. For the primary analysis, missing 
data due to lack of efficacy or AEs will be imputed as failures, and missing data without withdrawal or 
withdrawal due to reasons other than lack of efficacy or AEs will be imputed employing MI using randomized 
treatment-based MCMC methodology and is further explained in Section 13.   
Partial/missing start and end dates for AEs and concomitant medications will be imputed as follows: 
Partial/missing start date:  
• 
Dates with missing day only will be imputed as the 1st of the month unless the month and year are 
same as the month and year of first dose of study medication, in which case missing day will be 
imputed as the first dose day of study medication.  
• 
Dates with both day and month missing will be imputed as 1 Jan unless the year is same as the 
year of first dose of study medication, in which case missing day and month will be imputed as the 
first dose day and month of study medication. 
• 
Completely missing dates will be imputed as the first dose date of study medication unless the end 
date is on or before the first dose date of study medication, in which case missing date will be 
imputed as 1 Jan of the same year as the end date. 
Partial/missing end date:  


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• 
Dates with missing day only will be imputed as the last day of the month unless the month and year 
are the same as the month and year of the last dose of study medication, in which case missing 
day will be imputed as the last dose day of study medication.  
• 
Dates with both day and month missing will be imputed as 31 Dec unless the year is same as the 
year of the last dose of study medication, in which case missing day and month will be imputed as 
the last dose day and month of study medication.  
• 
If the ongoing flag is missing or “Yes” then the date will not be imputed unless death date is 
available, in which case the missing date will be imputed as the death date. If ongoing is “No” then 
the missing end date will be imputed as the last dose date. 
• 
If the imputed date is after the date of death, then the end date will be set equal to the date of 
death.  
The original dates will be displayed in data listings and the imputed dates will be used in derivations only 
(study day, treatment-emergence status, etc.). 
8.3 
Definition of Baseline 
Baseline measures are defined as the last measure prior to the initiation of study treatment. Change from 
Baseline will be calculated as Follow-up Visit measure – Baseline Visit measure. 
8.4 
Data Analysis Conventions 
All data analyses will be performed by SDC after the study is completed and the database has been locked 
and released for unmasking. Statistical programming and analyses will be performed using SAS® version 
9.4 or higher. Output will be provided in rich text format (RTF) for tables and portable document format 
(PDF) for tables, listings, and figures using landscape orientation.  
Summaries for continuous and ordinal variables will include the number of observations (n), arithmetic 
mean, standard deviation (SD), median, minimum, and maximum. Minima and maxima will be reported with 
the same precision as the raw values; means and medians will be presented to one additional decimal 
place than reported in the raw values. SDs will be presented to two additional decimal places than reported 
in the raw values. Summaries for discrete variables will include counts and percentages. All percentages 
will be rounded to one decimal place (i.e., XX.X%). Differences between the active treatment arm and 
placebo will be calculated as active minus placebo.  
All statistical tests will be one-sided with a significance level of 0.025 (α = 0.025) unless otherwise specified. 
Confidence intervals (CI) for differences between treatment arms as well as change from baseline will be 
two-sided at a 95% confidence level. All p-values will be rounded to 4 decimal places; p-values less than 
0.0001 will be presented as “<0.0001”; p-values greater than 0.9999 will be presented as “>0.9999.” 
Unless otherwise specified, summaries will be presented by treatment arm and, where appropriate, visit. 
Listings will be based on all randomized subjects unless otherwise specified, and sorted by subject number, 
visit/time point, and parameter as applicable. 


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8.5 
Adjustments for Multiplicity 
A closed hierarchical testing structure will be used where the analysis will be performed for the primary 
efficacy endpoint and, only if successful, the analysis will be performed for the three secondary efficacy 
endpoints using the Hochberg testing strategy. Specifically, if the null hypothesis, H01, is rejected at a one-
sided α of 0.025, the study will be considered a success for clinical cure and the family of three secondary 
efficacy endpoints will be tested using the Hochberg testing strategy with a familywise α of 0.025.  If the 
secondary endpoint with the largest p-value is significant at the 0.025 level, then all three secondary 
endpoints will be declared statistically significant. If the secondary endpoint with the largest p-value is not 
significant at the 0.025 level, then the secondary endpoint with the second largest p-value will be tested at 
the 0.0125 level. If this secondary endpoint is significant at the 0.0125 level, then it and the endpoint with 
the smallest p-value will be declared statistically significant. If not, the endpoint with the smallest p-value 
will be tested at the 0.0083 level. If the null hypothesis, H01 is not rejected at a one-sided α of 0.025 testing 
will stop and the three secondary efficacy hypotheses will not be tested. 
9. 
Disposition of Subjects and Protocol Deviations 
Subject disposition will be presented in terms of the numbers and percentages of subjects who were 
randomized to masked study medication with subcategories for those who did or did not dose with the 
masked study medication; those who were included in the FAS, COVID-19, PP, and Safety sets; and those 
who completed the study and discontinued from the study. Subjects who do not discontinue from the study 
will be considered study completers. Subject enrollment and disposition will be summarized by treatment 
arm and completion/discontinuation will be summarized by treatment arm and site. Percentages will be 
calculated using randomized subjects as the denominator unless otherwise specified.  
The reasons for premature study discontinuation will be summarized by treatment arm for all discontinued 
subjects. Percentages will be calculated using randomized subjects as the denominator. The reasons for 
study discontinuation that will be summarized include pregnancy, significant study treatment non-
compliance per PI determination, exclusion criterion, reasons relating to COVID-19, AE, and other. 
Withdrawal due to lack of efficacy will be summarized by treatment arm. A subject listing will be provided 
that includes the date of and reason for premature study discontinuation. 
The number and percentage of subjects with any deviation, major deviation, minor deviation, and the total 
number of deviations related to COVID-19 will be summarized by treatment arm for all randomized subjects. 
The protocol deviations that will be summarized include the following categories: informed consent, 
inclusion/exclusion and randomization, test article/study drug instillation and assignment at site, improper 
protocol procedures at site, site’s failure to report SAE/AE, visit out of window (missed, early, or late), 
subject non-compliance with test article, subject’s use of prohibited concomitant medication, subject’s 
failure to follow instructions, other, COVID-19: visit missed, COVID-19: assessment not performed, and 
COVID-19: other. A subject listing will be provided that includes the date of the deviation, the deviation 
code, the deviation description, classification of whether the deviation was judged to be COVID-19 related, 


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and the classifications of whether the deviations were judged to be major or minor and potential impact on 
the primary efficacy endpoint based on a masked review. 
In addition, subject listings will be provided that include informed consent date, inclusion and exclusion 
criteria violations, and exclusions from any population, subjects affected by COVID-19, and withdrawal due 
to lack of efficacy. Details of the study randomization, including randomization date, site, randomized 
treatment, and actual treatment, will also be included within a subject listing. 
Tables and listings for subject disposition and protocol deviations will be generated after the conclusion of 
Milestone 1 and will be re-run after the conclusion of Milestone 2, as described in Section 2.7. 
10. 
Demographic and Pretreatment Variables 
10.1 
Demographic Variables 
The demographic variables collected in this study include age, sex, childbearing potential, race, ethnicity, 
and iris color (right eye [OD] and left eye [OS]). Subjects can select more than one race, and they will be 
categorized as Multiple Race on summaries. Demographic variables will be summarized for the FAS, 
COVID-19 Analysis Set, PP set, and Safety set separately. Demographic variables will also be summarized 
by site for the FAS. 
Age (years) will be summarized by treatment arm and overall using continuous descriptive statistics as well 
as dichotomized into age categories of <65 years and ≥65 years. Age will be reported in years and 
calculated using the age at last birthday. 
The number and percentage of subjects will be presented by treatment arm and overall for sex, childbearing 
potential, race, ethnicity, and iris color (OD and OS). 
A subject listing that includes age, sex, race, ethnicity, and iris color will be provided. Childbearing potential 
will be listed with urine pregnancy results (see Section 15.2). 
Tables and listings for demographics and pre-treatment variables will be generated after the completion of 
Milestone 1, as described in Section 2.7. 
11. 
Medical History and Concomitant Medications 
Tables and listings for medical history will be generated after the completion of Milestone 1, as described 
in Section 2.7. 
As described in Section 2.7, tables, and listings for concomitant medications and procedures with a start 
date during the on-treatment interval will be generated after Milestone 1. These tables and listings will use 
all available data  and thus, concomitant medications and procedures with start dates during the on-
treatment interval and end dates after Day 43/57 will be shown in Milestone 1 tables and listings. After the 
completion of Milestone 2, the concomitant medication and procedures listings, and the on-treatment 
interval tables will be re-run to include the Day 90/post-treatment interval data. Additionally, separate tables 


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for concomitant medications and procedures with start dates during the post-treatment interval will be 
generated for Milestone 2. 
11.1 
Medical History 
Medical history will be coded using the Medical Dictionary for Regulatory Activities (MedDRA) Version 23.0. 
Ocular medical history will be summarized using discrete summary statistics and presented by treatment 
arm at the subject level by System Organ Class (SOC) and Preferred Term (PT) using the Safety set. Non-
ocular medical history will be similarly summarized at the subject level. Medical history will be summarized 
by PT and SOC at the subject level. A subject will only be counted once even if they have multiple histories 
for a given classification. All SOCs and PTs within an SOC will be sorted in decreasing frequency among 
all subjects. 
Listings of medical history will be generated separately for ocular and non-ocular data. 
11.2 
Concomitant Medications 
Concomitant medications will be coded using World Health Organization Drug Dictionary (WHODrug) 
Global (B3, March 2020) and summarized by therapeutic drug class (Anatomical Therapeutic Chemical 
[ATC] 4 classification) and preferred name. If the ATC 4 classification is not provided, the next lowest 
classification that is provided in the coding dictionary will be used. The preferred name will be defined as 
the active ingredient; if the active ingredient is not provided or includes more than two ingredients (e.g., 
multivitamins) then the drug name will be summarized as the preferred name. 
Concomitant medications are defined as those medications reported as having been taken (1) prior to 
initiation of study drug administration and continuing for any period of time following the first administration 
of study drug or (2) at any time following the first administration of study drug. 
Concomitant medications will be summarized using the Safety set. Medications will be tabulated for each 
treatment arm using frequencies and percentages. Subjects may have more than one medication per ATC 
text. At each level of subject summarization, a subject will be counted once if he/she reports one or more 
medications. Percentages will be based on the number of subjects in each treatment arm. The ATC classes 
and preferred names within an ATC class will be sorted in decreasing frequency among all subjects. Listings 
of concomitant medications will be generated separately for ocular and non-ocular data (as specified by the 
investigator), including indication, route of administration, start date, and end date or ongoing. 
11.3 
Concomitant Procedures 
Concomitant procedures will be coded using the Medical Dictionary for Regulatory Activities (MedDRA) 
Version 23.0 and summarized by SOC and PT.  
Concomitant procedures will be summarized using the Safety set. Procedures will be tabulated for each 
treatment arm using frequencies and percentages. Subjects may have more than one procedure per PT. 
At each level of subject summarization, a subject will be counted once if he/she reports one or more 
procedures. Percentages will be based on the number of subjects in each treatment arm. All SOCs and 


