Clinical trial NCT02795832
Conditions: Atopic Dermatitis
Sponsor: Ziarco Pharma Ltd
Start date: 2016-06-01; Completion date: 2017-03-01
Enrollment: 53 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
A Randomised, Adaptive Design, Double-Blind (3rd Party Open), Placebo Controlled, Sequential Group Study to Determine the Safety,Tolerability, Pharmacokinetics and Efficacy of Twice Daily Application of a Topical ZPL-5212372 (1.0% w/w) Ointment Administered for up to 2 Weeks in Adult Healthy Volunteers and Patients with Moderate to Severe Atopic Dermatitis

Detailed description:
This study was a randomised, adaptive design, double blind (3rd party open), placebo controlled, sequential group study in both healthy volunteers and patients with moderate to severe AD. This study was divided into 3 separate, sequential cohorts:

* Cohort 1 were to assess safety, toleration and pharmacokinetics (PK) with intensive monitoring as in-patients over 7 days of dosing in healthy volunteers.
* Cohort 2 were to assess safety, toleration and PK with intensive monitoring as in-patients over 7 days of dosing in moderate to severe AD patients.
* Cohort 3 were to assess efficacy, safety, toleration and PK as out-patients over 14 days of dosing in moderate to severe AD patients.

Subjects received 1.0% (w/w) ZPL-5212372 or matched placebo ointment topically, twice daily.

Arms: arm 1: ZPL-5212372 1% w/w Ointment BID arm 2: Placebo Ointment BID arm 3: ZPL-5212372 1% w/w Ointment BID arm 4: Placebo Ointment BID arm 5: ZPL-5212372 1% w/w OIntment BID arm 6: Placebo Ointment BID
Interventions: intervention 1: ZPL-5212372 1% w/w Ointment BID intervention 2: Placebo Ointment BID
Intervention descriptions: intervention 1: None intervention 2: None

Protocol documents:
Prot_001.pdf: 
CLINICAL STUDY PROTOCOL 
 A RANDOMISED, ADAPTIVE DESIGN, DOUBLE-BLIND (3RD PARTY 
OPEN), PLACEBO CONTROLLED, SEQUENTIAL GROUP STUDY TO 
DETERMINE THE SAFETY, TOLERABILITY, PHARMACOKINETICS 
AND EFFICACY OF TWICE DAILY APPLICATION OF A TOPICAL 
ZPL-5212372 (1.0% W/W) OINTMENT ADMINISTERED FOR UP TO 2 
WEEKS IN ADULT HEALTHY VOLUNTEERS AND PATIENTS WITH 
MODERATE TO SEVERE ATOPIC DERMATITIS 
Sponsor: 
Principal Investigator 
Sponsor Protocol No.: 
EudraCT No.: 
Study Drug Name: 
Development Phase: 
Date of Protocol: 
Protocol version: 
  Ziarco Pharma Ltd 
Innovation House, Discovery Park 
Ramsgate Road  
Sandwich 
Kent CT13 9ND 
United Kingdom 
 
 
 
 
 
 
ZPL521/101 / NCT02795832
2016-000376-26 
ZPL-5212372 
Phase 1/2a 
28 July 2016 
Final 2.0 
The study will be conducted according to the protocol and in compliance with Good 
Clinical Practice (GCP), with the Declaration of Helsinki, and with other applicable 
regulatory requirements. 
This document contains confidential information of Ziarco Pharma Ltd. 
Do not copy or distribute without written permission from the Sponsor. 
CONFIDENTIAL 








Ziarco 
Confidential 
Clinical Study Protocol 
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 Version 2.0 /28 July 2016 
 
PROTOCOL SYNOPSIS 
Title: 
A Randomised, Adaptive Design, Double-Blind (3rd Party Open), 
Placebo Controlled, Sequential Group Study to Determine the Safety, 
Tolerability, Pharmacokinetics and Efficacy of Twice Daily Application 
of a Topical ZPL-5212372 (1.0% w/w) Ointment Administered for up to 
2 Weeks in Adult Healthy Volunteers and Patients with Moderate to 
Severe Atopic Dermatitis 
 
Sponsor Study 
No: 
ZPL521/101 
 
Compound: 
ZPL-5212372 
 
Phase: 
Phase 1/2a 
 
Sponsor: 
Ziarco Pharma Ltd 
 
Principal 
Investigator: 
 
  
 
 
 
 
 
Objective(s): 
Primary 
Cohorts 1 and 2 
To assess the safety and tolerability, local and systemic, of ZPL-5212372 
administered as a topical ointment (containing 1.0% (w/w) concentration of 
ZPL-5212372) twice daily, for 1 week, to healthy subjects and patients with 
moderate to severe atopic dermatitis (AD). 
 
Cohort 3 
To evaluate the efficacy of ZPL-5212372 administered as a topical ointment 
(containing 1.0% (w/w) concentration of ZPL-5212372) twice daily, for 2 
weeks, to patients with moderate to severe AD. 
 
Secondary 
Cohorts 1 and 2 
To investigate the systemic pharmacokinetics of ZPL-5212372 following 
single and multiple dose topical applications of a 1.0% (w/w) concentration 
of ZPL-5212372 ointment, administered twice daily, for 1 week, to healthy 
subjects and patients with moderate to severe AD. 
 
Cohort 3 
To assess the safety and tolerability, local and systemic, of ZPL-5212372 
administered as a topical ointment (containing 1.0% (w/w) concentration of 
ZPL-5212372) twice daily, for 2 weeks, to patients with moderate to severe 
AD. 
 
 
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To investigate the trough concentrations of ZPL-5212372 in patients with 
moderate to severe AD following twice daily applications of ZPL-5212372 
administered as a topical ointment (containing 1.0% (w/w) concentration of 
ZPL 5212372). 
 
Exploratory 
Cohorts 1 to 3 
To explore the subjects perception of the Ziarco development ointment 
formulation using the Ziarco Ointment Questionnaire. 
 
Design: 
This is a randomised, adaptive design, double-blind (3rd party open), 
placebo controlled, sequential group study to determine the safety, 
tolerability, PK and efficacy of twice daily application of topical 
ZPL-5212372 (1.0% w/w) ointment administered as a 2:1 ratio for active 
and placebo groups. This study will consist of 3 sequential cohorts each 
separated by formal safety reviews. 
 
All subjects (both healthy volunteers and AD patients) will be required to 
attend a screening visit(s) to confirm suitability to enter the study.  In order 
to be eligible AD patients must have moderate-severe AD, defined as: 
• Eczema Area and Severity Index (EASI) of ≥9 and <48  
• Body surface area involvement (BSA) of between 10% and 40% 
• Investigator Global Assessment Score (IGA) of at least 3 (moderate 
disease) 
• Active disease for at least 6 months at the time of randomisation 
All AD patients will be required to switch to twice daily application of 
study emollient (instead of their usual emollient) for at least 7 days prior to 
randomisation (Day 1).  
Subjects will attend the clinic on Day 1.  If all inclusion/exclusion criteria 
are met subjects will be randomised to receive either topical 1.0% (w/w) 
ZPL-5212372 ointment or matching placebo ointment twice daily, for either 
7 days (Cohorts 1 and 2) or 14 days (Cohort 3). 
 
Cohort 1 will consist of 12 healthy adult volunteers, dosed twice daily for 7 
days as in-patients.  An initial sentinel group (n=3) will have study ointment 
applied over 10% BSA.  After 48 hours of dosing (i.e. 4 applications) the 
safety data will be reviewed. Provided that the study ointments are well 
tolerated the remainder of the cohort (n=9) may start dosing with coverage 
of 40% BSA with study ointments.  Assessments will be conducted on a 
daily basis as described in Sections 6 & 7.  Subjects will remain in the clinic 
until Day 9 assessments are complete. 
 
Cohort 2 will consist of 12 moderate to severe AD patients, dosed twice 
daily for 7 days as in-patients.  An initial sentinel group (n=3) will have 
study ointment applied over 10% BSA.  After 48 hours of dosing (i.e. 4 
applications) the safety data will be reviewed. Provided that the study 
ointments are well tolerated the remainder of the cohort (n=9) may start  
 
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dosing with coverage of 40% BSA with study ointments. Assessments will 
be conducted on a daily basis as described in Sections 6 & 7. Patients will 
remain in the clinic until Day 9 assessments are complete. 
 
Cohort 3 will consist of 30 moderate to severe AD patients, dosed twice 
daily for 14 days as out-patients.  Up to a maximum of 50% BSA of AD 
lesioned skin may be treated with study ointments.  Patients will return to 
the clinic following their Baseline visit (Day 1) on Day 5, Day 8, Day 10 
and Day 15 for the assessments described in Sections 6 & 7. 
All subjects (both healthy volunteers and AD patients) will be required to 
have a follow up visit 7-14 days following their last application of study 
ointment. 
 
Treatment: 
Each subject will be treated with 1.0% (w/w) ZPL-5212372 ointment or 
matching placebo, topically applied to the skin twice daily. 
Number of 
Subjects: 
Cohort 1 
Sufficient subjects will be screened to achieve approximately 12 
randomised and evaluable subjects.  
 
Cohort 2 
Sufficient patients will be screened to achieve approximately 12 randomised 
and evaluable patients. 
 
Cohort 3 
It is anticipated that approximately 45 patients will be screened to achieve 
approximately 30 randomised and evaluable patients. 
 
Population: 
 
Cohort 1  
Healthy volunteer adult subjects (18 to 55 years). 
 
Cohorts 2 and 3 
Adult subjects (18 to 65 years) with moderate to severe atopic dermatitis. 
 
Efficacy 
Endpoints: 
Cohort 3 Only 
Primary Endpoints: 
Percentage change from baseline in the Eczema Area and Severity Index 
(EASI) score  
Secondary Endpoints: 
Change from baseline in the Numerical Rating Score (NRS) for Pruritus 
(Worst Itch) over 24 hours  
Change from baseline in the NRS for Sleep Disturbance 
Change from baseline in Duration of Itching 
Change from baseline in Verbal Rating Score (VRS) for Pruritus 
Change from baseline in BSA 
Reduction in Investigators Global Assessment (IGA) Score (subject will be 
deemed to have had a response with a reduction from baseline of at least 2 
points) 
Patient Global Impression of Change (PGIC) 
 
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Safety 
Endpoints: 
 
Cohort 1  
Adverse events, local tolerance assessments, supine vital signs (blood 
pressure (BP) and pulse rate), 12-lead ECG, laboratory safety tests (clinical 
chemistry, haematology and urinalysis). 
 
Cohorts 2 and 3 
Adverse events, supine vital signs (BP and pulse rate), 12-lead ECG, 
laboratory safety tests (clinical chemistry, haematology and urinalysis). 
 
Exploratory 
Endpoints: 
Cohorts 1 to 3 
Subject responses to Ziarco Ointment Questionnaire. 
 
Pharmaco-
kinetic 
Endpoints: 
Cohorts 1 and 2 
ZPL-5212372 plasma concentrations Day 1 (sd): AUCτ, Cmax and Tmax 
ZPL-5212372 plasma concentrations Day 7 (ss): AUClast, AUCτ, Cmax, Tmax, 
and t½. 
Observed accumulation ratios based on AUCτ and Cmax 
 
Cohort 3 
Trough Plasma ZPL-5212372 concentrations 
 
Statistical 
Methods: 
 
Separate randomisation codes will be generated for each cohort. The 
randomisation for Cohort 3 will be stratified by EASI score (EASI ≤20 and 
EASI ≥20).  
 
All statistical testing for Cohort 3 efficacy data will be at the 5% level of 
significance (1-sided) and all point estimates for the comparison between 
treatment groups will be accompanied by 2-sided 90% confidence intervals. 
No adjustment will be made for multiplicity due to multiple 
endpoints/comparisons. No statistical testing will be performed with the 
data from Cohorts 1 and 2. 
For change from baseline endpoints baseline is defined as the last 
measurement available prior to the start of dosing on Day 1. 
Separate tables, listings and figures will be generated for each cohort and 
within each cohort endpoints will be summarized by treatment group and 
visit. 
Safety data (adverse events, vital signs, ECG and laboratory data) will be 
summarized by treatment group and visit (where applicable) using all 
subjects in the Safety Analysis Set – defined as all randomised subjects who 
received at least one dose of study treatment. The incidence of treatment 
emergent adverse events will be tabulated regardless of severity and by 
maximum severity. Those events regarded as possibly treatment related by 
the investigator will be summarized separately. Changes from baseline in 
laboratory parameters, vital signs and ECG data will be summarized in 
addition to absolute values.  
 
ZPL-5212372 concentrations (all cohorts) and parameters (Cohorts 1 and 2 
only) will be summarized using all subjects in the Pharmacokinetic Set - 
 
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defined as all ZPL-5212372 treated subjects in the Full Analysis Set who 
have at least one ZPL-5212372 concentration recorded. The ZPL-5212372 
pharmacokinetic parameters AUCinf, AUClast and t½ will be calculated using 
non-compartmental methods. Cmax and Tmax will be determined directly from 
the data. 
Individual profile and median plots will also be plotted. Median profiles for 
Cohorts 1 and 2 will be plotted by day.  
 
Though some efficacy assessments are included for Cohort 2 the data will 
only be listed and no formal comparisons will be made between the 
treatment groups. Consequently efficacy will only be assessed for Cohort 3. 
The Full Analysis Set (FAS) used for efficacy analyses will include all 
randomized subjects who received at least one dose of study treatment. A 
Per Protocol (PP) Analysis Set for a sensitivity analysis of the primary 
efficacy endpoint will include those subjects in the FAS who have not 
deviated from the protocol in such a way to influence their efficacy 
assessment. 
The primary efficacy endpoint is the percentage change from baseline to 
Week 2 in the EASI score. The percentage changes from baseline in the 
EASI score at Week 2 will be summarized and analysed by ANCOVA. The 
difference between treatment groups in the percentage changes from 
baseline will be estimated, along with its 90% confidence interval. Baseline 
will be included as a continuous covariate in addition to treatment.  
The proportion of subjects who achieve an EASI-50 response (50% 
reduction in EASI score from baseline) at Week 2 will be compared 
between the treatment groups. Group proportions and the difference 
between these proportions will be calculated along with the corresponding 
90% confidence interval.  These data will also be analysed using logistic 
regression, including treatment and baseline.  The odds ratio and 
accompanying 90% confidence interval will be presented. The proportion of 
subjects who achieve an EASI-75 response (75% reduction in EASI) and 
the proportion of subjects who are responders for IGA score (reduction from 
baseline of at least 2 points) at Week 2, will be analysed using the same 
analyses described for the EASI-50 response.  
 
The changes from baseline in body surface area, pruritus NRS score for 
worst itch, NRS score for sleep disturbance and duration of itching at Week 
2 will be analysed using the same analyses described for the primary 
analysis. The changes from baseline in VRS score will be summarized using 
categorical shift tables. The PGIC score will be summarized and analysed 
using logistic regression, presenting odds ratios and associated 90% 
confidence intervals for the comparison between treatment groups. 
 
Responses to the Ziarco ointment questionnaire will be summarized. 
 
 
 
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LIST OF STUDY PERSONNEL 
 
Sponsor 
 
 
 
 
 
 
Tel: 
 
 
 
 
 
Tel: 
 
 
Principal or Coordinating 
Investigator 
 
  
 
 
 
 
Tel: 
 
 
 
Serious Adverse Event 
Reporting 
  
 
 
 
 
Tel:      
 
Email : 
  
Fax :    
 
 
 
Clinical Supplies 
 
 
 
 
 
 
 
Telephone: 
  
Fax:            
   
 
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TABLE OF CONTENTS 
PROTOCOL SYNOPSIS .................................................................................................. 5 
LIST OF STUDY PERSONNEL .................................................................................... 10 
TABLE OF CONTENTS ................................................................................................ 11 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS .............................. 15 
1 
INTRODUCTION................................................................................................ 17 
1.1 
Indication ......................................................................................................... 17 
1.2 
Rationale for cPLA2 Inhibitors ...................................................................... 18 
Figure 1 - Arachidonic Acid Cascade: A Role for cPLA2α in Skin 
Pathophysiology ............................................................................................... 18 
1.3 
ZPL-5212372 Background .............................................................................. 19 
1.4 
Study Rationale ............................................................................................... 20 
1.5 
Dose Rationale ................................................................................................. 22 
1.6 
Risk-Benefit Assessment ................................................................................. 23 
2 
STUDY OBJECTIVES AND ENDPOINTS ...................................................... 24 
2.1 
Primary Objectives ......................................................................................... 24 
2.2 
Secondary Objectives ...................................................................................... 24 
2.3 
Exploratory Objectives ................................................................................... 24 
2.4 
Endpoints ......................................................................................................... 24 
3 
OVERALL DESIGN AND PLAN OF THE STUDY ....................................... 26 
3.1 
Overview .......................................................................................................... 26 
Table 1 - Summary of cohorts with overview of study ............................................ 26 
Figure 2 - Overall Study Design ................................................................................. 28 
Figure 3 - Study Design Cohort 1 (HV) ..................................................................... 29 
Figure 4 - Study Design Cohort 2 (AD Patients) ...................................................... 29 
Figure 5 - Study Design Cohort 3 (AD Patients) ...................................................... 30 
3.2 
Adaptive Categories and Features ................................................................. 30 
Table 2 - Investigational Medicinal Product/Dose ................................................... 31 
Table 3 - Timing/Scheduling ...................................................................................... 32 
Table 4 - Study Participants ....................................................................................... 32 
Table 5 - Assessments .................................................................................................. 32 
3.3 
Termination or Suspension of Study ............................................................. 33 
3.4 
Termination or Suspension of Investigational Sites ..................................... 33 
3.5 
Progression From Sentinel Group To Main Cohort .................................... 33 
3.6 
Safety Review Meeting .................................................................................... 33 
3.7 
Stopping Criteria ............................................................................................. 34 
3.7.1 
Cohort Stopping Criteria ...................................................................... 34 
3.7.2 
Individual Stopping Criteria ................................................................ 35 
3.8 
Planned Sample Size and Number of Study Centers ................................... 35 
4 
STUDY POPULATION ...................................................................................... 35 
4.1 
Inclusion Criteria: Cohort 1 ........................................................................... 36 
4.2 
Inclusion Criteria: Cohorts 2 and 3 ............................................................... 36 
4.3 
Exclusion Criteria: Cohort 1 .......................................................................... 37 
4.4 
Exclusion Criteria: Cohorts 2 and 3 .............................................................. 39 
 
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4.5 
Life Style Guidelines: Cohorts 1 and 2 .......................................................... 41 
4.6 
Life Style Guidelines: Cohort 3 ...................................................................... 42 
4.7 
Contraceptive Requirements .......................................................................... 42 
4.8 
Subject Withdrawal and Replacement .......................................................... 43 
4.9 
Temporary Discontinuations .......................................................................... 44 
5 
STUDY TREATMENT ....................................................................................... 44 
5.1 
Allocation to Treatment .................................................................................. 44 
5.2 
Breaking the Blind .......................................................................................... 44 
5.3 
Drug Supplies .................................................................................................. 45 
5.4 
Treatment Packaging and Labelling ............................................................. 45 
5.5 
Dispensing ........................................................................................................ 46 
5.6 
Drug Storage .................................................................................................... 46 
5.7 
Administration ................................................................................................. 46 
5.8 
Excessive Pharmacological Effects ................................................................ 48 
5.9 
Drug Accountability ........................................................................................ 48 
5.10 
Compliance (Cohort 3 only) ........................................................................... 49 
5.11 
Concomitant Medications ............................................................................... 49 
Table 6 - Prohibited Medications for Cohort 2 & 3 AD patients ............................ 51 
6 
STUDY CONDUCT ............................................................................................. 52 
6.1 
Screening .......................................................................................................... 52 
6.2 
Cohort 1 (Healthy Volunteers) ....................................................................... 53 
6.2.1 
Day 1 assessments................................................................................ 53 
6.2.2 
Days 2 to 6 Assessments ...................................................................... 54 
6.2.3 
Day 7 Assessments .............................................................................. 54 
6.2.4 
Day 8 Assessments .............................................................................. 55 
6.2.5 
Day 9 Assessments .............................................................................. 55 
6.3 
Cohort 2 (AD Patients) ................................................................................... 55 
6.3.1 
Day 1 assessments................................................................................ 56 
6.3.2 
Day 2 to 6 assessments ........................................................................ 57 
6.3.3 
Day 7 assessments................................................................................ 57 
6.3.4 
Day 8 assessments................................................................................ 57 
6.3.5 
Day 9 assessments................................................................................ 58 
6.4 
Cohort 3 (AD Patients) ................................................................................... 58 
6.4.1 
Baseline Visit (Day 1) .......................................................................... 58 
6.4.2 
Visits 2 to 4 (Days 5, 8 and 10) ........................................................... 59 
6.4.3 
Visit 5 Day 15 Or Early Termination .................................................. 59 
6.5 
Follow Up visit ................................................................................................. 60 
Table 7 - Follow up visit assessments for each cohort ............................................. 60 
6.6 
Early Termination Assessments ..................................................................... 60 
Table 8 - Early termination visit assessments for each cohort ................................ 61 
6.7 
Schedule of Activities ...................................................................................... 62 
Table 9 - Schedule of activities for Cohort 1 (Healthy Volunteers) ........................ 62 
Table 10 - Schedule of activities for Cohort 2 (Moderate to Severe AD Patients) 63 
Table 11 - Schedule of activities for Cohort 3 (Moderate to Severe AD Patients) 64 
7 
ASSESSMENTS ................................................................................................... 65 
7.1 
Physical Examination ...................................................................................... 65 
7.2 
Vital Signs Measurements .............................................................................. 65 
 
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7.3 
Electrocardiogram (ECG) Recording ........................................................... 65 
7.4 
Fitzpatrick Skin Type Assessment ................................................................. 66 
7.5 
Assessment of Local Toleration ..................................................................... 66 
7.6 
Skin Area Body Map ....................................................................................... 67 
7.7 
Eczema Area and Severity Index (EASI) ...................................................... 67 
7.8 
Body Surface Area (BSA) ............................................................................... 67 
7.9 
Investigator Global Assessment (IGA) .......................................................... 68 
7.10 
Ziarco Itch Questionnaire .............................................................................. 68 
7.10.1 
Numerical Rating Scale (NRS) for Pruritus (Itching).......................... 68 
7.10.2 
Duration of Itching ............................................................................... 68 
7.10.3 
Sleep Disturbance ................................................................................ 68 
7.10.4 
Verbal Rating Score ............................................................................. 68 
7.11 
Patient Global Impression of Change (PGIC) .............................................. 69 
7.12 
Ziarco Ointment Questionnaire ..................................................................... 69 
7.13 
Pharmacokinetic Blood Samples ................................................................... 69 
7.14 
Clinical Laboratory Safety Tests ................................................................... 70 
Table 12 – Safety Laboratory parameters ................................................................ 71 
7.15 
Blood Volume .................................................................................................. 71 
Table 13 - Approximate Blood Volume Required for Study ................................... 72 
7.16 
Safety ................................................................................................................ 72 
7.16.1 
Adverse Events .................................................................................... 72 
7.16.1.1 
Definitions ..................................................................................... 72 
7.16.1.2 
Assessment of Adverse Event ........................................................ 72 
7.16.1.3 
Recording Adverse Events ............................................................ 73 
7.16.1.4 
Reporting Serious Adverse Events ............................................... 74 
7.16.1.5 
Follow-up of Adverse Events ........................................................ 75 
8 
STATISTICAL METHODS ............................................................................... 75 
8.1 
Study Subjects ................................................................................................. 75 
8.1.1 
Randomisation ..................................................................................... 75 
8.1.2 
Disposition of Subjects ........................................................................ 75 
8.1.3 
Protocol Deviations .............................................................................. 75 
8.1.4 
Analysis Populations ............................................................................ 75 
8.2 
General Considerations .................................................................................. 76 
8.3 
Demographics, Baseline Characteristics and Concomitant Medications .. 76 
8.4 
Treatment Compliance ................................................................................... 76 
8.5 
Efficacy Analyses ............................................................................................. 76 
8.5.1 
Primary Endpoint ................................................................................. 77 
8.5.2 
Secondary Endpoints ........................................................................... 77 
8.5.3 
Exploratory Endpoints ......................................................................... 78 
8.6 
Safety Analyses ................................................................................................ 78 
8.7 
Pharmacokinetic Analyses .............................................................................. 78 
8.8 
Interim Analyses .............................................................................................. 78 
8.9 
Determination of Sample Size ........................................................................ 79 
9 
ETHICAL, LEGAL AND ADMINISTRATIVE ASPECTS ........................... 79 
9.1 
Data Quality Assurance .................................................................................. 79 
9.2 
Case Report Forms and Source Documentation .......................................... 79 
9.3 
Access to Source Data ..................................................................................... 79 
9.4 
Data Processing ............................................................................................... 80 
 
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9.5 
Archiving Study Records ................................................................................ 80 
9.6 
Good Clinical Practice .................................................................................... 81 
9.7 
Confidentiality ................................................................................................. 81 
9.8 
Informed Consent ............................................................................................ 81 
9.9 
Protocol Approval and Amendment .............................................................. 81 
9.10 
Duration of the Study ...................................................................................... 82 
9.11 
Premature Termination of the Study ............................................................ 82 
9.12 
Liability and Insurance ................................................................................... 82 
9.13 
Publication Policy ............................................................................................ 82 
10 
REFERENCE LIST ............................................................................................. 84 
11 
APPENDICES ...................................................................................................... 87 
APPENDIX 1 – CRITERIA FOR SAFETY VALUES OF POTENTIAL CLINICAL 
CONCERN ........................................................................................................... 88 
APPENDIX 2 – HEPATIC SAFETY ............................................................................. 89 
APPENDIX 3 – ECG VALUES OF POTENTIAL CLINICAL CONCERN ............. 90 
APPENDIX 4 – COHORT 1 10% BSA TREATMENT BODY MAP EXAMPLE ... 91 
APPENDIX 5 – COHORT 1 40% BSA TREATMENT BODY MAP EXAMPLE ... 92 
APPENDIX 6 – COHORT 2 10% BSA TREATMENT BODY MAP EXAMPLE ... 93 
APPENDIX 7 – COHORT 2 40% BSA TREATMENT BODY MAP EXAMPLE ... 94 
APPENDIX 8 – COHORT 3 BODY MAP EXAMPLE ............................................... 95 
APPENDIX 9 – ZIARCO ITCH QUESTIONNAIRE .................................................. 96 
APPENDIX 10 – BODY MAP ........................................................................................ 97 
APPENDIX 11 - EASI ..................................................................................................... 98 
APPENDIX 12 – % BODY SURFACE AREA GUIDANCE .................................... 100 
APPENDIX 13 – ZIARCO OINTMENT QUESTIONNAIRE .................................. 101 
APPENDIX 14 – PROTOCOL AMENDMENT 1 ...................................................... 102 
 
 
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LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS 
 AD 
ADR 
AE 
Atopic Dermatitis 
Adverse Drug Reaction 
Adverse event 
ALT 
ANCOVA 
AP 
Alanine aminotransferase  
Analysis of Covariance 
Alkaline phosphatase 
AST 
AUC 
BID 
BSA 
Aspartate aminotransferase  
Area Under the Curve (inf; last; t) 
Twice daily 
Body surface area 
CK 
Cmax 
 CMO 
 COX 
 cPLA2α  
Creatine Kinase 
Maximum observed concentration 
Chief Medical Officer 
Cyclo-oxygenase 
Cytosolic phospholipase A2α  
 CRA 
CRF 
Clinical Research Associate 
Case report form 
CS 
CRU 
DPI 
EASI 
ECG 
ECLIA 
Corticosteroids 
Clinical research unit 
Dry powder inhaler 
Eczema Area and Severity Index 
Electrocardiogram 
 Electro-chemiluminescence immunoassay 
EDC 
EMA 
FAS 
FBC 
FDA 
FIM 
FSH 
Electronic data capture 
European Medicines Agency 
Full analysis set 
Full blood count 
Food and Drug Administration 
First in man 
Follicle-stimulating hormone 
GCP 
Good Clinical Practice 
GGT 
Gamma-glutamyl transpeptidase 
Hb 
HbA1c 
HBcAb 
HBsAg 
hCG 
HIV 
HPA 
IC50 
ICD 
ICH  
IEC 
Haemoglobin 
Glycated haemoglobin 
Hepatitis B core Antibody 
Hepatitis B surface Antigen 
Human chorionic gonadotropin 
Human Immunodeficiency Virus 
Hypothalamic pituitary-adrenal 
Inhibitory concentration 
Informed consent document 
International Council on Harmonization 
Independent Ethics Committee 
IGA 
IUD 
IUS 
IWRS 
Investigator Global Assessment 
Intrauterine device 
Intrauterine hormone-releasing system 
Integrated Web-based Response System 
Kg 
Kilogram 
 
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 Version 2.0 /28 July 2016 
 
5-LO 
LFT 
LT 
MCH 
 MCV 
 MedDRA 
5-lipoxygenase 
Liver function tests  
Leukotrienes 
Mean corpuscular haemoglobin 
Mean corpuscular volume 
Medical Dictionary for Regulatory Activities 
mg 
mm Hg 
mL 
nM 
NOAEL 
NOEL 
NRS 
NSAID 
PAF 
PDF 
PG 
PGIC 
PI 
PK 
PMA 
PP 
QTcF 
QoL 
SAE 
SAP 
SC 
 
T½ 
TB 
TC 
TCI 
TNCB 
Tmax  
TXA2 
µg 
ULN 
Milligrams 
Millimeters of Mercury 
Milliliters 
Nanomole 
No Observed Adverse Effect Level 
No Observed Effect Level 
Numerical Rating Scale 
Non-steroidal anti-inflammatory drugs 
Platelet activating factor 
Portable Document Format 
Prostaglandins 
Patient Global Impression of Change 
Principal investigator 
Pharmacokineticµ 
Phorbol 12-myristate 13-acetate  
Per protocol 
Fridericia’s corrected QTc interval 
Quality of Life 
Serious adverse event 
 Statistical Analysis Plan 
 Subcutaneous 
 
 
 Half life 
 Total bilirubin 
Topical corticosteroids 
Topical calcineurin inhibitors 
2,4,6-Trinitrochlorobenzene  
Time at which the Cmax is observed 
Thromboxane A2 
Microgram 
Upper Limit of Normal 
UV 
UVA 
UVB 
VRS  
WBC 
WOCBP 
w/w 
 
Ultraviolet 
Ultraviolet A 
Ultraviolet B 
Verbal Rating Scale 
White Blood cells 
Women of child bearing potential 
Weight/weight 
 
 
 
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1 
INTRODUCTION 
1.1 
Indication 
Atopic dermatitis (AD) is a chronic, relapsing, inflammatory skin disease characterised by 
impaired epidermal barrier function, inflammatory infiltration, extensive pruritus and a 
clinical course defined by symptomatic flares and remissions. It is characterised by poorly 
defined erythema with oedema, vesiculation, and weeping in the acute stage and skin 
thickening (lichenification) in the chronic stage.1,2 These irritating symptoms result in a 
severely reduced quality of life (QOL). In particular, itching (pruritus) that continues 
throughout the day and worsens at night causes sleep loss and impacts everyday activities and 
psychosocial wellbeing.3 AD is commonly associated with other atopic and inflammatory 
disorders, such as asthma and allergic rhinitis. 
 
The disease occurs in all races and geographic locations, with the highest incidence in urban 
areas and developed countries.4,5 The prevalence of AD has at least doubled in industrial 
countries during the last three decades. Between 15% to 30% of children and 1.4% to 10.2% 
of adults are affected by AD.5,6,7,8 The majority of adults present with a chronic, recurrent 
course of the disease.9 
 
There are currently two models explaining the pathogenesis of AD. The predominant model 
describes AD as a result of impaired epidermal barrier function secondary to inherent 
structural and functional abnormalities of the skin. The disease evolves from the outside in, as 
a result of the abnormalities in the epidermal barrier.10 The second more traditional model 
describes AD primarily as a disorder of the immune system. In this model, environmental 
factors cause Langerhans cells, T-cells, and immune effector cells to modulate an 
inflammatory response.  
 
Clinical features of adult AD include diffuse lesions with an underlying background of 
erythema, skin dryness, intense pruritus, and sleep disturbances. The face is usually involved 
and a brown macular ring around the neck from localized deposition of amyloid is common. 
Xerosis is prominent and lichenification may be present. Visible skin lesions affect social 
interactions, psychological adjustment, work success, sexual relationships and overall quality 
of life.11 Presentation of major mood disturbances such as anxiety, depression, mood changes, 
self-esteem or stigmatization are prevalent in adult AD patients.12,13 Spouses of patients with 
AD also may feel the psychological burden of their partners' severe skin disease and can 
experience depressive or anxious symptoms.14 
 
Topical corticosteroids (TC) have been the mainstay of treatment for AD for several decades. 
Well known side-effects of TC treatment include skin thinning, and possible hypothalamic 
pituitary-adrenal (HPA) axis suppression.15,16 An alternative to TC are topical calcineurin 
inhibitors (TCIs). TCIs have been approved for the treatment of AD since 2000, the 
compounds work by inhibiting calcineurin, thus preventing the dephosphorylation activity of 
phosphatase and the production and release of inflammatory cytokines and T-cell 
proliferation. In January of 2006, the United States Food and Drug Administration (FDA) 
issued a black box warning on these compounds mentioning possible concerns about the 
potential of increased long term malignancy risk due to systemic immunosuppression and 
recommended that TCIs not be used as first-line therapy for AD.17  
 
This poses a significant healthcare challenge in treating patients with moderate to severe AD 
and who have chronic symptoms. Therefore there is a need for new treatment options in AD 
 
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that safely and effectively target both the inflammatory and pruritic components of this 
condition. 
1.2 
Rationale for cPLA2 Inhibitors 
Cytosolic phospholipase A2α (cPLA2α) releases arachidonic acid from phospholipid 
membrane and is the rate-limiting enzyme in the biosynthesis of chemical lipid mediators, 
including various prostaglandins (PGs), leukotrienes (LTs), thromboxaneA2 (TXA2) and 
platelet activating factor (PAF) as shown in Figure 1. 
 
Figure 1 - Arachidonic Acid Cascade: A Role for cPLA2α in Skin Pathophysiology 
 
 
 
It is well documented that PGs, LTs and TXA2 play important roles in inflammation, 
including recruitment of inflammatory cells, promotion of oedema by increasing vascular 
permeability and permitting leakage of plasma to the extravascular space.18 In AD, the levels 
of eicosanoids such as LTB4 and PGE2 are elevated in skin lesions, as compared to 
non-lesional skin in patients.19 PGE2 has been shown to induce vasodilation and itch in 
patients with AD.20 In addition, LTB4 and cysteinyl leukotrienes are involved in keratinocyte 
hyperproliferation, an important process in AD.21 
 
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Selective inhibitors of cPLA2α with distinct chemical structures have demonstrated 
significant anti-inflammatory activity in a variety of skin inflammation models. In a chronic 
mouse model of phorbol 12-myristate 13-acetate (PMA)-induced dermatitis, topical 
application of the cPLA2α inhibitor BMS-229724 significantly reduced oedema and 
neutrophil infiltration with efficacy comparable to topically-applied betamethasone.22 In a 21 
day 2,4,6-trinitrochlorobenzene (TNCB)-induced mouse ear inflammation model, the 
selective cPLA2α inhibitor RSC-3388 significantly inhibited oedema and inflammatory 
cytokine generation with efficacy comparable to the calcineurin inhibitor tacrolimus.23 The 
cPLA2α knockout mouse is highly resistant to inflammatory challenges and displayed 
significant reduction in both early and late phase allergen-induced cutaneous responses. 
ZPL-5212372 is a highly potent and selective inhibitor of cPLA2α that prevents the 
biosynthesis of these pro-inflammatory mediators. Ziarco plans to study ZPL-5212372 in a 
topical formulation for the treatment of patients with AD. 
1.3 
ZPL-5212372 Background 
ZPL-5212372 was formerly known as PF-05212372 and was being developed by Pfizer as an 
inhaled treatment for asthma, prior to its acquisition by Ziarco Pharma Limited in October 
2012. 
 
In vitro studies have shown ZPL-5212372 to be a potent (low nanomolar) inhibitor of 
cPLA2α in recombinant enzyme assays.  These assays also demonstrated the selectivity of the 
molecule as it failed to inhibit the closely related cPLA2ζ isoform or the physiologically 
related cyclo-oxygenase (COX) 1 and 2. ZPL-5212372 inhibited release of downstream 
prostaglandins, leukotrienes and thromboxanes in rat and human whole blood assays with 
50% inhibitory concentrations (IC50) ranging from 2.7 nM to 19.0 nM. The inhibition of 
multiple lipid mediators in these primary human cell systems supports the assumption that 
ZPL-5212372 will block the COX-1, COX-2 and 5-lipoxygenase (5-LO) pathways in man. 
 
Topically administered ZPL-5212372 was effective in blocking skin inflammation induced 
by PMA in the mouse ear oedema model. As ZPL-5212372 was originally developed for the 
asthma indications, in vivo studies have examined the effects of ZPL-5212372 inhibition of 
cPLA2α by inhaled delivery in a variety of lung inflammation models including mouse and 
sheep models of allergic asthma. Taken together, the clear anti-inflammatory effects 
demonstrated by ZPL-5212372 in models of skin and lung inflammation support the potential 
of ZPL-5212372 as a therapeutic agent to treat inflammatory skin diseases. 
 
Pre-clinically there were no safety pharmacology findings of clinical relevance with 
ZPL-5212372 and the results of genotoxicity studies indicate that ZPL-5212372 does not 
present a genotoxic risk to humans. In addition, ZPL-5212372 did not induce local toleration 
reactions in vitro in bovine eyes or sensitisation in vivo in guinea-pig skin either topically or 
intradermally. ZPL-5212372 was well tolerated in toxicology studies up to 2 weeks in 
duration in rats with once daily subcutaneous (SC) systemic administration and with twice 
daily topical administration to mini-pigs (0.5 mL/kg of vehicle or 0.3%, 1.0% or 3.0% (w/w) 
ZPL-5212372 ointment applied twice daily (BID) to 10% BSA). No safety findings were 
identified in either the 2 week rat SC or topical mini-pig studies, hence the NOAELs are 50 
mg/kg/day and 30 mg/kg/day (3.0% w/w ZPL-5212372 ointment), respectively.  In the rat 
following 50 mg/kg/day the Cmax plasma exposures at Day 13 were 5520 and 1740 ng/mL 
and the AUC0-t values were 44900 and 31200 ng.h/mL in males and females, respectively.  In 
 
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the mini-pig following 30 mg/kg/day the Cmax plasma exposures at Day 14 were 14.2 and 
44.7 ng/mL and the AUC0-t values were 394 and 908 ng.h/mL in males and females, 
respectively. 
To date, one Phase 1 clinical study has been conducted in 24 male healthy volunteers. In this 
study ZPL-5212372 was administered by dry powder inhaler (DPI) as part of the original 
asthma indication development program, supported by the 2 week inhalation toxicology 
studies. This study assessed the safety, toleration, plasma and urine pharmacokinetics and 
urine pharmacodynamics of ZPL-5212372 following single-dose inhalation administration of 
50 µg −10 mg of ZPL-5212372. Inhaled doses of ZPL-5212372 up to 10 mg were well 
tolerated. There were no deaths, serious adverse effects (SAEs) or severe adverse effects 
(AEs) associated with dosing of any treatments and no clinically significant changes in 
laboratory safety tests, vital signs or electrocardiograms (ECGs) were observed. There is no 
evidence at the present time of any dose relationship for AEs reported. Plasma 
concentration-time profiles of ZPL-5212372 indicate that systemic absorption following single 
inhaled dose administration is rapid (median Tmax 0.5 hours) with low systemic exposure 
(10 mg dose: Cmax 0.32 ng/mL and AUC0-last 1.32 ng.h/mL). Exposure increased 
dose-proportionally between the 5 and 10 mg dose groups. 
 
Full details of the pre-clinical and clinical data for ZPL-5212372 can be found in the single 
reference safety document, which for this study is the current ZPL-5212372 Investigator 
Brochure. 
1.4 
Study Rationale 
This study will be a randomised, adaptive design, double blind (3rd party open), placebo 
controlled, sequential group study in both healthy volunteers and patients with moderate to 
severe AD.  This study will be divided into 3 separate, sequential cohorts: 
 
• Cohort 1 will assess safety, toleration and pharmacokinetics (PK) with intensive 
monitoring as in-patients over 7 days of dosing in healthy volunteers. 
• Cohort 2 will assess safety, toleration and PK with intensive monitoring as in-patients 
over 7 days of dosing in moderate to severe AD patients. 
• Cohort 3 will assess efficacy, safety, toleration and PK as out-patients over 14 days of 
dosing in moderate to severe AD patients. 
 
Subjects will receive 1.0% (w/w) ZPL-5212372 or matched placebo ointment topically, twice 
daily. 
In total approximately 54 subjects (12 subjects each for Cohorts 1 and 2; 30 subjects for 
Cohort 3) will be enrolled into the study and subjects will be allocated active and placebo 
treatments in a 2:1 ratio in all 3 cohorts. 
 
To mitigate risk to participants in this study the cohorts will be recruited sequentially with 
progression through the cohorts dependent on adequate safety and PK data from the previous 
cohort. A decision process will be followed for each safety review and the findings at each 
review will be documented. Cohort 1, with healthy volunteers, will assess the safety, local 
toleration and plasma exposure with dosing twice daily for 7 days on normal skin. This 
cohort will start with a sentinel (n=3) group who will have study ointment applied to 10% 
BSA BID for 7 days.  If well tolerated, following safety assessments 48 hours after the first 
 
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application (i.e. 4 applications of study ointment), the remaining subjects (n=9) will begin 
dosing at 40% BSA, BID for 7 days.  If after a formal safety review of the Cohort 1 safety 
and PK data, the data indicate that the application of ointment can be considered safe and 
well tolerated, dosing of Cohort 2 will commence.  Cohort 2 will recruit moderate to severe 
AD patients in order to investigate safety, toleration and exposure following application of 
study ointment to AD diseased skin.  Cohort 2 will also include a sentinel (n=3) group who 
will only have study ointment applied to 10% BSA BID for 7 days.  Again if after 48 hours, 
after the first application of ointment, dosing is well tolerated in the sentinel group, the 
remainder of the cohort (n=9) will start with 40% BSA treated BID for 7 days with study 
ointment.  Subjects in Cohorts 1 and 2 will remain in the clinical research unit (CRU) for the 
duration of dosing and until 48 hours after the last study treatment to enable close monitoring.  
This will provide safety, toleration and PK information from both healthy and AD diseased 
skin treated with ZPL-5212372.  Subjects will only be recruited into Cohort 3 when the safety 
data from Cohorts 1 and 2 have been formally reviewed by the Safety Review Team. 
 
Cohort 3 (n=30) will recruit moderate to severe AD patients, who will be treated twice daily 
for 14 days as out-patients returning to the CRU for study assessments on days 5, 8, 10 and 
15.  Cohort 3 patients will be treated with study ointment up to 50% BSA to their AD 
lesioned skin.  The primary objective of Cohort 3 will be to assess the efficacy of ZPL-
5212372 over matched placebo ointment, in addition to obtaining 14 days safety and 
toleration data. Trough PK levels of ZPL-5212372 will also be collected.  This risk averse 
sequential adaptive design is being used as this will be the first time that ZPL-5212372 will 
be administered topically to human skin. From the data reviews and ongoing learning in the 
CRU, protocol defined adaptions may be made to the study between cohorts to minimize any 
risks to participants.  All adaptions outlined in this protocol are constrained by clear 
boundaries and stopping criteria.  All modifications to the in-life phase of the study will be 
documented and approved by Ziarco’s Chief Medical Officer (CMO) and the Principal 
Investigator (PI). 
 
No rescue medication will be provided in this study for AD patients due to the short duration 
of dosing (7 days for Cohort 2 and 14 days for Cohort 3).  Use of rescue medication in Cohort 
3 could potentially impact the efficacy data collected.  The severity of AD patients included 
in the study will be limited to moderate to severe patients with an Ezcema Area and Severity 
Index (EASI) score of ≤48 as AD patients with AD of a greater severity may not be able to 
tolerate the conditions in the study without rescue medication.  AD patients will be supplied 
with study emollient to be started 7 days prior to randomisation and to continue throughout 
their participation in the study.  AD patients in Cohort 2 will be resident in the CRU for the 
duration of treatment and will therefore have close contact with clinical staff to discuss any 
issues relating to their AD. Patients in Cohort 3 will regularly attend the CRU during 
treatment for study assessments and will be encouraged to contact the CRU between visits if 
they feel their AD is deteriorating. 
 
Placebo-controlled trials have been routinely utilised to test new molecular entities in the 
treatment of dermatological diseases, including AD. This trial is designed to show neither 
1.0% (w/w) ZPL-5212372 ointment or placebo has harmful effects when applied to human 
skin.  In addition Cohort 3 is designed to demonstrate efficacy of ZPL-5212372 over placebo. 
Placebo has been selected as the comparator in this study in order to establish the magnitude 
of change in clinical endpoints that occur spontaneously during the treatment period. Ethical 
analysis and international ethical guidance permit the use of placebo controls in randomised 
trials when there are compelling methodological reasons, and withholding treatment does not 
 
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pose a risk of serious harm to participants.24 The 2:1 randomisation has been selected to 
increase the number of subjects participating in the active arm, whilst including sufficient 
control subjects to make valid statistical comparisons. 
 
The 2 week maximum dosing duration has been selected based on the duration of completed 
toxicology studies, but is considered adequate to provide an indication of efficacy in this 
patient population. 
 
As AD affects males and females equally, women of child bearing potential (WOCBP) who 
are using highly effective methods of contraception will be included in this study as they 
represent an important section of the affected subjects.  The inclusion of WOCBP is 
consistent with both the recommendations of the Clinical Trials Facilitation Group (Clinical 
Trials Facilitation Group, 2014)25 following the Rome September 2014 meeting and also the 
European Medicines Agency (EMA) M3(R2) guidance.  M3(R2) states: 
“In all ICH regions, WOCBP can be included in repeated-dose Phase I and II trials before 
conduct of the female fertility study since an evaluation of the female reproductive organs is 
performed in the repeated-dose toxicity studies”. 
 
1.5 
Dose Rationale 
The dose selected for this study is a 1.0% (w/w) ZPL-5212372 topical ointment, to be applied 
twice daily for up to 2 weeks. 
 
The 1.0% (w/w) concentration of ZPL-5212372 has been selected to be 3 fold lower than the 
maximum concentration applied twice daily to mini-pigs for 14 days with no local toleration 
or systemic toxicity issues.  Currently there is no predicted efficacious dose level for 
topically administered ZPL-5212372 in humans.  At the present time the only human plasma 
exposure data for ZPL-5212372 is from the inhaled first in man (FIM) study conducted by 
Pfizer when ZPL-5212372 was being developed as a potential treatment for asthma.  
Systemic exposures of ZPL-5212372 following a single 10 mg inhaled dose resulted in an 
AUC0-12 1.32 ng.h/mL and Cmax 0.32 ng/mL.  The human systemic exposures following 
topical administration are also expected to be low and this is further supported by the much 
lower systemic exposures seen in mini-pig following topical administration compared to the 
systemic exposures in rats seen following SC administration of ZPL-5212372.  In addition 
the NOAELs in the 14 day repeat dose studies in both the rat (SC) and mini-pig (topically) 
were the highest dose levels administered.  PK data, in addition to safety data, will be 
collected in the initial clinical study with topically administered ZPL-5212372, hence both 
systemic exposure and safety can be carefully monitored.  Topical application of 
ZPL-5212372 to humans is expected to give low systemic exposure and plasma exposures 
will be monitored on an on-going basis to avoid systemic plasma exposures exceeding the 
mean AUC0-t value of 13550 ng.h/mL (the mean NOEL (15 mg/kg/day) systemic exposure 
seen in the rat after SC dosing for 13 days).  Exposures in humans following a topical dose of 
1.0% (w/w) ZPL-5212372 ointment are not expected to exceed the exposures seen in the 
mini-pig following the 30 mg/kg/day dose (mean AUC0-t 651 ng.h/mL). 
 
Pre-clinical studies indicate that ZPL-5212372 has a low potential for drug-drug interactions 
as either victim or perpetrator.  ZPL-5212372 is primarily hepatically cleared.  ZPL-5212372 
is highly bound to plasma protein, yet with very high rates of unbound clearance hence 
circulating systemic drug levels are low, ensuring high systemic safety from topical delivery.  
In addition the inhibitory effect of ZPL-5212372 has been shown to be reversible. 
 
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The combination of high potency, rapid clearance of unbound drug and hence low systemic 
burden makes ZPL-5212372 an ideal compound for topical delivery for the treatment of 
moderate to severe AD, both in terms of efficacy as well as systemic safety. The selected 
dose has been shown to have an adequate safety profile and be well tolerated and is not 
expected to exceed exposures associated with the NOAEL in preclinical toxicology studies.  
1.6 
Risk-Benefit Assessment 
Benefits 
There is preclinical data demonstrating that cPLA2α inhibition should be effective in 
relieving skin inflammation and pruritus, both of which are involved in AD.  Selective 
inhibitors of cPLA2α with distinct chemical structures have demonstrated significant 
anti-inflammatory activity in a variety of skin inflammation models.22,23 cPLA2α knockout 
mice are highly resistant to inflammatory challenges and displayed significant reduction in 
both early and late phase allergen-induced cutaneous responses. ZPL-5212372 is a selective 
cPLA2α inhibitor which in non-clinical pharmacology studies has been shown to potently 
inhibit a number of inflammatory-mediated processes both in the skin and lung. Hence it is 
believed that ZPL-5212372 has the potential to reduce the incidence and severity of both 
inflammation and pruritus in AD patients. 
 
Risks 
Previously ZPL-5212372 has been administered to healthy volunteers as single inhaled doses 
up to 10 mg with no safety or toleration issues. Adverse events were mild to moderate in 
severity and there have been no serious adverse events. 
 
Pre-clinically there were no safety pharmacology findings of clinical relevance with 
ZPL-5212372 and the results of genotoxicity studies indicate that ZPL-5212372 does not 
present a genotoxic risk to humans. In addition, ZPL-5212372 did not induce local toleration 
reactions in vitro in bovine eyes or in vivo sensitisation in guinea-pig skin either topically or 
intra-dermally.  ZPL-5212372 was well tolerated in toxicology studies up to 2 weeks in 
duration in rats with once daily subcutaneous (SC) systemic administration and with twice 
daily topical administration to mini-pigs (0.5 mL/kg of vehicle or 0.3%, 1.0% or 3.0% (w/w) 
ZPL-5212372 ointment applied BID to 10% BSA). No effects were identified in either the 2 
week rat SC or topical mini-pig studies hence the NOAELs are 50 mg/kg/day and 30 
mg/kg/day (3.0% w/w ZPL-5212372 ointment), respectively.  In the rat following 50 
mg/kg/day the Cmax plasma exposures at Day 13 were 5520 and 1740 ng/mL and the AUC0-t 
values were 44900 and 31200 ng.h/mL in males and females, respectively.  In the mini-pig 
following 30 mg/kg/day the Cmax plasma exposures at Day 14 were 14.2 and 44.7 ng/mL and 
the AUC0-t values were 394 and 908 ng.h/mL in males and females, respectively. 
 
In accordance with the latest European regulatory discussions and guidelines as outlined 
above, WOCBP are eligible for the study provided they are using a highly effective form of 
contraception.  A serum pregnancy test will be performed at screening.  In addition, regular 
urine pregnancy tests will be carried out on all WOCBP at randomisation and continuing until 
the follow up visit is complete, 7 to 14 days after application of the last dose of study 
ointment. 
 
 
 
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Conclusion 
In conclusion, the risk-benefit profile of ZPL-5212372 supports the continued development 
of this compound and confirms that there is minimal risk associated with the evaluation of 
this compound in an AD patient population.  However, as this is the first topical 
administration of ZPL-5212372 the study will be conducted initially in a cohort of healthy 
volunteer subjects followed by an in-house cohort of patients with moderate to severe AD 
prior to treating a cohort of moderate to severe AD patients as out-patients. 
 
2 
STUDY OBJECTIVES AND ENDPOINTS 
2.1 
Primary Objectives 
Cohorts 1 and 2 
To assess the safety and tolerability, local and systemic, of ZPL-5212372 administered as a 
topical ointment (containing 1.0% (w/w) concentration of ZPL-5212372) twice daily, for 1 
week, to healthy subjects and patients with moderate to severe AD. 
 
Cohort 3 
To evaluate the efficacy of ZPL-5212372 administered as a topical ointment (containing 
1.0% (w/w) concentration of ZPL-5212372) twice daily, for 2 weeks, to patients with 
moderate to severe AD. 
 
2.2 
Secondary Objectives 
Cohorts 1 and 2 
To investigate the systemic pharmacokinetics of ZPL-5212372 following single and multiple 
dose topical applications of a 1.0% (w/w) concentration of ZPL-5212372 ointment, 
administered twice daily, for 1 week, to healthy subjects and patients with moderate to severe 
AD. 
 
Cohort 3 
To assess the safety and tolerability, local and systemic, of ZPL-5212372 administered as a 
topical ointment (containing 1.0% (w/w) concentration of ZPL-5212372) twice daily, for 2 
weeks, to patients with moderate to severe AD. 
 
To investigate the trough concentrations of ZPL-5212372 in patients with moderate to severe 
AD following twice daily applications of ZPL-5212372 administered as a topical ointment 
(containing 1.0% (w/w) concentration of ZPL 5212372). 
 
2.3 
Exploratory Objectives 
Cohorts 1 to 3 
To explore the subjects perception of the Ziarco development ointment formulation using the 
Ziarco Ointment Questionnaire. 
 
2.4 
Endpoints 
Cohort 1 (Healthy Volunteers) 
 
 
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Primary Endpoints: Safety 
Adverse events, local tolerance assessments, supine vital signs (BP and pulse rate), 12-lead 
ECG, laboratory safety tests (clinical chemistry, haematology and urinalysis). 
Secondary Endpoints: Pharmacokinetics 
ZPL-5212372 plasma concentrations Day 1 (sd): AUCτ, Cmax and Tmax 
ZPL-5212372 plasma concentrations Day 7 (ss): AUClast, AUCτ, Cmax, Tmax, and t½. 
Observed accumulation ratios based on AUCτ and Cmax 
Exploratory Endpoints: 
Subject responses to Ziarco Ointment Questionnaire. 
Cohort 2 (Moderate to Severe AD patients) 
 
Primary Endpoints: Safety 
Adverse events, supine vital signs (BP and pulse rate), 12-lead ECG, laboratory safety tests 
(clinical chemistry, haematology and urinalysis). 
Secondary Endpoints: Pharmacokinetics 
ZPL-5212372 plasma concentrations Day 1 (sd): AUCτ, Cmax and Tmax 
ZPL-5212372 plasma concentrations Day 7 (ss): AUClast, AUCτ, Cmax, Tmax, and t½. 
Observed accumulation ratios based on AUCτ and Cmax 
Exploratory Endpoints: 
Patient responses to Ziarco Ointment Questionnaire. 
 
Cohort 3 (Moderate to Severe AD patients) 
 
Primary Endpoints: Efficacy 
Percentage change from baseline in the Eczema Area and Severity Index (EASI) score  
Secondary Endpoints: Efficacy 
Change from baseline in the Numerical Rating Score (NRS) for Pruritus (Worst Itch) over 24 
hours  
Change from baseline in the NRS for Sleep Disturbance 
Change from baseline in Duration of Itching 
Change from baseline in Verbal Rating Score (VRS) for Pruritus 
Change from baseline in BSA 
 
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Reduction in Investigators Global Assessment (IGA) Score (subject will be deemed to have 
had a response with a reduction from baseline of at least 2 points) 
Patient Global Impression of Change (PGIC) 
Secondary Endpoints: Safety 
Adverse events, supine vital signs (BP and pulse rate), 12-lead ECG, laboratory safety tests 
(clinical chemistry, haematology and urinalysis). 
Secondary Endpoints: Pharmacokinetics 
Trough Plasma ZPL-5212372 concentrations 
Exploratory Endpoints: 
Patient responses to Ziarco Ointment Questionnaire. 
 
3 
OVERALL DESIGN AND PLAN OF THE STUDY 
3.1 
Overview 
This is a randomised, adaptive design, double-blind (3rd party open), placebo controlled, 
sequential group study to determine the safety, tolerability, PK and efficacy of twice daily 
application of topical ZPL-5212372 (1.0% w/w) ointment administered for up to 2 weeks in 
adult healthy volunteers and in patients with moderate to severe atopic dermatitis. 
 
This study will consist of 3 sequential cohorts each separated by formal safety reviews.  The 
study will recruit cohorts sequentially and use sentinel subjects (n=3) in Cohorts 1 and 2 to 
minimize the risk to subjects and make it possible to modify specific features of the study 
dependent on emerging data; however the flexibility of the adaptive features will be 
constrained by clear boundaries/stopping criteria.  The main features of each cohort are 
summarized in Table 1 below: 
 
Table 1 - Summary of cohorts with overview of study 
 
Cohort 1 
Cohort 2 
Cohort 3 
Subject Population 
Healthy adults 
Moderate to severe AD 
patients 
Moderate to severe AD 
patients 
In-patients/Out-patients 
In-patients 
In-patients 
Out-patients 
 Total Number of subjects 
12 
12 
30 
Ratio Active:Placebo 
2:1 
2:1 
2:1 
Dosing Duration 
7 days 
7 days 
14 days 
Follow-up Visit (after last 
dose of study medication) 
7-14 days  
7-14 days 
7-14 days 
 
Progression from Cohort 1 to Cohort 3 will be dependent on the demonstration of adequate 
safety within the protocol defined boundaries. In Cohorts 1 and 2, complete data sets from a 
minimum of 4 active and 2 placebo subjects with 40% BSA treated with study ointment will 
need to be reviewed by the Safety Review Team in order to decide whether to proceed as 
planned to the next cohort.  In the event that it is decided not to proceed to the next cohort, 
the reviewers may decide to modify the study within the boundaries of the adaptive features 
and continue, to submit a substantial protocol amendment or suspend the study. In addition, 
 
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in order to ensure subject safety, Cohorts 1 and 2 will be split to have an initial sentinel group 
which will precede the remaining subjects in the cohort.  The sentinel group will consist of 3 
subjects (2:1 Active to Placebo) and these subjects will be dosed twice daily, to cover 10% 
BSA with study ointments for 7 days.  Provided that the study ointments are well tolerated 
after 48 hours (i.e. 4 applications) the remainder of the cohort (9 subjects; 2:1 Active to 
Placebo) may start dosing with coverage of 40% BSA BID for 7 days.  All subjects in 
Cohorts 1 and 2 will remain in the CRU for the duration of the study.  Cohort 3 will have AD 
affected skin dosed to a maximum of 50% BSA.  A 50% BSA has been selected as the 
maximum BSA to be dosed to ensure that the total BSA of AD affected skin is not 
excessively different from the maximum area of treated skin in Cohorts 1 and 2 (40% BSA).  
It also accounts for the fact that AD patients total BSA will vary over time due to flare ups of 
the disease for a multitude of reasons which cannot be controlled within the study, including 
due to withdrawal from their normal medication prior to randomisation. 
 
All subjects will have a screening visit to confirm suitability to enter the study, within 28 
days of Day 1.  AD patients in Cohorts 2 and 3 will be required to switch from their usual 
emollient to using the study emollient twice daily for at least 7 days prior to Day 1 and 
throughout their participation in the study.  Dosing will commence on Day 1 and continue for 
7 or 14 days, depending on which cohort subjects are allocated to.  Subjects in Cohorts 1 and 
2 will remain in the unit until Day 9 (48 hours after the last application of study ointment).  
Subjects in Cohort 3 will have their first application of ointment in the unit and continue 
dosing at home, returning to the unit for assessments on Days 5, 8, 10 and 15 (after the last 
ointment application at home on the evening of Day 14). 
 
All subjects will be required to attend the CRU for a follow-up visit 7-14 days after the last 
application of study ointment.  AD patients may recommence use with other topical or 
systemic treatments for their AD, if required, from either discharge from the CRU (Cohort 2) 
or the end of dosing (Cohort 3). 
 
This protocol will be adaptive and designed to enable knowledge gained from the previous 
cohort to be applied to subsequent cohorts.  Changes made will be within the boundaries of 
the adaptive elements with clear control mechanisms and guidance for staying within these 
boundaries. 
 
A summary of the study design is shown in Figures 2-5.  
 
 
 
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Figure 2 - Overall Study Design 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Decision to Proceed ± Modify Design 
within boundaries of protocol 
Study Suspended 
Approval of Substantial 
Protocol Amendment required 
Cohort 2 AD patients (n=3) 10% BSA 
Approval of Substantial 
Protocol Amendment required 
Decision to Proceed ± Modify Design 
within boundaries of protocol 
Study Suspended 
Cohort 1 HV (n=9) 40% BSA 
Safety and PK Review Meeting 
Safety and PK Review Meeting 
Cohort 3 AD patients (n=30) 
(10-50% BSA) 
Cohort 1 HV (n=3) 10% BSA 
Decision to Proceed if no local 
toleration issues seen after 48 hours of 
dosing ± Modify Design within 
boundaries of protocol 
Decision to Proceed if no local 
toleration issues seen after 48 hours of 
dosing ± Modify Design within 
boundaries of protocol 
Cohort 2 AD patients (n=9) 40% BSA 
 
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Figure 3 - Study Design Cohort 1 (HV) 
 
 
Figure 4 - Study Design Cohort 2 (AD Patients) 
 
Pre-Screening 
Baseline (Day 1) 
Randomisation (2:1 Active:Placebo) 
ZPL-5212372 1.0% (w/w) ointment BID 
for 7 days (either 10% or 40% BSA 
dosed) 
Placebo ointment BID for 7 days (either 
10% or 40% BSA dosed) 
Follow Up (7-14 days post last dose) 
Screening (up to 28 days before Day 1) 
Pre-Screening (where needed to withdraw 
concomitant medications) 
Study Emollient Use (Day -7 to -1) 
Baseline (Day 1) 
Randomisation (2:1 Active:Placebo) 
ZPL-5212372 1.0% (w/w) ointment BID 
for 7 days (either 10% or 40% BSA 
dosed) 
Placebo ointment BID for 7 days (either 
10% or 40% BSA dosed) 
 
Follow Up (7-14 days post last dose) 
Screening (up to 28 days before Day 1) 
 
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Figure 5 - Study Design Cohort 3 (AD Patients) 
 
 
A study schedule is provided in Tables 9-11 schedule of activities. 
 
3.2 
Adaptive Categories and Features 
The adaptive categories and individual features of this study along with the limits 
(boundaries) which define the maximal risk and minimum safety requirements associated 
with the features are described in Tables 2 to 5 below. 
 
 
Pre-Screening (where needed to withdraw 
concomitant medications) 
Study Emollient Use (Day -7 to -1) 
Baseline (Day 1) 
Randomisation (2:1 Active:Placebo) 
ZPL-5212372 1.0% (w/w) ointment BID 
for 14 days (10-50% BSA dosed) 
Placebo ointment BID for 14 days  
(10-50% BSA dosed) 
Follow Up (7-14 days post last dose) 
Screening (up to 28 days before Day 1) 
 
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Table 2 - Investigational Medicinal Product/Dose 
Adaptive study design 
category 
Adaptive Features 
Boundaries 
Dosing Regimen 
1. Dosing regimen may be 
adapted in accordance with 
PK, safety, tolerability - local 
and systemic data (as 
applicable) collected up to 
the decision making 
time-point.  
2. Permissibility of dosing 
regimen adaption within 
and/or between cohorts 
The term dosing regimen includes 
(1) the frequency of dosing, (2) 
the percentage BSA covered and 
(3) the duration of dosing. 
1. Maximum starting percent BSA dosed 
will be 10% BSA for Cohorts 1 & 2. 
2. Maximum total BSA dosed will be 40% 
BSA for Cohorts 1 & 2. 
3. Minimum percent BSA dosed will be 
10% BSA for Cohort 3. 
4. Maximum total BSA dosed will be 50% 
BSA for Cohort 3. 
5. Minimum dosing frequency will be once 
daily dosing. 
6. Maximum dosing frequency will be 
twice daily dosing. 
7. Maximum duration of dosing will be 14 
consecutive days. 
Study Ointment 
Application 
1. The volume of ointment 
applied to each subject may 
vary e.g. based on the 
condition of their skin, body 
size and gender. 
2. Subjects will have the same 
volume of ointment applied 
throughout the duration of the 
study. Any changes to a 
subjects recommended 
volume of study ointment 
will be documented. 
1. The minimum amount of study ointment 
to adequately and evenly cover the 
dosing area should be used for each 
subject. 
2. The minimum volume of study ointment 
that can be applied for each application 
is approximately 0.25 mL per 2% BSA 
covered i.e. 1.25 mL ointment for 10% 
BSA coverage; 5 mL for 40% BSA 
coverage & 6.25 mL for 50% BSA 
coverage. 
3. The maximum volume of study ointment 
that can be applied for each application 
is approximately 1 mL per 2% BSA 
covered i.e. 5 mL ointment for 10% BSA 
coverage; 20 mL for 40% BSA coverage 
& 25 mL for 50% BSA coverage. 
4. For Cohorts 1 and 2 the minimum 
amount of study ointment should be used 
for the first application. If there is not 
enough ointment to cover the dosing area 
additional study ointment can be applied 
(up to the maximum 1 mL per 2% BSA). 
5. For Cohort 3 the minimum amount of 
study ointment should be used for the 
first application in the CRU. If there is 
not enough ointment to cover the dosing 
area additional study ointment can be 
applied (up to the maximum 1 mL per 
2% BSA). 
Study Emollient 
Application 
1. Cohorts 2 and 3 only. 
Adaptable use of different 
methods of application for 
study ointment and study 
emollient 
2. Cohort 3 only. Alternate 
emollients other than those 
defined as ‘study emollients’ 
1. Use of study ointment before or after 
study emollient administration 
2. Duration of time between study ointment 
and study emollient applications between 
5 and 30 minutes. 
3. Stop the use of study emollient on areas 
treated with study ointment. 
 
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may be used. 
4. Stop the use of study emollient in the 
event of any hypersensitivity reactions 
caused by emollient. 
5. Change the emollient used from ‘study 
emollient’due to any potential or actual 
hypersensitivity reactions with the 
approval of the Ziarco CMO (Cohort 3 
only). 
Table 3 - Timing/Scheduling 
Adaptive study design 
category 
Adaptive Features 
Boundaries 
Visits 
1. The number and timing of the 
study in-life visits may be 
adjusted for Cohort 3. 
Excluding any unscheduled 
visits for subject safety. 
1. Minimum of 3 visits (excluding 
unscheduled visits) 
2. Maximum of 5 visits (excluding 
unscheduled visits) 
3. Visit windows may vary by ± 1 day 
2. Cohort 3 maybe subjected to 
short in-patient stay at 
beginning of treatment 
1. Maximum of 3 days in-patient stay to 
mitigate any local toleration issues and 
ensure appropriate dosing by patients 
Table 4 - Study Participants 
Adaptive study design 
category 
Adaptive Features 
Boundaries 
Subject number 
1. The number of subjects in 
Cohorts 1 and 2 may be 
decreased as there is no 
formal sample sizing for 
Cohorts 1 and 2. 
1. Upon completion of a cohort the 
minimum safety data requirements for 
progression between cohorts is complete 
datasets for 4 active and 2 placebo 
subjects treated with 40% BSA coverage 
of ointments. 
2. Minimum and maximum evaluable 
subjects for Cohorts 1 and 2 will be 6 
and 12. 
3. Ratio of 2 active :1 placebo will be used 
throughout the study. 
Table 5 - Assessments 
Adaptive study design 
category 
Adaptive Features 
Boundaries 
Assessments 
1. Safety/tolerability samples 
and assessments (such as 
dermal scoring, safety 
laboratory, vital signs, ECG’s 
etc). 
1. No changes to the timing and frequency 
of non-safety assessments is allowed.  
2. Timing and number of safety 
assessments may change based on 
clinical need. 
2. PK samples: Timing and 
number of PK samples may 
change based on the 
emerging PK profile. 
1. There will not be an increase in the 
number of PK samples taken but fewer 
samples may be taken. 
2. Timing of samples may be changed 
dependent on the emerging plasma 
concentration data. 
 