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PTs within an SOC will be sorted in decreasing frequency among all subjects. Listings of concomitant 
procedures will be generated separately for ocular and non-ocular data (as specified by the investigator). 
12. 
Dosing Compliance 
12.1 
Dosing Compliance 
Dosing compliance (% compliance) will be assessed by calculating the number of actual doses received 
and comparing that to the number of expected doses as follows: 
Compliance (%) = 
Number of Actual Doses Received x 100% 
Number of Expected Doses 
The number of actual doses received will be recorded in the CRF (first dose in Study Drug Instillation CRF 
and remaining doses in Study Drug Diary CRF). Two doses are expected each day except only one dose 
should be completed on the last day of the study. The number of expected doses that will be used for 
calculating compliance will be calculated as:  
2 x (Date of Last Dose – Date of First Dose) + 1 
for all subjects, regardless of study completion status. Date of last dose will be taken from the last date the 
study drug was taken from the subject diary or the study drug instillation page of the eCRF, whichever is 
later. In the case where drug was temporarily withdrawn, the number of days without study drug will be 
accounted for by subtracting 2 doses per day without study drug. 
A categorical dosing compliance variable will also be derived as non-compliant (<80%), compliant (≥80% 
and ≤125%), and over compliant (>125%). 
Dosing compliance (%) will be summarized with continuous descriptive statistics for each treatment arm, 
as well as by site, sex, age group, and race using the Safety set. The compliance category defined above 
will be summarized with discrete summary statistics.  
A subject listing of dosing compliance will also be produced after Milestone 1. 
13. 
Efficacy Analyses 
All efficacy analyses, as well as the generation of tables, listings, and figures, will be performed after the 
completion of Milestone 1, as described in Section 2.7.  
Study efficacy assessments performed include the following: 
• 
Assessment of collarette grade 
• 
Eyelash epilation and mite counts 
• 
Assessment of erythema grade 


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Collarette grade (score) is determined based on number of lashes with collarettes per eyelid (Table 3). The 
proportion of subjects cured in the upper eyelid of the analysis eye based on collarette grade will be 
summarized by visit and treatment arm. The percentage of subjects at each grade will be summarized by 
visit and treatment arm, as well as by site, sex, age group, and race. The mean collarette score and change 
from Baseline in collarette score will be summarized by visit and treatment arm. The proportion of subjects 
cured (Collarette Grade 0) in the upper eyelid of the analysis eye, and the change from baseline in the 
mean collarette score in the upper eyelid of the analysis eye by visit and treatment arm, will be displayed 
graphically by bar charts. 
Table 3. Collarette Grading 
Grade  Clinical Interpretation 
0 
0 to 2 lashes have collarettes per eyelid 
1 
3 to 10 lashes have collarettes per eyelid 
2 
More than 10 but less than ⅓ of lashes have collarettes per eyelid 
3 
⅓ or more but less than ⅔ of lashes have collarettes per eyelid 
4 
⅔ or more of lashes have collarettes per eyelid 
0.5-unit increments ARE NOT allowed 
Mite density in the analysis eye and change from baseline will be summarized by visit and treatment arm, 
as well as by site, sex, age group, and race. Mite density is the total number of mites counted divided by 
the total number of lashes epilated. The proportion of subjects with Demodex eradication will be 
summarized by visit and treatment arm, as well as by site, sex, age group, and race. The mean mite density 
and change from Baseline in collarette score will be summarized by visit and treatment arm. The proportion 
of subjects with Demodex eradication (Mite Density 0) in the analysis eye, and the change from baseline in 
mean mite density in the analysis eye by visit and treatment arm, will be displayed graphically by bar charts. 
Erythema of the eyelid margin is graded on a scale from 0 (normal) to 3 (severe erythema) (Table 4). The 
percentage of subjects at each grade level in the upper eyelid of the analysis eye will be summarized by 
visit and by treatment arm, as well as by site, sex, age group, and race. The mean erythema score and 
change from Baseline in collarette score will be summarized by visit and treatment arm. The proportion of 
subjects with erythema cure  (erythema grade 0) in the upper eyelid of the analysis eye, and the change 
from baseline in mean erythema score in the upper eyelid of the analysis eye by visit and treatment arm, 
will be displayed graphically with bar charts. 


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Table 4. Lid Margin Erythema Grading 
Grade 
Severity 
Clinical Interpretation 
0 
Normal 
Normal age-related lid coloration 
1 
Mild 
Pink capillary involvement along the lid edge, no patches of confluent 
capillary redness throughout the lid edge 
2 
Moderate 
Deep pink or red confluent capillary redness present locally along the lid 
edge 
3 
Severe 
Deep red, diffuse confluent capillary redness present along the lid edge 
0.5-unit increments ARE NOT allowed 
The proportion of subjects’ composite cure based on collarette of 0 and erythema score of 0 will be 
summarized by visit and by treatment arm, as well as by site, sex, age group, and race. The proportion of 
subjects with composite cure in the upper eyelid of the analysis eye by visit and treatment arm will be 
displayed graphically with bar charts. 
13.1 
Primary Analysis 
The primary efficacy endpoint is cure defined as collarette score of 0 for the upper eyelid of the analysis 
eye at Day 43. Descriptive statistics will be presented by treatment arm. Testing of the percentage of 
subjects cured at Day 43 based on the collarette grade will be completed using a difference of proportions 
test.  
The primary efficacy endpoint analysis will be conducted on the FAS, with intercurrent events handled as 
described in Section 2.6. Discontinuation of study medication and non-optimal compliance will be ignored. 
Missing data due to withdrawal due to lack of efficacy or AEs will be imputed as failure. Missing data without 
withdrawal or withdrawal due to reasons other than lack of efficacy or AEs will be imputed employing MI 
using randomized treatment-based MCMC methodology. The MCMC method simulates missing data under 
a missing at random assumption while accounting for the uncertainty in the imputed values. Missing values 
will be imputed with an arbitrary missing pattern. The procedure will use the posterior mode from the EM 
algorithm as the starting values for the MCMC method. The following SAS® code will be used to generate 
the multiple imputation datasets: 
PROC MI DATA=INDATA SEED=19107 NIMPUTE=20 OUT=OUT1 MINIMUM=0 MAXIMUM=4 ROUND=1; 
 
 
BY TRT; 
 
 
MCMC INITIAL=EM; 
 
 
VAR BASE DAY08 DAY15 DAY22 DAY43; 
RUN; 
where 
- INDATA is the name of the input dataset 
- OUT1 is the name of the output dataset 
- TRT is the name of the treatment group variable 
- BASE is the baseline assessment 
- DAY08 is the assessment at Day 8 
- DAY15 is the assessment at Day 15 
- DAY22 is the assessment at Day 22 
- DAY43 is the assessment at Day 43 


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The MI datasets will be used to calculate cure based on a collarette score of 0 at Day 43. The following 
SAS® code will then be used to calculate the difference of proportions for each imputed dataset and 
combine the analyses across imputations: 
PROC SORT DATA=OUT1; BY _IMPUTATION_; RUN; 
where 
- OUT1 is the name of the input dataset 
- _IMPUTATION_ is the imputation number 
and  
PROC FREQ DATA=OUT1; 
BY _IMPUTATION_ ; 
TABLES TRT*CURE / RISKDIFF CL; 
ODS OUTPUT RISKDIFFCOL2=PROP; 
RUN; 
where 
- OUT1 is the name of the input dataset 
- _IMPUTATION_ is the imputation number 
- TRT is the name of the treatment group variable 
- CURE is the indicator for cure based on a collarette score of 0 at Day 43 
- PROP is the name of the output dataset 
and 
PROC MIANALYZE DATA=PROP; 
 
 
WHERE ROW=’Difference’; 
 
 
MODELEFFECTS RISK; 
 
 
STDERR ASE; 
 
 
ODS OUTPUT PARAMETERESTIMATES=PEST; 
RUN; 
where 
- PROP is the name of the input dataset 
Statistical comparison between treatment arms will be one-sided using an α of 0.025.  
In addition to the primary analysis, sensitivity analyses as described in Section 13.1.1 will be conducted to 
assess the endpoint under different assumptions of missingness.  
Furthermore, the primary endpoint of the proportion of subjects cured based on a collarette score of 0 will 
be analyzed at all visits in the upper eyelid of the analysis eye, the upper eyelid of the fellow eye, the lower 
eyelid of the analysis eye, and the lower eyelid of the fellow eye using observed data only. Separate 
stratified analyses will be conducted for the upper eyelid of the analysis eye by site, sex, age group, and 
race on the FAS using observed data only, unless p-values are not calculable due to sparsity of sample 
size limitations. In addition, if the COVID-19 Analysis Set consists of ≤95% of study subjects, the analysis 
will be conducted for the upper eyelid of the analysis eye at Day 43 on the COVID-19 Analysis Set with 
observed data only. These analyses will be conducted using similar SAS® code as described in Section 
13.1.1.3. 
13.1.1 SENSITIVITY ANALYSES 
The following sensitivity analyses will be conducted on the primary efficacy endpoint. 


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13.1.1.1 
SENSITIVITY ANALYSIS I 
The primary efficacy analysis will be performed on the FAS set using the difference of proportions test. The 
hypothetical strategy introduced in Section 2.6 will be used for the intercurrent event, where missing data 
will be imputed assuming missing not at random (imputing from the vehicle treatment arm). Discontinuation 
of study medication and non-optimal compliance will be ignored. The following SAS® code will be used to 
generate the MI datasets: 
PROC MI DATA=INDATA SEED=43623 NIMPUTE=20 OUT=MDATA MINIMUM=0 MAXIMUM=4 ROUND=1; 
 
 
BY TRT; 
 
 
MCMC IMPUTE=MONOTONE; 
 
 
VAR BASE DAY08 DAY15 DAY22 DAY43; 
RUN; 
PROC MI DATA=MDATA SEED=346238 NIMPUTE=1 OUT=OUT1 MINIMUM = . 0 0 0 0 0 MAXIMUM 
= . 4 4 4 4 4 ROUND = . 1 1 1 1 1; 
BY _IMPUTATION_; 
CLASS TRT; 
MONOTONE REG(DAY08 = BASE/DETAILS); 
MONOTONE REG(DAY15 = BASE DAY08/DETAILS); 
MONOTONE REG(DAY22 = BASE DAY08 DAY15/DETAILS); 
MONOTONE REG(DAY43 = BASE DAY08 DAY15 DAY22/DETAILS); 
MNAR MODEL(BASE DAY08 DAY15 DAY22 DAY43 / MODELOBS=(TRT=’Vehicle’)); 
VAR BASE DAY08 DAY15 DAY22 DAY43; 
RUN; 
13.1.1.2 
SENSITIVITY ANALYSIS II 
The primary efficacy analysis will be performed on the FAS set using the difference of proportions test. The 
hypothetical strategy introduced in Section 2.6 will be used for the intercurrent event, where missing data 
will be imputed assuming missing at random (imputing from the same treatment arm as the subject with 
missing data) using similar code as provided in Section 13.1 for the primary efficacy analysis. 
Discontinuation of study medication and non-optimal compliance will be ignored.  
13.1.1.3 
SENSITIVITY ANALYSIS III 
The primary efficacy analysis will be performed on the PP set with observed data only using the difference 
of proportions test. In addition, a Fisher’s exact test will be performed. The following SAS® code will be 
used to generate the analyses:  
ODS OUTPUT CHISQ = OUT_CHISQ FISHERSEXACT = OUT_F; 
PROC FREQ data = INDATA; 
      TABLES TRT * OTCM / FISHER EXPECTED OUTEXPECT OUT = OUT_EXP; 
RUN; 
Where 
- INDATA is the name of the input dataset 
- TRT is the name of the treatment group variable 
- OTCM is the indicator for cure based on a collarette score of 0 at Day 43 
- OUT_CHISQ is the name of the output dataset that will contain the p-value for 
the difference of proportions test when all expected cell counts are at least 5  
- OUT_F is the name of the output dataset that will contain the p-value for the 
Fisher’s exact test and the difference of proportions test when any expected cell 
count is < 5 
- OUT_EXP is the name of the output dataset that will contain the expected cell 
counts 


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13.1.1.4 
SENSITIVITY ANALYSIS IV 
The primary efficacy analysis will be performed on the FAS set with observed data only using the difference 
of proportions test. In addition, a Fisher’s exact test will be performed. Analyses will be conducted using 
similar SAS® code as described in Section 13.1.1.3. 
13.1.1.5 
SENSITIVITY ANALYSIS V 
The primary efficacy analysis will be performed on the FAS set using the difference of proportions test. The 
hypothetical strategy introduced in Section 2.6 will be used for the intercurrent event where missing data 
will be imputed as failure. Discontinuation of study medication and non-optimal compliance will be ignored. 
In addition, a Fisher’s exact test will be performed.  
13.2 
Secondary Analyses 
The secondary efficacy endpoints are the proportion of subjects with Demodex eradication (mite density of 
0 mites/lash) at Day 43; the proportion of subjects cured based on a composite collarette and erythema 
score of 0 at Day 43; and the proportion of subjects cured based on an erythema score of 0 at Day 43. All 
secondary endpoint analyses will be conducted on the FAS.  
As specified in Section 8.2, missing data without withdrawal or withdrawal due to reasons other than lack 
of efficacy or AEs will be imputed employing MI using randomized treatment-based MCMC methodology. 
The following SAS® code will be used to generate the MI datasets: 
PROC MI DATA=INDATA SEED=10719 NIMPUTE=20 OUT=OUT1 MINIMUM=0 MAXIMUM=4 ROUND=1; 
 
 
BY TRT; 
 
 
MCMC INITIAL=EM; 
 
 
VAR BASE DAY08 DAY15 DAY22 DAY43; 
RUN; 
 
The MI datasets will be used to calculate cure based on each respective secondary endpoint. The following 
SAS® code will then be used to calculate the difference of proportions for each imputed dataset and 
combine the analyses across imputations: 
PROC SORT DATA=OUT1; BY _IMPUTATION_; RUN; 
PROC FREQ DATA=OUT1; 
BY _IMPUTATION_ ; 
TABLES TRT*CURE / RISKDIFF CL; 
ODS OUTPUT RISKDIFFCOL2=PROP; 
RUN; 
PROC MIANALYZE DATA=PROP; 
 
 
WHERE ROW=’Difference’; 
 
 
MODELEFFECTS RISK; 
 
 
STDERR ASE; 
 
 
ODS OUTPUT PARAMETERESTIMATES=PEST; 
RUN; 
Comparisons will be one-sided using an α of 0.025. 


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13.2.1 SECONDARY EFFICACY ENDPOINTS 
The first secondary efficacy endpoint is the proportion of subjects cured at Day 43, as defined by Demodex 
eradication (mite density of 0 mites/lash) in the analysis eye. Descriptive statistics will be presented by 
treatment arm. 
Analysis will be conducted on the FAS, with intercurrent events handled as described in Section 2.6. 
Discontinuation of study medication and non-optimal compliance will be ignored. Missing data due to 
withdrawal due to lack of efficacy or AEs will be imputed as failure. Missing data without withdrawal or 
withdrawal due to reasons other than lack of efficacy or AEs will be imputed employing MI using randomized 
treatment-based MCMC methodology as described in Section 13.1. Testing of the percentage of subjects 
cured at Day 43 based on the Demodex eradication will be completed using a difference of proportions test.  
In addition, a sensitivity analysis will be conducted in which all missing data will be imputed as failure. The 
difference of proportions test and Fisher’s exact test will be used in this sensitivity analysis. Furthermore, 
the proportion of subjects cured based on Demodex eradication will be analyzed at all visits in both the 
analysis and fellow eye using observed data only. Stratified analyses will be conducted for the upper eyelid 
of the analysis eye by site, sex, age group, and race on the FAS using observed data only, unless p-values 
are not calculable due to sparsity or sample size limitations. In addition, if the COVID-19 Analysis Set 
consists of ≤95% of study subjects, the analysis will be conducted for the upper eyelid of the analysis eye 
at Day 43 on the COVID-19 Analysis Set with observed data only. These analyses will be conducted using 
similar SAS® code as described in Section 13.1.1.3. 
The second secondary efficacy endpoint is the proportion of subjects cured at Day 43, as defined by a 
composite score of 0 based on collarette and erythema scores in the analysis eye. Descriptive statistics will 
be presented by treatment arm. 
Analysis will be conducted on the FAS, with intercurrent events handled as described in Section 2.6. 
Discontinuation of study medication and non-optimal compliance will be ignored. Missing data due to 
withdrawal due to lack of efficacy or AEs will be imputed as failure. Missing data without withdrawal or 
withdrawal due to reasons other than lack of efficacy or AEs will be imputed employing MI using randomized 
treatment-based MCMC methodology as described in Section 13.1. Testing of the percentage of subjects 
cured at Day 43 based on a composite score of 0 will be completed using a difference of proportions test.  
In addition, a sensitivity analysis will be conducted in which all missing data will be imputed as failure. The 
difference of proportions test and Fisher’s exact test will be used in this sensitivity analysis. Furthermore, 
the proportion of subjects cured based on a composite score of 0 for collarette and erythema scores will be 
analyzed at all visits in both the analysis and fellow eye, and both the upper and lower eyelid, using 
observed data only. Stratified analyses will be conducted for the upper eyelid of the analysis eye by site, 
sex, age group, and race on the FAS using observed data only, unless p-values are not calculable due to 
sparsity or sample size limitations. In addition, if the COVID-19 Analysis Set consists of ≤95% of study 
subjects, the analysis will be conducted for the upper eyelid of the analysis eye at Day 43 on the COVID-


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19 Analysis Set with observed data only. These analyses will be conducted using similar SAS® code as 
described in Section 13.1.1.3. 
The third secondary efficacy endpoint is the proportion of subjects cured at Day 43, as defined by an 
erythema score of 0 in the analysis eye. Descriptive statistics will be presented by treatment arm. 
Analysis will be conducted on the FAS, with intercurrent events handled as described in Section 2.6. 
Discontinuation of study medication and non-optimal compliance will be ignored. Missing data due to 
withdrawal due to lack of efficacy or AEs will be imputed as failure. Missing data without withdrawal or 
withdrawal due to reasons other than lack of efficacy or AEs will be imputed employing MI using randomized 
treatment-based MCMC methodology as described in Section 13.1. Testing of the percentage of subjects 
cured at Day 43 based on an erythema cure of 0 will be completed using a difference of proportions test.  
In addition, a sensitivity analysis will be conducted in which all missing data will be imputed as failure. The 
difference of proportions test and Fisher’s exact test will be used in this sensitivity analysis. Furthermore, 
the proportion of subjects based on an erythema cure of 0 will be analyzed at all visits in both the analysis 
and fellow eye, and both the upper and lower eyelid, using observed data only. Stratified analyses will be 
conducted for the upper eyelid of the analysis eye by site, sex, age group, and race on the FAS using 
observed data only, unless p-values are not calculable due to sparsity or sample size limitations. In addition, 
if the COVID-19 Analysis Set consists of ≤95% of study subjects, the analysis will be conducted for the 
upper eyelid of the analysis eye at Day 43 on the COVID-19 Analysis Set with observed data only. These 
analyses will be conducted using similar SAS® code as described in Section 13.1.1.3. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  




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of relationship to study drug or seriousness of the event and should be categorized with possible values: 
“Mild”, “Moderate”, or “Severe”. 
The relationship of each AE to the study drug should be determined by the Investigator with possible values: 
“Definitely Related”, “Potentially Related”, and “Not Related”. 
Definitely and potentially related TEAEs are considered treatment-related TEAEs.  
Drop instillation-related AEs are AEs that are determined by the investigator to be related to the instillation 
of the study drug. 
An overall summary will be presented that includes the number of events and the number and percentage 
of subjects who experienced at least one TEAE by treatment arm and over all subjects. This summary will 
also include breakdowns of TEAEs further categorized as ocular or non-ocular; treatment-emergent serious 
adverse events (TE-SAEs) for ocular, non-ocular, and combined; TEAEs by maximal severity; treatment-
related TEAEs by relatedness; drop instillation-related TEAEs; TEAEs leading to premature treatment 
discontinuation; TEAEs leading to temporary treatment discontinuation; and TEAEs leading to death. 
Ocular and non-ocular TEAEs will be summarized using discrete summary statistics and presented by 
treatment arm at the subject and event level by SOC and PT. If a subject reports the same PT multiple 
times, that PT will only be counted once within that SOC. As with the PT, if a subject reports multiple 
conditions within the same SOC, that SOC will only be counted once. In the summary, SOCs will be 
summarized in ascending alphabetical order; PTs will be listed in order of descending frequency for all 
subjects within each SOC.  
Separate summaries will be provided for the following categories of TEAEs:  
• 
Ocular TEAEs by SOC and PT 
• 
Non-ocular TEAEs by SOC and PT 
• 
Ocular TEAEs by SOC, PT, and Maximal Severity 
• 
Non-ocular TEAEs by SOC, PT, and Maximal Severity 
• 
Ocular Treatment Related TEAEs by SOC and PT 
• 
Non-ocular Treatment Related TEAEs by SOC and PT 
• 
Ocular TEAEs by SOC, PT, Maximal Severity, and Strongest Relationship to Study Drug 
• 
Non-ocular TEAEs by SOC, PT, Maximal Severity, and Strongest Relationship to Study Drug 
• 
TEAEs That Led to Premature Treatment Discontinuation 
• 
TEAEs that Led to Temporary Treatment Discontinuation 
• 
Treatment-emergent SAEs 
Summaries of TEAEs by maximal severity will be presented for ocular AEs and non-ocular AEs separately. 
The number of subjects with any TEAEs (along with percentages) will be tabulated by SOC and PT within 