 
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As the study progresses and safety, toleration and PK data become available the adaptive 
features of the study may need modifying to improve subject safety and reduce associated 
risks.  Any decisions to modify any adaptive features within the boundaries described in the 
protocol will be documented and approved (signed) by the CMO prior to any implementation 
of the modifications.  If the protocol requires changes which are outside the boundaries 
described in the protocol then a protocol amendment will be written. Any associated study 
documents e.g. informed consent document (ICD), will be updated and re-approved as 
appropriate. 
3.3 
Termination or Suspension of Study  
The study will be completed as planned unless one or more of the following criteria are 
satisfied that require temporary suspension or early termination of the study. 
• New information or other evaluation regarding the safety or efficacy of the study 
medication that indicates a change in the known risk/benefit profile for the compound, 
such that the risk/benefit is no longer acceptable for subjects participating in the 
study. 
• Significant violation of Good Clinical Practice (GCP) that compromises the ability to 
achieve the primary study objectives or compromises subject safety. 
 
3.4 
Termination or Suspension of Investigational Sites 
A study site may be terminated prematurely or suspended if the site (including the 
investigator) is found in significant violation of GCP, the protocol or the contractual 
agreement or is unable to ensure adequate performance of the study. 
 
In the event that the sponsor, the IEC or regulatory authority elects to terminate or suspend 
the study or the participation of an investigational site, a study-specific procedure for early 
termination or suspension will be provided by the sponsor; the procedure will be followed by 
applicable investigational sites during the course of termination or study suspension. 
 
3.5 
Progression From Sentinel Group To Main Cohort 
Cohorts 1 and 2 both contain an initial sentinel group (n=3) who will only have 10% BSA 
treated with study ointment BID for 7 days. After 48 hours (i.e. 4 consecutive applications) of 
study dosing, if no local toleration (Cohort 1 healthy volunteers) or no persistent dermal AE’s 
(Cohort 2 AD patients) are recorded the remaining n=9 participants in the cohort may start 
study treatment with 40% BSA being dosed BID for 7 days.  The investigator will review all 
the sentinel group’s blinded safety data and the investigator’s findings and recommendation 
will be communicated to Ziarco by email.  Based on the investigators findings Ziarco will, 
either confirm via email that the remainder of the cohort may commence dosing to 40% BSA 
or, if there is any concern from either the investigator or the Ziarco clinical team, a 
teleconference will be arranged and the data from the sentinel group will be discussed.  The 
outcome and decision of the meeting will be documented and the minutes will be signed by 
both the investigator and the Ziarco CMO. 
 
3.6 
Safety Review Meeting 
Blinded review of subject safety data (AEs, local toleration assessments, ECGs, laboratory 
safety tests and vital signs) and pharmacokinetic data will be performed by the Safety Review 
 
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Team in a formal safety review meeting at the end of Cohorts 1 and 2 to identify any 
potential safety signals and decide whether progression of the study to the next cohort is 
appropriate.  There will be sufficient time between cohorts to allow for a thorough review of 
data and decision-making. The decision will be documented. 
 
During the study, subject safety will be assessed through physical examination, vital signs, 
ECGs, local toleration assessments (Cohort 1 only), safety clinical laboratory tests and 
adverse event monitoring. 
 
Continuation to the next cohort will proceed unless study drug-related intolerance or AEs are 
identified through safety monitoring. Progression to Cohort 2 will not proceed if 5 or more 
Cohort 1 subjects, dosed over 40% BSA, have persistent (on at least 3 consecutive occasions) 
local toleration reactions of ≥ 5 total score in the modified Draize local toleration assessment. 
Progression to Cohort 3 will not proceed if 5 or more Cohort 2 AD patients, dosed over 40% 
BSA, report persistent (on at least 3 consecutive occasions) skin related AE’s potentially 
linked to study ointment application as opposed to changes in their normal pattern of AD 
disease.  This includes if a patients’ AD worsens beyond tolerability. 
 
The maximum human plasma exposure level has been set at 13550 ng.hr/mL which is the no 
observed effect level (NOEL) exposure in rats following subcutaneous systemic dosing 
(AUC0-t). Progression to the next cohort will not proceed if the mean area under the 
concentration-time curve (AUCτ) PK data in the previous cohort exceeds 13550 ng.hr/mL or 
if any of the safety stopping criteria are met. 
 
Further details on the operation of the Safety Review Meetings will be defined in a separate 
procedures document. 
 
3.7 
Stopping Criteria 
3.7.1 Cohort Stopping Criteria 
Formal safety review meetings will be held following each cohort to assess the safety, 
toleration and pharmacokinetic data collected and decide whether to proceed to the next 
cohort. 
 
• PK Stopping Criteria: Progression to the next cohort will not proceed if the mean 
AUCτ from existing cohort data exceeds 13550 ng.h/mL. 
• Skin Toleration Stopping Criteria: Progression to Cohort 2 will not proceed if 5 or 
more Cohort 1 subjects, dosed over 40% BSA, have persistent (on at least 3 
consecutive occasions) local toleration reactions of ≥ 5 total score in the Draize local 
toleration assessment. Progression to Cohort 3 will not proceed if 5 or more Cohort 2 
AD patients, dosed over 40% BSA, report persistent (on at least 3 consecutive 
occasions) skin related AE’s potentially linked to study ointment application as 
opposed to changes in their normal pattern of AD disease. This includes if a patients’ 
AD worsens beyond tolerability. 
• Safety Stopping Criteria: Dose escalation will not proceed if any of the following 
occur in a ZPL-5212372-treated subject: 
• Severe adverse reaction(s) indicating study drug-related intolerance. 
 
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• Clinically significant ECG abnormalities (such as arrhythmias) or effects on other 
vital signs indicating dose-related intolerance. 
• Clinically significant laboratory abnormalities (see Appendix 1 and 2) indicating 
dose-related intolerance. 
• Other findings that, at the joint discretion of the CMO and investigator, indicate 
dose escalation should be halted. 
If any of the above occurs, prompt medical treatment should be provided as appropriate and 
adverse events promptly reported to the Sponsor via telephone or email. 
 
After satisfactory review of the available pharmacokinetic, safety, and tolerability data 
progression to the next cohort will occur if the administration to the previous cohort was well 
tolerated.  Adaptive features may be discussed and modifications made based on the 
safety/PK or learnings from the cohort.  Any modifications within the boundaries specified in 
the protocol will not require a protocol amendment, but will be documented in the minutes of 
the safety review meeting. 
 
3.7.2 Individual Stopping Criteria 
• If a healthy volunteer subject has a persistent (on at least 3 consecutive occasions) 
grade 3 or greater score in erythema, oedema or desquamation, application of study 
ointment will be discontinued and only safety and PK evaluations will be performed 
for the remaining scheduled study visits until the AE is resolved. 
• If, in the opinion of the investigator, a patient’s AD worsens beyond tolerability the 
patient will have their study medication discontinued and will be followed up until 
the worsening of their AD has resolved and is under control. 
3.8 
Planned Sample Size and Number of Study Centers 
For Cohorts 1 and 2 it is envisaged that sufficient subjects will be screened to achieve 
approximately 12 randomised and evaluable subjects (i.e. sufficient subjects completing 
study treatment to meet the requirements of the data review before proceeding to the next 
cohort). Recruitment for Cohorts 1 and 2 will be from a single centre in the United Kingdom. 
For Cohort 3 it is envisaged that approximately 45 subjects will be screened to achieve 
approximately 30 randomised and evaluable subjects. Recruitment for Cohort 3 will be from 
across approximately 4 centers in the United Kingdom. See Section 8.9 for a discussion of 
sample size. 
 
4 
STUDY POPULATION 
Upon screening, each subject will receive a 3 digit subject number which will be used 
throughout the study.  Randomisation will occur on Day 1, after all screening procedures 
have been performed and eligibility for the study confirmed. 
 
Any subjects who terminate their study participation for any reason, regardless of whether 
study drug was applied or not, will retain their subject number. 
 
 
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Subjects will be randomised on a 2:1 basis to topical 1.0% (w/w) ZPL-5212372 ointment or 
matching placebo ointment applied twice daily. For the randomisation of subjects, the 
investigator will use an Integrated Web-based Response System (IWRS). The IWRS will use 
a computer-generated randomisation schedule to assign subjects to a treatment sequence. For 
Cohort 3 the randomisation will be stratified based on baseline EASI score (either EASI ≤20 
or EASI >20) to ensure an even distribution of EASI and associated BSA affected across both 
treatment groups. 
To fulfil the study objectives it is essential that appropriate subjects are enrolled.  The 
following eligibility criteria are designed to select subjects for whom protocol treatment is 
considered appropriate.  Subject eligibility should be reviewed and documented by an 
appropriate member of the site study team.  The study population will consist of a cohort of 
adult healthy volunteers and 2 cohorts of adult patients with moderate to severe AD. 
4.1 
Inclusion Criteria: Cohort 1 
Healthy volunteer subjects for Cohort 1 must meet all of the following inclusion criteria to be 
eligible for inclusion in the study 
1 
Healthy males or females, aged between 18 and 55 years, inclusive (healthy is defined as 
no clinically relevant abnormalities identified by a detailed medical history, full physical 
examination, including blood pressure and pulse rate measurement, 12-lead ECG and 
clinical laboratory tests). 
2 
Females should be either of non-child-bearing potential (as specified in Section 4.7) or 
must agree to use highly effective methods of contraception (as specified in Section 4.7). 
Males with partners who are WOCBP must also use contraception (as specified in 
Section 4.7) 
3 
Body weight of ≥ 50 kg. 
4 
Body Mass Index of ≤34.9 kg/m2. 
5 
Subjects must have a Fitzpatrick Skin Type26 of between I to III to be included in the 
study. 
6 
Willing and able to comply with scheduled visits, treatment plan, laboratory tests and 
other study procedures. 
7 
Evidence of a personally signed and dated informed consent form, indicating that the 
subject has been informed of all pertinent aspects of the study. 
4.2 
Inclusion Criteria: Cohorts 2 and 3 
Moderate to severe AD patients for Cohorts 2 and 3 must meet all of the following inclusion 
criteria to be eligible for inclusion in the study 
1 Males and females aged 18-65 years inclusive with physician documented history or 
diagnosis of atopic dermatitis for at least 6 months prior to screening. AD should be 
diagnosed by the Eichenfield revised criteria of Hanifin and Rajka.1 
2 Females should be either of non-child-bearing potential (as specified in Section 4.7) or 
must agree to use highly effective methods of contraception (as specified in Section 4.7). 
 
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Males with partners who are WOCBP must also use contraception (as specified in Section 
4.7) 
3 Body weight of ≥ 50 kg. 
4 Body Mass Index of ≤34.9 kg/m2. 
5 Eczema Area and Severity Index (EASI) of ≥9 and <48 at Screening and an EASI of ≥12 
and <48 at Day 1. 
6 An Investigator’s Global Assessment (IGA) score ≥ 3 at both Screening and Day 1. 
7 Atopic dermatitis affecting between ≥10 to <40% BSA at Screening and ≥10% to <50% 
BSA on Day 1. 
8 Patients must be willing to stop applying their daily emollients and instead use the study 
emollient twice daily from at least 7 days prior to randomisation and throughout their 
participation in the study. 
9 Willing and able to comply with scheduled visits, treatment plan, laboratory tests and 
other study procedures. 
10 Evidence of a personally signed and dated informed consent form, indicating that the 
subject has been informed of all pertinent aspects of the study. 
4.3 
Exclusion Criteria: Cohort 1 
Healthy volunteer subjects for Cohort 1 presenting with any of the following will not be 
included in the study: 
 
1 
Evidence or history of clinically significant haematological, renal, endocrine, pulmonary, 
gastrointestinal, cardiovascular, hepatic, psychiatric, neurologic, or allergic disease 
(including drug allergies, but excluding untreated, asymptomatic, seasonal allergies at 
time of dosing). 
2 
Have tattoos covering areas of skin to be dosed with study ointment. 
3 
Subjects who are hirsute in areas of skin to be dosed with study ointment. 
4 
Subjects who are unwilling to stop hair removal by any means (including shaving, 
waxing or depilatory creams) to skin areas to be dosed with study ointment for 2 weeks 
prior to randomisation and throughout the duration of the study. 
5 
Have concomitant skin disease or infection (e.g. acne, impetigo) or presence of skin 
comorbidities in the study area to be dosed that may interfere with study assessments. 
6 
Hypersensitivity to any excipients in the study ointment formulation, study emollient 
 or study shower cream 
 
7 
Hypersensitivity to any laundry detergents. 
 
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8 
Use of a tanning booth/parlour/sunbathing (including excessive exposure to sunlight) or 
use of tanning products within 4 weeks before start of the study and for the duration of 
their participation in the study. 
9 
History of sensitivity to nonsteroidal anti-inflammatory drug(s) (NSAIDs).  
 
10 Clinically significant cardiac disease or ECG abnormalities, including: 
• Predominant heart rhythm other than normal sinus rhythm 
• AV block greater than first degree 
• Resting pulse rate >100 or <50 bpm 
• 
Electrocardiographic evidence or a history of myocardial infarction 
• Electrocardiographic evidence of conduction system abnormality 
• Subjects with pre-randomisation evidence of QTc prolongation (defined as >450 
msec). 
The PI should decide whether ECG abnormalities other than those listed are clinically 
significant and should exclude the subject from enrolment. 
11 Subjects with a personal or family history of prolonged QTc interval. 
12 Clinically significant liver function tests (LFTs; AST, ALT, total Bilirubin) >1.5 X upper 
limit of normal (ULN).  Subjects with LFTs >1.5 X ULN and <2 X ULN may be retested 
once. 
 
13 Positive HIV, HBsAg and HBcAb result or positive anti-hepatitis C virus serology (as 
determined by multi-antigen EIA) 
14 Pregnant and nursing females. 
15 Donation or intent to donate blood, or blood components during the study or within one 
month after completion of the study.  
16 Subjects who have received treatment with an investigational drug within 3 months prior 
to screening. 
17 Subjects with a known history (of at least 2 years) of drug abuse. 
18 A positive urine drug screen. 
19 Subjects who are smokers (including subjects who use nicotine substitutes). 
20 History of regular alcohol consumption exceeding 21 drinks/week in males and 14 
drinks/week in females (1 drink = 150 mL of wine or 360 mL of beer or 45 mL of spirits) 
within 6 months of screening. 
21 Subjects using potent CYP3A4 inhibitors (i.e, cimetidine, clarithromycin, cyclosporine, 
danazol, dexamethasone, diethyldithiocarbamate, erythromycin, fluconazole, fluoxetine, 
grapefruit juice, ketoconazole, metronidazole, norfloxacin, omeprazole, ranitidine, 
sertraline, valproic acid) 
 
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22 Inability to comprehend or unwilling to follow the study requirements including 
restrictions on treatments, attendance at out-subject clinic visits and participation in 
laboratory testing as called for by the protocol. 
23 Any other severe acute or chronic medical or psychiatric condition or laboratory 
abnormality that may increase the risk associated with study participation or 
investigational product administration or may interfere with the interpretation of study 
results and, in the judgment of the investigator in discussion with the Ziarco CMO, 
would make the subject inappropriate for entry into this study. 
4.4 
Exclusion Criteria: Cohorts 2 and 3 
Moderate to severe AD patients presenting with any of the following will not be included in 
the study: 
 
1 AD of such severity (EASI >48) that the subject could not comply with the demands of 
the study and/or the subject is not a suitable candidate for a placebo-controlled study. 
2 Have concomitant skin disease or infection (e.g. acne, impetigo) or presence of skin 
comorbidities in the study area to be dosed that may interfere with study assessments. 
3 Patients who are hirsute in areas of skin to be dosed with study ointment. 
4 Patients who are unwilling to stop hair removal by any means (including shaving, waxing 
or depilatory creams) to skin areas to be dosed with study ointment for 2 weeks prior to 
randomisation and throughout the duration of the study. 
5 Hypersensitivity to any excipients in the study ointment formulation, study emollient or 
study shower cream  
6 Hypersensitivity to any laundry detergents (Cohort 2 only). 
7 Have received phototherapy (e.g. UVA, UVB or PUVA therapy), or systemic therapy 
(e.g. immunosuppressants [such as cyclosporine, azathioprine, methotrexate], cytostatics) 
known or suspected to have an effect on AD, within 4 weeks of the start of the study.  All 
other biologics should not have been used within 3 months of the start of study. 
8 Have received systemic corticosteroids (e.g. oral, intravenous, intraarticular, rectal) 
within 4 weeks of the start of the study. Subjects on a stable maintenance dose (over the 
preceding 3 months) of inhaled or intranasal CS may participate. 
9 Patients treated with oral antihistamines or topical calcineurin inhibitors or topical 
steroids within 7 days of starting study; intranasal antihistamines for the treatment of 
allergic rhinitis are acceptable. 
10 Use of a tanning booth/parlour/sunbathing (including excessive exposure to sunlight) or 
use of tanning products within 4 weeks before start of the study and for the duration of 
their participation in the study. 
11 Have used antiseptic treatments (e.g. bleach baths, potassium permanganate etc.) within 4 
weeks before the start of the study.  (Patients who have recently used antiseptic treatment 
 
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may be rescreened at a later date if they wish to participate in the study and agree to stop 
antiseptic treatment.) 
12 History of sensitivity to nonsteroidal anti-inflammatory drug(s) (NSAIDs). 
13 Patients who have evidence of significant concomitant clinical disease that could interfere 
with the conduct or safety of this study, based upon a complete medical history, full 
physical examination, and a 12-lead resting ECG and laboratory safety tests.  For 
example the presence of any other acute and/or unstable clinical disease, other than AD, 
such as: 
• Poorly controlled Type I or Type II diabetes. 
• Seizure disorder or epilepsy. 
• Cerebrovascular accident. 
• Active or latent infection (eg, hepatitis). 
• Cancer (other than cutaneous basal cell) in the last 5 years. 
• Congestive heart failure. 
• Coronary artery disease. 
• Renal insufficiency and/or serum creatinine >1.5 X upper limit of normal (ULN). 
Subjects with serum Creatinine <2 X ULN may be retested once. 
• A major surgical operation during the 30 days prior to screening. 
 
Stable well-controlled chronic conditions such as controlled hypertension [excluding 
those on ß-blockers, calcium channel blockers (Class I or II)], thyroid disease, well-
controlled Type 1 or Type 2 diabetes, hypercholesterolemia, gastroesophageal reflux, or 
depression under control with medication (other than tricyclic antidepressants), are 
acceptable provided the symptoms and medications would not be predicted to 
compromise safety or interfere with the tests and interpretations of this study. 
14 Clinically significant cardiac disease or ECG abnormalities, including: 
• Predominant heart rhythm other than normal sinus rhythm 
• AV block greater than first degree 
• Resting pulse rate >100 or <50 bpm 
• 
Electrocardiographic evidence or a history of myocardial infarction 
• Electrocardiographic evidence of conduction system abnormality 
• Subjects with pre-randomisation evidence of QTc prolongation (defined as >450 
msec). 
The PI should decide whether ECG abnormalities other than those listed are clinically 
significant and should exclude the subject from enrolment. 
15 Patients with a personal or family history of prolonged QTc interval. 
16 Clinically significant liver function tests (LFTs; AST, ALT, total Bilirubin) Subjects with 
LFTs >1.5 X ULN and <2 X ULN may be retested once. 
 
17 Positive HIV, HBsAg and HBcAb result or positive anti-hepatitis C virus serology (as 
determined by multi-antigen EIA) 
18 Pregnant and nursing females. 
 
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19 Donation or intent to donate blood, or blood components during the study or within one 
month after completion of the study.  
20 Patients who have received treatment with an investigational drug within 3 months prior 
to screening. 
21 Patients with a known history (of at least 2 years) of drug abuse. 
22 A positive urine drug screen. 
23 Patients who are smokers (including patients who use nicotine substitutes). (Cohort 2 
only) 
24 History of regular alcohol consumption exceeding 21 drinks/week in males and 14 
drinks/week in females (1 drink = 150 mL of wine or 360 mL of beer or 45 mL of spirits) 
within 6 months of screening. 
25 Patients using potent CYP3A4 inhibitors (i.e, cimetidine, clarithromycin, cyclosporine, 
danazol, dexamethasone, diethyldithiocarbamate, erythromycin, fluconazole, fluoxetine, 
grapefruit juice, ketoconazole, metronidazole, norfloxacin, omeprazole, ranitidine, 
sertraline, valproic acid 
26 26 Inability to comprehend or unwilling to follow the study requirements including 
restrictions on treatments use of study emollient, attendance at out-subject clinic visits 
and participation in laboratory testing as called for by the protocol. 
27 Any other severe acute or chronic medical or psychiatric condition or laboratory 
abnormality that may increase the risk associated with study participation or 
investigational product administration or may interfere with the interpretation of study 
results and, in the judgment of the investigator in discussion with the Ziarco CMO, would 
make the subject inappropriate for entry into this study. 
4.5 
Life Style Guidelines: Cohorts 1 and 2 
The following life-style guidelines affect all subjects in Cohorts 1 and 2: 
 
Meals and Dietary Restrictions 
Water may be consumed without restriction.  Non-caffeinated drinks (except grapefruit 
or grapefruit-related citrus fruit juices - see below) may be consumed with meals and the 
evening snack. 
Subjects will be fasted from midnight. 
Breakfast will be provided approximately 1 hour before the morning application except 
on Days 1 and Day 7 when no food will be provided until after the 4 hours post dose 
assessments have been completed. 
Lunch will be provided approximately 4 hours after the morning application. 
Dinner will be provided approximately 9 to 10 hours after the morning application. 
An evening snack will be permitted. 
 
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Subjects will not be allowed to eat or drink grapefruit or grapefruit-related citrus fruits 
(eg, Seville oranges, pomelos) from 7 days prior to the first dose of study medication 
until collection of the final pharmacokinetic blood sample. 
While in the CRU, the total daily nutritional composition should be approximately 50% 
carbohydrate, 35% fat and 15% protein.  The daily caloric intake per subject should not 
exceed approximately 3200 kcal. 
 
Alcohol and Caffeine  
Subjects will abstain from alcohol for 24 hours prior to admission to the CRU and 
continue abstaining from alcohol until collection of the final pharmacokinetic sample.  
Subjects may undergo an alcohol breath test at the discretion of the investigator. 
Subjects will abstain from caffeine-containing products for 24 hours prior to the start of 
dosing until collection of the final pharmacokinetic sample. 
 
Activity 
Subjects will abstain from unaccustomed strenuous exercise (e.g., heavy lifting, weight 
training, calisthenics, aerobics) for at least 48 hours prior to each blood collection for 
clinical laboratory tests.  Regular routine exercise and walking at a normal pace will be 
permitted. 
Subjects should have a normal day time awake, night time asleep cycle throughout the 
study. 
 
Showering 
Subjects will briefly shower (approximately 5 minutes) once in the morning at least 30 
minutes prior to the first application of study ointment using 
 
 
 
Clothing 
Subjects will be provided with cotton T-shirts and trousers to wear during the study. 
 
4.6 
Life Style Guidelines: Cohort 3 
Patients in Cohort 3 will be recommended to use old clothes or test the ointment on a 
small, unobtrusive area of clothing as this is a developmental formulation which has not 
been tested for compatibility with different types of fabric. 
4.7 
Contraceptive Requirements 
Contraceptive requirement for the study have been defined using the recent recommendations 
of the Clinical Trials Facilitation Group, Rome September 2014 meeting and also the EMA 
M3(R2) guidance.25 
 
Females 
 
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Females of child-bearing potential must agree to use highly effective methods of 
contraception from screening to 60 days after the last study dose.  Highly effective methods 
are: 
• combined (estrogen and progestogen containing) hormonal contraception associated 
with inhibition of ovulation administered by oral or intravaginal route 
• oral, injectable or implantable progestogen-only hormonal contraception associated 
with inhibition of ovulation 
• intrauterine device (IUD) 
• intrauterine hormone-releasing system ( IUS) 
• bilateral tubal occlusion 
• vasectomised partner 
• true sexual abstinence, where this is in line with the usual lifestyle of the subject; 
periodic (calendar, ovulation, symptothermal or post-ovulation) abstinence and 
withdrawal are not acceptable methods of contraception. 
 
Females are considered to be of non-child-bearing potential if they fulfill at least one of the 
following criteria: 
• Postmenopausal, defined as 45 to 65 years of age and amenorrheic for at least 2 years 
PLUS have a serum follicle stimulating hormone (FSH) level within the laboratory’s 
reference range for postmenopausal women, at screening. 
• have undergone a documented hysterectomy and/or bilateral oophorectomy. 
 
Males with partners who are WOCBP 
Males with partners who are WOCBP must agree to use contraception from the start of 
dosing study ointment until 120 days after the last dose of study ointment. 
 
4.8 
Subject Withdrawal and Replacement 
Subjects will be discontinued from the investigational product and potentially from the study 
in the following circumstances: 
1 
Enrollment in other clinical studies involving investigational products or enrollment in 
other types of clinical research judged not to be scientifically or medically compatible 
with this study. 
2 
The subject requests to be discontinued from the study. 
3 
Use of prohibited concomitant medications. 
4 
Sponsor decision to stop the study or to stop the subject’s participation in the study for 
medical, safety, pharmacokinetic or regulatory, or other reasons consistent with 
applicable laws, regulations, and good clinical practices (GCP). 
5 
A severe or serious adverse event occurs that the investigator considers is related to 
study drug. 
 
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6 
Any criteria for laboratory safety or vital sign values of Potential Clinical Concern 
(Appendix 1). 
7 
Any criteria for Hepatic Safety (Appendix 2). 
8 
Any criteria for ECG Values of Potential Clinical Concern (Appendix 3). 
Subjects may have their study drug discontinued if they have abnormal laboratory results 
which are considered to be clinically significant as assessed by the investigator.  This 
decision should be made in the best interest of the subject and follow up should be conducted 
using standard clinical practices.  Subjects may also be temporarily discontinued from study 
drug while AEs and/or laboratory test abnormalities are being investigated and/or at the 
request of the sponsor’s CMO.  The investigator should discuss with Ziarco’s CMO any 
subject for whom temporary or permanent discontinuation of study drug is being proposed. 
Subjects who discontinue early from the study should be asked to complete all the efficacy 
and safety assessments scheduled for the end of study assessment. 
Subjects may withdraw from the study at any time without penalty and for any reason 
without prejudice to their future medical care.  In all cases, the reason(s) for withdrawal, 
including the primary reason, must be recorded on the case report form (CRF). If a subject is 
prematurely withdrawn from the study for any reason, the investigator must make every 
effort to perform the evaluations described for the early termination/follow up visit (in clinic) 
and a further follow up contact (either by telephone or in clinic) 7 to 14 days later.  Subjects 
who withdraw due to lack of efficacy (Cohort 3) or adverse events which may be attributed to 
double blind treatment will not be replaced.  
Cohort 3 patients who withdraw due to personal reasons, poor compliance, other protocol 
deviations or are lost to follow up may be replaced until at least 30 evaluable patients have 
completed the study. 
4.9 
Temporary Discontinuations 
A subject who misses one or more doses due to personal circumstances, or has an AE (not 
related to study drug), may be allowed to restart dosing after discussion between the 
investigator and CMO. 
 
5 
STUDY TREATMENT 
5.1 
Allocation to Treatment 
All subjects will be allocated a unique 3-digit subject number, by the IWRS, that will be 
recorded in the CRF and on all subject related study documentation.  The IWRS will use a 
computer generated randomisation schedule to assign subjects to a treatment sequence on a 
2:1 basis and will stratify subjects in Cohort 3 based on the EASI score to ensure an even 
distribution of severities across the treatment groups. 
5.2 
Breaking the Blind 
This study is double-blind third party open i.e. the study will be subject- and investigator-
blinded.  Details of personnel who will be able to review unblinded subjects’ data during 
study will be held in a separate document by Ziarco. 
 
 
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At the initiation of the study, the study site will be instructed on the method for breaking the 
blind.  The study blind should not be broken except in a medical emergency, where 
knowledge of the study drug received would affect the treatment of the emergency or 
obtaining this information is a regulatory requirement. If time permits, the investigator should 
notify the Sponsor prior to contacting IWRS. All calls resulting in an unblinding event will be 
reported by the IWRS to the Sponsor. 
 
If the blind is broken, the date, time and reason must be recorded in the subject’s CRF 
Blood specimens will be obtained from all subjects for pharmacokinetic analysis to maintain 
the study blind at the investigator site.  The study monitor will remain blinded to treatment 
until the conclusion of the study.  To minimize the potential for bias, treatment randomisation 
information will be kept confidential by Ziarco personnel and will not be released to the 
investigator, investigator site personnel, or the study monitor until the conclusion of the 
study. 
 
ZPL-5212372 will be provided as single dose strength of a 1.0% (w/w) concentration of 
ointment.  Placebo will be provided as visually matched ointment.  All study drugs will be 
supplied in identical pots and will be similar in colour and appearance, thereby maintaining 
double-blind conditions for the subject and investigator. 
5.3 
Drug Supplies 
 
 to all Cohort 1 and 2 subjects, for use when 
showering each morning whilst resident in the CRU. 
 
will be supplied as study emollient to all AD patients (Cohorts 2 and 3 only) for use prior to 
randomisation and throughout the study by MAC. 
 
will be supplied as an alternate study emollient to any AD patients (Cohort 3 only) who have 
a potential (or known) hypersensitivity reaction to 
 for use prior to randomisation 
and throughout the study by MAC. 
 
and supplied with a certificate of analysis. 
Matching placebo ointment will be manufactured by 
 
and supplied with a certificate of analysis. 
ZPL-5212372 and placebo will be administered as a topical ointment.  Both ointments 
contain the same excipients; placebo ointment has been manufactured in the same way except 
for the addition of 1.0% (w/w) ZPL-5212372.  
5.4 
Treatment Packaging and Labelling 
Ointment will be packaged in
 
 and will be labelled and released in accordance with 
the Clinical Trials Directive 2001/20/EC and GMP Directive 2003/94/EC for Investigational 
Medicinal Products. Each jar will contain 100 g of ointment.  Supplies will be identified by 
 
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the batch number, expiry date and a unique 5-digit jar code which will be used by the IWRS 
to identify the content and allocate jars to study subjects. 
 
5.5 
Dispensing 
 
 
 
 
For Cohort 3 study medication will be supplied to the subject as:  
• Randomisation Visit (Day 1): 4 glass jars containing either 1.0% (w/w) ZPL-5212372 
ointment or matching placebo ointment, plus an additional glass jar containing 1.0% 
(w/w) ZPL-5212372 ointment or matching placebo ointment, to be used as spare 
medication, in the event of late visits or lost/damaged container, to ensure continuous 
dosing. 
• On Visit 3 (Day 8): empty or partially used glass jars containing either 1.0% (w/w) 
ZPL-5212372 ointment or matching placebo ointment will be replaced as required to 
enable dosing till the end of the study based on the patients usage. If required the 
spare ointment supply will also be replaced as necessary. 
Cohort 3 patients will be supplied with an adequate quantity (including some spares) of 
 and sealable plastic bags to store and return used 
 for disposal at the CRU.  In addition, patients will be supplied with an adequate 
quantity of disposable gloves to allow carers to apply the study ointment onto hard to reach 
areas of their body as required.   
 
Cohort 3 patients will be instructed by the unit staff to only open and use one jar of study 
ointment at a time. 
5.6 
Drug Storage 
Study medication should be stored at 15 to 25°C, in a securely locked area, accessible to 
authorised personnel only in the CRU Pharmacy.  For Cohort 3 patients study ointment 
should be stored at 15 to 25°C (i.e. room temperature) and kept out of the reach of children.  
Ointment remaining more than 14 days after opening should not be used. 
5.7 
Administration 
Subjects in Cohorts 1 and 2 will have twice daily applications of either 1.0% (w/w) 
ZPL-5212372 ointment or matching placebo ointment approximately 12 hours apart.  The 
morning dose will be administered between 07:00 to 10:00 hours and the evening dose 
approximately 12 hours later between 19:00 – 22:00 hours. Study ointment will be dispensed 
from the pharmacy (one jar per subject) to clinical staff.  Clinical staff will consult the 
subjects ‘body map’ to confirm whether 10 or 40% BSA will have application of ointment.  
For 10% BSA approximately 
 
 and for 40% BSA approximately 
 
 by clinic staff at the subject’s bedside.  If the initial volume 
of ointment is inadequate to cover the dosing area, additional ointment maybe applied up to a 
maximum of 1 mL/2% BSA (i.e. 5 mL for 10% BSA or 20 mL for 40% BSA). 
 
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Study Ointment Administration 
• Healthy volunteer subjects in Cohort 1 in the 10% BSA sentinel group will have study 
ointment applied by clinic staff directly onto their skin 
 onto the top 
half of their back (see Appendix 4 for example body map).  Subjects in the 40% BSA 
group will have study ointment applied by clinic staff directly onto their skin 
 
 onto their back and the fronts and backs of their upper legs (see Appendix 5 
for example body map).  Clinical staff will gently spread the study ointment over the 
entire dosing area with a gloved hand to ensure a thin, even coverage of ointment.  No 
study emollient will be applied to Cohort 1 healthy volunteers to ensure local 
toleration of the study ointment can be fully assessed. 
 