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each SOC by treatment arm. To count the number of subjects with any TEAEs, if a subject has multiple 
TEAEs coded to the same PT, the subject will be counted once under the maximal severity.  
All AEs and TEAEs will be presented in a subject listing. TEAEs that led to premature treatment 
discontinuation, TEAEs that led to temporary treatment discontinuation and AEs that led to study 
discontinuation will be listed separately. SAEs will be presented in a separate listing. In addition, a separate 
listing will be generated for drop instillation-related AEs. 
As described in Section 2.7, tables, and listings for adverse events with a start date during the on-treatment 
interval will be generated after Milestone 1. These tables and listings will use all available data and thus, 
adverse events with start dates during the on-treatment interval and end dates after Day 43/57 will be 
shown in Milestone 1 tables and listings. After the completion of Milestone 2, the adverse events listings, 
and the on-treatment interval tables will be re-run to include the Day 90/post-treatment interval data. 
Additionally, separate tables for adverse events with start dates during the post-treatment interval will be 
generated for Milestone 2. 
14.3 
Corrected Distance Visual Acuity (CDVA) 
Corrected distance visual acuity will be evaluated near the beginning of each study visit (i.e., prior to slit 
lamp examination). Subjects should use their most recent correction to attain their CDVA; if they forget their 
spectacles, their prescription can be placed in a trial frame or a pinhole can be used with the Early 
Treatment Diabetic Retinography Study (ETDRS) Fast procedure under standard illumination. The subject 
will be asked to focus on an ETDRS visual acuity chart 4 meters away, checking one eye at a time. The 
number of letters read correctly will be used to compute the subject’s logarithm of the minimum angle of 
resolution (logMAR) score for CDVA. The procedure used will be consistent with the recommendations 
provided for using the ETDRS-Fast method. 
The observed and change from baseline CDVA will be summarized for each eye using continuous 
descriptive statistics by visit and treatment arm. The proportion of subjects at each visit that had ≥ 0.22 
logMAR change from baseline will be summarized separately for improvement (change from baseline of ≤ 
-0.22 logMAR) or worsening (change from baseline of ≥ 0.22 logMAR) by treatment and eye. A subject 
listing of visual acuity will also be produced. This listing will include a variable that indicates if a subject had 
a visual acuity logMAR change from baseline (worsening or improvement). 
As described in Section 2.7, tables and listings for CDVA will be generated after the completion of Milestone 
1 for the on-treatment interval (excluding Day 90) and will be re-run and updated to include Day 90 data 
after the completion of Milestone 2. 
14.4 
Slit-Lamp Biomicroscopy Examination 
At each study visit, a slit-lamp biomicroscopy examination will be performed. The following structures will 
be assessed for pathology defined as normal, abnormal (not clinically significant [NCS]), or abnormal 
(clinically significant [CS]): 


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• 
Cornea  
• 
Conjunctiva 
• 
Anterior chamber 
• 
Iris 
• 
Eyelids and lashes 
The results will be summarized using counts and percentages for each treatment arm and for all actively 
treated subjects at each visit for each eye. Percentages will be based on the number of eyes in each 
treatment arm with responses. Shift tables for the slit-lamp biomicroscopy parameters will be provided 
comparing each follow-up visit to baseline. A subject listing of the slit-lamp biomicroscopy parameters will 
also be produced. 
As described in Section 2.7, tables and listings for slit-lamp biomicroscopy examinations will be generated 
after the completion of Milestone 1 for the on-treatment interval (excluding Day 90) and will be re-run and 
updated to include Day 90 data after the completion of Milestone 2. 
14.5 
Corneal Staining 
Corneal fluorescein staining will be assessed at the Day 1 (baseline), 8, 15, 22, 43, and 57 study visits 
using the National Eye Institute (NEI)/Industry Scale. The NEI/Industry Scale uses a standardized grading 
system of 0 to 3 for each of the five areas on each cornea. Grade 0 will be specified when no staining is 
present. The maximum score is 15. The following will be assessed: 
• 
Inferior 
• 
Superior 
• 
Central 
• 
Temporal 
• 
Nasal 
The above measures will be summarized using counts and percentages for each treatment arm at each 
visit. Percentages will be based on the number of eyes in each treatment arm with responses. 
In addition, the observed values and change from baseline for total corneal staining, sum of the above 
regions, will be summarized for each eye using continuous descriptive statistics for each treatment arm. A 
subject listing of the corneal staining will also be produced. 
As described in Section 2.7, tables and listings for corneal staining will be generated after Milestone 1 and 
corneal staining listings will only be re-run after Milestone 2 if performed during an unscheduled visit during 
the post-treatment interval.  
14.6 
Intraocular Pressure (IOP) 
At the Day 1 (baseline), 43, and 57 study visits, IOP will be assessed by applanation tonometry in each 
eye. Results will be taken from a single measurement and will be recorded in mmHg. 


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The IOP values and changes from baseline for each eye will be summarized using continuous descriptive 
statistics by visit and eye for each treatment arm. A subject listing of IOP will also be produced. This listing 
will include a variable that indicates if a subject had an IOP increase from baseline of ≥6 or ≥10 mmHg. 
As described in Section 2.7, tables and listings for IOP will be generated after Milestone 1 and IOP listings 
will only be re-run after Milestone 2 if performed during an unscheduled visit during the post-treatment 
interval.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
15. 
Other Analyses 
 
 
 
 
 
 
 





Prot_003.pdf: 
Randomized, Controlled, Multicenter, Double-Masked, 
Parallel, Phase 3 Trial to Evaluate the Safety and 
Efficacy of TP-03 for the Treatment of Demodex 
Blepharitis (Saturn-2)  
Protocol Number: TRS-010 
IND Sponsor: Tarsus Pharmaceuticals, Inc. 
Funded by: Tarsus Pharmaceuticals, Inc. 
 
9-Dec-2021
Registration No: NCT04784091


Table of Contents 
STATEMENT OF COMPLIANCE ............................................................................................................................. 1 
1 
PROTOCOL SUMMARY ............................................................................................................................... 1 
1.1 
Synopsis ................................................................................................................................................ 1 
1.2 
Schema ................................................................................................................................................. 3 
1.3 
Schedule of Activities (SoA) ............................................................................................................... 6 
2 
INTRODUCTION ............................................................................................................................................ 6 
2.1 
Study Rationale .................................................................................................................................... 6 
2.2 
Background........................................................................................................................................... 7 
2.3 
Risk/Benefit Assessment .................................................................................................................... 9 
2.3.1 
Known Potential Risks ..................................................................................................... 9 
2.3.2 
Known Potential Benefits ................................................................................................ 9 
2.3.3 
Assessment of Potential Risks and Benefits ................................................................ 9 
3 
OBJECTIVES AND ENDPOINTS .............................................................................................................. 10 
4 
STUDY DESIGN ........................................................................................................................................... 12 
4.1 
Overall Design .................................................................................................................................... 12 
4.2 
Scientific Rationale for Study Design.............................................................................................. 12 
4.3 
Justification for Dose ......................................................................................................................... 12 
4.4 
End of Study Definition ..................................................................................................................... 13 
5 
STUDY POPULATION ................................................................................................................................ 13 
5.1 
Inclusion Criteria ................................................................................................................................ 13 
5.2 
Exclusion Criteria ............................................................................................................................... 14 
 
 
5.4 
Screen Failures .................................................................................................................................. 16 
5.5 
Strategies for Recruitment and Retention ...................................................................................... 16 
6 
STUDY TREATMENT .................................................................................................................................. 16 
6.1 
Study Treatment(s) Administration.................................................................................................. 16 
6.1.1 
Study Treatment Description ........................................................................................ 16 
6.1.2 
Dosing and Administration ............................................................................................ 18 
6.2 
Preparation/Handling/Storage/Accountability ................................................................................ 18 
6.2.1 
Acquisition and Accountability ...................................................................................... 18 
6.2.2 
Formulation, Appearance, Packaging, and Labeling ................................................ 18 
6.2.3 
Product Storage and Stability ....................................................................................... 19 
6.2.4 
Preparation ...................................................................................................................... 19 
6.3 
Participant ID Assignment and Measures to Minimize Bias: Randomization and Masking.... 19 
6.4 
Study Treatment Compliance .......................................................................................................... 19 
6.5 
Concomitant Therapy ........................................................................................................................ 20 
6.5.1 
Rescue Medicine ............................................................................................................ 20 
7 
STUDY TREATMENT DISCONTINUATION AND PARTICIPANT DISCONTINUATION ................. 20 
7.1 
Discontinuation of Study Treatment ................................................................................................ 20 
7.2 
Participant Discontinuation from the Study .................................................................................... 21 
7.3 
Lost to Follow-Up ............................................................................................................................... 21 
8 
STUDY ASSESSMENTS AND PROCEDURES ..................................................................................... 21 
8.1 
Efficacy Assessments ....................................................................................................................... 21 
8.2 
Safety and Other Assessments ....................................................................................................... 22 
8.3 
Adverse Events and Serious Adverse Events ............................................................................... 22 
8.3.1 
Definition of Adverse Events (AE) ............................................................................... 22 


  
8.3.2 
Definition of Serious Adverse Events (SAE) .............................................................. 22 
8.3.3 
Classification of an Adverse Event .............................................................................. 23 
8.3.4 
Time Period and Frequency for Event Assessment and Follow-Up ....................... 25 
8.3.5 
Adverse Event Reporting .............................................................................................. 25 
8.3.6 
Serious Adverse Event Reporting ............................................................................... 26 
8.3.7 
Reporting Events to Participants ................................................................................. 26 
8.3.8 
Events of Special Interest ............................................................................................. 26 
8.3.9 
Reporting of Pregnancy ................................................................................................ 27 
9 
STATISTICAL CONSIDERATIONS .......................................................................................................... 27 
9.1 
Statistical Hypotheses ....................................................................................................................... 27 
9.2 
Sample Size Determination .............................................................................................................. 29 
9.3 
Populations for Analyses .................................................................................................................. 30 
9.4 
Statistical Analyses............................................................................................................................ 30 
9.4.1 
General Approach .......................................................................................................... 30 
9.4.2 
Analysis of the Primary Efficacy Endpoint(s) ............................................................. 30 
9.4.3 
Analysis of the Secondary Endpoint(s) ....................................................................... 31 
 
 
9.4.5 
Safety Analyses .............................................................................................................. 31 
 
 
 
 
 
 
 
 
9.4.10 
Final Report .................................................................................................................... 32 
10 
SUPPORTING DOCUMENTATION AND OPERATIONAL CONSIDERATIONS .............................. 32 
10.1 
Regulatory, Ethical, and Study Oversight Considerations ........................................................... 32 
10.1.1 
Informed Consent Process ........................................................................................... 32 
10.1.2 
Confidentiality and Privacy ........................................................................................... 34 
10.1.3 
Future Use of Stored Specimens and Data ............................................................... 35 
10.1.4 
Key Roles and Study Governance .............................................................................. 35 
10.1.5 
Safety Oversight ............................................................................................................. 35 
10.1.6 
Clinical Monitoring .......................................................................................................... 35 
10.1.7 
Quality Assurance and Quality Control ....................................................................... 35 
10.1.8 
Data Handling and Record Keeping ............................................................................ 36 
10.1.9 
Protocol Deviations ........................................................................................................ 37 
10.1.10 
Publication and Data Sharing Policy ........................................................................... 37 
10.1.11 
Conflict of Interest Policy .............................................................................................. 38 
10.2 
Additional Considerations ................................................................................................................. 38 
10.3 
Abbreviations ...................................................................................................................................... 38 
10.4 
Protocol Amendment History ........................................................................................................... 40 
11 
REFERENCES ............................................................................................................................................. 41 
12 
APPENDIX A: EXAMS AND PROCEDURES .......................................................................................... 43 
 
 
12.2 
Corrected Distance Visual Acuity .................................................................................................... 43 
12.3 
Slit Lamp Biomicroscopy .................................................................................................................. 45 
12.3.1 
Collarette Grading .......................................................................................................... 45 
12.3.2 
Lid Margin Erythema Grading ...................................................................................... 45 
12.3.3 
Corneal Fluorescein Staining ....................................................................................... 46 




Saturn-2 Trial 
 
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1 
STATEMENT OF COMPLIANCE 
 
The trial will be carried out in accordance with International Conference on Harmonisation Good Clinical 
Practice (ICH GCP) and the following:  
 
• 
United States (US) Code of Federal Regulations (CFR) applicable to clinical studies (45 CFR Part 46, 21 
CFR Part 50, 21 CFR Part 56, and 21 CFR Part 312)  
 
Investigators and clinical trial site staff who are responsible for the conduct, management, or oversight 
of clinical trials have completed Human Subjects Protection and ICH GCP Training. 
 