• AD patients in Cohort 2 in the 10% BSA sentinel group will have study ointment 
applied by clinic staff directly onto 10% of their AD affected skin 
  In 
some patients this will be all their AD affected skin and in others it may only be a 
portion of their AD affected skin (see Appendix 6 for example body map). Patients in 
the 40% BSA group will have study ointment applied by clinic staff directly onto all 
their AD affected skin.  In some patients this will be all their AD affected skin and in 
others it maybe their total AD affected skin does not cover 40% BSA hence additional 
non-AD lesion skin will also need to be covered (see Appendix 7 for example body 
map).  Individual patient body maps will need to be followed on each dosing 
occasion.  Clinical staff will gently spread the study ointment over the entire dosing 
area with a gloved hand to ensure a thin, even coverage of ointments.  After 15 
minutes the study emollient will be applied by the patient to the same areas treated 
with the study ointment and any additional body areas the patient routinely applies 
emollient to.  Patients will need to wash their hands immediately after application of 
study emollient in case of contamination with study ointment. 
 
• AD patients in Cohort 3 will apply twice daily (approximately 12 hours apart) 
applications of either 1.0% (w/w) ZPL-5212372 ointment or matching placebo 
ointment at home.  At the patients randomisation visit the extent of their AD will be 
assessed and a recommended amount (approximately 1.25 to 5 mL/10% BSA) of 
ointment for use on their AD affected skin will be recorded on their individual body 
map (see Appendix 8 for example body map).  Patients will be instructed not to use 
ointment on non-AD affected skin. Study ointment will be dispensed from the 
pharmacy (three jars per subject) to clinical staff.  The first application of study 
ointment will be applied by the patient in the clinic under the supervision and 
direction of the clinical staff i.e. the amount to apply, body area to cover using the 
body map as a guide and how to 
 
 will be explained, as this will vary from patient to patient.  Study emollient 
will be applied at least 15 minutes AFTER application of study ointment to all study 
ointment treated areas and any additional body areas the patient usually applies 
emollient to.  Thereafter patients will be advised to dispense and apply the study 
ointment and emollient in the same manner for the remainder of the study at home.  
Patients will be supplied with a copy of their individual body map and dosing 
instructions.  Patients will record their study ointment and emollient applications on a 
paper dosing diary which they will bring to the clinic on visits for compliance 
assessments by the clinic staff (this information will not be databased). On study visit 
days, patients should NOT apply their morning application of study ointment and 
 
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emollient.  Instead they should bring their remaining study ointment and emollient to 
the clinic and the study ointment and emollient will be applied in the clinic after the 
visits study assessments have been performed under the supervision of clinic staff. 
 
In all cases, once recorded the areas of skin to be treated with study ointment will be treated 
consistently for the duration of the study regardless of any changes in the size or extent of 
AD affected skin areas during the study period. 
 
Regardless of the location of an AD patient’s lesioned skin no study ointment will be 
applied to areas on the face or scalp above the jaw line. 
 
Emollients 
The emollient 
 will be supplied for this study for AD patients in Cohorts 2 and 3.  
In addition an alternate emollient will be available for Cohort 3 patients who have a potential 
for any hypersensitivity skin reactions to 
  Any emollients and bathing products 
used by the patient should be documented at screening. Patient emollients may be used up 
until 7 days prior to Day 1 and then patients must stop using their current emollient and 
switch to using the study emollient in the same manner as their previous emollient.  From 
randomisation patients must use study emollient at least 15 minutes after application of study 
ointment.  Patients should use study emollient on all skin areas which have been treated with 
study ointment and in addition any areas which the patient would treat as part of their normal 
skin care routine.  Cohort 2 patients must continue using study emollient twice daily until 
discharge from the CRU on Day 9 and Cohort 3 patients must use study emollient twice daily 
until they have completed their visit on Day 15. 
 
Shower Cream 
 
subjects, for use when showering each morning whilst resident in the CRU. 
5.8 
Excessive Pharmacological Effects 
No mechanistically specific antidotes for ZPL-5212372 are available and standard supportive 
measures should be used in the case of excessive pharmacological effects. 
5.9 
Drug Accountability 
The investigator is responsible for maintaining accurate study drug accountability records 
throughout the study. 
 
The IWRS will be used to allocate jars of study drug to all study subjects.  In addition, details 
of jars dispensed to patients and applications of study medication will be documented in 
accountability forms, the CRF and the patient diaries. 
 
Cohort 1 and 2 (in-patients): Jars of study ointment allocated by the IWRS will be 
documented on the accountability log.  Jars will be weighed and the weight recorded. Clinical 
staff, and the checker, will need to confirm and document in the CRF that both the correct 
volume (mL) of ointment and the correct jar of ointment is used on each dosing occasion. 
Empty jars will be weighed and the weight recorded and at the end of dosing any partially 
used jars will be weighed and the weight recorded.  At the end of the subject’s participation 
 
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in the study the approximate total amount of study ointment used by the subject over 7 days 
will then be calculated. Each dose administered will also be documented in the CRF. 
 
Cohort 3 (out-patients): Jars of study ointment allocated by the IWRS and will be weighed, at 
the time of dispensing, and the weight recorded on the accountability log.  The initial dose 
applied in the clinic will be taken from one of the patient’s allocated jars and the application 
documented in the CRF. Clinical staff, and the checker, will need to confirm and document in 
the CRF that both the correct volume (mL) of ointment and the correct jar of ointment is used 
on the initial dosing occasion. The patient must bring all study medication (including empty 
containers) to the clinic at each visit, so that compliance can be confirmed. At the end of the 
study all jars allocated to the patient will be weighed and the weight recorded on the drug 
accountability log and used to calculate the approximate total amount of study ointment used 
by the patient over the 14 days treatment period. 
 
Any supplies, including placebo remaining at the end of study will be returned to the Sponsor 
or their representative or destroyed locally on behalf of the Sponsor and the destruction fully 
documented. 
 
5.10 Compliance (Cohort 3 only) 
At each clinic visit patient compliance with the study ointment regimen will be monitored by 
recording any empty containers and weighing any partially used containers and by reviewing 
the patients dosing diary.  Patients failing to take 2 or more doses in a 7 day period will be 
reminded of the importance of complying with the dosing requirements.  If a patient 
consistently fails to take their medication, the investigator should ask the patient about 
compliance and following this discussion should alert the Ziarco CMO and study monitor and 
a decision will be made as to whether the subject should be withdrawn for non-compliance. 
 
5.11 Concomitant Medications 
Any medication the subject takes other than the study drug is considered a concomitant 
medication. All concomitant medications must be recorded in the CRF. The following 
information must be recorded in the CRF for each concomitant medication: generic name, 
route of administration, start date, stop date and indication.  
 
At screening, subjects will be asked what concomitant medications they are currently taking.  
At each subsequent study visit, subjects will be asked what medications they have taken since 
the last clinic visit. 
 
Any medications taken prior to the start of dosing will be documented as prior medications.  
Any medications taken after the first dose of study medication will be documented as 
concomitant medications. 
The type of emollient or bathing products used by the AD patients (Cohorts 2 and 3) should 
be captured during screening. For AD patients study emollient will be used from 7 days prior 
to dosing and throughout the remainder of the study until either discharge from the CRU on 
Day 9 (Cohort 2) or following their visit on Day 15 (Cohort 3). 
 
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AD patients may not use the medications and treatments listed in Table 6 within the indicated 
time interval prior to the start of dosing and these medications must be withheld for the 
duration of the study, and where possible until completion of the follow up visit.  However 
AD patients may commence use with other topical or systemic treatments for their AD if 
required from either discharge from the CRU (Cohort 2) or the end of dosing (Cohort 3). 
 
 
 
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Table 6 - Prohibited Medications for Cohort 2 & 3 AD patients 
Medication 
Time interval prior to Day 1 
Biologic treatments for AD 
3 months 
UVA, UVB or PUVA therapy 
4 weeks 
Oral calcineurin inhibitors and immunosuppressants 
(e.g. cyclosporine, azathioprine, methotrexate ) 
4 weeks 
Systemic corticosteroids (all potencies) 
4 weeks 
Oral antihistamines and leukotriene inhibitors and 
tricyclic antidepressants 
1 week 
Topical steroids (any potency) 
1 week 
Topical calcineurin inhibitors (tacrolimus, 
pimecrolimus) 
1 week 
Cytostatics known or suspected to have an effect on 
AD 
4 weeks 
Allergen Immunotherapy 
4 weeks 
Investigational drug 
3 months from start of 
screening 
Potent CYP3A4 inhibitors (e.g. cimetidine, 
clarithromycin, cyclosporine, danazol, dexamethasone, 
diethyldithiocarbamate, erythromycin, fluconazole, 
fluoxetine, grapefruit juice, ketoconazole, 
metronidazole, norfloxacin, omeprazole, ranitidine, 
sertraline, valproic acid) 
1 week 
Oral antibiotics are allowed for up to 2 weeks for disease-associated superficial skin 
infections, but subjects should have completed any course prior to the start of Day 1.  Note 
that active skin infection is an exclusion criterion. 
 
The following concomitant procedures are prohibited during study participation:  
 
• Elective major surgical procedures  
• Use of artificial tanning treatments or products 
 
 
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6 
STUDY CONDUCT 
This is a multicenter study in the United Kingdom, Cohorts 1 and 2 will be conducted at a 
single site and Cohort 3 will be conducted at up to 4 study sites. 
Schedules of assessments are presented in Section 6.7 Tables 9 to 11.  Details of study 
assessments are provided in Section 7. 
Cohort 3 visits should occur on the scheduled day ± 1 day, except screening and follow-up, 
where some flexibility is allowed. All times should be recorded using the 24-hour clock (e.g., 
23:20 not 11:20 pm). 
6.1 
Screening 
All subjects will be given both a written and verbal description of the study. Subjects should 
be given adequate time to think about whether they want to participate and to ask questions. 
The investigator (or an appropriate delegate at the investigator site) will obtain written 
consent from each subject prior to commencing any study related activities or assessments. 
Following consent, all subjects will undergo screening in the 28 days prior to the start of the 
study (Day 1), to ensure that they meet the selection criteria for the study. 
At screening all subjects will be allocated a unique 3-digit subject number by the IWRS, 
which will be recorded in the CRF and used for the duration of the study. 
The following procedures will be performed: 
• Subject demography 
• Complete medical history (including medication [prescription and non-prescription 
drugs/treatments, topical products, vitamins and dietary supplements taken in the last 
4 weeks], alcohol and tobacco use) 
• Full physical examination (including height and weight) 
• Fitzpatrick skin type assessment (for Cohort 1 only must be I to III for a subject to be 
eligible) 
• Duplicate vital signs (supine blood pressure and pulse rate) 
• Single 12-lead ECG for Cohorts 1 and 2. Triplicate 12-lead ECG for Cohort 3. 
• Breath alcohol testing 
• Blood and urine samples for: 
• 
Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
• 
Blood for Hepatitis B/C  
• 
HIV serology 
• 
Serum for Hb1Ac 
• 
Serum FSH test (post-menopausal females only) 
• 
Serum pregnancy test (pre-menopausal women only) 
• 
Urine drug screen 
• EASI, IGA and BSA assessment for Cohorts 2 and 3 only (AD patients) 
Provided all inclusion and exclusion criteria are met the subject may start the wash out period 
(if applicable). 
 
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To prepare for study participation, subjects will be reminded of the study restrictions and 
prohibited concomitant medications.  AD patients will be provided with study emollient and 
reminded to start using study emollient twice daily at least 7 days prior to Day 1 and 
throughout the study. 
6.2 
Cohort 1 (Healthy Volunteers) 
Cohort 1 will be split with a lead (n=3) sentinel group which will only have 10% BSA treated 
with study ointments.  If after 48 hours of treatment (4 applications) the study ointments are 
well tolerated then the remaining subjects of the cohort (n=9) may start treatment with 40% 
BSA being treated with study ointments.  Apart from this difference in percentage BSA 
treated all subjects in Cohort 1 will have the following assessments conducted in the 
designated timeframe. 
Subjects will be admitted to the Unit on the morning of Day 1 or the evening before (if 
required) and will remain in the Unit until Day 9 following collection of the 48 hours post 
dose blood sample. 
6.2.1 Day 1 assessments 
On Day 1 the following assessments will be carried out prior to randomisation and 
dosing: 
• Review of concomitant medications, including emollient use (may be conducted the 
evening before) 
• Review changes in the subject’s medical history since screening (may be conducted 
the evening before) 
• Brief physical examination (may be conducted the evening before) 
• Completion of body map of skin areas to record the skin area to be treated with study 
ointments (may be conducted the evening before) 
• Breath alcohol testing (may be conducted the evening before) 
• Triplicate 12-lead ECG 
• Duplicate vital signs (supine blood pressure and pulse rate) 
• Assessment of skin area to be treated for any abnormalities e.g. erythema 
• Review of baseline adverse events 
• Obtain blood and urine samples: 
• 
Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
• 
Blood sample for baseline reference.  Serum to be stored frozen at the MAC 
laboratory through to completion of the study for possible use as a baseline 
reference should additional safety laboratory tests or markers of disease 
activity be indicated. 
• 
Urine for pregnancy testing (pre-menopausal women only) (may be conducted 
the evening before) 
• 
Urine drug screen (may be conducted the evening before) 
Provided the subject meets all the study entry criteria they will be randomised to 
treatment.  The IWRS will allocate jars of study ointment to be dispensed to the subject 
 
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and the appropriate study ointment either 1.0% (w/w) ZPL-5212372 or matched placebo 
will be 
 by the unit staff, as described in 
section 5.7, immediately prior to administration.  Particular care must be taken to avoid 
contamination of the area to be used for blood sampling. 
After dosing the following assessments will be carried out at 1, 2, 4, 8 and 12 hours after 
the morning application: 
• 
Duplicate supine blood pressure and pulse rate 
• 
Triplicate 12-lead ECG  
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
• 
Local toleration assessment (12 hours post dose only) 
Following the 12 hour blood sample and assessment of local toleration, the second 
application of study ointment will be administered by the clinical staff.  After dosing the 
following assessments will be carried out at 13, 16 and 24 hours after the first application 
of ointment: 
• 
Duplicate supine blood pressure and pulse rate (no collection at 16 hours) 
• 
Triplicate12-lead ECG (no collection at 16 hours) 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
 
Assess symptoms by spontaneous reporting of adverse events and by asking the subjects 
to respond to a non-leading question such as “How do you feel?” 
6.2.2 Days 2 to 6 Assessments 
On Days 2 to 6 inclusive, subjects will be dosed at 12 hourly intervals (twice per day) and 
the following assessments will be carried out prior to each study ointment application: 
• 
Duplicate supine blood pressure and pulse rate 
• 
Triplicate 12-lead ECG 
• 
Local toleration assessment 
• 
Obtain blood and urine samples for  
• 
Safety laboratory tests (haematology, clinical chemistry and urinalysis) - Day 
4 prior to first application only 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
 
Assess symptoms by spontaneous reporting of adverse events and by asking the subjects 
to respond to a non-leading question such as “How do you feel?” 
6.2.3 Day 7 Assessments 
On Day 7 the following assessments will be carried out pre-dose and 1, 2, 4, 8, 12 and 16 
hours after the final application of study ointment on the morning of Day 7: 
• 
Duplicate supine blood pressure and pulse rate (no collection at 16 hours) 
• 
Triplicate 12-lead ECG (no collection at 16 hours) 
 
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• 
Local toleration assessment (immediately prior to final administration of study 
ointment only) 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
 
Assess symptoms by spontaneous reporting of adverse events and by asking the subjects 
to respond to a non-leading question such as “How do you feel?” 
6.2.4 Day 8 Assessments 
On Day 8 the following assessments will be carried out at 24 and 36 hours after the final 
application of study ointment on the morning of Day 7: 
• 
Duplicate supine blood pressure and pulse rate 
• 
Triplicate 12-lead ECG 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
• 
Local toleration assessment (at 24 hours after final administration of study ointment 
only) 
• 
Completion of Ziarco Ointment Questionnaire 
• 
Assess symptoms by spontaneous reporting of adverse events and by asking the 
subjects to respond to a non-leading question such as “How do you feel?” 
6.2.5 Day 9 Assessments 
On Day 9 the following assessments will be carried out at 48 hours after the final 
application of study ointment on the morning of Day 7: 
• 
Brief physical exam 
• 
Local toleration assessment 
• 
Duplicate supine blood pressure and pulse rate 
• 
Triplicate 12-lead ECG 
• 
Obtain blood and urine samples for  
• 
Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
• 
Assess symptoms by spontaneous reporting of adverse events and by asking the 
subjects to respond to a non-leading question such as “How do you feel?” 
On Day 9 after the 48 hour post dose assessments have been completed subjects may 
leave the Unit at the discretion of the supervising clinical staff and will be asked to return 
for a follow up visit 7-14 days following the final application of study ointment. 
6.3 
Cohort 2 (AD Patients) 
Cohort 2 will be split with a lead (n=3) sentinel group which will only have 10% BSA treated 
with study ointments.  If after 48 hours of treatment (4 applications) the study ointments are 
well tolerated then the remaining patients in the cohort (n=9) may start treatment with up to 
40% BSA being treated with study ointments.  Apart from this difference in percentage BSA 
treated all subjects in Cohort 2 will have the following assessments conducted in the 
designated timeframe. 
 
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Patients will attend the Unit on the morning of Day 1 or the evening before (if required) and 
will remain in the Unit until Day 9 following the 48 hours post dose blood sample. 
6.3.1 Day 1 assessments 
On Day 1 the following assessments will be carried out prior to randomisation and 
dosing: 
• EASI, IGA and BSA assessments.  For inclusion in the study, an EASI score of 
between 12 and 48, an IGA score of ≥3 and BSA affected by AD of ≥10% and ≤ 50% 
must be recorded (may be conducted the evening before) 
• Review of concomitant medications (may be conducted the evening before) 
• Review changes in the subject’s medical history since screening (may be conducted 
the evening before) 
• Brief physical examination (may be conducted the evening before) 
• Completion of body map of skin areas affected by AD and identification and 
recording of skin area to be treated (may be conducted the evening before) 
• Breath alcohol testing (may be conducted the evening before) 
• Triplicate12-lead ECG 
• Duplicate vital signs (supine blood pressure and pulse rate) 
• Review of baseline adverse events 
• Obtain blood and urine samples: 
• 
Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
• 
Blood sample for baseline reference.  Serum to be stored frozen at the MAC 
laboratory through to completion of the study for possible use as a baseline 
reference should additional safety laboratory tests or markers of disease 
activity be indicated. 
• 
Urine for pregnancy testing in pre-menopausal women only (may be 
conducted the evening before) 
• 
Urine drug screen (may be conducted the evening before) 
Provided the patient meets all the study entry criteria they will be randomised to 
treatment.  The IWRS will allocate jars of study ointment to be dispensed to the patient 
and the appropriate study ointment either 1.0% (w/w) ZPL-5212372 or matched placebo 
will be 
 by the unit staff, as described in 
section 5.7, immediately prior to administration.  Particular care must be taken to avoid 
contamination of the area to be used for blood sampling.  At least 15 minutes after study 
ointment application study emollient should be applied. 
After dosing the following assessments will be carried out at 1, 2, 4, 8 and 12 hours after 
the after the morning application: 
• 
Duplicate supine blood pressure and pulse rate 
• 
Triplicate 12-lead ECG  
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
 
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 Version 2.0 /28 July 2016 
 
Following the 12 hour blood sample the second application of study ointment and 
emollient will be administered by the clinical staff.  After dosing the following 
assessments will be carried out at 13, 16 and 24 hours after the first application of 
ointment: 
• 
Duplicate supine blood pressure and pulse rate (no collection at 16 hours) 
• 
Triplicate 12-lead ECG (no collection at 16 hours) 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
 
Assess symptoms by spontaneous reporting of adverse events and by asking the patients 
to respond to a non-leading question such as “How do you feel?” 
6.3.2 Day 2 to 6 assessments 
On Days 2 to 6 inclusive subjects will be dosed at 12 hourly intervals (twice per day) and 
the following assessments will be carried out prior to each study ointment application: 
• 
Duplicate supine blood pressure and pulse rate 
• 
Obtain blood and urine samples for  
• 
Safety laboratory tests (haematology, clinical chemistry and urinalysis) - Day 
4 prior to first application only 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
 
Assess symptoms by spontaneous reporting of adverse events and by asking the patients 
to respond to a non-leading question such as “How do you feel?” 
6.3.3 Day 7 assessments 
On Day 7 the following assessments will be carried out pre-dose and 1, 2, 4, 8, 12 and 16 
hours after the final application of study ointment on the morning of Day 7: 
• 
Duplicate supine blood pressure and pulse rate (no collection at 16 hours) 
• 
Triplicate 12-lead ECG (no collection at 16 hours) 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
• 
EASI, IGA and BSA assessments (on a single occasion only) 
 
Assess symptoms by spontaneous reporting of adverse events and by asking the patients 
to respond to a non-leading question such as “How do you feel?” 
6.3.4 Day 8 assessments 
On Day 8 the following assessments will be carried out at 24 and 36 hours after the final 
application of study ointment on the morning of Day 7: 
• 
Duplicate supine blood pressure and pulse rate 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
• 
Triplicate 12-lead ECG (24 hours post dose only)Completion of Ziarco Ointment 
Questionnaire 
 
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Assess symptoms by spontaneous reporting of adverse events and by asking the patients 
to respond to a non-leading question such as “How do you feel?” 
6.3.5 Day 9 assessments 
On Day 9 the following assessments will be carried out at 48 hours after the final 
application of study ointment on the morning of Day 7: 
• 
Brief physical exam 
• 
Duplicate supine blood pressure and pulse rate 
• 
Obtain blood and urine samples for  
• 
Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
Assess symptoms by spontaneous reporting of adverse events and by asking the patients 
to respond to a non-leading question such as “How do you feel?” 
On Day 9 after the 48 hour post dose assessments have been completed patients may 
leave the Unit at the discretion of the supervising clinical staff and will be asked to return 
for a follow up visit 7-14 days following the final application of study ointment. 
6.4 
Cohort 3 (AD Patients) 
6.4.1 Baseline Visit (Day 1) 
AD patients will attend the Unit on the morning of Day 1 when the following assessments 
will be carried out prior to randomisation and dosing: 
• EASI, BSA and IGA assessments.  For inclusion in the study, an EASI score of 
between 12 and 48, an IGA score of ≥3 and BSA affected by AD of ≥10% and ≤50% 
must be recorded. 
• Completion of the Ziarco Itch Questionnaire 
• Review of concomitant medications 
• Review changes in the subject’s medical history since screening 
• Brief physical examination 
• Duplicate vital signs (supine blood pressure and pulse rate) 
• Review of baseline adverse events 
• Completion of body map of skin areas affected by AD and identification and 
recording of skin area to be treated 
• Breath alcohol testing 
• Obtain blood and urine samples: 
• 
Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
• 
Blood sample for pharmacokinetic analysis of ZPL-5212372 
• 
Blood sample for baseline reference.  Serum to be stored frozen at the MAC 
through to completion of the study for possible use as a baseline reference 
should additional safety laboratory tests or markers of disease activity be 
indicated. 
• 
Urine for pregnancy testing (pre-menopausal women only) 
 
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• 
Urine drug screen  
Provided the patient meets all the study entry criteria they will be randomised to 
treatment.  The IWRS will allocate jars of study ointment to be dispensed to the patient 
and the appropriate study ointment either 1.0% (w/w) ZPL-5212372 or matched placebo.  
The patient will be shown how to dispense the study ointment and administer the study 
ointment by the clinical staff.  At least 15 minutes after study ointment application study 
emollient should be applied under clinical staff’s supervision. 
Patients may then leave the Unit at the discretion of the supervising clinical staff after 
being provided with enough study medication and study emollient for at least one week. 
Patients will be reminded of the study restrictions and also instructed on study medication 
dosing requirements and concomitant medications use and study emollient use. 
 
6.4.2 Visits 2 to 4 (Days 5, 8 and 10) 
Patients will attend the Unit on Days 5, 8 and 10.  On these days the patient will be asked 
not to apply the morning dose of ointment or study emollient at home, but to apply it at the 
clinic under instruction of the clinic staff after the following assessments have been carried 
out: 
 
• Review study medication compliance 
• Duplicate supine blood pressure and pulse rate pre-dose 
• EASI, BSA and IGA assessments. 
• Completion of the Ziarco Itch Questionnaire 
• Blood sample for pharmacokinetic analysis of ZPL-5212372. 
• Review concomitant medications, including emollient use 
• Assess symptoms by spontaneous reporting of adverse events and by asking the 
patients to respond to a non-leading question such as “How do you feel?” 
Once all pre-dose activities have been completed the patient will apply both study 
ointment and study emollient under supervision of clinical staff, provided the dose was not 
already taken prior to the visit. If the patient has applied the morning dose of ointment 
prior to the visit then the approximate time of dosing should be recorded on the CRF. 
 
Patients may then leave the Unit at the discretion of the supervising clinical staff after 
being provided with enough study medication and emollient, for the following week.  
Subjects will be reminded of the study restrictions and concomitant medications use and 
study emollient use. 
 
6.4.3 Visit 5 Day 15 Or Early Termination 
Patients will have applied their last study ointment application on the evening of Day 14, 
the day prior to the Day 15 clinic visit. Patients will attend the Unit on Day 15 when the 
following assessments will be carried out:  
 
• Brief physical examination  
• Duplicate supine blood pressure and pulse rate 
 
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• EASI, BSA and IGA assessments 
• Completion of the Ziarco Itch Questionnaire 
• Completion of the PGIC Questionnaire 
• Completion of the Ziarco Ointment Questionnaire 
• Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
• Blood sample for pharmacokinetic analysis of ZPL-5212372 (the date and time of the 
last dose of study drug should be recorded in the CRF) 
• Collect study medication (including emollient) 
• Drug compliance check 
• Review concomitant medications, including emollient usage 
• Assess symptoms by spontaneous reporting of adverse events and by asking the 
patients to respond to a non-leading question such as “How do you feel?” 
Patients may then leave the Unit at the discretion of the supervising clinical staff and will 
be asked to return for a follow up visit 7-14 days following the final application of study 
ointment. 
6.5 
Follow Up visit  
Subjects will return to the Unit 7-14 days following the last application of study ointment for 
a follow up visit. The assessments conducted at the follow up visit will vary depending in 
which cohort the subject participated. Table 7 summarises the assessments to be conducted 
for each cohort:  
 
Table 7 - Follow up visit assessments for each cohort 
Assessment 
Cohort 1 
(Healthy 
Volunteers) 
Cohort 2 
(AD Patients) 
Cohort 3 
(AD 
Patients) 
Brief physical examination 
Yes 
Yes 
Yes 
Single 12-lead ECG 
Yes 
Yes 
Yes 
Duplicate supine blood pressure and pulse rate. 
Yes 
Yes 
Yes 
Safety laboratory tests (haematology, clinical chemistry 
and urinalysis) 
Yes 
Yes 
Yes 
Urine pregnancy testing (pre-menopausal women only) 
Yes 
Yes 
Yes 
EASI, BSA and IGA assessments 
No 
Yes 
Yes 
Ziarco Itch Questionnaire 
No 
No 
Yes 
Review concomitant medications, (including emollient 
usage if applicable) 
Yes 
Yes 
Yes 
Review of adverse events 
Yes 
Yes 
Yes 
6.6 
Early Termination Assessments 
Subjects who discontinue early from the study should, if possible, have early termination 
assessments performed.  For patients in Cohort 3 they should have an early termination visit 
take place as soon as possible after the patient stops taking study drug. The observations and 
procedures summarized in Table 8 should be performed. The assessments conducted will 
vary depending in which cohort the subject participated. For the completion of 
questionnaires, the subject should be requested to answer with respect to how they were on 
their last dosing day. 
 
 
 
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Table 8 - Early termination visit assessments for each cohort 
Assessment 
Cohort 1 
(Healthy 
Volunteers) 
Cohort 2 
(AD Patients) 
Cohort 3 
(AD 
Patients) 
Brief physical examination 
Yes 
Yes 
Yes 
Single 12-lead ECG 
Yes 
Yes 
No 
Duplicate supine blood pressure and pulse rate. 
Yes 
Yes 
Yes 
Safety laboratory tests (haematology, clinical chemistry 
and urinalysis) 
Yes 
Yes 
Yes 
Urine pregnancy testing (pre-menopausal women only) 
No 
No 
Yes 
Blood sample for pharmacokinetic analysis of 
ZPL-5212372 
Yes 
Yes 
Yes 
Assessment of local toleration 
Yes 
No 
No 
EASI, BSA and IGA assessments 
No 
Yes 
Yes 
Ziarco Itch Questionnaire 
No 
No 
Yes 
PGIC 
No 
No 
Yes 
Ziarco Ointment Questionnaire 
No 
No 
Yes 
Review concomitant medications, (including emollient 
usage if applicable) 
Yes 
Yes 
Yes 
Review of adverse events 
Yes 
Yes 
Yes 
 
After the early termination visit, the subject should be instructed to return for the follow up 
visit which should be scheduled for 7-14 days after their last application of study ointment. 
 
 
 
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6.7 
Schedule of Activities 
Table 9 - Schedule of activities for Cohort 1 (Healthy Volunteers) 
Activities 
Screening1 
Day 1 
 
Day 2 to Day 6 
 
Day 7 
 
Day 8 
Day 9/ Early 
Termination 
Follow up 7-14 
days post last dose 
In-patient in CRU  
 
X2-------------------------------------------------------------------------------------------------------------X 
 
Informed Consent 
X 
 
 
 
 
 
 
Demographics  
X 
 
 
 
 
 
 
Medical history 
X 
X3 
 
 
 
 
 
Physical exam4 ( inc ht/wt) 
X 
X 
 
 
 
X 
X 
12-lead ECG5 
X 
X 
X 
X 
X 
X 
X 
Vital signs (BP and PR)6 
X 
X 
X 
X 
X 
X 
X 
Breath alcohol and drug screen 
X 
X 
 
 
 
 
 
Pregnancy Test7 
X 
X 
 
 
 
 
X 
Serum FSH8 
X 
 
 
 
 
 
 
Safety labs (inc urinalysis) 
X 
X9 
X10 
 
 
X 
X 
Hepatitis B/C, HbA1c, HIV 
X 
 
 
 
 
 
 
Fitzpatrick skin type assessment 
X 
 
 
 
 
 
 
Body Map Recorded 
 
X 
 
 
 
 
 
PK blood sample11 
 
X 
X 
X 
X 
X 
 
Assessment of local toleration12 
 
X 
X 
X 
X 
X 
 
Ointment Questionnaire 
 
 
 
 
X 
 
 
Randomisation 
 
X 
 
 
 
 
 
Dosing13 
 
X----------------------------------------------------X 
 
 
 
Concomitant & prior 
medications  
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
AE collection and review12 
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
 
1 
Screening may occur over more than 1 day. 
2 
Subjects may come into the CRU on the evening before and some pre-dose activities may be completed. 
3 
Review changes in the subject’s medical history since screening. 
4 
Full Physical examination at Screening, a brief examination on all other occasions (guided by AEs, but to include, as a minimum, general appearance, heart, lungs and skin). 
5 
Single ECG assessments at screening, follow up and early termination (if applicable).  Triplicate ECG assessments will be performed at pre-dose, 1, 2, 4, 8, 12, 13 (Day 1 only) and 24 hours post-morning dose on Day 1 
and Day 7, prior to each dose on Days 2 to 6, and at 36 hours (Day 8) and at 48 hours (Day 9) post last dose. 
6 
Duplicate, supine only, vital sign assessments will be performed at Screening, pre-dose, 1, 2, 4, 8, 12, 13 (Day 1 only) and 24 hours post-morning dose on Day 1 and Day 7, prior to each dose on Days 2 to 6, and at 36 hours 
(Day 8) and at 48 hours (Day 9) post last dose, follow up and at early termination (if applicable). 
7 
Serum pregnancy test at Screening, urine at all other specified time points; for women of child-bearing potential only. 
8 
Serum FSH testing for post-menopausal females 45-60 years only. 
9 
An additional blood sample will be taken at Baseline and stored frozen at MAC, as a reference sample, in case extra tests are required.  
10 An additional safety lab sample will be taken on Day 4. 
11 Blood samples for analysis of ZPL-5212372 will be taken prior to dosing and at 1, 2, 4, 8, 12, 13 (Day 1 only), 16 and 24 hours post-morning dose on Day 1 and Day 7, prior to each dose on Days 2 to 6, and at 36 hours 
(Day 8) and at 48 hours (Day 9) post last dose. 
12 Assessment of local toleration on treated skin will be conducted prior to each dosing occasion and at 24and 48 hours post last dose. 
13 Dosing twice daily, 12 hours apart from Days 1 to 6, the last application of study medication will be the morning of Day 7. 
14 Adverse events will be collected from the signing of the consent form until the follow up visit. SAE’s collected up to 30 days post last dose. 
 