The protocol, informed consent form(s), recruitment materials, and all participant materials will be 
submitted to the Institutional Review Board (IRB) for review and approval.  Approval of both the 
protocol and the consent form must be obtained before any participant is enrolled.  Any amendment to 
the protocol will require review and approval by the IRB before the changes are implemented to the 
study.  In addition, all changes to the consent form will be IRB-approved; a determination will be made 
regarding whether a new consent needs to be obtained from participants who provided consent using a 
previously approved consent form. 
 
1 
 
PROTOCOL SUMMARY 
  
1.1 
SYNOPSIS  
 
Title: 
Randomized, Controlled, Multicenter, Double-Masked, Parallel, Phase 3 
Trial to Evaluate the Safety and Efficacy of TP-03 for the Treatment of 
Demodex Blepharitis (Saturn-2) 
  
Test Article: 
TP-03, lotilaner ophthalmic solution, 0.25% 
 
Study Description and 
Hypotheses: 
This Phase 3 study is a randomized, controlled, multicenter, double-
masked, parallel trial to compare the safety and efficacy of TP-03 to 
vehicle control for the treatment of Demodex blepharitis.  The 
hypothesis for the study is the proportion of participants cured at Day 43 
with treatment by TP-03, 0.25%, is greater than the proportion cured by 
treatment with vehicle.    
  
Control Article: 
Vehicle of TP-03 
 
Study Design: 
Prospective, randomized, double-masked, multicenter, two-arm, parallel 
vehicle-controlled study 
 
Objective: 
To demonstrate the safety and efficacy of TP-03, 0.25%, for the 
treatment of Demodex blepharitis 














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8 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
   
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Saturn-2 Trial 
 
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2.3 
RISK/BENEFIT ASSESSMENT   
 
2.3.1 KNOWN POTENTIAL RISKS  
Potential risks for participation in this study include mild: ocular irritation (inflammation), conjunctival 
hyperemia, ocular discomfort, ocular burning, ocular soreness, transitory pain, and ocular surface 
staining following instillation of TP-03 or vehicle.  Other potential risks include a mild, transitory 
medicinal taste following instillation of TP-03 or vehicle.  
 
2.3.2 KNOWN POTENTIAL BENEFITS  
Potential benefits for those participants randomized to treatment with TP-03 include a reduction in 
Demodex density and improvement in the signs of Demodex blepharitis.  Participants randomized to the 
control arm may not receive any direct benefit. 
 
2.3.3 ASSESSMENT OF POTENTIAL RISKS AND BENEFITS  
 
 
 
  The 
benefits of treatment with an efficacious anti-infective that can eradicate Demodex folliculorum is 
believed to outweigh the potential risks. 
 








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13 
  
 
 
 
 
 
4.4 
END OF STUDY DEFINITION 
 
A participant is considered to have completed the study if he or she has completed the last visit.  The 
end of the study is defined as completion of the last visit by all participants at all sites. 
 
5 
STUDY POPULATION 
 
5.1 
INCLUSION CRITERIA 
 
Screening is defined as the study visit where baseline assessments are performed prior to first 
administration of the study drug. 
 
Participants who participated in the Saturn-1 clinical trial should not participate in the Saturn-2 clinical 
trial.  Participants who meet all of the following criteria may be enrolled.   
 
1. Male or female, aged ≥18 years of age 
2. Be willing to sign the informed consent and deemed capable of complying with the 
requirements of the study protocol 
3. Has blepharitis*  
4. Meet all of the following criteria in at least one eye: 
a. Have more than 10 lashes with collarettes present on the upper lid (score of 2 or 
greater)  
b. Have at least mild erythema of the upper eyelid margin  
c. Have a Demodex density, upper and lower eyelids combined, of 1.5 or more mites per 
lash 
Note:  All three criteria (a to c) must be met in the same eye.  Only one eye must qualify for the 
participant to be eligible. 
 
 
 
6. Stated willingness to comply with all study procedures and availability for the duration of the 
study  
 
 
1 Perks D. (2015) AHC 2224920: 13 Week Oral (Gavage) Administration Toxicity Study in the Rat Followed by a 4-
Week Treatment-free Period. (Report No. 8275617). 
 
 


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* The blepharitis diagnosis can be made on the date of screening. 
 
 
 
 
 
 
 
 
 
5.2 
EXCLUSION CRITERIA 
 
An individual who meets any of the following criteria will be excluded from participation in this study: 
 
1. Have had prescription: systemic, nasal, inhaled, ocular topical or topical anti-inflammatory 
steroid treatment; antibacterial treatment; or antiparasitic treatment within 14 days of 
Screening  
2. Have had topical tea tree oil or hypochlorous acid treatment of the ophthalmic area within 14 
days of Screening or unwilling to forego the use of these treatments during the study 
3. Have used lid hygiene products (e.g., lid scrubs) within 14 days of Screening or unwilling to 
forego the use of lid hygiene products during the study  
4. Have initiated treatment with an ocular topical prostaglandin analogue within 30 days of 
Screening or have any plans to change or discontinue treatment during the study  
5. Have used a prostaglandin analogue to promote eyelash growth within 30 days of Screening or 
any plans to initiate treatment during the study  
6. Have used artificial eyelashes, eyelash extensions or had other cosmetic eyelash or eyelid 
procedures (e.g., eyeliner tattooing, eyelash tinting, eyelash curling perm, etc.) within 7 days of 
Screening or be unwilling to forego their use during the study  
 
 
  
8. Have an acute ocular infection or have an active ocular inflammation other than blepharitis  
 
 
 
  
11. Have hypersensitivity to lotilaner or any of the formulation components  
12. Have participated in any clinical trial with an investigational drug or device within 30 days of 
Screening or concurrent enrollment in such a trial with the exception of treatment with the 
investigational drug or device having been completed at least three months prior 
13. Be pregnant or lactating at the time of Screening 
 
 
  




Saturn-2 Trial 
 
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16 
 
  
 
5.4 
SCREEN FAILURES 
 
Screen failures are defined as participants who consent to participate in the clinical trial but are not 
subsequently randomly assigned to the study treatment or entered in the study.  A minimal set of 
screen failure information is required to ensure transparent reporting of screen failure participants to 
meet the Consolidated Standards of Reporting Trials (CONSORT) publishing requirements and to 
respond to queries from regulatory authorities.  Minimal information includes demography, screen 
failure details, eligibility criteria, and any serious adverse event (SAE). 
 
 
 
 
 
5.5 
STRATEGIES FOR RECRUITMENT AND RETENTION 
 
It is anticipated that at least 500 patients will be screened in order to reach target enrollment.  The 
source of participants is expected to come primarily from outpatient clinics and the general public.  As 
Demodex positive patients are widely available, participants are expected to come from in-office visits of 
participating investigators.  In the general public, participants may be identified by their symptoms (e.g., 
itching and burning of the lids margins) and can be reached through print advertisements (e.g., 
newspaper, magazine advertisement, flyer, poster, etc.); social media; radio or television advertisement; 
and/or telephone screening script.   
 
As the risk to participants is considered minimal, employees of the study site and family members of site 
staff may be invited to participate in the clinical trial.  Site staff who work on the clinical trial are not 
eligible to participate. 
 
To retain participants, participants may receive visit reminders and some reimbursement for their time 
and travel expenses. 
 
In the event prohibited medications / treatments are indicated for care, participants will not be 
discontinued. 
 
6 
STUDY TREATMENT 
 
6.1 
STUDY TREATMENT(S) ADMINISTRATION 
  
6.1.1 STUDY TREATMENT DESCRIPTION 
 
TP-03 
Lotilaner is the investigational product substance.  Lotilaner is an isoxazoline and inhibits the γ-
aminobutyric acid (GABA)-gated chloride channels (GABACls) of arthropods.  The GABA-mediated 
chloride influx leads to hyperpolarization of the cellular membrane and generates an inhibitory 






Saturn-2 Trial 
 
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19 
 
6.2.3 PRODUCT STORAGE AND STABILITY 
All preparations of both the study drug and vehicle control product should be stored at 15° to 30°C.  
There are no light or humidity requirements.  Participants will be instructed to store the study drug at 
room temperature in a climate-controlled environment (15° to 30 °C) avoiding extreme heat or cold. 
 
6.2.4 PREPARATION 
 
Not applicable. 
 
6.3 
PARTICIPANT ID ASSIGNMENT AND MEASURES TO MINIMIZE BIAS: RANDOMIZATION 
AND MASKING 
A balanced (1:1) randomization will be used for this study.  A computer-generated blocked 
randomization schedule will be generated by 
.  
 will use the randomization 
schedule to package the study drug.  The labelled study drug will be shipped to the US distribution 
center for distribution to the clinical sites. 
 
 
 
 
 
 
If a potential study participant has provided written informed consent and met all eligibility criteria, the 
participant is considered eligible to participate.  If the potential participant elects to participate in the 
study, they can continue with the Day 1 procedure or be scheduled to return for Day 1.  At Day 1, they 
will be dispensed the next available study drug bottle from the current site inventory by the Interactive 
Response Technology (IRT) system 
, which uses block 
stratification by site.  The participant will receive their assigned study drug at the completion of the Day 
1 visit and the participant will be considered enrolled in the study.  The participant will be assigned a 
unique identification number consisting of their site number followed by a hyphen and then the 
screening number.   
Throughout the study, both the participant and site personnel performing study assessments will remain 
masked to the study medication.  The active and vehicle control ophthalmic solutions will be 
indistinguishable in appearance and will be distributed in bottles identical in appearance. 
Treatment assignments should only be unmasked if needed for proper care of the participant.  Sites will 
be provided instructions on the procedure to be followed to unmask the treatment assignment for a 
participant.  Inadvertent unmasking will be reported to the sponsor and IRB as required. 
 
 
6.4 
STUDY TREATMENT COMPLIANCE 
 


Saturn-2 Trial 
 
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20 
Participants will be instructed on proper instillation and storage of the study drug and provided with 
written instructions.  To assess adherence to the protocol regarding administration of the study product, 
participants will be instructed to record in a dosing compliance diary on a daily basis.  Dosing compliance 
will be assured by an in-office review of a dosing diary.  
 
 
  Participants will 
be reinstructed on dosing compliance and this will be documented in the source documents. 
 
6.5 
CONCOMITANT THERAPY 
 
For this protocol, a prescription medication is defined as a medication that can be prescribed only by a 
properly authorized/licensed clinician.  Medications to be recorded in the source document are 
concomitant prescription medications, over-the-counter medications, supplements, and relevant 
treatments and/or procedures (e.g., lid scrubs, tea tree oil, etc.).  All concomitant medications including 
over-the-counter medications and nutritional supplements used within the 30 days prior to the first visit 
will be documented.  Any changes to concomitant medication use or initiation of new medications will 
be reported throughout the study. 
 
For the duration of the study, any therapy considered necessary for the participant’s welfare may be 
given to the participant at the discretion of the investigator.  Initiation of new medications or changes to 
concomitant medications should be recorded in the source document. 
 