 
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Table 10 - Schedule of activities for Cohort 2 (Moderate to Severe AD Patients) 
Activities 
Screening1 
Washout 
Period2 
Day 1 
 
Day 2 to Day 6 
 
Day 7 
 
Day 8 
Day 9/ Early 
Termination 
Follow up 7-14 days 
post last dose 
In-patient in CRU 
 
 
X3-------------------------------------------------------------------------------------------------X 
 
Informed Consent 
X 
 
 
 
 
 
 
 
Demographics  
X 
 
 
 
 
 
 
 
Medical history 
X 
 
X4 
 
 
 
 
 
Physical exam5 (incl ht/wt) 
X 
 
X 
 
 
 
X 
X 
12-lead ECG6 
X 
 
X 
 
X 
 
X 
X 
Vital signs (BP and PR)7 
X 
 
X 
X 
X 
X 
X 
X 
Breath alcohol and drug screen 
X 
 
X 
 
 
 
 
 
Pregnancy Test8 
X 
 
X 
 
 
 
 
X 
Serum FSH9 
X 
 
 
 
 
 
 
 
Safety labs (inc urinalysis) 
X 
 
X10 
X11 
 
 
X 
X 
Hepatitis B/C, HbA1c, HIV 
X 
 
 
 
 
 
 
 
Fitzpatrick skin type assessment 
X 
 
 
 
 
 
 
 
Study emollient use12 
 
X------------------------------------------------------------------------------------------------------------------------X 
 
Body Map Recorded 
 
 
X 
 
 
 
 
 
PK blood sample13 
 
 
X 
X 
X 
X 
X 
 
EASI/IGA/BSA14 
X 
 
X 
 
X 
 
 
X 
Ointment Questionnaire 
 
 
 
 
 
X 
 
 
Randomisation 
 
 
X 
 
 
 
 
 
Dosing15 
 
 
X----------------------------------------X 
 
 
 
Concomitant & prior medications  
X------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
AE collection and review16 
X------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
 
1 
Screening may occur over more than 1 day. 
2 
Washout period from AD medication of between 1 week and 1 month dependent on each patient’s current AD medication.  Study Emollient should be started 7 days before Day 1. 
3 
Patients may come into the CRU on the evening before and some pre-dose activities may be completed. 
4 
Review changes in the subject’s medical history since screening. 
5 
Full Physical examination at Screening, a brief examination on all other occasions (guided by AEs, but to include, as a minimum, general appearance, heart, lungs and skin). 
6 
Single ECG assessment at screening, follow up and early termination (if applicable).  Triplicate ECG assessments will be performed at pre-dose, 1, 2, 4, 8, 12, 13 (Day 1 only) and 24 hours post-morning dose on Day 
1 and Day 7. 
7 
Duplicate, supine only, vital sign assessments will be performed at Screening,  pre-dose, 1, 2, 4, 8, 12, 13 (Day 1 only) and 24 hours post-morning dose on Day 1 and Day 7, prior to each dose on Days 2 to 6, and at 
36 hours (Day 8) and at 48 hours (Day 9) post last dose and at follow up. 
8 
Serum pregnancy test at Screening, urine at all other specified time points; for women of child-bearing potential only. 
9 
Serum FSH testing for post-menopausal females 45-60 years only. 
10 
An additional blood sample will be taken at Baseline and stored frozen at the MAC, as a reference sample, in case extra tests are required.  
11 
An additional safety lab sample will be taken on Day 4. 
12 
Study emollient to be used from day -7 onwards until discharge from the CRU on day 9. 
13 
Blood samples for analysis of ZPL-5212372 will be taken prior to dosing and at 1, 2, 4, 8, 12, 13 (Day 1 only), 16 and 24 hours post-morning dose on Day 1 and Day 7, prior to each dose on Days 2 to 6, and at 36 
hours (Day 8) and at 48 hours (Day 9) post last dose. 
14 
EASI/IGA/BSA assessed at screening and prior to randomisation to ensure inclusion criteria are met.  Assessed again at the end of the study (or early termination) to document severity and extent of AD following 
study medication. 
15 
Dosing twice daily, 12 hours apart from Days 1 to 6, the last application of study medication will be the morning of Day 7. 
16 
Adverse events will be collected from the signing of the consent form until the follow up visit. SAE’s collected up to 30 days post last dose. 
 
 
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Table 11 - Schedule of activities for Cohort 3 (Moderate to Severe AD Patients) 
Activities 
Screening1 
Washout 
Period2 
Baseline Visit 1 
(Day 1) 
Visit 2 
(Day 5) 
Visit 3 
(Day 8) 
Visit 4 
(Day 10) 
Visit 5 
(Day 15)/ Early 
Termination 
Follow up 7-14 
days post last 
dose 
Informed Consent 
X 
 
 
 
 
 
 
 
Demographics  
X 
 
 
 
 
 
 
 
Medical history 
X 
 
X3 
 
 
 
 
 
Physical exam4 ( incl ht/wt) 
X 
 
X 
 
 
 
X 
X 
12-lead ECG5 
X 
 
 
 
 
 
 
X 
Vital signs (BP and PR)6 
X 
 
X 
X 
X 
X 
X 
X 
Breath alcohol and drug screen 
X 
 
X 
 
 
 
 
 
Pregnancy Test7 
X 
 
X 
 
 
 
X 
X 
Serum FSH8 
X 
 
 
 
 
 
 
 
Safety labs (inc urinalysis) 
X 
 
X9 
 
 
 
X 
X 
Hepatitis B/C, HbA1c, HIV 
X 
 
 
 
 
 
 
 
Fitzpatrick skin type assessment 
X 
 
 
 
 
 
 
 
Study emollient use10 
 
X---------------------------------------------------------------------------------------------------------------------------X 
 
Body Map Recorded 
 
 
X 
 
 
 
 
 
PK blood sample11 
 
 
X 
X 
X 
X 
X 
 
EASI/BSA/IGA 
X 
 
X 
X 
X 
X 
X 
X 
Ziarco Itch Questionnaire12 
 
 
X 
X 
X 
X 
X 
X 
PGIC 
 
 
 
 
 
 
X 
 
Ointment Questionnaire 
 
 
 
 
 
 
X 
 
Randomisation 
 
 
X 
 
 
 
 
 
Dosing13 
 
 
X-------------------------------------------------------------------------X 
 
 
Concomitant & prior medications  
X------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
AE collection and review13 
X------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
 
1. 
Screening may occur over more than 1 day.  
2. 
Washout period from AD medication of between 1 week and 1 month dependent on each patient’s current AD medication. Study Emollient should be started 7 days before Day 1. 
3. 
Review changes in the patient’s medical history since screening. 
4. 
Full Physical examination at Screening, a brief examination at all other visits (guided by AEs, but to include, as a minimum, general appearance, heart, lungs and skin). 
5. 
A triplicate ECG assessment will be performed at Screening and a single ECG at Follow Up. 
6. 
Duplicate, supine only, vital sign assessments to be performed at Screening and pre-dose on days 1, 5, 8, 10, 15 and follow-up. 
7. 
Serum pregnancy test at Screening, urine at all other specified time points; for women of child-bearing potential only. 
8. 
Serum FSH testing for post-menopausal females 45-60 years only. 
9. 
An additional blood sample will be taken at Baseline and stored frozen at the central lab, as a reference sample, in case extra tests are required.  
10. Study emollient to be used from day -7 onwards until completion of visit on day 15. 
11. Blood samples for analysis of ZPL-5212372 will be taken prior to dosing on days 1, 5, 8 and 10 and an approximately 12 hours post last dose sample will be taken on Day 15. 
12. Pruritus NRS, VRS, duration and impact of itch on sleep disturbance for the previous 24 hour period at clinic visits (Days 1, 5, 8, 10 and 15) by clinical staff asking the patients to complete the Ziarco Itch 
Questionnaire (Appendix 9). 
13. To be dosed topically twice daily at home except on study visit days when the morning dose will be applied in the clinic after all other study assessments have been completed. Medication should be applied in the 
morning (preferably after showering) and approximately 12 hours later in the evening.  Study medication should be applied approximately 15 minutes prior to using the Study Emollient. Subjects will apply their 
last dose of study medication at home in the evening of day 14 prior to their visit to the clinic on day 15. 
14. Adverse events will be collected from the signing of the consent form until the follow up visit. SAE’s collected up to 30 days post last dose.  
 
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7 
ASSESSMENTS 
Where time-points coincide, assessments should be taken in the following order: 
• 12-lead ECG, vital signs and then blood for PK closest to the required time (± 
10% from nominal time) 
7.1 
Physical Examination 
All physical examinations will be carried out by trained medical personnel.  At 
screening the subject will have a full physical examination, to include an assessment 
of head, neck, heart, lungs, abdomen, skin, peripheral circulation, joints and general 
appearance.  Any abnormalities should be recorded in the study CRF. At all other 
visits subjects will have a brief physical examination, which should include general 
appearance, heart, lungs and skin.  Other systems examined should be determined by 
clinical findings and any adverse events reported. 
7.2 
Vital Signs Measurements 
Blood pressure (BP, systolic and diastolic mmHg) and pulse (beats per minute) will 
be measured in duplicate (approximately 2 minutes between measurements). 
All measurements will be assessed with the subject in a supine position.  Subjects 
should be supine for at least 5 minutes before taking the measurement.  Blood 
pressure will be measured from the same arm throughout the study and recorded to 
the nearest mmHg.  Ideally this will be the subject’s dominant arm unless this arm is 
affected by AD and study ointment application could pose a potential contamination 
issue.  To minimise any contamination issue the blood pressure cuff should be wiped 
clean with an alcohol wipe between subjects. 
Pulse should be measured in the brachial/radial artery for at least 30 seconds. 
7.3 
Electrocardiogram (ECG) Recording 
Single or triplicate (3 recordings over a 10 minute period) ECG recordings will be 
measured (see Section 6.7 schedule of activities for details).  Measurements will be 
taken after the subject has rested for at least 10 minutes in a supine position using a 
12-lead ECG machine. 
To ensure safety of the subjects, a qualified individual at the investigator site will 
make comparisons to the baseline measurements.  If the QTcF is ≥60 msec from 
baseline; or an absolute QTcF value is ≥500 msec for any ECG, the ECG intervals 
should be inspected carefully to ensure that the RR interval has been recorded 
correctly, and a single ECG will be repeated at 5-minute intervals up to 3 times to 
confirm any increased QT interval values.  If the values are consistently high, the 
subject will be withdrawn from the study, allowed to return home (so long as no other 
clinical findings preclude this) and will be instructed to return for the follow up visit 
7-14 days later. 
In some cases, it may be appropriate to repeat abnormal ECGs to rule out improper 
lead placement as contributing to the ECG abnormality.  It is important that leads are 
placed in the same positions each time in order to achieve precise ECG recordings.  If 
a machine-read QTcF value is prolonged, as defined above, repeat measurements may 
 
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not be necessary if a qualified physician’s interpretation determines that the QTcF 
values are acceptable for the individual. 
7.4 
Fitzpatrick Skin Type Assessment 
All subjects will have their skin type assessed using the Fitzpatrick Skin Type 
assessment26 and recorded in the CRF at screening.  For Cohort 1 subjects only, 
subjects must have a Skin Type of between I to III to meet the inclusion criteria for 
the study. 
 
Fitzpatrick skin type classification: 
Type I – White; very fair; red or blonde hair; blue eyes; freckles (Always burns, never 
tans) 
Type II – White; fair; red or blonde hair; blue, hazel or green eyes (Usually burns, 
tans with difficulty) 
Type III – Cream white; fair with any eye or hair colour (Sometimes mild burn, 
gradually tans) 
Type IV – Light brown (Rarely burns, tans with ease) 
Type V – Dark brown (Very rarely burns, tans very easily) 
Type VI – Black (Never burns, tans very easily) 
 
7.5 
Assessment of Local Toleration 
For Cohort 1 subjects only.  All subjects will have skin area to be dosed assessed for 
local toleration using a modified Draize Score28 prior to application of study ointment 
and until discharge from the CRU. 
 
Erythema and Eschar Formation 
Grade 
No erythema  
0 
Very slight erythema (barely perceptible)  
1 
Well defined erythema  
2 
Moderate to severe erythema  
3 
Severe erythema (beet redness) to slight eschar formation (injuries in depth)  
4 
 
Oedema Formation 
Grade 
No oedema  
0 
Very slight oedema (barely perceptible) 
1 
Slight oedema (edges of area well defined by definite raising) 
2 
Moderate oedema (raised approximately 1 mm) 
3 
Severe oedema (raised more than 1 mm and extending beyond the area of exposure) 4 
 
Desquamation 
Grade 
No desquamation 
0 
Mild desquamation (dry skin) 
1 
Moderate desquamation (flaky skin) 
2 
Severe desquamation (skin cracking) 
3 
 
 
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For each assessment the grading value for erythema and eschar formation, oedema 
formation and desquamation will be recorded and a total of the three gradings will 
also be recorded.  Individual subject stopping criteria will be based on a score of ≥ 3 
for any of the parameters on 3 consecutive occasions.  Cohort 1 stopping criteria will 
be based on a total score of ≥ 5 on 3 consecutive occasions in 5 subjects dosed over a 
40% BSA. 
7.6 
Skin Area Body Map 
For Cohort 1 healthy volunteer subjects: schematic body map (Front and Back views) 
will be used on Day 1 prior to randomisation to map the area of skin to be treated with 
study ointments to ensure consistency of the percentage body area to be treated. 
For Cohort 2 and 3 AD patients: the areas of skin affected by AD and the approximate 
percentage of BSA affected by AD will be recorded on a schematic body map (Front 
and Back views) on Day 1 prior to randomisation for each patient.  The body map will 
also be used to indicate the area of skin which will be dosed for the duration of the 
study (Appendix 10).  For Cohort 2 patients, depending on whether they are in the 
10% BSA treated sentinel group or the remaining 40% BSA treated group, it may be 
not all their AD lesioned skin will treated with study ointment (See Appendix 6 for an 
example) or that skin which is not currently affected by AD lesions will need to be 
treated with study ointment (see Appendix 7 for an example).  However in all cases, 
once recorded the areas of skin to be treated with study ointment will be treated 
consistently for the duration of the study regardless of any changes in the size or 
extent of AD affected skin areas during the study period. 
7.7 
Eczema Area and Severity Index (EASI) 
For Cohort 2 and 3 AD patients only.  The Eczema Area and Severity Index (EASI) 27 
(see Appendix 11) is a validated tool used to measure the severity and extent of atopic 
eczema.  The total score incorporates the extent of body regions affected and the 
intensity of a representative area of eczema. The approximate percentages affected by 
eczema are calculated for each region. 
The EASI score will be used to determine eligibility for the study and any changes in 
AD severity/extent following study medication (Cohort 2) as well as assessing 
efficacy (Cohort 3 only). 
The investigator will be provided with further detail of how to calculate the EASI 
score at the site initiation visits. 
EASI must be ≥9 and ≤48 at screening and ≥12 and ≤48 on Day 1 for the patient to be 
eligible for inclusion in the study. 
7.8 
Body Surface Area (BSA) 
For Cohorts 2 and 3 AD patients only.  Assessment of the percentage of a patient’s 
body surface area affected by AD will be made by best estimates (see Appendix 12 
for guidance) of the Investigator and recorded on the individual patients body map.  
Affected BSA must be ≥ 10% and ≤40% at screening and must be ≥ 10% and ≤50% 
on Day 1 for the patient to be randomised. 
 
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7.9 
Investigator Global Assessment (IGA) 
For Cohorts 2 and 3 AD patients only.  An overall assessment of the severity of AD 
will be made, by the investigator, using the Investigator Global Assessment at each 
visit. IGA scores take values on a 5-point scale from 0-4, where 0=clear to 4=severe 
disease.  
0=clear (no inflammatory signs of AD),  
1=almost clear (just perceptible erythema, and just perceptible papulation/infiltration) 
2=mild disease (mild erythema, and mild papulation/infiltration) 
3=moderate disease (moderate erythema, and moderate papulation/infiltration) 
4=severe disease (severe erythema, and severe papulation/infiltration) 
 
At screening and Day 1 this must be ≥ 3 for the patient to be eligible. 
7.10 Ziarco Itch Questionnaire 
For Cohort 3 AD patients only.  Ziarco has developed an Itch Questionnaire to 
capture both Numerical and Verbal rating Scales for Pruritus and Sleep Disturbance 
(Appendix 9). 
 
Pruritus NRS, VRS, duration of itch and impact of itch on sleep disturbance for the 
previous 24 hour period will be assessed on clinic visits by clinical staff asking the 
patients to complete the Ziarco Itch Questionnaire. 
7.10.1 
Numerical Rating Scale (NRS) for Pruritus (Itching) 
The Pruritus NRS is an assessment tool that will be used to assess the subject’s worst 
itch as a result of AD in the previous 24 hours. 
They will be asked the following question: 
On a scale of 0 (No Itching) to 10 (Itching as bad as you can imagine), please 
rate the WORST itching that you felt over the last 24 hours. 
 
7.10.2 
Duration of Itching 
Patients will be asked the following question to determine their duration of itching: 
Over the last 24 hours approximately how many hours, if any, did you itch?  
 
7.10.3 
Sleep Disturbance 
Patients will be asked the following question to determine the level of sleep 
disturbance due to itching: 
On a scale of 0 (No sleep disturbance) to 10 (Awake all night), please rate how 
much your sleep was disturbed by itch last night.  
 
7.10.4 
Verbal Rating Score 
Patients will be asked to rate their itch over the last 24 hours using a list of adjectives 
describing different levels of symptom intensity: 
 
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Over the last 24 hours how would you rate your itch? 
no itch 
mild 
moderate  
severe 
 
7.11 Patient Global Impression of Change (PGIC) 
For Cohort 3 AD patients only. At the end of treatment (Day 15) or early termination 
visit, the patient will be asked to rate their degree of improvement (or worsening) of 
their AD compared to before the start of treatment with study drug, using a 7-point 
scale, standardized PGIC.  
 
Since the start of the study (dosing), my overall status is: 
1 □ Very Much Improved 
2 □ Much Improved 
3 □ Minimally Improved 
4 □ No change 
5 □ Minimally Worse 
6 □ Much Worse 
7 □ Very Much Worse 
 
7.12 Ziarco Ointment Questionnaire 
At the end of the treatment phase each subject will be asked a series of questions 
about their impressions of the study ointment they have used.  Information regarding 
the ointment will be used to learn the practicalities of handling the ointment in a 
clinical setting for subsequent cohorts and clinical development.  The full list of 
questions contained in the Ziarco Ointment Questionnaire can be found in Appendix 
13. 
7.13 Pharmacokinetic Blood Samples 
Blood samples for determination of ZPL-5212372 concentrations will be collected 
either via a direct venepuncture or via an indwelling cannula whichever is more 
appropriate and will reduce potential contamination from application of topical study 
medication. 
A 5.0 mL sample, to provide a minimum of 2 mL plasma for pharmacokinetic 
analysis, will be collected into an appropriately labeled tube containing lithium 
heparin.  Blood samples will be centrifuged within 1 hour of collection at 1700 g and 
4°C for 10 minutes. Plasma will be transferred to appropriate labelled screw-capped 
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Plasma samples will be transferred to the bioanalytical laboratory on dry ice to 
maintain frozen conditions. Shipment details for the analytical laboratory will be 
provided to the sites prior to shipping any samples. 
Samples will be analysed using a validated analytical method.  After completion of 
the study plasma samples may also be analysed for metabolites and further evaluation 
of the assay performance. 
7.14 Clinical Laboratory Safety Tests 
 
Blood and urine samples will be collected for laboratory safety tests for all cohorts at 
the times detailed in Section 6. 
Details of the urine and blood sampling procedures and subsequent storage and 
shipment can be found in the ZPL-5212372 TDL Trials Laboratory manual.  Sites will 
be provided with lab kits from TDL Trials laboratory. Details of parameters to be 
tested are listed in Table 12. 
 
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Table 12 – Safety Laboratory parameters 
 
Biochemistry 
Sodium 
Potassium 
Calcium 
Urea 
Uric acid 
Creatinine 
Glucose 
Triglycerides 
Total Bilirubin 
Total protein 
Albumin 
Alkaline phosphatase 
Aspartate transferase (AST) 
Alanine transferase (ALT) 
Gamma glutamyl transferase (GGT) 
Total cholesterol 
hCG (screening, WOCBP only) 
FSH (screening, post-menopausal females only) 
Haematology  
Red cell count 
Haemoglobin 
Packed cell volume (hematocrit) 
Mean cell volume (MCV) 
Mean cell haemoglobin (MCH) 
Mean cell haemoglobin concentration (MCHC) 
Platelet count 
White cell count 
Neutrophils 
Lymphocytes 
Monocytes 
Eosinophils 
Basophils 
 
Serological Markers (Screening only) 
Hepatitis B surface antigen (HBsAg) 
Hepatitis B core antibody (HbcAb) 
Anti-hepatitis C virus serology (by  ECLIA) 
Human immunodeficiency virus (HIV) 
Glycosylated haemoglobin (HbA1c) 
Urinalysis 
pH, protein, glucose, ketone and blood 
Any significant abnormalities in the urinalysis should be investigated via microscopy and the findings 
reported as a comment in the CRF.  
Pregnancy testing (to be measured at the clinic, using kits supplied) 
Drug Screen - Cocaine, cannabinoids, opiates, barbiturates, benzodiazepines, methadone and 
amphetamines (Screening and Day 1 only) 
Breath Alcohol Test 
Screening and Day 1 only 
 
An extra blood sample (10 mL) will be taken pre-dose on Day 1 and the serum will be 
stored frozen at the central laboratory as a reference sample in the event that further 
tests or re-tests are required. 
7.15 Blood Volume 
The approximate volume of blood to be taken from each subject during the study is 
detailed in Table 13. Additional blood may be required for repeats of safety laboratory 
tests. 
 
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Table 13 - Approximate Blood Volume Required for Study 
Test 
Number of 
Samples 
Volume (mL) 
Total (mL) 
Haematology 
5 
5 
25 
Biochemistry 
5 
5 
25 
Hep B/C/HbA1c/HIV 
1 
5 
5 
PK samples (Cohorts 1 and 2) 
28 
5 
140 
PK samples (Cohort 3) 
5 
5 
25 
Baseline serum sample 
1 
10 
10 
Total volume for Cohorts 1/2 
 
 
205 
Total volume for Cohort 3 
 
 
90 
 
7.16 Safety  
7.16.1 
Adverse Events 
7.16.1.1 Definitions 
An AE (or adverse event) is any untoward medical event that occurs in a subject and 
does not necessarily have a causal relationship with study treatment. An AE can 
therefore be any unfavorable and unintended sign (including an abnormal laboratory 
finding), symptom, or disease temporally associated with the use of an investigational 
product, whether or not related to the product. 
A treatment-emergent AE will be defined as an AE that begins or that worsens in 
severity after at least one dose of study drug (ZPL-5212372 or placebo ointment) has 
been administered. 
An adverse drug reaction (ADR) is any untoward and unintended response in a 
subject to an investigational medicinal product which is related to any dose 
administered to that subject. 
An unexpected adverse reaction is an adverse reaction the nature and severity of 
which is not consistent with the information about the medicinal product in the 
Investigator's Brochure. 
All AEs, including intercurrent illnesses, occurring during the study will be 
documented in the CRF. Concomitant illnesses, which existed prior to entry into the 
study, will not be considered AEs unless they worsen during the treatment period. 
Pre-existing conditions will be recorded in the CRF as Medical History. 
7.16.1.2 Assessment of Adverse Event 
Each AE will be assessed by the investigator or delegated physician with regard to the 
following categories. 
 
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Seriousness 
A serious AE (SAE) (or serious ADR or unexpected serious adverse reaction) is 
defined as any untoward medical occurrence that at any dose: 
• Results in death 
• Is life-threatening 
This means that the subject is at risk of death at the time of the event; it does 
not mean that the event hypothetically might have caused death if it were more 
severe. 
• Requires or prolongs subject hospitalization 
• Results in persistent or significant disability or incapacity 
• Is a congenital anomaly or birth defect 
• Is an important medical event(s) that may not be immediately life-threatening 
or result in death or hospitalization but that may jeopardize the subject or 
require intervention to prevent one of the above outcomes. Examples of such 
medical events include allergic bronchospasm requiring intensive treatment in 
an emergency room or at home, blood dyscrasias or convulsions that do not 
result in subject hospitalization, or the development of drug dependency or 
drug abuse. 
Note: Cases of potential drug-induced liver injury as assessed by laboratory test 
values (“Hy’s Law Cases”) are also reportable.  If a subject develops abnormal values 
in aspartate transaminase (AST) or alanine transaminase (ALT) or both, concurrent 
with abnormal elevations in total bilirubin and no other known cause of liver injury, 
that event would be classified as a Hy’s Law Case.  In this clinical study, the term 
SAE will be understood to also include Hy’s Law Cases. 
Medical and scientific judgment should be exercised in deciding whether a case is 
serious and whether expedited reporting is appropriate. 
Severity 
The intensity of each AE must be assessed by the investigator using one of the 
following categories, and recorded in the CRF: 
• Mild: An AE that does not interfere with usual activities; 
• Moderate: An AE that interferes with usual activities; 
• Severe: An AE that prevents usual activities 
Causality 
The investigator will assess the causality / relationship between the study drug and the 
AE and record that assessment in the CRF. Causality will be shown as Probably 
related, Possibly related, Not related or Related. 
 
7.16.1.3 Recording Adverse Events 
Adverse event reporting will extend from signing the informed consent until 
completion of final visit/follow-up telephone call. Adverse events occurring after the 
 
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end of the study must be reported if the investigator considers there is a causal 
relationship with the study drug. All AEs, regardless of the relationship to study drug, 
will be recorded in the CRF. 
  
All AE reports should contain a brief description of the event, date of onset, date of 
resolution, intensity, treatment required, relationship to study drug, action taken with 
the study drug, outcome, and whether the event is classified as serious. 
 
7.16.1.4 Reporting Serious Adverse Events 
All SAEs that occur during the study and up to 30 days after receiving the last dose of 
study drug, whether considered to be associated with the study drug or not, must be 
reported within 24 hours by telephone or fax to the study pharmacovigilance team. 
The minimum information required for an initial report is: 
• Sender of report (name, address of investigator) 
• Subject identification (screening/randomisation number, initials, NOT 
subject name) 
• Protocol number 
• Description of SAE 
• Causality assessment, if possible 
 
However, as far as possible all points on the SAE form should be covered in the initial 
report and the completed SAE form faxed to the pharmacovigilance team (details 
below). The completed SAE should be sent to the address below and Ziarco Chief 
Medical Officer informed by email. In addition, the event must be documented in the 
CRF. 
 
 
  
 
 
 
 
Tel: 
 
Email : 
 
Fax : 
 
 
 
After receipt of the initial report, the pharmacovigilance team will review the 
information and, if necessary, contact the investigator, to obtain further information 
for assessment of the event.  
 will be responsible for all 
information processing and reporting according to local legal requirements.  
 
 
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7.16.1.5 Follow-up of Adverse Events 
All AEs experienced by a subject, irrespective of the suspected causality, will be 
monitored until the event has resolved, any abnormal laboratory values have returned 
to baseline or stabilized at a level acceptable to the investigator and the Ziarco Chief 
Medical Officer, until there is a satisfactory explanation for the changes observed, or 
until the subject is lost to follow-up. 
 
8 
STATISTICAL METHODS  
Before the study database is locked, a statistical analysis plan (SAP) will be finalized 
providing detailed methods for the analyses outlined below.  
Any deviations from the planned analyses will be described and justified in the final 
clinical study report.  
Separate tables, listings and figures will be generated for each cohort. 
 
8.1 
Study Subjects  
8.1.1 
Randomisation  
Separate randomisation codes will be generated for each cohort. Computer generated 
pseudo random codes will assign subjects to treatment. 
 
The randomisation for Cohort 3 will be stratified by EASI score (EASI ≤20 and EASI 
>20). 
 
8.1.2 
Disposition of Subjects  
The number and percentage of subjects entering and completing each phase of the 
study will be presented for each treatment group. Reasons for withdrawal pre- and 
post-randomisation will also be summarized.  
 
8.1.3 
Protocol Deviations  
Deviations from the protocol, including violations of inclusion/exclusion criteria, will 
be categorized as “minor” or “major” in cooperation with the Sponsor. Deviations 
will be assessed prior to the study database being locked. 
 
8.1.4 
Analysis Populations  
Full Analysis Set: 
All randomised subjects who received at least one dose 
of study treatment 
Safety Set: 
All randomised subjects who received at least one dose 
of study treatment 
Per Protocol Set: 
(Cohort 3 only) 
All subjects in the Full Analysis Set who have not 
violated any inclusion or exclusion criteria and/or 
deviated from the protocol, in a way that could influence 
their efficacy assessment. Precise reasons for excluding 
subjects from the population will be fully defined and 
documented prior to locking the study database. 
Pharmacokinetic Set: 
All ZPL-5212372 treated subjects in the Full Analysis 
 
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Set who have at least one ZPL-5212372 concentration 
recorded (including concentrations below the limit of 
quantification) 
The Full Analysis Set (FAS) will be used in the analyses of efficacy endpoints.  
A secondary analysis of the primary endpoint for Cohort 3 may also be performed 
based upon the Per Protocol (PP) Set.  
All safety and pharmacokinetic analyses will be based upon the Safety and 
Pharmacokinetic sets respectively. 
 
8.2 
General Considerations  
All statistical testing for Cohort 3 data will be at the 5% level of significance (1-sided) 
and all point estimates for the comparison between treatment groups will be 
accompanied by 2-sided 90% confidence intervals. No adjustment will be made for 
multiplicity due to multiple endpoints/comparisons. No statistical testing will be 
performed with the data from Cohorts 1 and 2.  
All endpoints will be summarized by treatment group and visit. Continuous data will 
be summarized using descriptive statistics (e.g. mean and standard deviation) and 
categorical data will be summarized using frequency tables (counts and percentages). 
For change from baseline endpoints baseline is defined as the last measurement 
available prior to the start of dosing on Day 1. 
All possible efforts will be made to ensure that subjects complete all the required 
assessments. As missing data may potentially bias the outcome of the statistical 
analyses and the subsequent estimation of the magnitude of the treatment effect, 
different strategies will be applied to provide a balanced assessment of treatment 
efficacy. The handling of missing data for each endpoint and sensitivity analyses will 
be detailed in the statistical analysis plan prior to the study database being locked.  
Any outliers that are detected during the review of the data will be investigated. 
Methods for dealing with outliers will be defined in the SAP or in an addendum to the 
SAP. 
8.3 
Demographics, Baseline Characteristics and Concomitant Medications  
Demographics, baseline characteristics (Cohorts 2 and 3 only), medical history and 
concomitant medication data will be summarized for each treatment group and overall 
within each cohort. 
 
8.4 
Treatment Compliance  
As Cohorts 1 and 2 will receive their study ointment as in-patients non-compliance 
will be reported by the site staff through protocol deviations.  For Cohort 3 
compliance will be assessed through reviewing the patient diary and the amount of 
study ointment used. 
 
8.5 
Efficacy Analyses  
As Cohort 1 includes healthy volunteers no efficacy data will be collected. 
 
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Though some efficacy assessments are included for Cohort 2 the data will only be 
listed and no formal comparisons will be made between the treatment groups. 
Consequently efficacy will only be assessed for Cohort 3. Hence the following 
analyses apply to Cohort 3 data. 
 
8.5.1 
Primary Endpoint  
The primary efficacy endpoint is the percentage change from baseline to Week 2 in 
the EASI score. 
The primary analysis will be performed using the FAS. A secondary analysis of the 
primary endpoint may be conducted using the PP analysis set, if sufficiently different 
to the FAS, to examine the robustness of the primary analysis results. 
The percentage changes from baseline in the EASI score at Week 2 will be 
summarized by treatment group and analysed by ANCOVA. The difference between 
treatment groups in the percentage changes from baseline will be estimated, along 
with its 90% confidence interval. Baseline will be included as a continuous covariate 
in addition to treatment. This analysis will be repeated for the sensitivity analyses 
where different approaches will be adopted for handling missing data. 
Sub-groups will include the levels of the stratification factor. Other sub-groups will be 
specified in the SAP. 
 
8.5.2 
Secondary Endpoints  
The proportion of subjects who achieve an EASI-50 response (50% reduction in EASI 
score from baseline) at Week 2 will be compared between the treatment groups. 
Group proportions and the difference between these proportions will be calculated 
along with the corresponding 90% confidence interval.  These data will also be 
analysed using logistic regression, including treatment and baseline.  The odds ratio 
and accompanying 90% confidence interval will be presented. 
 
The proportion of subjects who achieve an EASI-75 response (75% reduction in 
EASI) and the proportion of subjects who are responders for IGA score (reduction 
from baseline of at least 2 points) at Week 2, will be analysed using the same analyses 
described for the EASI-50 response. The changes from baseline in IGA score will also 
be summarized using categorical shift tables. 
 
The changes from baseline in body surface area, pruritus NRS score for worst itch, 
NRS score for sleep disturbance and duration of itching at Week 2 will be analysed 
using the same analyses described for the primary analysis.  
 
The changes from baseline in VRS score will be summarized using categorical shift 
tables.  
 
The PGIC score will be summarized and analysed using logistic regression, 
presenting odds ratios and associated 90% confidence intervals for the comparison 
between treatment groups. 
 
Secondary endpoints will be analysed using the Full Analysis Set. 
 
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8.5.3 
Exploratory Endpoints  
Responses to the Ziarco ointment questionnaire will be summarized by treatment 
group. 
 
8.6 
Safety Analyses  
Safety data will be summarized by treatment group for each cohort using all subjects 
in the Safety Analysis Set.  
No imputation will be used for handling missing data, with the exception of 
conservative approaches taken for missing adverse event information (e.g. intensity). 
Details of such conventions will be documented in the SAP prior to the study database 
being locked.  
Adverse events will be coded using the MedDRA dictionary. The incidence of all 
treatment emergent adverse events will be tabulated by treatment group and by 
treatment group and severity. Those events regarded as possibly treatment related by 
the investigator will be summarized separately.  
Changes from baseline in laboratory parameters, vital signs and ECG data will be 
summarized by visit for each treatment group.  
For Cohort 1 only, local toleration assessed by the Draize score, will be summarized 
by visit for each treatment group. The changes from baseline in Draize score will also 
be summarized using categorical shift tables. 
 
8.7 
Pharmacokinetic Analyses 
ZPL-5212372 concentrations (all cohorts) and parameters (Cohorts 1 and 2 only) will 
be summarised for each cohort using all subjects in the Pharmacokinetic Set.  
 
Individual profile and median plots will also be plotted. Median profiles for Cohorts 1 
and 2 will be plotted by day.  
 
The ZPL-5212372 pharmacokinetic parameters AUCτ, AUClast and t½ will be 
generated using non-compartmental methods. Cmax and Tmax will be determined 
directly from the data.  Where possible, the apparent terminal elimination phase rate 
constant (kel) will be calculated by linear regression of the log-linear plasma 
concentration-time curve.  Only those time points judged to describe the terminal log-
linear decline will be used in the regression.  The apparent terminal elimination half-
life (t½) will be calculated as follows: 
t½ = ln2/kel 
8.8 
Interim Analyses  
The study will have data reviews conducted between each cohort (see section 3). 
 
 
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8.9 
Determination of Sample Size  
The sample sizes for Cohorts 1 and 2 are not based on formal statistical testing. Their 
sample sizes are considered sufficient to obtain suitable safety, tolerability and PK 
data prior to starting Cohort 3. 
 
For Cohort 3 the sample size of 30 subjects has been determined on the basis of 
comparing the mean ZPL-5212372 Week 2 percentage change from baseline in the 
EASI score to that for placebo. With 20 subjects in the ZPL-5212372 group and 10 
subjects in the placebo group, there is 80% power of obtaining a statistically 
significant result, assuming that there is an improvement of at least 30% with 
ZPL-5212372 compared to placebo and a standard deviation for the percentage 
changes from baseline between subjects of 30%. This is based on the analyses 
described and a 1-sided test at the 5% level of significance.  
 
9 
ETHICAL, LEGAL AND ADMINISTRATIVE ASPECTS 
9.1 
Data Quality Assurance 
The Sponsor (or its delegate) will conduct a site visit to verify the qualifications of 
each investigator, inspect the site facilities, and inform the investigator of 
responsibilities and the procedures for ensuring adequate and correct documentation. 
 
The investigator is required to prepare and maintain adequate and accurate case 
histories designed to record all observations and other data pertinent to the study for 
each study participant. All information recorded on the CRFs for this study must be 
consistent with the subjects’ source documentation (i.e., medical records). 
 
9.2 
Case Report Forms and Source Documentation 
All data obtained during this study should be entered in the CRFs promptly. All 
source documents from which CRF entries are derived should be placed in the 
subject’s medical records. Measurements for which source documents are usually 
available include laboratory assessments, and ECG recordings.  
 
The original CRFs for each subject will be checked against source documents at the 
study site by the study monitor. 
  
After review by the study monitor, completed CRFs will be marked as complete and 
verified.  
 Data Management will review the CRFs within 
the electronic data capture (EDC) system. Where data is discrepant, Data 
Management will raise queries for the site to resolve within the EDC. 
 
9.3 
Access to Source Data 
During the course of the study, a monitor will make site visits to review protocol 
compliance, compare CRFs and individual subject’s medical records, assess drug 
accountability, and ensure that the study is being conducted according to pertinent 
regulatory requirements. CRF entries will be verified with source documentation. The 
review of medical records will be performed in a manner to ensure that subject 
confidentiality is maintained. 
 
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Checking of the CRFs for completeness and clarity, and cross-checking with source 
documents will be required to monitor the progress of the study. The investigator 
should be available at each monitoring visit to answer any queries. Moreover, 
regulatory authorities of certain countries, IECs, and/or the Sponsor’s Clinical Quality 
Assurance Group may wish to carry out such source data checks and/or on-site audit 
inspections. Direct access to source data will be required for these inspections and 
audits; they will be carried out giving due consideration to data protection and 
medical confidentiality. The investigator assures the Sponsor of the necessary support 
at all times. 
 
9.4 
Data Processing 
The site will be supplied with the following data collection tool: a web browser 
address for EDC that has been fully validated and conforms to 21CFR11 
requirements. The trained investigator site staff will enter the data required by the 
protocol into the CRFs from source documents (e.g., medical records and 
study-specific data capture tools as needed) using the 
 supplied data collection 
tool.  All information in the CRFs must be traceable to these source documents. Data 
recorded directly into the CRFs will be defined before study start and the CRFs will 
be considered the source data. Clinical Research Associates and a Data Manager will 
review CRFs entered by investigational staff for completeness and accuracy. 
Automatic quality programs check for data discrepancies in the CRFs and the 
resulting queries will be notified to the investigational site using an electronic data 
query process within the EDC. Designated investigator site staff are required to 
respond to queries and make any necessary changes to the data. Details of the data 
correction process will be specified in the Data Management Plan. After database 
lock, the investigator will receive a CD-ROM of the subject CRFs (PDF) for 
archiving at the investigational site. 
 
A validated, electronic database will be employed from the EDC system. An audit 
trail of all changes to this database, including the date, reason for the data change and 
who made the change, will be maintained within the same database. The audit trail 
will be part of the archived data at the end of the study. 
 
9.5 
Archiving Study Records 
Adequate records as required by ICH GCP and FDA CFR, will be maintained for the 
study.  This will include subject medical records, investigator logs, CRFs, laboratory 
reports, work sheets, signed SICFs, drug dispensing records, adverse experience 
reports, information regarding subjects’ discontinuation and electronic data. Essential 
documents should be retained until at least 2 years after the last approval of a 
marketing application in an ICH region and until there are no pending or 
contemplated marketing applications in an ICH region or at least 2 years have elapsed 
since the final discontinuation of clinical development of the investigational product. 
These documents should be retained for a longer period. However, if required by the 
applicable regulatory requirements or by an agreement with the sponsor. It is the 
responsibility of the sponsor to inform the investigator/institution as to when these 
documents no longer need to be retained. 
 
 
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9.6 
Good Clinical Practice 
The procedures set out in this study protocol are designed to ensure that the Sponsor 
and investigator abide by the principles of the Good Clinical Practice guidelines of the 
International Conference on Harmonization (ICH), and of the Declaration of Helsinki 
(2013). The study also will be carried out in keeping with local legal requirements. 
 
9.7 
Confidentiality 
All study findings and documents will be regarded as confidential. The investigator 
and members of his/her research team must not disclose such information without 
prior written approval from the Sponsor. 
 
The anonymity of participating subjects must be maintained. Subjects will be 
identified on CRFs and other documents submitted to 
 by their subject number 
and/or birth year, not by name. Documents not to be submitted to 
 that identify 
the subject (e.g., the signed informed consent) must be maintained in confidence by 
the investigator. 
 
9.8 
Informed Consent 
Before each subject is admitted to the study, informed consent will be obtained from 
the subject (or his/her legally authorized representative) according to the regulatory 
and legal requirements of the participating country (i.e., the Declaration of Helsinki, 
ICH GCP, and other applicable local regulations). This consent form must be dated 
and retained by the investigator as part of the study records. The investigator will not 
undertake any investigation specifically required only for the clinical study until valid 
consent has been obtained. The terms of the consent and when it was obtained must 
also be documented in the CRF. 
 
If a protocol amendment is required, the informed consent form may need to be 
revised to reflect the changes to the protocol. If the consent form is revised, it must be 
reviewed and approved by the appropriate IEC, and signed by all subjects 
subsequently enrolled in the study as well as those currently enrolled in the study. 
 
9.9 
Protocol Approval and Amendment 
Before the start of the study, the study protocol and/or other relevant documents will 
be approved by the IEC/Competent Authorities, in accordance with local legal 
requirements. The Sponsor must ensure that all ethical and legal requirements have 
been met before the first subject is enrolled in the study. 
 
This protocol is to be followed exactly. To alter the protocol, amendments must be 
written, receive approval from the appropriate personnel, and receive IEC/Competent 
Authority approval prior to implementation (if appropriate). Administrative changes 
(not affecting the subject benefit/risk ratio) may be made without the need for a 
formal amendment. All amendments will be distributed to all protocol recipients, with 
appropriate instructions. 
 
 
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9.10 Duration of the Study 
The planned duration of the study for each subject will be approximately 6 to 8 weeks 
(assuming 4 weeks screening, 1-2 weeks treatment [Cohort 1 and 2: 1 week of 
treatment or Cohort 3: 2 weeks of treatment] and 1-2 weeks follow-up).  The study 
will close when all subjects have completed the final follow-up visit. 
 
9.11 Premature Termination of the Study 
If the investigator or the Sponsor becomes aware of conditions or events that suggest 
a possible hazard to subjects if the study continues, the study may be terminated after 
appropriate consultation between the relevant parties. The study may also be 
terminated early at the Sponsor’s discretion in the absence of such a finding. 
Conditions that may warrant termination include, but are not limited to: 
• The discovery of an unexpected, significant, or unacceptable risk to the 
subjects enrolled in the study; 
• Failure to enroll subjects at an acceptable rate; 
• A decision on the part of the Sponsor to suspend or discontinue development 
of the drug. 
 
9.12 Liability and Insurance 
The Sponsor will take out reasonable third-party liability insurance cover in 
accordance with all local legal requirements. The civil liability of the investigator, the 
persons instructed by him and the hospital, practice or institute in which they are 
employed and the liability of the Sponsor with respect to financial loss due to personal 
injury and other damage that may arise as a result of the carrying out of this study are 
governed by the applicable law. 
 
The Sponsor will arrange for subjects participating in this study to be insured against 
financial loss due to personal injury caused by the pharmaceutical products being 
tested or by medical steps taken in the course of the study.  
 
9.13 Publication Policy 
By signing the study protocol, the investigator agrees with the use of results of the 
study for the purposes of national and international registration, publication and 
information for medical and pharmaceutical professionals. If necessary, the 
authorities will be notified of the investigator's name, address, qualifications and 
extent of involvement. 
 
After study completion, data from the entire study will be considered for reporting at 
a scientific meeting and for publication in a scientific journal.  Ziarco will coordinate 
these activities and will work with the Principal Investigator to determine how the 
manuscript is written and edited, the number and order of authors, the publication to 
which it will be submitted and other related issues. Authorship will be based on 
criteria stipulated by leading clinical journals (e.g. contribution to one or more areas 
of study design, data analysis and interpretation, manuscript preparation and review, 
etc.). 
 
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An investigator shall not publish any data (poster, abstract, paper, etc.) without having 
consulted with the Sponsor in advance. 
 
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10 REFERENCE LIST 
 
1. Eichenfield LF (2004). Consensus guidelines in diagnosis and treatment of 
atopic dermatitis. Allergy, 59 (Suppl. 78): 86-92. 
 
2. Williams HC (1994). The UK Working Party's Diagnostic criteria for atopic 
dermatitis I: Derivation of a minimum set of discriminators for atopic 
dermatitis. Br. J. Dermatol, 131: 383-396. 
 
3. Kini SP (2011). The impact of pruritus on quality of life: the skin equivalent 
of pain. Arch. Dermatol, 147(10): 1153-1156. 
 
4. Shaw T, Currie G, Koudelka C & Simpson E (2003) Eczema Prevalence in the 
United States: Data from the 2003 National Survey of Children’s Health. J 
Invest. Dermatol, 131, 67–73 
 
5. Beikert F, Langenbruch A, M.A. Radtke M & Augustin M (2013) Willingness 
to pay and quality of life in patients with rosacea. JEADV 27:734–738 
 
6. Williams H (2006). How epidemiology has challenged 3 prevailing concepts 
about atopic dermatitis. J Allergy Clin Immunol, 118(1): 209-213. 
 
7. Orfali R, Shimizu M, Takaoka R, Zaniboni M, Ishizaki A, Costa A, Tiba A, 
Sato M & Aoki V (2013) Atopic dermatitis in adults: clinical and 
epidemiological considerations. Rev Assoc Med Bras. 59 (3):270–275 
 
8. Abuabara K & Margolis D (2013) Do children really outgrow their eczema, or 
is there more than one eczema? J Allergy Clin Immunol 132:1139-40. 
 
9. Pugliarello S, Cozzi A, Gisondi P & Girolomoni G (2011) Phenotypes of 
atopic dermatitis. JDDG 9:12–20 
 
10. Elias P & Steinhoff M (2008) “Outside-to-Inside” (and Now Back to 
“Outside”) Pathogenic Mechanisms in Atopic Dermatitis. J Invest Dermatol. 
128(5): 1067–1070. 
 
11. Coghi S, Bortoletto M, Sampaio S, Franco de Andrade Junior H & Aoki V 
(2007) Quality of life is severely compromised in adult patients with atopic 
dermatitis in brazil, especially due to mental components. CLINICS 62 
(3):235-42 
 
12. Ginsburg I, Prystowsky J, Kornfeld D & Wolland H (1993) Role of emotional 
factors in adults with atopic dermatitis. lnt J Dermatol 32:656-660 
 
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13. Schmid-Ott G, Kuensebeck H, Jaeger B, Werfel, T, Frahm, K, Ruttman J, 
Kapp A & Lamprecht F (1999) Validity study for the Stigmatization 
experience in Atopic Dermatitis and Psoriatic patients. Acta Derm Venereal. 
79: 443-447. 
 
14. Bender B (2002) Psychologic dysfunction associated with atopic dermatitis. 
Immunology & allergy clinics of North America 22 (1) 43-53. 
 
15. Sidbury R & Poorsattar S (2006) Pediatric atopic dermatitis: shouldwe treat it 
differently? Dermatologic Therapy, 19, 83 –90 
 
16. Thomas KS, Dean T, O’Leary C, Sach TH, Koller K, Frost A, Pallett I, Crook 
AM, Meredith S, Nunn AJ, Burrows N, Pollock I, Graham-Brown R, O'Toole 
E, Potter D & Williams HC (2011) A Randomised Controlled Trial of Ion-
Exchange Water Softeners for the Treatment of Eczema in Children. PLoS 
Med 8 (2): 1-11 
 
17. Prucha H, Schnopp C, Akdis C, Lauener R, Wollenberg A, Ring J & 
Traidl-Hoffmann C (2013) Pimecrolimus, a topical calcineurin inhibitor used 
in the treatment of atopic eczema. Expert Opin. Drug Metab. Toxicol. 
9(11):1507-1516 
 
18. Hewson CA, Patel S, Calzette L, Campwala H, Havard S, Luscombe E, Clarke 
PA, Peachell PT, Matera MG, Cazzola M, Page C, Abraham WM, Williams 
CM, Clark JD, Liu WL, Clarke NP & Yeadon M (2012) Preclinical evaluation 
of an inhibitor of cytosolic phospholipase A2a for the treatment of asthma. J 
Pharm Exp Ther. 340: 656-665. 
 
19. Fogh K, Herlin T & Kragballe K (1989). Eicosanoids in skin of patients with 
atopic dermatitis: prostaglandin E2 and leukotriene B4 are present in 
biologically active concentrations. J Allergy Clin Immunol, 83 (2 Pt 1): 450-
455. 
 
20. Neisius U, Olsson R, Rukwied R, Lischetzki G & Schmlez M (2002). 
Prostaglandin E2 induces vasodilation and pruritus, but no protein 
extravasation in atopic dermatitis and controls. J Am Acad Dermatol, 47 (1): 
28-32. 
 
21. Sadik CD, Sezin T & Kim ND (2013). Leukotrienes orchestrating allergic skin 
inflammation. Exp Dermatol, 22 (11): 705-709. 
 
22. Burke JR, Davern LB, Stanley PL, Gregor KR, Banville J, Remillard R, 
Russell JW, Brassil PJ, Witmer MR, Johnson G, Tredup JA & Tramposch KM 
 
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Ziarc o  
C o nfi de ntial  
Cli nical St u d y Pr ot oc ol  
Z P L 5 2 1/ 1 0 1  
 Versi o n  2. 0 / 2 8 J ul y 2 0 1 6  
 
( 2 0 0 1). B M S- 2 2 9 7 2 4 is a ti g ht- bi n di n g i n hi bit or of c yt os olic p h os p h oli pas e 
A 2 t hat acts at t he li pi d/ water i nterface a n d p ossesses a nti -i nfla m mat or y 
acti vit y i n s ki n i nfla m mati o n m o dels. J P E T , 2 9 8: 3 7 6- 3 8 5. 
 
2 3.  Ya ma m ot o M, Har u n a T, I m ura K, Hi kita I, F ur u e Y, Hi gas hi n o K, Ga h ara Y, 
De g uc hi M, Yas ui K & Ari m ura A ( 2 0 0 8). I n hi bit or y effect of a p ot e nt a n d 
selecti ve c yt os olic p h os p h oli pase A 2
 i n hi bit or R S C
- 3 3 8 8 o n s ki n 
i nfla m mati o n i n mice. P h ar m ac ol o gy,  8 1: 3 0 1- 3 1 1. 
 
2 4.  Mill u m J & Gra d y C “ T h e Et hics of Place b o -c o ntr olle d trials: met h o d ol o gical 
j ustificati o ns” C o nte m p Cli n Tri als  3 6: 5 1 0- 5 1 4 ( 2 0 1 3). 
 
2 5.  Cli nical  Trials  Facilitati o n  Gr o u p.  ( 2 0 1 4).  Rec o m me n dati o ns  relate d  t o 
c o ntrace pti o n  
a n d  
pre g n a nc y  
testi n g  
i n  
cli nical  
trials. 
htt p:// w w w. h ma.e u/filea d mi n/ dateie n/ H u ma n _ Me dici nes/ 0 1 -
A b o ut _ H M A/ W or ki n g _ Gr o u ps/ C T F G/ 2 0 1 4 _ 0 9 _ H M A _ C T F G _ C o ntrace pti o n.
p df  
htt p:// w w w.ic h. or g/filea d mi n/ P u blic _ We b _ Site/I C H _ Pr o d ucts/ G ui deli nes/ M ul
ti disci pli nar y/ M 3 _ R 2/ Ste p 4/ M 3 _ R 2 _ _ G ui deli ne. p df 
 
2 6.  F itz patric k T B ( 1 9 8 8) T h e vali dit y a n d practicalit y of s u n reacti ve s ki n t y p es I 
t hr o u g h t o VI. Arc h Der m at ol  1 2 4: 8 6 9- 8 7 1. 
2 7.  Ha nifi n J M, T h urst o n M, O m ot o M, C herill R, T ofte SJ & Grae b er M  ( 2 0 0 1). 
T he ecze ma area a n d se v erit y i n de x ( E A SI): ass ess me nt of relia bilit y i n at o pic 
der matitis. E A SI E v al uat or Gr o u p. . Ex p Der m at ol. , Fe b; 1 0( 1): 1 1- 8. 
 
2 8.  Draize J H , W o o dar d G & Cal ver y  H ( 1 9 4 4) Met h o ds f or t he st u d y of irritati o n 
a n d  t o xicit y  of  s u bsta nces  a p plie d  t o picall y  t o  t he  s ki n  a n d  m u c o us 
me m bra nes. J P h ar m ac ol Ex p T her 8 2: 3 7 7- 3 9 0. 
 
 
 
 
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11 APPENDICES 
 
 
 
 
 
  
 
 
 
 
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APPENDIX 1 – CRITERIA FOR SAFETY VALUES OF POTENTIAL 
CLINICAL CONCERN 
Haematology 
 
Hemoglobin 
<0.8 x baseline 
Hematocrit 
<0.8 x baseline 
WBC 
<2.5 or >17.5 x 103/mm3 
Platelets 
<75 or >700 x 103/mm3 
Chemistry 
 
Total bilirubin 
>1.5 times the upper limit of the reference range 
AST 
>3 times upper limit of the reference range 
ALT 
>3 times upper limit of the reference range 
GGT 
>3 times upper limit of the reference range 
Alk Phosphatase 
>3 times upper limit of the reference range 
Creatinine 
>1.3 times upper limit of the reference range 
Urea 
>1.3 times upper limit of the reference range 
Glucose, fasting 
<0.6 or >1.5 times the limits of the reference range 
Uric acid 
>1.2 times upper limit of the reference range 
Sodium 
<0.95 or >1.05 times the limits of the reference range 
Potassium 
<0.9 or >1.1 times the limits of the reference range 
Calcium 
<0.9 or >1.1 times the limits of the reference range 
Albumin 
<0.8 or >1.2 times the limits of the reference range 
Total protein 
<0.8 or >1.2 times the limits of the reference range 
Urinalysis 
 
Urine WBC 
≥6/HPF 
Urine RBC 
≥6/HPF 
Vital Signs 
 
Pulse Rate 
Supine/Sitting: <40 or >120 bpm     Erect: <40 or >140 bpm 
Blood Pressure 
Systolic ≥30 mm Hg change from baseline in same posture 
 
Systolic <90 mm Hg 
 
Diastolic ≥20 mm Hg change from baseline in same posture 
 
Diastolic <50 mm Hg 
 
 
 
 
 
 
 
 
 
 
 
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APPENDIX 2 – HEPATIC SAFETY 
Baseline Liver Function Test (LFT) Requirements 
Before including a subject in the study they must have: 
Negative HBsAg, HBcAb and anti-hepatitis C virus serology 
No clinical evidence of hepatic disease 
During the study - if 
 
Follow up for marked abnormality 
HBsAg, HBcAb, anti-HAV (IgM and IgG), AST, ALT 
AP (including isoenzymes), TB (direct and indirect), one stage prothrombin time, 
serum proteins and, if clinically warranted, cytomegalovirus, toxoplasma and 
heterophile antibody titres.  LFTs and FBC repeated WEEKLY until they return either 
to baseline or to a level deemed acceptable by the investigator and the Ziarco CMO. 
 
GGT and AP fractionation 
{ 
Direct/indirect bilirubin 
Coombs Test 
Reticulocyte count 
 
CK + isoenzymes 
Yes 
Continue as 
per protocol 
No 
Deteriorates 
Discontinue drug 
Phone Ziarco CMO 
Follow up 
Resolved 
No further action 
Continue as per 
protocol 
No 
Repeat LFTs 
Further tests as shown 
Save some serum 
Phone Ziarco CMO  
Results confirmed  
Suggest liver origin 
Yes 
ALT >1.5 ULN 
AST >1.5 ULN 
AP >1.5 ULN 
TB >1.2 ULN 
Marked 
Minimal 
AP >1.5 ULN 
TB >1.5 ULN 
AST >1.5 ULN 
ALT >1.5ULN 
Allocate abnormality 
ALT >3 ULN 
AST >3 ULN 
AP >1.5 ULN 
T B>2 ULN 
ALT, AST, TB, AP 
Full Blood Count 
Weekly for 4 weeks 
 
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APPENDIX 3 – ECG VALUES OF POTENTIAL CLINICAL CONCERN 
Interval Durations of Potential Clinical Concern 
PR Interval  
 
≥300 msec; ≥25% increase when baseline >200 msec; 
 
 
 
Increase ≥50% when baseline ≤200 msec 
QRS Interval   
≥200 msec; ≥25% increase when baseline >100 msec 
 
 
 
≥50% increase when baseline ≤100 msec 
QT/QTcF Interval  
≥500 msec; an increase of ≥60 msec from baseline 
 
Rhythms of Potential Clinical Concern 
-Asymptomatic Marked Sinus Bradycardia (rate <35 bpm) 
-Asymptomatic Supraventricular Couplets, Atrial Bigeminy lasting >30 seconds 
-Asymptomatic Ventricular Couplets, Ventricular Bigeminy lasting >30 seconds 
-Asymptomatic Type I Second Degree (Wenckebach) AV Block of >30 seconds 
duration 
-Asymptomatic Frequent Premature Ventricular Complexes (PVCs) (≥200/24 hours) 
-Asymptomatic Frequent Premature Atrial Complexes (PACs) (≥240/24 hours) 
 
Adverse Experiences 
-Symptomatic Marked Sinus Bradycardia (rate <40 bpm) 
-Asymptomatic Sinus Pause >3 seconds without an escape beat 
-Asymptomatic Atrial Flutter or Fibrillation, subcategorized by ventricular response 
rate:  controlled = rate <120 bpm; rapid = rate >120 bpm 
-Asymptomatic Supraventricular Tachycardia ≥3 beats (rate >120 bpm) 
-Asymptomatic Nonsustained Ventricular Rhythms ≥3 beats, but duration of 
<30 seconds, including idioventricular rhythm (rate <40 bpm), accelerated 
idioventricular rhythm (40< x<100) and monomorphic/polymorphic ventricular 
tachycardia >100 bpm (such as Torsade des pointes) 
-Asymptomatic Type II Second Degree (Mobitz) AV Block 
-Asymptomatic Complete (third degree) Heart Block 
 
Serious Adverse Experiences 
-Sustained Ventricular Arrhythmias (>30 seconds duration) 
-Ventricular Fibrillation 
-At the discretion of the investigator, any arrhythmia classified as an adverse 
experience 
 
 
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APPENDIX 4 – COHORT 1 10% BSA TREATMENT BODY MAP 
EXAMPLE 
Site: 
 
Subject 
Initials: 
 
Date: 
 
 
Subject 
Number: 
 
 
 
Cohort 1 (HV) – 10% BSA Treatment Example 
 
Please use the diagram below to mark the approximate location and size of the areas affected by atopic 
dermatitis in RED and record the areas to be treated with Study Ointment with cross hatches in 
BLACK.  Record the estimated BSA affected by atopic dermatitis (if applicable) and also the estimated 
BSA to be treated with Study Ointment (to nearest whole number). 
 
BSA Affected by AD_____0________  %  
BSA Treated with Study Ointment ____10________  % 
 
 
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APPENDIX 5 – COHORT 1 40% BSA TREATMENT BODY MAP 
EXAMPLE 
Site: 
 
Subject 
Initials: 
 
Date: 
 
 
Subject 
Number: 
 
 
 
Cohort 1 (HV) – 40% BSA Treatment Example 
 
Please use the diagram below to mark the approximate location and size of the areas affected by atopic 
dermatitis in RED and record the areas to be treated with Study Ointment with cross hatches in 
BLACK.  Record the estimated BSA affected by atopic dermatitis (if applicable) and also the estimated 
BSA to be treated with Study Ointment (to nearest whole number). 
 
BSA Affected by AD_____0________  %  
BSA Treated with Study Ointment ____40________  % 
 
 
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APPENDIX 6 – COHORT 2 10% BSA TREATMENT BODY MAP 
EXAMPLE 
Site: 
 
Subject 
Initials: 
 
Date: 
 
 
Subject 
Number: 
 
 
 
Cohort 2 (AD Patients) – 10% BSA Treatment Example 
 
Please use the diagram below to mark the approximate location and size of the areas affected by atopic 
dermatitis in RED and record the areas to be treated with Study Ointment with cross hatches in 
BLACK.  Record the estimated BSA affected by atopic dermatitis (if applicable) and also the estimated 
BSA to be treated with Study Ointment (to nearest whole number). 
 
BSA Affected by AD_____20_______  %  
BSA Treated with Study Ointment ____10________  % 
 
 
 
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APPENDIX 7 – COHORT 2 40% BSA TREATMENT BODY MAP 
EXAMPLE 
Site: 
 
Subject 
Initials: 
 
Date: 
 
 
Subject 
Number: 
 
 
 
 
Cohort 2 (AD Patients) – 40% BSA Treatment Example 
 
Please use the diagram below to mark the approximate location and size of the areas affected by atopic 
dermatitis in RED and record the areas to be treated with Study Ointment with cross hatches in 
BLACK.  Record the estimated BSA affected by atopic dermatitis (if applicable) and also the estimated 
BSA to be treated with Study Ointment (to nearest whole number). 
 
BSA Affected by AD_____20_______  %  
BSA Treated with Study Ointment ____40_______  % 
 
 
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APPENDIX 8 – COHORT 3 BODY MAP EXAMPLE 
Site: 
 
Subject 
Initials: 
 
Date: 
 
 
Subject 
Number: 
 
 
 
Cohort 3 (AD Patients) –Example 
 
Please use the diagram below to mark the approximate location and size of the areas affected by atopic 
dermatitis in RED and record the areas to be treated with Study Ointment with cross hatches in 
BLACK.  Record the estimated BSA affected by atopic dermatitis (if applicable) and also the estimated 
BSA to be treated with Study Ointment (to nearest whole number). 
 
BSA Affected by AD_____20_______  %  
BSA Treated with Study Ointment ____20_______  % 
 
 
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APPENDIX 9 – ZIARCO ITCH QUESTIONNAIRE 
1. On a scale of 0 (No Itching) to 10 (Itching as bad as you can imagine), please rate the WORST 
itching that you felt over the last 24 hours.  
 
 
 
 
 
 
 
 
 
 
 
 
 
        0              1              2              3              4              5              6              7              8              9              10 
No Itching                                                                                                                                              Itching as bad  
                                                                                                                                                                 as you can 
                                                                                                                                                                     imagine 
 
 
2. Over the last 24 hours approximately how many hours, if any, did you itch?    ___  hours  
 
3.  On a scale of 0 (No sleep disturbance) to 10 (Awake all night), please rate how much your sleep was 
disturbed by itch last night.  
 
 
        0              1              2              3              4              5              6              7              8              9              10 
No sleep                                                                                                                                                     Awake all 
Disturbance                                                                                                                                                   night 
 
4. Over the last 24 hours how would you rate your itch?  
 
No itch 
Mild 
Moderate 
Severe 
 
 
 
 
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APPENDIX 10 – BODY MAP 
Site: 
 
Subject 
Initials: 
 
Date: 
 
 
Subject 
Number: 
 
 
Please use the diagram below to mark the approximate location and size of the areas affected by atopic 
dermatitis in RED and record the areas to be treated with Study Ointment with cross hatches in 
BLACK.  Record the estimated BSA affected by atopic dermatitis (if applicable) and also the estimated 
BSA to be treated with Study Ointment (to nearest whole number). 
 
BSA Affected by AD______________  %  
 
BSA Treated with Study Ointment ______________  % 
 
 
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APPENDIX 11 - EASI 
An EASI score is a tool used to measure the severity and extent of atopic eczema 
(Eczema Area and Severity Index).  
The body is divided into 4 sections of total skin area: head including neck (10%); 
arms including extremities (20%); trunk (30%) and legs including extremities (40%). 
Each area is scored and the 4 scores are combined into the final EASI 
Within each area, the severities of eczema are determined for four clinical signs: 
erythema (E), oedema/papulation (O), excoriation (X) and lichenification (L). 
Severity parameters are measured on a scale of 0 to 3, from none to severe.  Half 
scores may be used. 
 
 
 
For guidance, the following definitions may be used: 
 
Severity 
Erythema 
Oedema/Papulation 
Excoriation 
Lichenification 
None 
No evidence of 
erythema 
 
No evidence of 
oedema/papulation 
 
No evidence of 
excoriation 
 
No evidence of 
lichenification 
 
Mild 
Faintly 
detectable, pink 
 
Barely perceptible 
elevation 
Scant, superficial 
excoriations 
Slight thickening 
of the skin with 
skin markings 
minimally 
exaggerated 
Moderate 
Clearly 
distinguishable 
dull red 
 
Clearly perceptible 
elevation but not 
prominent 
Many superficial 
and /or some 
deeper 
excoriations 
Clearly thickened 
skin with 
exaggerated skin 
markings and/or 
some prurigo 
nodules 
Severe 
Deep dark or 
fiery bright red 
 
Prominent elevation 
Diffuse extensive 
superficial and 
/or many deep 
excoriations 
Prominent skin 
thickening with 
exaggerated skin 
markings creating 
deep furrows 
and/or many 
prurigo nodules 
For each section, the percentage area of skin involved with eczema is estimated and 
transformed into an extent grade from 0 to 6: 
• 
0% of involved area, grade: 0 
• 
1-9% of involved area, grade: 1 
• 
10-29% of involved area, grade: 2 
• 
30-49% of involved area, grade: 3 
• 
50-69% of involved area, grade: 4 
• 
70-89% of involved area, grade: 5 
Score 
Grade 
0 
None 
1 
Mild 
2 
Moderate 
3 
Severe 
 
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• 
90-100% of involved area, grade: 6 
The sum of all four severity parameters is calculated for each section of skin and then 
multiplied by the weight of the respective section (0.1 for head, 0.2 for arms, 0.3 for 
trunk and 0.4 for legs).  This value is then multiplied by the extent score for that body 
area. 
 
The EASI score is calculated using the following formula: 
EASI = [0.1 x (Ehead+ Ohead+Xhead + Lhead) x Extenthead] + [0.2 x Earms+ Oarms+Xarms + 
Larms) x Extentarms] + [0.3 x (Etrunk + Otrunk  + Xtrunk +Ltrunk) x Extenttrunk] + [0.4 x (Elegs 
+Olegs +Xlegs +Llegs) x Extentlegs] 
 
The EASI value can range from 0-72 
 
Body 
Region 
Erythema 
(E) (0-3) 
Oedema/ 
papulation 
(O) (0-3) 
Excoriation 
(X) (0-3) 
Lichenification 
(L) (0-3) 
Extent 
Grade for 
Region (0-6) 
Multiplier 
Score per 
Body 
Region 
Head 
(           + 
+ 
+ 
) 
x 
x 0.1 
= 
Arms 
(           + 
+ 
+ 
) 
x 
x 0.2 
= 
Trunk 
(           + 
+ 
+ 
) 
x 
x 0.3 
= 
Legs 
(           + 
+ 
+ 
) 
x 
x 0.4 
= 
The final EASI score is the sum of the 4 region scores: 
 
 
 
 
 
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APPENDIX 12 – % BODY SURFACE AREA GUIDANCE 
 
 
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APPENDIX 13 – ZIARCO OINTMENT QUESTIONNAIRE 
 
1. Did you find the study ointment easier to apply than your usual topical treatment 
or emollients? 
 
□ Yes 
 
 
□ No  
 
□ No Difference 
 
2. On a scale of 1 to 5 where 1 is Very Good and 5 is Very Poor how would you rate 
the study ointment for the following: 
 
Texture? 
□ 1 Very Good       □ 2 Good       □ 3 Indifferent       □ 4 Poor       □ 5 Very Poor 
 
Smell? 
□ 1 Very Good       □ 2 Good       □ 3 Indifferent       □ 4 Poor       □ 5 Very Poor 
 
Absorption into your skin? 
□ 1 Very Good       □ 2 Good       □ 3 Indifferent       □ 4 Poor       □ 5 Very Poor 
 
3. Would you be happy to wear your own clothes after application of the ointment if 
your clothes were in contact with the skin which had been treated with study 
ointment? 
 