6.5.1 RESCUE MEDICINE 
  
No rescue medication is available for this indication. 
 
7 
STUDY TREATMENT DISCONTINUATION AND PARTICIPANT DISCONTINUATION 
 
7.1 
DISCONTINUATION OF STUDY TREATMENT 
 
Discontinuation from use of the study treatment does not mean discontinuation from the study, and 
remaining study procedures should be completed as indicated by the study protocol.  If a clinically 
significant finding is identified after enrollment, the investigator or qualified designee will determine if 
any change in participant management is needed.  Any new clinically relevant finding will be reported as 
an AE.  Participants to be discontinued for AE(s) will be followed until the event is resolved or 
considered medically stable by the investigator. 
 
The data to be collected at the time of study treatment discontinuation may include the following: 
• 
An AE assessment and performance of safety assessments including visual acuity, IOP, slit lamp 
biomicroscopy, endothelial cell density, hematology, blood chemistry, drug concentration 
analysis and urinalysis  
• 
Any follow-up evaluations required per investigator determination 
 
Temporary discontinuation of the study treatment could include the onset of such conditions as an 
allergic conjunctivitis or signs or symptoms of eyelid allergy (e.g., injection, swelling, itching, etc.).  Per 
investigator discretion, the study treatment could be discontinued until such a time that the allergic 


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conjunctivitis has shown sufficient improvement such that the study treatment could be re-started.  Re-
challenging with the study treatment could help to determine if the allergic response is related to the 
study drug.    
The principal investigator reserves the right to discontinue the study at his/her site for any safety, 
ethical or administrative reason at any time. 
7.2 
PARTICIPANT DISCONTINUATION FROM THE STUDY 
 
Participants are free to discontinue from participation in the study at any time upon request.  An 
investigator may discontinue a participant from the study for the following reasons: 
 
• 
Pregnancy 
• 
If any clinical AE or other medical condition or situation occurs such that continued participation 
in the study would not be in the best interest of the participant 
• 
If the participant meets an exclusion criterion (either newly developed or not previously 
recognized) that precludes further study participation 
• 
Reasons relating to COVID-19 
• 
Other (specify reason) 
 
The reason for participant discontinuation from the study will be recorded on the exit eCRF.  Participants 
who sign the informed consent form and are randomized but do not receive the study treatment will 
not be replaced.  Participants who sign the informed consent form, are randomized and receive the 
study treatment and subsequently discontinue, or are discontinued from the study, will not be replaced.   
 
7.3 
LOST TO FOLLOW-UP 
 
A participant will be considered lost to follow-up if he or she fails to return for their scheduled visit 
within the visit window and is unable to be contacted by the study site staff.  
 
The following actions must be taken if a participant fails to return to the clinic for a required study visit: 
• 
The site will attempt to contact the participant and reschedule the missed visit as soon as 
possible, counseling the participant on the importance of continuing the administration of the 
study drop and of maintaining the assigned visit schedule.  In addition, the site should ascertain 
if the participant wishes to and/or should continue in the study. 
• 
Before a participant is deemed lost to follow-up, the investigator or designee will make every 
effort to regain contact with the participant (where possible, three telephone calls and, if 
necessary, a certified letter to the participant’s last known mailing address or local equivalent 
methods).  These contact attempts should be documented in the participant’s medical record or 
study file.  
• 
Should the participant continue to be unreachable, he or she will be considered to have 
discontinued from the study with a primary reason of lost to follow-up. 
 
8 
STUDY ASSESSMENTS AND PROCEDURES 
  
8.1 
EFFICACY ASSESSMENTS  




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adverse events include drug deficiencies that might have led to an SAE if a) suitable action had not been 
taken or b) intervention had not been made or c) if circumstances had been less fortunate. 
 
Events that are considered sight-threatening in the opinion of the investigator will be considered SAEs. 
 
A planned hospitalization for a pre-existing condition without a serious deterioration in health is not 
considered to be an SAE.  Hospitalization for less than 24 hours is not considered an SAE unless the 
precipitating event was unanticipated and also related to the investigational drug. 
  
 
8.3.3 CLASSIFICATION OF AN ADVERSE EVENT 
  
8.3.3.1 SEVERITY OF EVENT 
 
The following guidelines will be used to describe the severity of AEs.  
 
• 
Mild – Events require minimal or no treatment and do not interfere with the participant’s daily 
activities.  
• 
Moderate – Events result in a low level of inconvenience or concern with the therapeutic 
measures.  Moderate events may cause some interference with functioning. 
• 
Severe – Events interrupt a participant’s usual daily activity and may require systemic drug 
therapy or other treatment.  Severe events are usually potentially life-threatening or 
incapacitating.   
 
Of note, the term “severe” does not necessarily equate to “serious.”  Severity is a measurement of 
intensity.  Thus, a severe reaction is not necessarily a SAE.  For example, a headache may be severe in 
intensity, but would not be serious unless it met one or more of the criteria for a serious adverse event. 
 
8.3.3.2 RELATIONSHIP TO STUDY TREATMENT 
 
All AEs must have their relationship to study treatment assessed by the clinician who examines and 
evaluates the participant based on temporal relationship and his/her clinical judgment.  The degree of 
certainty about causality will be graded using the categories below.  In a clinical trial, the study product 
must always be suspect.  
 
• 
Definitely Related – Evidence indicates a reasonable temporal sequence of the event with the 
study drug administration exists or that the association of the event with study drug 
administration is unknown and the event is not reasonably supported by other conditions such 
that: 
o There is a clinically plausible time sequence between onset of the AE and study treatment 
administration, and/or 
o There is a biologically plausible mechanism for study treatment causing or contributing to 
the AE, and 
o The AE cannot be reasonably attributed to concurrent/underlying illness, other drugs or 
procedures. 
• 
Potentially Related – Evidence indicates a possible relationship to the study drug such that: 


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o The event occurs within a reasonable period of time relative to the treatment that makes a 
causal relationship possible, but plausible explanations may also be provided by other 
causes such as other drugs, products, chemicals, underlying disease, environment, etc. 
o The event is possibly related to the study treatment. 
• 
Not Related – Evidence indicates no plausible direct relationship to the study drug such that: 
o A clinically plausible temporal sequence is inconsistent with the onset of the AE and drug 
administration, and/or 
o A causal relationship is considered biologically implausible 
o The AE can be attributed to concurrent/underlying illness, other drugs or procedures. 
 
8.3.3.3 EXPECTEDNESS  
  
The investigator will be responsible for determining whether an AE is expected or unexpected.  An AE 
will be considered unexpected if the nature, severity, or frequency of the event is not consistent with 
the risk information previously described for the study treatment.  Expectedness should be determined 
if the AE is characterized as related to treatment and is serious (met one or more of the criteria for an 
SAE). 
 
8.3.3.4 OUTCOME 
The clinical outcome of an AE will be characterized as follows: 
• 
Resolved without sequelae 
• 
Resolved with sequelae (specify) 
• 
Death 
• 
Ongoing (i.e., continuing at time of study discontinuation) 
• 
Unknown 
 
8.3.3.5 TREATMENT OR ACTION TAKEN 
The clinical treatment or action taken from an AE will be characterized as follows: 
• 
None 
• 
Medical intervention 
• 
Surgical intervention 
• 
Unknown 
• 
Other (specify) 
 
8.3.3.6 ACTION TAKEN WITH INVESTIGATIONAL PRODUCT 
The action taken with the IP will be characterized as follows: 
• 
None 
• 
Drug temporarily withdrawn 
• 
Drug discontinued 
• 
Unknown 


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8.3.4 TIME PERIOD AND FREQUENCY FOR EVENT ASSESSMENT AND FOLLOW-UP 
 
The occurrence of an AE or SAE may come to the attention of study personnel during study visits and 
interviews of a study participant presenting for medical care, or upon review by a study monitor.  At 
study visits, study personnel should pose general questions to the participant to ascertain if the 
participant experienced any issues prior to the visit.  
 
All AEs, including local and systemic reactions not meeting the criteria for SAEs, will be captured on the 
appropriate eCRF.  Information to be collected includes event description, date of onset, clinician’s 
assessment of severity, relationship to study product (assessed only by those with the training and 
authority to make a diagnosis), and date of resolution/stabilization of the event.  All AEs occurring while 
in the study must be documented appropriately regardless of relationship.   
 
Any medical condition that is present at the time that the participant is screened will be considered as 
Baseline and not reported as an AE.  However, if the study participant’s condition deteriorates at any 
time during the study, it will be recorded as an AE.  
 
Changes in the severity of an AE will be documented to allow an assessment of the duration of the event 
at each level of severity to be performed.   
 
The investigator or designee will record all reportable events with start dates occurring any time after 
study drug is first administered until 7 (for non-serious AEs) or 30 days (for SAEs) after the last day of 
study participation.  At each study visit, the investigator will inquire about the occurrence of AE/SAEs 
since the last visit.  Events will be followed for outcome information until resolution or stabilization. 
 
Whenever possible, recognized medical terms should be used when recording AEs.  Colloquialisms 
and/or abbreviations should not be used.  Only one medical concept, preferably a diagnosis instead of 
individual symptoms, should be recorded as the event (e.g., record congestive heart failure rather than 
dyspnea, rales and cyanosis).  However, if a constellation of signs and/or symptoms cannot be medically 
characterized as a single diagnosis or syndrome at the time of reporting, each individual sign or 
symptom should be recorded as a separate AE.  A diagnosis that is subsequently established should be 
reported as follow-up information.   
 
Adverse events occurring secondary to other events (e.g., sequelae) should be identified by the primary 
cause.  A "primary" event, if clearly identifiable, should represent the most accurate clinical term to 
record as the AE event term.  For example: Orthostatic hypotension  fainting and fall to floor  head 
trauma  neck pain.  The primary event is orthostatic hypotension and the sequelae are head trauma 
and neck pain. 
 
Centralized safety oversight for this multi-site study will be provided by the Medical Monitor.    
 
8.3.5 ADVERSE EVENT REPORTING  
 
The investigator must record nonserious adverse events and submit on the eCRF in a timely manner.  
 




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8.3.9 REPORTING OF PREGNANCY  
 
Women of Childbearing Potential (WOCBP) includes any females who have experienced menarche and 
who have not undergone successful surgical sterilization (hysterectomy, bilateral tubal ligation or 
bilateral oophorectomy) or are not postmenopausal at least 12 months since last menses.  WOCBP will 
be required to use designated methods of birth control during the course of the study.  All women who 
are pregnant, nursing an infant or planning a pregnancy during the duration of this study will be 
excluded from participation.  
 
If a participant or investigator suspects that the participant may be pregnant prior to study drug 
administration, the study drug should be withheld until the results of pregnancy testing are available.  If 
pregnancy is confirmed, the participant should not administer the study drug and should not be enrolled 
in the study. 
 
If a female participant becomes pregnant during the study, use of the study drug should be discontinued 
immediately.  The investigator will notify the sponsor (or designee) immediately after the pregnancy is 
confirmed.  The investigator will (1) obtain a consent from the female participant for pregnancy follow-
up and (2) follow the progress of the pregnancy to term.  The investigator should document the 
outcome of the pregnancy and provide a copy of the documentation to the sponsor or designee.  The 
sponsor or sponsor’s designee will be responsible for reporting to the IRB and regulatory agencies as 
appropriate. 
 