□ Yes 
 
 
□ No  
 
□ Don’t Know 
 
4. Any other thoughts or feedback on the study ointment? 
 
 
 
 
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APPENDIX 14 – PROTOCOL AMENDMENT 1 CHANGES 
Protocol Final version 2.0 / 28 July 2016 contains protocol amendment 1 changes 
 
Summary of Changes 
 
Rationale for Substantial Amendment 1: The purpose of this amendment is to 
document a change to the study emollient for Cohort 3 patients as a consequence of 
unforeseen hypersensitivity reactions to the 
 study emollient by some 
patients in Cohort 2. 
 
Listing of changes made 
 
Section 3.2 Adaptive Categories and Features Table 2 Study Emollient 
Application 
Additional text 
Adaptive features: Cohort 3 only. Alternate emollients other than those defined as 
‘study emollients’ may be used. 
Boundaries: Stop the use of study emollient in the event of any hypersensitivity 
reactions caused by emollient. 
Change the emollient used from ‘study emollient’ due to any potential or actual 
hypersensitivity reactions with the approval of the Ziarco CMO (Cohort 3 only). 
 
Section 4.2 Inclusion Criteria: Cohorts 2 and 3 Criteria 8 
Original text 
 
 daily from at least 7 days prior to randomisation 
and throughout their participation in the study. 
 
New Text 
Patients must be willing to stop applying their daily emollients and instead use the 
study 
emollient twice daily from at least 7 days prior to randomisation and throughout their 
participation in the study. 
 
Section 4.4 Exclusion Criteria: Cohorts 2 and 3 Criteria 5 
Original text 
Hypersensitivity to any excipients in the study ointment formulation, study emollient 
 or study shower cream 
 
 
New Text 
Hypersensitivity to any excipients in the study ointment formulation, study emollient 
or study shower cream. 
 
Section 5.3 Drug Supplies 
Additional text 
Epaderm 3 in 1 ointment manufactured by
 
 will be supplied as an alternate study emollient to any AD patients (Cohort 
3 only) who have a potential (or known) hypersensitivity reaction to 
 for 
use prior to randomisation and throughout the study by MAC. 
 
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Section 5.7 Administration Emollients 
Original text 
The emollient 
 will be supplied for this study for AD patients in Cohorts 2 
and 3.  Any emollients and bathing products used by the patient should be 
documented at screening. Patient emollients may be used up until 7 days prior to Day 
1 and then patients must stop using their current emollient and switch to using the 
study emollient 
 in the same manner as their previous emollient.  From 
randomisation patients must use study emollient at least 15 minutes after application 
of study ointment.  Patients should use study emollient on all skin areas which have 
been treated with study ointment and in addition any areas which the patient would 
treat as part of their normal skin care routine.  Cohort 2 patients must continue using 
study emollient twice daily until discharge from the CRU on Day 9 and Cohort 3 
patients must use study emollient twice daily until they have completed their visit on 
Day 15. 
 
New Text 
The emollient 
 will be supplied for this study for AD patients in Cohorts 2 
and 3.  In addition an alternate emollient will be available for Cohort 3 patients who 
have a potential for any hypersensitivity skin reactions to 
  Any emollients 
and bathing products used by the patient should be documented at screening. Patient 
emollients may be used up until 7 days prior to Day 1 and then patients must stop 
using their current emollient and switch to using the study emollient in the same 
manner as their previous emollient.  From randomisation patients must use study 
emollient at least 15 minutes after application of study ointment.  Patients should use 
study emollient on all skin areas which have been treated with study ointment and in 
addition any areas which the patient would treat as part of their normal skin care 
routine.  Cohort 2 patients must continue using study emollient twice daily until 
discharge from the CRU on Day 9 and Cohort 3 patients must use study emollient 
twice daily until they have completed their visit on Day 15. 
 
Rationale for Non-Substantial Amendment: will be made to Exclusion criteria 23 
which excludes smokers from the study to only apply this exclusion to the Cohort 2 
in-patient cohort rather than the Cohort 3 the out-patient cohort. 
 
Listing of changes made 
 
Section 4.4 Exclusion Criteria: Cohorts 2 and 3 Criteria 23 
Original text 
Patients who are smokers (including patients who use nicotine substitutes). 
 
New Text 
Patients who are smokers (including patients who use nicotine substitutes). (Cohort 2 
only) 
 
Additional minor corrections/clarifications  
 
List of abbreviations and definitions of terms 
Original text 
EIA 
 
 Elisa immunoassay 
 
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New Text 
ECLIA 
Electro-chemiluminescence immunoassay 
Reason 
An oversight in the original protocol resulted in the wrong method assay for hepatitis 
C serology being listed.  This updates the abbreviation. 
 
Section 4.4 Exclusion Criteria: Cohorts 2 and 3 Criteria 24 onwards 
Numbering of exclusion criteria corrected to remove duplication of number 24. 
 
Section 6.1 Screening 
Original text 
• Blood and urine samples for: 
• Safety laboratory tests (haematology and clinical chemistry) 
New Text 
• Blood and urine samples for: 
• Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
Reason 
To ensure consistency on safety laboratory tests being conducted with Section 6.7 
Schedule of Activities 
 
Section 6.2.1 Day 1 assessments 
Original text 
• Obtain blood and urine samples: 
• Safety laboratory tests (haematology and clinical chemistry) 
New Text 
• Obtain blood and urine samples: 
• Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
Reason 
To ensure consistency on safety laboratory tests being conducted with Section 6.7 
Schedule of Activities 
 
Section 6.2.2 Days 2 to 6 Assessments 
Original text 
• Obtain blood and urine samples for: 
• Safety laboratory tests (Day 4 prior to first application only) 
New Text 
• Obtain blood and urine samples for  
• Safety laboratory tests (haematology, clinical chemistry and urinalysis) - Day 
4 prior to first application only 
Reason 
To ensure consistency on safety laboratory tests being conducted with Section 6.7 
Schedule of Activities 
 
 
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Section 6.2.5 Day 9 Assessments 
Original text 
• Obtain blood and urine samples for: 
• Safety laboratory tests  
New Text 
• Obtain blood and urine samples for  
• Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
Reason 
To ensure consistency on safety laboratory tests being conducted with Section 6.7 
Schedule of Activities 
 
Section 6.3.1 Day 1 assessments 
Original text 
• Obtain blood and urine samples: 
• Safety laboratory tests (haematology and clinical chemistry) 
New Text 
• Obtain blood and urine samples: 
• Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
Reason 
To ensure consistency on safety laboratory tests being conducted with Section 6.7 
Schedule of Activities 
 
Section 6.3.2 Day 2 to 6 assessments 
Original text 
• Obtain blood and urine samples for: 
• Safety laboratory tests (Day 4 prior to first application only) 
New Text 
• Obtain blood and urine samples for  
• Safety laboratory tests (haematology, clinical chemistry and urinalysis) - Day 
4 prior to first application only 
Reason 
To ensure consistency on safety laboratory tests being conducted with Section 6.7 
Schedule of Activities 
 
Section 6.3.4 Day 8 assessments 
Original text 
• Duplicate supine blood pressure and pulse rate 
• Blood sample for pharmacokinetic analysis of ZPL-5212372 
• Completion of Ziarco Ointment Questionnaire 
New Text 
• Duplicate supine blood pressure and pulse rate 
 
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• Blood sample for pharmacokinetic analysis of ZPL-5212372 
• Triplicate 12-lead ECG (24 hours post dose only) 
• Completion of Ziarco Ointment Questionnaire 
Reason 
To ensure consistency on ECGs being conducted with Section 6.7 Schedule of 
Activities 
 
Section 6.4.1 Baseline Visit (Day 1) 
Original text 
• Completion of body map of skin areas affected by AD and identification and 
recording of skin area to be treated 
• Obtain blood and urine samples: 
o Safety laboratory tests (haematology and clinical chemistry) 
o Blood sample for pharmacokinetic analysis of ZPL-5212372 
o Blood sample for baseline reference.  Serum to be stored frozen at the 
MAC through to completion of the study for possible use as a baseline 
reference should additional safety laboratory tests or markers of 
disease activity be indicated. 
o Urine for pregnancy testing (pre-menopausal women only) 
 
New Text 
• Completion of body map of skin areas affected by AD and identification and 
recording of skin area to be treated 
• Breath alcohol testing 
• Obtain blood and urine samples: 
o Safety laboratory tests (haematology, clinical chemistry and urinalysis) 
o Blood sample for pharmacokinetic analysis of ZPL-5212372 
o Blood sample for baseline reference.  Serum to be stored frozen at the 
MAC through to completion of the study for possible use as a baseline 
reference should additional safety laboratory tests or markers of 
disease activity be indicated. 
o Urine for pregnancy testing (pre-menopausal women only) 
o Urine drug screen  
Reason 
To ensure consistency on ECGs being conducted with Section 6.7 Schedule of 
Activities 
 
Section 6.7 Schedule of Activities Table 9 Footnote 12 
Original text 
Assessment of local toleration on treated skin will be conducted prior to each dosing 
occasion and at 24, 36 and 48 hours post last dose. 
 
New Text 
 
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Assessment of local toleration on treated skin will be conducted prior to each dosing 
occasion and at 24 and 48 hours post last dose. 
 
Reason 
To ensure consistency on local toleration assessments being conducted with Section 
6.2.4  
 
Section 6.7 Schedule of Activities Table 10 Footnote 12 
Original text 
Study emollient to be used from day -6 onwards until discharge from the CRU on day 
9. 
 
New Text 
Study emollient to be used from day -7 onwards until discharge from the CRU on day 
9. 
 
Reason 
To ensure consistency with the rest of the protocol. 
 
Section 6.7 Schedule of Activities Table 11 Footnote 10 
Original text 
Study emollient to be used from day -6 onwards until day 14 final application of 
ointment. 
 
New Text 
Study emollient to be used from day -7 onwards until completion of visit on day 15. 
 
Reason 
To ensure consistency with the rest of the protocol. 
 
Section 7.13 Pharmacokinetic Blood Samples 
Original text 
A 5.0 mL sample, to provide a minimum of 2 mL plasma for pharmacokinetic 
analysis, will be collected into an appropriately labeled tube containing lithium 
heparin.  Blood samples will be centrifuged within 15 minutes of collection at 1700 g 
and 4°C for 10 minutes. Plasma will be transferred to appropriate labelled 
screw-capped polypropylene tubes and stored at approximately -20°C within 1 hour 
of collection. 
Plasma samples will be transferred to the bioanalytical laboratory on dry ice to 
maintain frozen conditions. A sample handling document and shipment details for the 
analytical laboratory will be provided to the sites at study initiation. 
Samples will be analyzed using a validated analytical method.  After completion of 
the study plasma samples may also be analyzed for metabolites and further evaluation 
of the assay performance. 
New Text 
A 5.0 mL sample, to provide a minimum of 2 mL plasma for pharmacokinetic 
analysis, will be collected into an appropriately labeled tube containing lithium 
heparin.  Blood samples will be centrifuged within 1 hour of collection at 1700 g and 
4°C for 10 minutes. Plasma will be transferred to appropriate labelled screw-capped 
polypropylene tubes and stored at approximately -20°C within 1 hour of collection.  
 
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Plasma samples will be transferred to the bioanalytical laboratory on dry ice to 
maintain frozen conditions. Shipment details for the analytical laboratory will be 
provided to the sites prior to shipping any samples. 
Samples will be analysed using a validated analytical method.  After completion of 
the study plasma samples may also be analysed for metabolites and further evaluation 
of the assay performance. 
Reason 
Updated information regarding centrifugation post clinical assay validation and 
correction of minor typographical errors. 
 
Section 7.14 Clinical Laboratory Safety Tests Table 12 
Original text 
Anti-hepatitis C virus serology (by multi-antigen EIA) 
New Text 
Anti-hepatitis C virus serology (by ECLIA) 
Reason 
Update a minor error in original protocol. 
 
Section 7.16.1.2 Assessment of Adverse Event Causality 
Original text 
The investigator will assess the causality / relationship between the study drug and the 
AE and record that assessment in the CRF. Causality will be shown as Probably 
related, Possibly related, or Not related. 
New Text 
The investigator will assess the causality / relationship between the study drug and the 
AE and record that assessment in the CRF. Causality will be shown as Probably 
related, Possibly related, Not related or Related. 
Reason 
Update a minor error or omission in original protocol and for consistency with SAE 
form. 
 
Section 7.16.1.4 Reporting Serious Adverse Events 
Original text 
The original SAE should be sent to the address below and Ziarco Chief Medical 
Officer informed by email. In addition, the event must be documented in the CRF. 
New Text 
The completed SAE should be sent to the address below and Ziarco Chief Medical 
Officer informed by email. In addition, the event must be documented in the CRF. 
Reason 
Updated for clarity to ensure no delays in reporting SAE’s as emailed or faxed copies 
are acceptable to ensure reporting timelines are met. 
 
Section 7.16.1.4 Reporting Serious Adverse Events 
Original text 
After receipt of the initial report, the pharmacovigilance team will review the 
information and, if necessary, contact the investigator, to obtain further information 
for assessment of the event.  
 will be responsible for all 
information processing and reporting according to local legal requirements. Where 
necessary, investigators will inform the authorities in their own countries. 
New Text 
 
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After receipt of the initial report, the pharmacovigilance team will review the 
information and, if necessary, contact the investigator, to obtain further information 
for assessment of the event.  
 will be responsible for all 
information processing and reporting according to local legal requirements.  
Reason 
Updated for clarity to ensure no confusion on regulatory reporting responsibilities. 
 
Section 7.16.1.4 Reporting Serious Adverse Events 
Original text 
After receipt of the initial report, the pharmacovigilance team will review the 
information and, if necessary, contact the investigator, to obtain further information 
for assessment of the event.  
 will be responsible for all 
information processing and reporting according to local legal requirements. Where 
necessary, investigators will inform the authorities in their own countries. 
New Text 
After receipt of the initial report, the pharmacovigilance team will review the 
information and, if necessary, contact the investigator, to obtain further information 
for assessment of the event.  
 will be responsible for all 
information processing and reporting according to local legal requirements.  
Reason 
Updated for clarity and consistency with SAE reporting plan to ensure no confusion 
on regulatory reporting responsibilities. 
 
Section 8.1.1. Randomisation 
Original text 
The randomisation for Cohort 3 will be stratified by EASI score (EASI ≤20 and EASI 
≥20). 
New Text 
The randomisation for Cohort 3 will be stratified by EASI score (EASI ≤20 and EASI 
>20). 
Reason 
Correct a mistake in the original protocol. 
 
Section Appendix 11 - EASI 
Original Text 
Within each area, the severities of eczema are determined for four clinical signs: 
erythema (E), oedema/papulation (O), excoriation (X) and lichenification (L). 
Severity parameters are measured on a scale of 0 to 3, from none to severe. 
New Text 
Within each area, the severities of eczema are determined for four clinical signs: 
erythema (E), oedema/papulation (O), excoriation (X) and lichenification (L). 
Severity parameters are measured on a scale of 0 to 3, from none to severe.  Half 
scores may be used. 
Reason 
To highlight the fact that half scores can be used in the calculation of the EASI and 
for consistency with the IWRS system where the EASI data is collected for the study. 
 
 
 
Page 109 of 109 
 

SAP_000.pdf: 
Statistical Analysis Plan
Page 1 of 43 
Sponsor Name: 
Ziarco Pharma Ltd 
Protocol Number: 
ZPL521/101 / NCT02795832
Protocol Title: 
A Randomised, Adaptive Design, Double-Blind (3rd 
Party Open), Placebo Controlled, Sequential Group 
Study To Determine The Safety, Tolerability, 
Pharmacokinetics And Efficacy Of Twice Daily 
Application Of A Topical ZPL-5212372 (1.0% W/W) 
Ointment Administered For Up To 2 Weeks In Adult 
Healthy Volunteers And Patients With Moderate To 
Severe Atopic Dermatitis 
Protocol Version and Date: 
1.0 (8th March 2016) 
2.0 (28th July 2016) 
Protocol Amendment numbers 
and dates: 
N/A 
Author(s): 
SAP Version: 
Final v2.0 
SAP Version Date: 
16th March 2017
Notice of Confidential and Proprietary Information: 
This document is confidential. It contains proprietary information of Ziarco Pharma Ltd. Any 
viewing or disclosure of such information that is not authorized in writing by Ziarco Pharma Ltd is 
strictly prohibited. Such information may be used solely for the purpose of reviewing or 
performing this study. 


 
 
   
 
 


Ziarco Pharma Ltd 
 
Date: 16th March 2017 
ZPL521/101                                                                          
Statistical Analysis Plan 
 
 
SAP Version v2.0     
 
Page 3 of 43 
TABLE OF CONTENTS 
1. 
GLOSSARY OF ABBREVIATIONS .............................................................................. 6 
2. 
INTRODUCTION ..................................................................................................... 8 
2.1. 
Responsibilities ........................................................................................................................ 8 
3. 
STUDY OVERVIEW ................................................................................................. 8 
3.1. 
Study Objectives ...................................................................................................................... 8 
3.1.1. 
Primary Objectives ............................................................................................................... 8 
3.1.2. 
Secondary Objectives ........................................................................................................... 8 
3.1.3. 
Exploratory Objectives .......................................................................................................... 9 
3.2. 
Study Design ............................................................................................................................ 9 
3.3. 
Determination of Sample Size ................................................................................................. 10 
3.4. 
Treatment Assignment and Blinding ....................................................................................... 10 
3.5. 
Administration of Study Medication ....................................................................................... 11 
3.6. 
Study Flowchart ..................................................................................................................... 13 
4. 
ENDPOINTS ......................................................................................................... 17 
4.1. 
Primary Efficacy Endpoint ....................................................................................................... 17 
4.2. 
Secondary Efficacy Endpoints ................................................................................................. 17 
4.3. 
Exploratory Endpoint ............................................................................................................. 19 
4.4. 
Safety Endpoints .................................................................................................................... 19 
4.5. 
Pharmacokinetic Endpoints .................................................................................................... 19 
5. 
ANALYSIS SETS ..................................................................................................... 21 
5.1. 
Screened Population .............................................................................................................. 21 
5.2. 
Randomised Set ..................................................................................................................... 21 
5.3. 
Full Analysis Set (FAS) ............................................................................................................ 21 
5.4. 
Per Protocol (PP) Analysis Set ................................................................................................. 21 
5.5. 
Safety Analysis Set ................................................................................................................. 21 


Ziarco Pharma Ltd 
 
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5.6. 
Pharmacokinetic (PK) Analysis Set .......................................................................................... 22 
5.7. 
Protocol Deviations ................................................................................................................ 22 
6. 
GENERAL ASPECTS FOR STATISTICAL ANALYSIS .................................................... 22 
6.1. 
General Methods ................................................................................................................... 22 
6.2. 
Missing Data .......................................................................................................................... 23 
6.2.1. 
Efficacy Data ...................................................................................................................... 23 
6.2.2. 
Pharmacokinetic Data ........................................................................................................ 24 
6.2.3. 
Safety Data ........................................................................................................................ 24 
6.3. 
Visit Windows ........................................................................................................................ 25 
6.4. 
Stratification .......................................................................................................................... 25 
6.5. 
Subgroups ............................................................................................................................. 25 
7. 
DEMOGRAPHIC, OTHER BASELINE CHARACTERISTICS AND MEDICATION .............. 25 
7.1. 
Patient Disposition and Withdrawals ...................................................................................... 25 
7.2. 
Demographic and Other Baseline Characteristics ..................................................................... 26 
7.3. 
Medical History and Concomitant Diseases ............................................................................. 27 
7.4. 
Prior and concomitant Medications ........................................................................................ 27 
8. 
EFFICACY ............................................................................................................. 28 
8.1. 
Primary Efficacy Endpoint and Analysis ................................................................................... 28 
8.1.1. 
Sensitivity Analyses of the Primary Endpoint ........................................................................ 28 
8.1.2. 
Additional Analyses of the Primary Endpoint ........................................................................ 29 
8.2. 
Secondary Endpoints and Analysis .......................................................................................... 29 
8.2.1. 
EASI-50 and EASI-75 ........................................................................................................... 29 
8.2.2. 
EASI Components ............................................................................................................... 30 
8.2.3. 
Investigators Global Assessment (IGA) ................................................................................. 30 
8.2.4. 
Numerical Rating Scale (NRS) .............................................................................................. 30 
8.2.5. 
Verbal Rating Score (VRS) ................................................................................................... 31 
8.2.6. 
Patient Global Impression of Change (PGIC) ......................................................................... 31 
8.2.7. 
Body Surface Area (BSA) ..................................................................................................... 31 
8.3. 
Exploratory Endpoint ............................................................................................................. 31 
8.3.1. 
Ziarco Ointment Questionnaire ........................................................................................... 31 
9. 
ANALYSIS OF PHARMACOKINETICS ...................................................................... 31 
9.1. 
Concentration data ................................................................................................................ 31 


Ziarco Pharma Ltd 
 
Date: 16th March 2017 
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9.2. 
Pharmacokinetic Parameter Data ........................................................................................... 32 
10. 
SAFETY ............................................................................................................ 32 
10.1. 
Draize Score ........................................................................................................................... 32 
10.2. 
Extent of Exposure ................................................................................................................. 32 
10.3. 
Adverse Events ...................................................................................................................... 33 
10.4. 
Laboratory Evaluations .......................................................................................................... 34 
10.5. 
Vital Signs .............................................................................................................................. 35 
10.6. 
ECG ....................................................................................................................................... 36 
10.7. 
Physical Examination ............................................................................................................. 37 
11. 
INTERIM ANALYSES .......................................................................................... 37 
12. 
CHANGE FROM ANALYSIS PLANNED IN PROTOCOL ........................................... 38 
13. 
REFERENCE LIST ............................................................................................... 38 
14. 
INDEX OF TABLES ............................................................................................. 39 
15. 
INDEX OF LISTINGS ........................................................................................... 42 
16. 
INDEX OF FIGURES ........................................................................................... 43 
 
 
 


Ziarco Pharma Ltd 
 
Date: 16th March 2017 
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1. 
GLOSSARY OF ABBREVIATIONS 
Abbreviation 
Description 
AD 
Atopic Dermatitis 
AE 
Adverse Event 
ANCOVA 
Analysis of Covariance 
ATC 
Anatomic Therapeutic Chemical 
AUC 
Area Under the Curve 
BLQ 
Below Limit of Quantification 
BSA 
Body Surface Area 
CI 
Confidence Interval 
CRU 
Clinical Research Unit 
CSR 
Clinical Study Report 
CV 
Coefficient of Variation 
EASI 
Eczema Area and Severity Index 
ECG 
Electrocardiograms 
FAS 
Full Analysis Set 
IGA 
Investigator Global Assessment 
IMP 
Investigational Medicinal Product 
IWRS 
Integrated Web-based Response System 
LOQ 
Limit of Quantification 
LLOQ 
Lower Limit of Quantification 
LOCF 
Last Observation Carried Forward 
LS 
Least Square 
MCMC 
Markov Chain Monte Carlo 
MedDRA 
Medical Dictionary for Regulatory Activities 
NRS 
Numerical Rating Scale 
QTcF 
Heart Rate Corrected QT interval, using Fridericia’s formula 
PGIC 
Patient Global Impression of Change 
PK 
Pharmacokinetics 
PP 
Per Protocol 


Ziarco Pharma Ltd 
 
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Abbreviation 
Description 
PT 
Preferred Term 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SD 
Standard Deviation 
SOC 
System Organ Class 
 
 
TEAE 
Treatment Emergent Adverse Event 
TFLs 
Tables, Figures, Listings 
VRS 
Verbal Rating Score 
WHO 
World Health Organization 
 
 
 


Ziarco Pharma Ltd 
 
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2. 
INTRODUCTION 
The purpose of this statistical analysis plan (SAP) is to ensure that the data listings, summary 
tables and figures which will be produced, and the statistical methodologies that will be used, are 
complete and appropriate to allow valid conclusions regarding the study objectives.  Any 
amendments to the SAP will be made prior to database lock. Any additional analyses not 
described in the final SAP or deviations from the final SAP will be documented in the clinical study 
report (CSR). 
2.1. 
RESPONSIBILITIES 
 will perform the statistical analyses and are responsible for the 
production and quality control of all tables, figures and listings (TFLs).  Pharmacokinetic (PK) 
parameters will be derived by 
 and provided to 
 for presentation.  
3. 
STUDY OVERVIEW 
3.1. 
STUDY OBJECTIVES 
3.1.1. 
Primary Objectives 
Cohorts 1 and 2 
To assess the safety and tolerability, local and systemic, of ZPL-5212372 administered as a topical 
ointment (containing 1.0% [w/w] concentration of ZPL-5212372) twice daily, for 1 week, to 
healthy subjects and patients with moderate to severe atopic dermatitis (AD). 
Cohort 3 
To evaluate the efficacy of ZPL-5212372 administered as a topical ointment (containing 1.0% 
[w/w] concentration of ZPL-5212372) twice daily, for 2 weeks, to patients with moderate to 
severe AD. 
3.1.2. 
Secondary Objectives 
Cohorts 1 and 2 
To investigate the systemic pharmacokinetics of ZPL-5212372 following single and multiple dose 
topical applications of a 1.0% [w/w] concentration of ZPL-5212372 ointment, administered twice 
daily, for 1 week, to healthy subjects and patients with moderate to severe AD. 
Cohort 3 
To assess the safety and tolerability, local and systemic, of ZPL-5212372 administered as a topical 
ointment (containing 1.0% [w/w] concentration of ZPL-5212372) twice daily, for 2 weeks, to 
patients with moderate to severe AD. 
To investigate the trough concentrations of ZPL-5212372 in patients with moderate to severe AD 


Ziarco Pharma Ltd 
 
Date: 16th March 2017 
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following twice daily applications of ZPL-5212372 administered as a topical ointment (containing 
1.0% [w/w] concentration of ZPL-5212372). 
3.1.3. 
Exploratory Objectives 
Cohorts 1 to 3 
To explore the subject’s perception of the Ziarco development ointment formulation using the 
Ziarco Ointment Questionnaire. 
3.2. 
STUDY DESIGN 
This is a randomised, adaptive design, double-blind (3rd party open), placebo controlled, 
sequential group study in adult healthy volunteers and in patients with moderate to severe AD. 
The study consists of 3 sequential cohorts each separated by formal safety reviews conducted by 
the Safety Review Team.  The study will recruit cohorts sequentially and use sentinel subjects 
(n=3) in Cohorts 1 and 2 to minimize the risk to subjects and make it possible to modify specific 
features of the study dependent on emerging data; however the flexibility of the adaptive 
features will be constrained by clear boundaries/stopping criteria (refer to Protocol Sections 3.2 
and 3.7).  The main features of each cohort are summarised in Table 1 below: 
Table 1: Overview of Cohorts 
 
Cohort 1 
Cohort 2 
Cohort 3 
Subject Population 
Healthy adults 
Moderate to severe 
AD patients 
Moderate to severe 
AD patients 
In-patients/Out-patients 
In-patients 
In-patients 
Out-patients 
Total Number of subjects 
12 
12 
30 
Ratio Active:Placebo 
2:1 
2:1 
2:1 
Dosing Duration 
7 days 
7 days 
14 days 
Follow-up Visit (after last 
dose of study medication) 
7-14 days 
7-14 days 
7-14 days 
 
Progression from Cohort 1 to Cohort 3 will be dependent on the demonstration of adequate 
safety within the protocol defined boundaries. In Cohorts 1 and 2, complete data sets from a 
minimum of 4 active and 2 placebo subjects with 40% body surface area (BSA) treated with study 
ointment will need to be reviewed by the Safety Review Team in order to decide whether to 
proceed as planned to the next cohort.  In the event that it is decided not to proceed to the next 
cohort, the reviewers may decide to: 
 
modify the study within the boundaries of the adaptive features and continue,  


Ziarco Pharma Ltd 
 
Date: 16th March 2017 
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 
to submit a substantial protocol amendment, or  
 
suspend the study.  
In addition, in order to ensure subject safety, Cohorts 1 and 2 will be split to have an initial 
sentinel group which will precede the remaining subjects in the cohort.  The sentinel group will 
consist of 3 subjects (2:1 Active to Placebo) and these subjects will be dosed twice daily, to cover 
10% BSA with study ointments for 7 days.  Provided that the study ointment is well tolerated after 
48 hours (i.e. 4 applications) the remainder of the cohort (9 subjects; 2:1 Active to Placebo) may 
start dosing with coverage of 40% BSA BID for 7 days.  Cohort 3 will have AD affected skin dosed 
to a maximum of 50% BSA.   
All subjects will have a screening visit to confirm suitability to enter the study, within 28 days of 
Day 1 (first day of dosing).  AD patients in Cohorts 2 and 3 will be required to switch from their 
usual emollient to using the study emollient twice daily for at least 7 days prior to Day 1 and 
throughout their participation in the study.  Dosing will commence on Day 1 and continue for 7 or 
14 days, depending on which cohort subjects are allocated to.  Subjects in Cohorts 1 and 2 will 
remain in the clinical research unit (CRU) until Day 9 (48 hours after the last application of study 
ointment).  Subjects in Cohort 3 will have their first application of ointment in the unit and 
continue dosing at home, returning to the unit for assessments on Days 5, 8, 10 and 15 (after the 
last ointment application at home on the evening of Day 14). 
All subjects will be required to attend the CRU for a follow-up visit 7-14 days after the last 
application of study ointment.  AD patients may recommence use with other topical or systemic 
treatments for their AD, if required, from either discharge from the CRU (Cohort 2) or the end of 
dosing (Cohort 3). 
3.3. 
DETERMINATION OF SAMPLE SIZE 
The sample sizes for Cohorts 1 and 2 are not based on formal statistical testing. Their sample sizes 
are considered sufficient to obtain suitable safety, tolerability and PK data prior to starting Cohort 
3. 
For Cohort 3 the sample size of 30 subjects has been determined on the basis of comparing the 
mean ZPL-5212372 Week 2 percentage change from baseline in the Eczema Area and Severity 
Index (EASI) score to that for placebo. With 20 subjects in the ZPL-5212372 group and 10 subjects 
in the placebo group, there is 80% power of obtaining a statistically significant result, assuming 
that there is an improvement of at least 30% with ZPL-5212372 compared to placebo and a 
standard deviation for the percentage changes from baseline between subjects of 30%. This is 
based on the analyses described and a 1-sided test at the 5% level of significance.  
3.4. 
TREATMENT ASSIGNMENT AND BLINDING 
Upon screening, each subject will receive a 3-digit number, by the Integrated Web-based 
Response System (IWRS), which will be used throughout the study. Screened subjects who drop 
out of the study before randomisation will retain their number. 


Ziarco Pharma Ltd 
 
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Subjects will be randomised on a 2:1 basis to topical ZPL-5212372 or placebo, twice daily. For the 
randomisation of subjects, the investigator will use the IWRS. The IWRS will use a computer-
generated randomisation schedule to assign subjects to treatment.  In Cohort 3 subjects will be 
stratified by baseline EASI score (≤20 and >20), to ensure an even distribution of severities across 
both treatment groups. 
The study is double-blind third party open i.e. the study will be subject- and investigator-blinded.  
Details of personnel who will be able to review unblinded subjects’ data during the study will be 
held in a separate document by Ziarco. 
The study blind should not be broken except in a medical emergency, where knowledge of the 
study drug received would affect the treatment of the emergency, or obtaining this information is 
a regulatory requirement. If time permits, the investigator should notify the Sponsor prior to 
contacting IWRS. All calls resulting in an unblinding event will be reported by the IWRS to the 
Sponsor.  If the blind is broken, the date, time and reason will be recorded in the subject’s CRF. 
3.5. 
ADMINISTRATION OF STUDY MEDICATION 
Subjects in Cohorts 1 and 2 will have twice daily topical applications of either 1.0% (w/w) 
ZPL-5212372 ointment or matching placebo ointment approximately 12 hours apart.  The 
morning dose will be administered between 07:00 to 10:00 hours and the evening dose 
approximately 12 hours later between 19:00 – 22:00 hours. Study ointment will be dispensed 
from the pharmacy (one jar per subject) to clinical staff.  Clinical staff will consult the subject’s 
body map to confirm whether 10 or 40% BSA will have application of ointment.  For 10% BSA 
approximately 
 and for 
40% BSA approximately 
 by 
clinic staff at the subject’s bedside.  If the initial volume of ointment is inadequate to cover the 
dosing area, additional ointment may be applied up to a maximum of 1 mL/2% BSA (i.e. 5 mL for 
10% BSA or 20 mL for 40% BSA). 
Subjects in Cohort 3 will apply twice daily (approximately 12 hours apart) applications of either 
1.0% (w/w) ZPL-5212372 ointment or matching placebo ointment at home.  At the randomisation 
visit the extent of their AD will be assessed and a recommended amount (approximately 1.25 to 5 
mL/10% BSA) of ointment for use on their AD affected skin will be recorded on their individual 
body map.  Subjects will be instructed not to use ointment on non-AD affected skin. Study 
ointment will be dispensed from the pharmacy (three jars per subject) to clinical staff.  The first 
application of study ointment will be applied by the subject in the clinic under the supervision and 
direction of the clinical staff i.e. the amount to apply, body area to cover using the body map as a 
guide and how to draw up the correct amount of study ointment 
 will be explained, as 
this will vary from subject to subject.  
Emollients  
The emollient 
 will be supplied for this study for AD subjects in Cohorts 2 and 3.  In 
addition an alternate emollient will be available for Cohort 3 subjects who have a potential for 


Ziarco Pharma Ltd 
 
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any hypersensitivity skin reactions to 
 (Protocol version 2.0). Any emollients and bathing 
products used by the subject should be documented at screening. Subject emollients may be used 
up until 7 days prior to Day 1 and then subjects must stop using their current emollient and switch 
to using the study emollient in the same manner as their previous emollient.  From randomisation 
subjects must use study emollient at least 15 minutes after application of study ointment.  
Subjects should use study emollient on all AD affected skin areas which have not been treated 
with study ointment and in addition any areas which the subject would treat as part of their 
normal skin care routine (Protocol Section 3.2 Table 2 adaptive feature).  Cohort 2 subjects must 
continue using study emollient twice daily until discharge from the CRU on Day 9 and Cohort 3 
subjects must use study emollient twice daily until they have completed their visit on Day 15. 
Shower Cream 
 
subjects, for use when showering each morning whilst resident in the CRU. 
 