 
9 
STATISTICAL CONSIDERATIONS  
 
9.1 
STATISTICAL HYPOTHESES 
 
This study is designed to demonstrate the superiority of TP-03 to vehicle control. 
   
• 
Population:  Participants with Demodex blepharitis as defined by the enrollment criteria 
 
• 
Primary Efficacy Endpoint:  Cure based on a collarette score of 0 for the upper eyelid of the 
analysis eye at Day 43  
 
• 
Secondary Efficacy Endpoints:   
o Demodex mite eradication (mite density of 0 mites/lash) from the analysis eye at Day 43 
o Composite cure based on collarette score (0) and erythema score (0) for the upper 
eyelid of the analysis eye at Day 43 
o Erythema cure based on erythema score (0) for the upper eyelid of the analysis eye at 
Day 43 
 
• 
 
o 
 
 
 
 
 
 
 


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• 
Hypotheses: 
The primary efficacy hypothesis is:  
H01: The proportion of participants cured at Day 43, based on a collarette score of 0 for the 
upper eyelid of the analysis eye, by treatment with TP-03, 0.25%, is ≤ the proportion cured by 
treatment with vehicle.  
H11: The proportion of participants cured at Day 43, based on a collarette score of 0 for the 
upper eyelid of the analysis eye, by treatment with TP-03, 0.25%, is > the proportion cured by 
treatment with vehicle.  
The secondary efficacy hypotheses are:  
Demodex Eradication 
H02: The proportion of participants with Demodex eradicated at Day 43 (mite density of 0 
mites/lash) in the analysis eye by treatment with TP-03, 0.25%, is ≤ the proportion with 
Demodex eradicated by treatment with vehicle.  
H12: The proportion of participants with Demodex eradicated at Day 43 (mite density of 0 
mites/lash) in the analysis eye by treatment with TP-03, 0.25%, is > the proportion with 
Demodex eradicated by treatment with vehicle.  
Composite of Collarette and Erythema Cure 
H03: The proportion of participants with a composite cure, collarette score of 0 and erythema 
score of 0, for the upper eyelid of the analysis eye at Day 43 by treatment with TP-03, 0.25%, is ≤ 
the proportion with a composite cure by treatment with vehicle.  
H13: The proportion of participants with a composite cure, collarette score of 0 and erythema 
score of 0, for the upper eyelid of the analysis eye at Day 43 by treatment with TP-03, 0.25%, is > 
the proportion with a composite cure by treatment with vehicle.  
Erythema Cure 
H04: The proportion of participants with an erythema cure, score of 0, for the upper eyelid of the 
analysis eye at Day 43 by treatment with TP-03, 0.25%, is ≤ the proportion with an erythema 
cure by treatment with vehicle.  
H14: The proportion of participants with an erythema cure, score of 0, for the upper eyelid of the 
analysis eye at Day 43 by treatment with TP-03, 0.25%, is > the proportion with an erythema 
cure by treatment with vehicle.  
 
 


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9.2 
SAMPLE SIZE DETERMINATION 
 
A total sample size of 300 subjects (150 subjects per arm) yields approximately 99% power to establish 
superiority of TP-03 to vehicle in the participants meeting the primary efficacy endpoint assuming a 
response rate of 80.0% in TP-03 and 15.8% for vehicle treatments, respectively using Pearson chi-
squared to test a 1-sided hypothesis with a significance level of 0.025.  Response rates for sample size 
calculations are based upon a prior OUS clinical study of TP-03, 0.25%, for the treatment of Demodex 
blepharitis. 
 
Based on the results of a more recent Phase 2b/3 study, TRS-009 (Saturn-1), with a response rate of 
44.0% for the TP-03 treatment group and 7.4% for the vehicle treatment group, a total sample size of 
300 subjects still provides greater than 99% power. 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
The study will be considered a success and TP-03, 0.25%, will be considered superior to vehicle in clinical 
cure if H01 is rejected in favor of H11.  With this sample size, the study will have > 95% power to claim 
success for clinical cure.  A closed hierarchical testing structure will be used such that the analysis will 
first be performed for the primary efficacy endpoint. 
 
 
 
 
 
 
 
 
 
 
 
  
 
Accounting for a 30% discontinuation, approximately 209 subjects per arm (approximately 418 total) will 
be randomized.  The planned enrollment is much greater than typically planned to achieve 300 


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completed subjects due to concerns regarding the impact of COVID-19 and the possibility that up to two 
sites are unable to complete the study. 
 
9.3 
POPULATIONS FOR ANALYSES 
 
The analysis datasets will include the following:  
• 
Full Analysis Set – The full analysis set will include all randomized participants.  Participants in 
the FAS will be analyzed as randomized. 
• 
COVID-19 Analysis Set - The COVID-19 analysis set includes all randomized subjects, but excludes 
subjects that discontinued due to COVID-19 complications, subjects that would have 
discontinued due to COVID-19 had they remained on study, and subjects that had significant 
COVID-19-related protocol deviations.  Subjects that would have discontinued due to COVID-19 
had they remained on study will be determined and documented prior to unmasking.  Protocol 
deviations will be assessed prior to unmasking for their significance based on their impact to 
primary endpoint data.  Sensitivity analyses of the primary efficacy and key secondary analyses 
will be performed on the COVID-19 population and subjects will be analyzed as randomized if at 
least 5% of subjects met these criteria. 
• 
Per Protocol Set – The per-protocol (PP) set will include participants in the FAS who do not have 
significant protocol deviations that effect the primary endpoint analyses.  Protocol deviations 
will be assessed prior to unmasking.  Participants in the PP set will be analyzed as treated.  
• 
Safety Set – The safety set will include all participants who have received at least one dose of 
the investigational product.  Participants in the safety population will be analyzed as treated.  
 
9.4 
STATISTICAL ANALYSES 
 
9.4.1 GENERAL APPROACH 
 
A formal Statistical Analysis Plan (SAP) will be completed prior to unmasking of the study data.  If there 
are any discrepancies between the protocol and the SAP, the SAP will take precedence.  
Quantitative/continuous data will be summarized using descriptive statistics (n, mean, standard 
deviation, median, minimum, and maximum).  Qualitative/categorical data will be summarized using 
frequencies and percentages.  Statistical testing, unless otherwise indicated, will be performed at a 1-
sided 0.025 significance level.  
 
When applicable, two-sided 95% confidence intervals will be reported.  Baseline will be considered the 
last non-missing measure prior to initiation of study therapy.  Change from baseline will be calculated as 
follow-up visit minus baseline. 
 
9.4.2 ANALYSIS OF THE PRIMARY EFFICACY ENDPOINT(S) 
 
The primary efficacy endpoint analysis will be conducted with the FAS, with intercurrent events handled 
as described in the missing data section.  Descriptive statistics will be presented by treatment arm.  
Testing of the percentage of subjects cured at Day 43 based on the collarette grade will be completed 
using a difference of proportions test. 
 
 


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Multiplicity Adjustment  
A closed hierarchical testing structure will be used where the analysis will be performed for the primary 
efficacy endpoint and, only if successful, the analysis will be performed for the three secondary efficacy 
endpoints using the Hochberg testing strategy.  Specifically, if the null hypothesis, H01, is rejected at a 
one-sided α of 0.025, the study will be considered a success for clinical cure and the family of three 
secondary efficacy endpoints will be tested using the Hochberg testing strategy with a familywise α of 
0.025.  If the secondary endpoint with the largest p-value is significant at the 0.025 level, then all three 
secondary endpoints will be declared statistically significant.  If the secondary endpoint with the largest 
p-value is not significant at the 0.025 level, then the secondary endpoint with the next largest p-value 
will be tested at the 0.0125 level.  If this secondary endpoint is significant at the 0.0125 level, then it and 
the remaining secondary variable will be declared statistically significant.  If not, the remaining 
secondary will be tested at the 0.0083 level.  If the null hypothesis, H01 is not rejected at a one-sided α of 
0.025, testing will stop and the three secondary efficacy hypotheses will not be tested. 
  
Missing Data  
The primary efficacy analysis will be conducted with missing data and intercurrent events handled in the 
following manner:  
• 
Discontinuation of study medication and non-optimal compliance is ignored [treatment policy 
strategy].  
• 
Withdrawal due to lack of efficacy or adverse events: missing data will be imputed as failure 
[hypothetical strategy]. 
• 
Missing data without withdrawal or withdrawal due to reasons other than lack of efficacy or 
adverse events: missing data will be imputed employing Multiple Imputation (MI) using 
randomized treatment-based Markov Chain Monte Carlo (MCMC) methodology [hypothetical 
strategy].  
 
Sensitivity analysis on the primary efficacy endpoint will be performed as specified in the Statistical 
Analysis Plan (SAP).   
 
9.4.3 ANALYSIS OF THE SECONDARY ENDPOINT(S) 
 
The statistical approach for the secondary efficacy endpoints will be the same as for the primary efficacy 
endpoint analysis. 
 
 
 
 
 
 
 
9.4.5 SAFETY ANALYSES 
 
All safety analyses will be conducted using the safety set.  Safety measures will be summarized for each 
treatment arm and visit (as appropriate) using descriptive statistics.  Adverse events will be coded using 


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the Medical Dictionary for Regulatory Activities (MedDRA).  Each MedDRA preferred term will be 
counted once only for a given participant.  The severity, frequency, relationship of AEs to study 
treatment, expectedness if serious, duration, and outcome will be presented either in a table or listing.  
Adverse events leading to premature discontinuation from the study treatment and serious treatment-
emergent AEs will be presented either in a table or a listing. 
 
 
 
 
 
 
 
 
  
 
  
 
 
 
 
 
   
 
 
 
 
 
 
9.4.10 FINAL REPORT 
 
Once all study visits through Day 43 are complete and all associated study data has been received and 
corrected, the study data will be locked.  Once the data has been locked and all exclusions from the PP 
analysis population and participants who would have discontinued due to COVID-19 had they remained 
on study have been determined, the treatment assignments will be unmasked for analysis.  A complete 
analysis of the results will be performed and a final report will be prepared.  When the last of the Day 90 
visits have been completed and all associated study data has been received and corrected, the analysis 
of the endothelial cell density results will be performed.  A report of the endothelial cell density results 
will be prepared and the report will be submitted to the NDA. 
 
10 SUPPORTING DOCUMENTATION AND OPERATIONAL CONSIDERATIONS 
  
10.1 REGULATORY, ETHICAL, AND STUDY OVERSIGHT CONSIDERATIONS 
 
10.1.1 INFORMED CONSENT PROCESS 
 
 
10.1.1.1 CONSENT AND OTHER INFORMATIONAL DOCUMENTS PROVIDED TO PARTICIPANTS 
 


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Consent forms describing in detail the study treatment, study procedures, and risks are given to the 
participant and written documentation of informed consent is required prior to starting study 
treatment.  Consent materials submitted with this protocol could include questionnaires, printed or 
web-based advertisement materials, instruction sheets, and phone scripts. 
 
10.1.1.2 CONSENT PROCEDURES AND DOCUMENTATION 
 
Informed consent is a process that is initiated prior to the individual’s agreeing to participate in the 
study and continues throughout the individual’s study participation.  Consent forms will be Institutional 
Review Board (IRB)-approved and the participant will be asked to read and review the document.  The 
investigator will explain the research study to the participant and answer any questions that may arise.  
A verbal explanation will be provided in terms suited to the participant’s comprehension of the 
purposes, procedures, and potential risks of the study and of their rights as research participants.  
Participants will have the opportunity to carefully review the written consent form and ask questions 
prior to signing.  The participants should have the opportunity to discuss the study with their family or 
surrogates or think about it prior to agreeing to participate.  The participant will sign the informed 
consent document prior to any procedures being done specifically for the study.  Participants must be 
informed that participation is voluntary and that they may discontinue from the study at any time, 
without prejudice.  A copy of the informed consent document will be given to the participants for their 
records.  The informed consent process will be conducted and documented in the source document 
(including the date), and the form signed, before the participant undergoes any study-specific 
procedures.  The rights and welfare of the participants will be protected by emphasizing to them that 
the quality of their medical care will not be adversely affected if they decline to participate in this study.   
 
During the informed consent process, potential study participants will be informed that they have a 50% 
chance of being assigned to the vehicle control arm. 
 
To obtain consent from speakers of languages other than English, the consent and applicable study-
related materials will be translated into a language understandable to the participant.  A member of the 
research team or non-family member interpreter will be available to interpret the initial and ongoing 
informed consent discussion for the participant.  In case of emergencies, a member of the research 
team who is fluent in the participant’s language will be available or the research team will have 24-hour 
access to a translation service that can sufficiently communicate with the participant. 
 
 
 
• 
 
 
 
 
• 
 
 
 
 
 
 
10.1.1.3 STUDY DISCONTINUATION AND CLOSURE 
 


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This study may be temporarily suspended or prematurely terminated if there is sufficient reasonable 
cause.  Possible reasons for termination or temporary suspension of the study could include: 
 
• 
Study closure based on sponsor/funder decision, regulatory or other oversight bodies 
• 
Review of serious, unexpected, and related AEs 
• 
Noncompliance with protocol requirements 
• 
Determination of unexpected, significant or unacceptable risk to participants due to COVID-19 
     
Written notification documenting the reason for study suspension or termination will be provided by 
the suspending or terminating party to study participants, the investigator, the Investigational New Drug 
(IND) sponsor and regulatory authorities as required.  If the study is prematurely terminated or 
suspended, the principal investigator (PI) will promptly inform study participants, the Institutional 
Review Board (IRB), and sponsor, and will provide the reason(s) for the termination or suspension.  
Study participants will be contacted, as applicable, and be informed of changes to the study visit 
schedule. 
 
The study may resume once concerns about safety, protocol compliance, and data quality are 
addressed, and satisfy the sponsor, IRB and/or Food and Drug Administration (FDA). 
 
10.1.2 CONFIDENTIALITY AND PRIVACY  
 
Participant confidentiality and privacy is strictly held in trust by the participating investigators, their 
staff, and the sponsor.  This confidentiality is extended to cover testing of biological samples in addition 
to the clinical information relating to participants.  Therefore, the study protocol, documentation, data, 
and all other information generated will be held in strict confidence.  No information will be shared with 
any parties other than those specifically identified in the ICF/HIPAA signed by the subject, without 
written permission from the subject.  
 
All research activities will be conducted in as private a setting as possible. 
 
The study monitor, other authorized representatives of the sponsor, representatives of the Institutional 
Review Board (IRB), regulatory agencies or pharmaceutical company supplying study product may 
inspect all documents and records required to be maintained by the investigator, including but not 
limited to, medical records (office, clinic, or hospital) and pharmacy records for the participants in this 
study.  The clinical study site will permit access to such records. 
 
The study participant’s contact information will be securely stored at each clinical site for internal use 
during the study.  At the end of the study, all records will continue to be kept in a secure location for as 
long a period as dictated by the reviewing IRB, institutional policies, or sponsor requirements. 
 
Study participant research data, which is for purposes of statistical analysis and scientific reporting, will 
be transmitted to and stored 
 
).  This will not 
include the participant’s contact or identifying information.  Rather, individual participants and their 
research data will be identified by a unique study identification number and their initials.  The study 
data entry and study management systems used by clinical sites and by 
 research staff will be 
secured and password protected.  At the end of the study, all study databases will be de-identified and 
archived at a location specified by the sponsor. 


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10.1.4 KEY ROLES AND STUDY GOVERNANCE 
 
   
 
  
  
 
 
 
 
Study team roles and responsibilities of those involved in the conduct, management, or oversight of the 
trial will be established.  
10.1.5 SAFETY OVERSIGHT 
 
Safety oversight will be under the direction of a Medical Monitor, an individual having appropriate 
expertise.  The Medical Monitor will be independent from study conduct and free of conflict of interest. 
 
10.1.6 CLINICAL MONITORING 
 
Clinical site monitoring is conducted to ensure that the rights and well-being of trial participants are 
protected, that the reported trial data are accurate, complete, and verifiable, and that the conduct of 
the trial is in compliance with the currently approved protocol/amendment(s), with International 
Conference on Harmonisation Good Clinical Practice (ICH GCP), and with applicable regulatory 
requirement(s).   
 
• 
Monitoring for this study will be performed by
.  
• 
Details of clinical site monitoring are documented in a Clinical Monitoring Plan (CMP).  The CMP 
describes in detail who will conduct the monitoring, at what frequency monitoring will be done, 
at what level of detail monitoring will be performed, and the distribution of monitoring reports. 
 
10.1.7 QUALITY ASSURANCE AND QUALITY CONTROL 


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Each clinical site will perform internal quality management of study conduct, data and biological 
specimen collection, documentation and completion.   
 
Quality control (QC) procedures will be implemented beginning with the data entry system and data QC 
checks that will be run on the database will be generated.  Any missing data or data anomalies will be 
communicated to the site(s) for clarification/resolution. 
 
Following written Standard Operating Procedures (SOPs), the monitors will verify that the clinical trial is 
conducted and data are generated and biological specimens are collected, documented (recorded), and 
reported in compliance with the protocol, International Conference on Harmonisation Good Clinical 
Practice (ICH GCP), and applicable regulatory requirements (e.g., Good Manufacturing Practices (GMP)).  
 
The investigational site will provide direct access to all trial-related sites, source data/documents, and 
reports for the purpose of monitoring and auditing by the sponsor, and inspection by local and 
regulatory authorities. 
 
10.1.8 DATA HANDLING AND RECORD KEEPING  
  
10.1.8.1 DATA COLLECTION AND MANAGEMENT RESPONSIBILITIES  
 
Data collection is the responsibility of the clinical trial staff at the site under the supervision of the site 
investigator.  The investigator is responsible for ensuring the accuracy, completeness, legibility, and 
timeliness of the data reported.   
  
Study participation should be captured in a participant’s medical record.  This is to ensure that anyone 
who would access the patient medical record has adequate knowledge that the patient is participating 
in a clinical trial. 
  
All source documents should be completed in a neat, legible manner to ensure accurate interpretation 
of data.  Source data are all information, original records of clinical findings, observations, or other 
activities in a clinical trial necessary for the reconstruction and evaluation of the trial.  Electronic source 
data are data initially recorded in electronic form.  Examples of source data include, but are not limited 
to, hospital records, clinical and office charts, laboratory notes, memoranda, participants’ memory aids 
or evaluation checklists, pharmacy dispensing records, audio recordings of counseling sessions, recorded 
data from automated instruments, copies or transcriptions certified after verification as being accurate 
and complete, microfiches, photographic negatives, microfilm or magnetic media, x-rays, and 
participant files and records kept at the pharmacy, at the laboratories, and medico-technical 
departments involved in the clinical trial.  Hardcopies of the study visit worksheets may be provided for 
use as source document worksheets for recording data for each participant enrolled in the study.   
  
Clinical data (including AEs and concomitant medications) and clinical laboratory data will be entered 
into 
, a 21 CFR Part 11-compliant data capture system 
managed by SDC.  The data system includes password protection and internal quality checks, such as 
automatic range checks, to identify data that appear inconsistent, incomplete, or inaccurate.  Clinical 
data will be entered directly from the source documents.  Data recorded in the electronic case report 


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form (eCRF) derived from source documents should be consistent with the data recorded on the source 
documents. 
  
Adverse events will be coded using MedDRA and concomitant medications will be coded using 
WHODrug for summarization and reporting.  
 
10.1.8.2 STUDY RECORDS RETENTION  
 
Study documents should be retained for a minimum of 2 years after the last approval of a marketing 
application and until there are no pending or contemplated marketing applications or until at least 2 
years have elapsed since the formal discontinuation of clinical development of the study 
treatment.  These documents should be retained for a longer period, however, if required by local 
regulations.  No records will be destroyed without the written consent of the sponsor.  It is the 
responsibility of the sponsor to inform the investigator when these documents no longer need to be 
retained. 
 
10.1.9 PROTOCOL DEVIATIONS  
 
A protocol deviation is any noncompliance with the clinical trial protocol, International Conference on 
Harmonisation Good Clinical Practice (ICH GCP), or Manual of Procedures requirements.  The 
noncompliance may be either on the part of the participant, the investigator, or the study site staff.  As 
a result of deviations, corrective actions are to be developed by the site and implemented promptly.  
 
These practices are consistent with ICH GCP:  
• 
4.5 Compliance with Protocol, sections 4.5.1, 4.5.2, and 4.5.3  
• 
5.1 Quality Assurance and Quality Control, section 5.1.1  
• 
5.20 Noncompliance, sections 5.20.1, and 5.20.2.  
 
It is the responsibility of the site investigator to use continuous vigilance to identify and report 
deviations within approximately 7 working days of identification of the protocol deviation, or within 
approximately 7 working days of the scheduled protocol-required activity.  All deviations must be 
addressed in study source documents and reported to the sponsor or its representative.  Relevant 
protocol deviations must be sent to the reviewing Institutional Review Board (IRB) per their policies.  
The site investigator is responsible for knowing and adhering to the reviewing IRB requirements.   
 
10.1.10 
PUBLICATION AND DATA SHARING POLICY 
 
This study will be conducted in accordance with the following publication and data sharing policies and 
regulations: 
 
This study will comply with the Clinical Trials Registration and Results Information Submission rule.  As 
such, this trial will be registered at ClinicalTrials.gov, and results information from this trial will be 
submitted to ClinicalTrials.gov.  In addition, every attempt will be made to publish results in peer-
reviewed journals.   
 
 
 




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IOP 
Intraocular pressure 
IP 
Investigational product 
IUPAC 
International Union of Pure and Applied Chemistry 
IRB 
Institutional Review Board 
IRT 
Interactive Response Technology 
LDPE 
Low-density polyethylene 
logMAR 
Logarithm of the minimum angle of resolution 
MedDRA 
Medical Dictionary for Regulatory Activities 
μL 
Microliter 
MD 
Doctor of Medicine 
NADA 
New Animal Drug Application 
NEI 
National Eye Institute 
NOAEL 
No observed adverse effect level 
OD 
Doctor of Optometry 
OUS 
Outside United States 
pH 
Potential hydrogen 
PI 
Principal Investigator 
PP 
Per protocol 
QC 
Quality Control 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SDC 
Statistics & Data Corporation 
SMO 
Site Management Organization 
SOA 
Schedule of Activities 
TID 
Three times a day 
TP-03 
Lotilaner ophthalmic solution 
TRS 
Tarsus 
US 
United States 
WOCBP 
Women of Childbearing Potential 
 
 
 


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10.4 PROTOCOL AMENDMENT HISTORY 
 
Version 
Date 
Description of Change  
Brief Rationale 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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