 


Ziarco Pharma Ltd 
 
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3.6. 
STUDY FLOWCHART 
Schedule of Activities For Cohort 1 (Healthy Volunteers) 
Activities 
Screening1 
Day 1 
 
Day 2 to Day 6 
 
Day 7 
 
Day 8 
Day 9/ Early Termination 
Follow up 7-14 days post last 
dose 
In-patient in CRU  
 
X2-------------------------------------------------------------------------------------------------------------X 
 
Informed Consent 
X 
 
 
 
 
 
 
Demographics  
X 
 
 
 
 
 
 
Medical history 
X 
X3 
 
 
 
 
 
Physical exam4 ( inc ht/wt) 
X 
X 
 
 
 
X 
X 
12-lead ECG5 
X 
X 
X 
X 
X 
X 
X 
Vital signs (BP and PR)6 
X 
X 
X 
X 
X 
X 
X 
Breath alcohol and drug screen 
X 
X 
 
 
 
 
 
Pregnancy Test7 
X 
X 
 
 
 
 
X 
Serum FSH8 
X 
 
 
 
 
 
 
Safety labs (inc urinalysis) 
X 
X9 
X10 
 
 
X 
X 
Hepatitis B/C, HbA1c, HIV 
X 
 
 
 
 
 
 
Fitzpatrick skin type assessment 
X 
 
 
 
 
 
 
Body Map Recorded 
 
X 
 
 
 
 
 
PK blood sample11 
 
X 
X 
X 
X 
X 
 
Assessment of local toleration12 
 
X 
X 
X 
X 
X 
 
Ointment Questionnaire 
 
 
 
 
X 
 
 
Randomisation 
 
X 
 
 
 
 
 
Dosing13 
 
X----------------------------------------------------X 
 
 
 
Concomitant & prior medications  
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
AE collection and review14 
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
1 
Screening may occur over more than 1 day. 
2 
Subjects may come into the CRU on the evening before and some pre-dose activities may be completed. 
3 
Review changes in the subject’s medical history since screening. 
4 
Full Physical examination at Screening, a brief examination on all other occasions (guided by AEs, but to include, as a minimum, general appearance, heart, lungs and skin). 
5 
Single ECG assessments at screening, follow up and early termination (if applicable).  Triplicate ECG assessments will be performed at pre-dose, 1, 2, 4, 8, 12, 13 (Day 1 only) and 24 hours post-morning dose on Day 1 and 
Day 7, prior to each dose on Days 2 to 6, and at 36 hours (Day 8) and at 48 hours (Day 9) post last dose. 
6 
Duplicate, supine only, vital sign assessments will be performed at Screening, pre-dose, 1, 2, 4, 8, 12, 13 (Day 1 only) and 24 hours post-morning dose on Day 1 and Day 7, prior to each dose on Days 2 to 6, and at 36 
hours (Day 8) and at 48 hours (Day 9) post last dose, follow up and at early termination (if applicable). 
7 
Serum pregnancy test at Screening, urine at all other specified time points; for women of child-bearing potential only. 
8 
Serum FSH testing for post-menopausal females 45-60 years only. 
9 
An additional blood sample will be taken at Baseline and stored frozen at MAC, as a reference sample, in case extra tests are required.  
10 An additional safety lab sample will be taken on Day 4. 
11 Blood samples for analysis of ZPL-5212372 will be taken prior to dosing and at 1, 2, 4, 8, 12, 13 (Day 1 only), 16 and 24 hours post-morning dose on Day 1 and Day 7, prior to each dose on Days 2 to 6, and at 36 hours 
(Day 8) and at 48 hours (Day 9) post last dose. 
12 Assessment of local toleration on treated skin will be conducted prior to each dosing occasion and at 24 and 48 hours post last dose. 
13 Dosing twice daily, 12 hours apart from Days 1 to 6, the last application of study medication will be the morning of Day 7. 
14 Adverse events will be collected from the signing of the consent form until the follow up visit. SAE’s collected up to 30 days post last dose. 


Ziarco Pharma Ltd 
 
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Schedule of Activities For Cohort 2 (Moderate To Severe AD Patients) 
Activities 
Screening1 
Washout Period2 
Day 1 
Day 2 to Day 6 
Day 7 
Day 8 
Day 9/ Early Termination 
Follow up 7-14 days post last dose 
In-patient in CRU 
 
 
X3-------------------------------------------------------------------------------------------------X 
 
Informed Consent 
X 
 
 
 
 
 
 
 
Demographics  
X 
 
 
 
 
 
 
 
Medical history 
X 
 
X4 
 
 
 
 
 
Physical exam5 (incl ht/wt) 
X 
 
X 
 
 
 
X 
X 
12-lead ECG6 
X 
 
X 
 
X 
 
X 
X 
Vital signs (BP and PR)7 
X 
 
X 
X 
X 
X 
X 
X 
Breath alcohol and drug screen 
X 
 
X 
 
 
 
 
 
Pregnancy Test8 
X 
 
X 
 
 
 
 
X 
Serum FSH9 
X 
 
 
 
 
 
 
 
Safety labs (inc urinalysis) 
X 
 
X10 
X11 
 
 
X 
X 
Hepatitis B/C, HbA1c, HIV 
X 
 
 
 
 
 
 
 
Fitzpatrick skin type assessment 
X 
 
 
 
 
 
 
 
Study emollient use12 
 
X------------------------------------------------------------------------------------------------------------------------X 
 
Body Map Recorded 
 
 
X 
 
 
 
 
 
PK blood sample13 
 
 
X 
X 
X 
X 
X 
 
EASI/IGA/BSA14 
X 
 
X 
 
X 
 
 
X 
Ointment Questionnaire 
 
 
 
 
 
X 
 
 
Randomisation 
 
 
X 
 
 
 
 
 
Dosing15 
 
 
X----------------------------------------X 
 
 
 
Concomitant & prior medications  
X------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
AE collection and review16 
X------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
1 
Screening may occur over more than 1 day. 
2 
Washout period from AD medication of between 1 week and 1 month dependent on each patient’s current AD medication.  Study Emollient should be started 7 days before Day 1. 
3 
Patients may come into the CRU on the evening before and some pre-dose activities may be completed. 
4 
Review changes in the subject’s medical history since screening. 
5 
Full Physical examination at Screening, a brief examination on all other occasions (guided by AEs, but to include, as a minimum, general appearance, heart, lungs and skin). 
6 
Single ECG assessment at screening, follow up and early termination (if applicable).  Triplicate ECG assessments will be performed at pre-dose, 1, 2, 4, 8, 12, 13 (Day 1 only) and 24 hours post-morning dose on Day 1 
and Day 7. 
7 
Duplicate, supine only, vital sign assessments will be performed at Screening,  pre-dose, 1, 2, 4, 8, 12, 13 (Day 1 only) and 24 hours post-morning dose on Day 1 and Day 7, prior to each dose on Days 2 to 6, and at 
36 hours (Day 8) and at 48 hours (Day 9) post last dose and at follow up. 
8 
Serum pregnancy test at Screening, urine at all other specified time points; for women of child-bearing potential only. 
9 
Serum FSH testing for post-menopausal females 45-60 years only. 
10 
An additional blood sample will be taken at Baseline and stored frozen at the MAC, as a reference sample, in case extra tests are required.  
11 
An additional safety lab sample will be taken on Day 4. 
12 
Study emollient to be used from day -7 onwards until discharge from the CRU on day 9. 
13 
Blood samples for analysis of ZPL-5212372 will be taken prior to dosing and at 1, 2, 4, 8, 12, 13 (Day 1 only), 16 and 24 hours post-morning dose on Day 1 and Day 7, prior to each dose on Days 2 to 6, and at 36 
hours (Day 8) and at 48 hours (Day 9) post last dose. 


Ziarco Pharma Ltd 
 
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14 
EASI/IGA/BSA assessed at screening and prior to randomisation to ensure inclusion criteria are met.  Assessed again at the end of the study (or early termination) to document severity and extent of AD following 
study medication. 
15 
Dosing twice daily, 12 hours apart from Days 1 to 6, the last application of study medication will be the morning of Day 7. 
16 
Adverse events will be collected from the signing of the consent form until the follow up visit. SAE’s collected up to 30 days post last dose. 
 


Ziarco Pharma Ltd 
 
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Schedule of Activities For Cohort 3 (Moderate To Severe AD Patients) 
Activities 
Screening1 
Washout 
Period2 
Baseline Visit 1 
(Day 1) 
Visit 2 
(Day 5) 
Visit 3 
(Day 8) 
Visit 4 
(Day 10) 
Visit 5 
(Day 15)/ Early Termination 
Follow up 7-14 days 
post last dose 
Informed Consent 
X 
 
 
 
 
 
 
 
Demographics  
X 
 
 
 
 
 
 
 
Medical history 
X 
 
X3 
 
 
 
 
 
Physical exam4 ( incl ht/wt) 
X 
 
X 
 
 
 
X 
X 
12-lead ECG5 
X 
 
 
 
 
 
 
X 
Vital signs (BP and PR)6 
X 
 
X 
X 
X 
X 
X 
X 
Breath alcohol and drug screen 
X 
 
X 
 
 
 
 
 
Pregnancy Test7 
X 
 
X 
 
 
 
X 
X 
Serum FSH8 
X 
 
 
 
 
 
 
 
Safety labs (inc urinalysis) 
X 
 
X9 
 
 
 
X 
X 
Hepatitis B/C, HbA1c, HIV 
X 
 
 
 
 
 
 
 
Fitzpatrick skin type assessment 
X 
 
 
 
 
 
 
 
Study emollient use10 
 
X---------------------------------------------------------------------------------------------------------------------------X 
 
Body Map Recorded 
 
 
X 
 
 
 
 
 
PK blood sample11 
 
 
X 
X 
X 
X 
X 
 
EASI/BSA/IGA 
X 
 
X 
X 
X 
X 
X 
X 
Ziarco Itch Questionnaire12 
 
 
X 
X 
X 
X 
X 
X 
PGIC 
 
 
 
 
 
 
X 
 
Ointment Questionnaire 
 
 
 
 
 
 
X 
 
Randomisation 
 
 
X 
 
 
 
 
 
Dosing13 
 
 
X-------------------------------------------------------------------------X 
 
 
Concomitant & prior medications  
X------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
AE collection and review14 
X------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X 
1. 
Screening may occur over more than 1 day.  
2. 
Washout period from AD medication of between 1 week and 1 month dependent on each patient’s current AD medication. Study Emollient should be started 7 days before Day 1. 
3. 
Review changes in the patient’s medical history since screening. 
4. 
Full Physical examination at Screening, a brief examination at all other visits (guided by AEs, but to include, as a minimum, general appearance, heart, lungs and skin). 
5. 
A triplicate ECG assessment will be performed at Screening and a single ECG at Follow Up. 
6. 
Duplicate, supine only, vital sign assessments to be performed at Screening and pre-dose on days 1, 5, 8, 10, 15 and follow-up. 
7. 
Serum pregnancy test at Screening, urine at all other specified time points; for women of child-bearing potential only. 
8. 
Serum FSH testing for post-menopausal females 45-60 years only. 
9. 
An additional blood sample will be taken at Baseline and stored frozen at the central lab, as a reference sample, in case extra tests are required.  
10. Study emollient to be used from day -7 onwards until completion of visit on day 15. 
11. Blood samples for analysis of ZPL-5212372 will be taken prior to dosing on days 1, 5, 8 and 10 and an approximately 12 hours post last dose sample will be taken on Day 15. 
12. Pruritus NRS, VRS, duration and impact of itch on sleep disturbance for the previous 24 hour period at clinic visits (Days 1, 5, 8, 10 and 15) by clinical staff asking the patients to complete the Ziarco Itch 
Questionnaire (Appendix 9). 
13. To be dosed topically twice daily at home except on study visit days when the morning dose will be applied in the clinic after all other study assessments have been completed. Medication should be applied in 
the morning (preferably after showering) and approximately 12 hours later in the evening.  Study medication should be applied approximately 15 minutes prior to using the Study Emollient. Subjects will apply 
their last dose of study medication at home in the evening of day 14 prior to their visit to the clinic on day 15. 
14. Adverse events will be collected from the signing of the consent form until the follow up visit. SAE’s collected up to 30 days post last dose.  


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4. 
ENDPOINTS 
As Cohort 1 includes healthy volunteers no efficacy data will be collected.  Though some efficacy 
assessments are included for Cohort 2 the data will only be listed and no formal comparisons will 
be made between the treatment groups.  Consequently efficacy endpoints will only be assessed 
for Cohort 3. 
4.1. 
PRIMARY EFFICACY ENDPOINT 
The primary efficacy endpoint for Cohort 3 is the percentage change from baseline to Week 2 in 
the EASI score.  
The EASI is a validated tool used to measure the severity and extent of atopic eczema (refer to 
Protocol Appendix 11 – EASI for descriptions of the ratings and the algorithm used to calculate the 
EASI score).  The total score incorporates the extent of body regions affected and the intensity of 
a representative area of eczema. The approximate percentages affected by eczema are calculated 
for each region. A higher score indicates more severe disease. 
EASI is assessed at all visits for Cohorts 2 and 3, and must be ≥9 and ≤48 at screening and ≥12 and 
≤48 on Day 1 for the patient to be eligible for inclusion in the study. 
4.2. 
SECONDARY EFFICACY ENDPOINTS 
The secondary efficacy endpoints for Cohort 3 are: 
EASI-50 
EASI-50 response (yes/no). This is defined as a ≥50% reduction from baseline in EASI score at 
Week 2. 
EASI-75 
EASI-75 response (yes/no). This is defined as a ≥75% reduction from baseline in EASI score at 
Week 2. 
EASI Components 
A score for the EASI components (erythema [E], oedema/papulation [O], excoriation [X] and 
lichenification [L]) will be calculated across the body regions in a similar way to the calculation of 
the EASI total score.  For example: 
Erythema=[0.1xEheadxExtenthead]+ [0.2xEarmsxExtentarms]+ [0.3xEtrunkxExtenttrunk]+[0.4xElegsxExtentlegs] 
The secondary endpoints associated with Cohort 3 are percentage change from baseline for each 
of the EASI components (erythema, oedema/papulation, excoriation and lichenification). 
Investigator Global Assessment (IGA) 
An overall assessment of the severity of AD will be made, by the investigator, using the IGA at 


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each visit for Cohorts 2 and 3. IGA scores take values on a 5-point scale from 0-4, where 0=clear to 
4=severe disease.  At screening and Day 1 IGA must be ≥ 3 for the subject to be eligible. 
The following secondary endpoints will be assessed for IGA for Cohort 3: 
 
A subject will be considered as having IGA success if they achieve a score of ‘Clear’ or 
‘Almost clear’; note, as subjects require a score of ≥ 3 to enter the study they must have a 
reduction of ≥ 2 from baseline to achieve success 
 
A subject will be considered as having an IGA response if they achieve a score of ‘Clear’ or 
‘Almost clear’, or a reduction of ≥ 2 from baseline  
Numerical Rating Scale (NRS) 
The Pruritus NRS is an assessment tool that will be used to assess the subject’s worst itch as a 
result of AD in the previous 24 hours. Subjects in Cohort 3 will be required to complete the Ziarco 
Itch questionnaire at each visit from baseline to follow-up. They will be asked the question “On a 
scale of 0 (No Itching) to 10 (Itching as bad as you can imagine), please rate the WORST itching 
that you felt over the last 24 hours.” 
To determine the duration of itching, subjects will be asked “Over the last 24 hours approximately 
how many hours, if any, did you itch?”.  To determine the level of sleep disturbance due to 
itching, subjects will be asked “On a scale of 0 (No sleep disturbance) to 10 (Awake all night), 
please rate how much your sleep was disturbed by itch last night”. 
 
Subjects will be asked to rate their itch over the last 24 hours using a verbal rating score (VRS).  
This is a list of adjectives describing different levels of symptom intensity - no itch, mild, moderate 
and severe. 
The secondary endpoints for Cohort 3 associated with NRS are: 
 
Change from baseline in the NRS for pruritus (worst itch) at Week 2 
 
Average change from baseline in NRS for pruritus (worst itch) over Days 8, 10 and 15 
 
Change from baseline in the NRS for sleep disturbance at Week 2  
 
Change from baseline in total duration of itching at Week 2 
 
Change from baseline in the VRS for pruritus at Week 2. 
Patient Global Impression of Change (PGIC)   
At the end of treatment (Day 15 or Early Termination Visit) subjects in Cohort 3 will be asked to 
rate their degree of improvement (or worsening) of their AD compared to before the start of 
treatment with study drug, using a 7-point scale.  The scale ranges from 1 (very much approved) 
to 7 (very much worse). 
The PGIC will be dichotomized into responders, defined as responses of ‘Very Much Improved’, 
‘Much Improved’ or ‘Minimally improved’ (1, 2 or 3 on the scale above) and non-responders (4, 5, 
6, or 7 on the scale above, plus missing data).  
 
 


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Body Surface Area (BSA) 
Assessment of the percentage of a subject’s BSA affected by AD will be made by best estimates of 
the Investigator at each visit for subjects in Cohorts 2 and 3. Affected BSA must be ≥ 10% and 
≤40% at screening and must be ≥ 10% and ≤50% on Day 1 for the patient to be randomised. 
The endpoints associated with BSA for Cohort 3 are change from baseline and percentage change 
from baseline at Week 2. 
4.3. 
EXPLORATORY ENDPOINT 
Ziarco Ointment Questionnaire 
This is assessed at the end of the treatment phase for each cohort.  Subjects will be asked a series 
of questions about their impressions of the study ointment they have used.  Information 
regarding the ointment will be used to learn the practicalities of handling the ointment in a 
clinical setting for subsequent cohorts and clinical development (refer to Protocol Appendix 13 – 
Ziarco Ointment Questionnaire for a full list of questions). 
 
4.4. 
SAFETY ENDPOINTS 
The safety endpoints assessed for all 3 cohorts are: 
 
Adverse events 
 
Vital signs (blood pressure, pulse rate, and temperature) –change from baseline at each 
visit, maximum and minimum on-treatment measurement 
 
12-lead Electrocardiograms (ECG)  - change from baseline at each visit and maximum on-
treatment measurement 
 
Laboratory safety tests (chemistry, haematology, and urinalysis) – change from baseline 
and percentage change from baseline 
Modified Draize Score 
For Cohort 1 subjects only, local toleration will be assessed by the modified Draize score prior to 
application of study ointment and until discharge from the CRU.  For each assessment the grading 
value for erythema and eschar formation, oedema formation and desquamation will be recorded 
and the total of the three gradings will also be recorded.  Grading is on a 5-point scale from 0-4, 
where 0=none to 4=severe.   
4.5. 
PHARMACOKINETIC ENDPOINTS 
Plasma ZPL-5212372 concentrations will be determined for all 3 cohorts. All concentration values 
will be reviewed prior to analysis and values that appear to be unexpectedly high due to potential 
contamination will be excluded and the reason(s) documented. 
The safety review meeting for Cohort 1 data showed insufficient data to enable pharmacokinetic 
(PK) parameters to be calculated for Cohort 1, due to the very low and sporadic systemic 
exposure in a small number of subjects who had study ointment applied over 40% BSA.   


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PK parameters will be derived for Cohort 2 for Days 1 and 7.  The ZPL-5212372 pharmacokinetic 
parameters AUCτ, AUClast and t½ will be generated using non-compartmental methods. Cmax and 
Tmax will be determined directly from the data.  The parameters are: 
Day 1: 
AUCτ 
The area under the serum concentration versus time curve, from time 0 to 12 
hours, as calculated using the linear trapezoidal method when concentrations 
are increasing and the logarithmic trapezoidal method when concentrations 
are decreasing. 
Cmax 
Maximum observed drug concentration. 
Tmax 
Time of the maximum serum concentration. If the maximum value occurs at 
more than one time point, Tmax is defined as the first time point with this value. 
Day 7: 
AUCτ 
The area under the serum concentration versus time curve, from time 0 to 12 
hours, as calculated using the linear trapezoidal method when concentrations 
are increasing and the logarithmic trapezoidal method when concentrations 
are decreasing. 
AUClast 
The area under the serum concentration versus time curve, from time 0 to last 
available time point (48 hours), as calculated using the linear trapezoidal 
method when concentrations are increasing and the logarithmic trapezoidal 
method when concentrations are decreasing. 
Cmax 
Maximum observed drug concentration. 
Tmax 
Time of the maximum serum concentration. If the maximum value occurs at 
more than one time point, Tmax is defined as the first time point with this value.  
Kel 
Where possible, the apparent terminal elimination phase rate constant (kel) will 
be calculated by linear regression of the log-linear plasma concentration-time 
curve.  Only those time points judged to describe the terminal log-linear 
decline will be used in the regression. Values for this parameter will only be 
reported for an individual if a minimum of 3 measurable concentration-time-
points during the log-linear portion of the terminal elimination phase, 
excluding Cmax, are available 
t½ 
Apparent first-order terminal elimination half-life will be calculated as ln2/kel 
RAUCτ 
Accumulation ratio calculated as Day 7 AUCτ / Day 1 AUCτ 
RCmax 
Accumulation ratio calculated as Day 7 Cmax / Day 1 Cmax 


Ziarco Pharma Ltd 
 
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5. 
ANALYSIS SETS 
5.1. 
SCREENED POPULATION 
The screened population will include all subjects who signed the informed consent form and are 
screened for participation in this study. This population will only be used for the purposes of 
describing the subject disposition. 
5.2. 
RANDOMISED SET 
The randomised set will include all randomised subjects. This population will only be used for the 
purposes of describing the subject disposition. 
5.3. 
FULL ANALYSIS SET (FAS) 
The FAS will include all randomised subjects who receive at least one application of 
investigational medical product (IMP). Subjects will be included in their randomised treatment 
group even if they receive the wrong IMP. This population will be used for the primary efficacy 
analysis and most other analyses for efficacy and exploratory endpoints. 
5.4. 
PER PROTOCOL (PP) ANALYSIS SET  
The PP analysis set is only applicable for Cohort 3.  It will include all subjects in the FAS who have 
not violated any inclusion or exclusion criteria and/or deviated from the protocol, in a way that 
could influence their efficacy assessment. For example subjects will be excluded if they: 
 
have poor IMP compliance, for example deviated from BID application over the 14 days 
and/or jar returned without being used. The IMP compliance will be assessed by a clinical 
review of dosing data and protocol deviations 
 
receive the incorrect IMP 
Subjects will be included in their ‘as treated’ treatment group. Note that this will be the same as 
the randomised treatment because receiving the wrong IMP is a major deviation. 
Data listings, including protocol deviations (Section 5.6), will be provided to identify subjects to be 
excluded from the PP analysis set. Precise reasons for excluding subjects will be documented prior 
to locking the database. 
5.5. 
SAFETY ANALYSIS SET 
The safety analysis set will include all subjects who receive at least 1 application of IMP. The 
safety analysis set summaries will be based on the actual treatment received for the whole 
duration of the study. The rules for handling data from subjects who receive study medication 
from the incorrect jar during the study will be decided on a case by case basis before database 


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lock.  This analysis set will be used for all safety analyses. 
5.6. 
PHARMACOKINETIC (PK) ANALYSIS SET 
The PK analysis set will include all subjects in the FAS who received ZPL-521 with any valid post-
baseline ZPL-521 plasma concentration result, including values below the lower limit of 
quantification (LLOQ). The PK analysis set summaries will be based on the actual treatment 
received for the whole duration of the study. The rules for handling data from subjects who 
receive study medication from the incorrect jar during the study will be decided on a case by case 
basis before database lock. 
5.7. 
PROTOCOL DEVIATIONS 
All protocol deviations will be reviewed and documented before database lock. Protocol 
deviations will be recorded by the site staff, study monitors and medical monitor reviewers.  They 
may also be identified through programmable checks of the data. 
Key protocol deviations include: 
 
Violations of inclusion and exclusion criteria. This includes unknown violations at 
enrolment and on-study violations, like taking a prohibited medication  
 
Poor IMP compliance  
 
Subject receiving the incorrect IMP  
6. 
GENERAL ASPECTS FOR STATISTICAL ANALYSIS 
6.1. 
GENERAL METHODS 
All statistical tests for Cohort 3 will be 1-sided at the 0.05 level of significance and accompanied 
by a 2-sided 90% CI. There will not be any adjustments for multiple comparisons.  No statistical 
testing will be performed with the data from Cohorts 1 and 2. 
All summaries and listings will be presented by treatment group and by cohort. For Cohorts 1 and 
2, adverse events and pharmacokinetic data will be presented by treatment group and BSA 
treated (10%, 40% and overall) to assess whether BSA impacts this data.  
Only on-treatment records will be summarised for post-baseline visits, except follow-up, unless 
stated otherwise.  A record is considered on-treatment if it is on or before the date of last study 
medication plus 1 day.  
Continuous variables will be summarised using the number of observations (n), mean, standard 
deviation (SD), 95% Confidence interval (CI) of mean, median, minimum, and maximum. 
Geometric mean and coefficient of variation (CV) will be included for PK parameters, where 
appropriate.  Categorical variables will be summarised using frequencies and percentages.  The 
number of missing values will be presented in summary tables.  The missing count will not include 


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subjects who have discontinued at earlier visits. 
Unless otherwise specified, the estimated mean and median for a set of values should be printed 
out to 1 more decimal place than the original values, and standard deviations should be printed 
out to 2 more decimal places than the original values. The minimum and maximum should report 
the same number of decimal places as the original values. The coefficient of variation will always 
be reported as a percentage to 1 decimal place. Percentages will be displayed with 1 decimal 
place; except percentages will not be presented when the count is zero and 100% will be 
presented as an integer. 
Unless stated otherwise, baseline will be defined as the last non-missing measurement prior to 
administration of the first dose of IMP.  
In by-visit summary tables the baseline will be summarised using all available data, but also for 
each visit using only the baseline data from subjects with available data at the visit; hence the 
mean change from baseline will equal the mean visit value – mean baseline value.   
All data will be listed.  Data listings will present study days in addition to dates, where study day is 
derived as (assessment date – first day of dosing + 1). The first day of dosing will be identified as 
Study Day 1. 
All statistical analyses will be performed using SAS® (Version 9.2 or higher, SAS Institute Inc., Cary, 
NC, USA).  
6.2. 
MISSING DATA 
All possible efforts will be made to ensure that subjects complete all the required assessments. As 
missing data may potentially bias the outcome of the statistical analyses and the subsequent 
estimation of the magnitude of the treatment effect, different strategies will be applied to 
provide a balanced assessment of treatment efficacy.  
Raw data will be reported in subject listings, but where imputed data are also presented (e.g. 
study start and stop days) they will be identified as imputed values in the listings. 
Specific approaches for handling missing data are described below. Other missing data will not be 
imputed.  
6.2.1. 
Efficacy Data 
Missing efficacy data for Cohort 3 may result from: 
 
subject discontinuing from the study 
 
for data collected at the site the assessment may have been missed 
For the sensitivity analysis of the primary efficacy endpoint missing values will be imputed using 
Markov Chain Monte Carlo (MCMC) multiple imputation methods. Means and variances from the 


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available data will be used to determine the MCMC starting values; imputed values will be 
selected with maximum likelihood methods. Initially, 200 MCMC runs will be generated before 
selecting the first imputed data set. An additional 24 data sets will be generated, separated by 
100 iterations of the MCMC. The number of data sets may be increased if the analyses are not 
found to be sufficiently stable with respect to standard errors and p-values. Randomisation seed 
numbers will be specified for the MCMC run to ensure consistency and repeatability. These 
imputations will be performed separately within each treatment group. 
The MCMC imputation assumes that data are missing at random.  
To assess the influence of missing data several sensitivity analyses will be conducted for the 
primary efficacy endpoint, including a last observation carried forward (LOCF) analysis and worst 
case imputations when the missingness is considered informative; e.g. subject withdrew due to 
lack of efficacy. Details regarding which approaches will be used are in Section 8.1.1. 
Missing categorical data will generally be considered as a non-response when dichotomizing 
responses for logistic modelling. Specific details are described in Section 8. 
6.2.2. 
Pharmacokinetic Data 
Concentrations of ZPL-5212372 reported as <LOQ will be taken as zero in the calculation of 
descriptive statistics.  The data will be presented as <LOQ in the listing, with LOQ replaced by the 
relevant LOQ value. 
For the calculation of PK parameters the following rules will apply: 
 
the value will be taken as zero for all values <LOQ before the first quantifiable 
concentration 
 
the value will be taken as missing for all values <LOQ after the last quantifiable 
concentration 
 
for values between quantifiable concentrations the pharmacokineticist will decide 
whether the value should be considered as missing or taken as zero base on the 
plausibility of a <LOQ value in the context of the whole concentration time profile.  The 
pharmacokineticist will document this decision along with the justification. 
Missing AUC and Cmax values due to all concentrations recorded as <LOQ will be set to zero for the 
calculation of descriptive statistics, except for geometric mean where values will be treated equal 
to LOQ.   
6.2.3. 
Safety Data 
AEs with missing classification will be assumed to be the following: 
 
missing causality will be taken as treatment related 


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 
missing seriousness will be taken as serious 
 
missing severity will be taken as severe 
Handling of missing dates is detailed in the relevant sections below. 
6.3. 
VISIT WINDOWS 
There is no requirement for visit windows for Cohorts 1 and 2 as subjects will remain in the CRU 
for the duration of the study, so all assessments will be presented and analysed according to the 
visit assessment recorded. 
For Cohort 3, all assessments will be presented and analysed according to the visit assessment 
recorded. Due to the short study duration visit windows will not be used. If possible, any early 
termination visits will be assigned to the closest study visit if they are on-treatment. Prior to 
database lock the data will be reviewed to identify any outlying visits and make decisions on how 
to deal with any issues. All decisions will be documented prior to database lock. 
6.4. 
STRATIFICATION 
No stratification is used for the randomisation in Cohorts 1 and 2. 
The randomisation for Cohort 3 is stratified by baseline EASI (≤20 or >20). This will be checked and 
any subjects who were misallocated to the wrong strata will be summarised according to the 
group they should have been assigned to.  Misallocations will be listed. 
6.5. 
SUBGROUPS 
The following subgroups will be considered for Cohort 3 only: 
 
Stratification subgroup: baseline EASI (≤20 or >20) 
 
7. 
DEMOGRAPHIC, OTHER BASELINE CHARACTERISTICS AND 
MEDICATION 
7.1. 
PATIENT DISPOSITION AND WITHDRAWALS 
The following will be tabulated over all screened subjects (the number of subjects in the screened 
set will be used as the denominator in the calculation of percentages): 
 
Number of subjects screened 
 
Number (%) of subjects who enter the washout period 
 
Number (%) of subjects who withdraw prior to the washout period 
 
Number (%) of subjects who withdraw prior to randomisation 
 
Number (%) subjects withdrawing before randomisation by reason 
 


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 
Number (%) of subjects randomised 
The following will be tabulated for each treatment group (the number of subjects in the 
Randomised set will be used as the denominator in the calculation of percentages): 
 
Number of subjects randomised 
 
Number (%) of subjects who complete the treatment period 
 
Number (%) of subjects who withdraw during the treatment period 
 
Number (%) subjects withdrawing during treatment period by reason 
 
 
Number (%) of subjects who complete the post-dose study follow-up  
 
Number (%) of subjects who withdraw between last dose of study medication and the 
post-dose follow-up visit. 
 
Number (%) subjects who withdraw during follow-up by reason  
In addition the number (%) of subjects in each analysis set will also be tabulated. 
7.2. 
DEMOGRAPHIC AND OTHER BASELINE CHARACTERISTICS 
Demography (gender, race, ethnicity, age [years], weight [kg], height [cm] and BMI [kg/m2]) will 
be summarised overall and by treatment group for the FAS.  Age, weight, height and BMI will be 
summarised using summary statistics for continuous variables. Gender, race, ethnicity and 
Fitzpatrick Skin Type Classification will be summarised using summary statistics for categorical 
variables. 
Age = (Day 1 date - date of birth + 1)/365.25 truncated to complete years. 
For Cohorts 2 and 3, the following baseline characteristics will be summarised using descriptive 
statistics by treatment group and overall, for subjects in the FAS; except for EASI (categorical), 
VRS and IGA which will present counts and percentages: 
 
Duration of atopic dermatitis (years) 
 
EASI 
 
EASI (categorical): ≤20 and >20 
 
% BSA Affected  
 
IGA score 
 
NRS for worst pruritus (over 24 hours)* 
 
duration of itching (over 24 hours)* 
 
NRS for sleep disturbance* 
 
VRS for pruritus (over 24 hours)* 
* Only summarised for Cohort 3. 
Duration of atopic dermatitis in years will be calculated from the date of onset recorded on 
medical history relative to Day 1 i.e. Day 1 date - onset date divided by 365.25. 


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7.3. 
MEDICAL HISTORY AND CONCOMITANT DISEASES 
Medical history will be coded using the latest Medical Dictionary for Regulatory Activities 
(MedDRA) Version available. Summaries will be presented for the FAS by System Organ Class 
(SOC) and Preferred Term (PT) with counts and percentages for each treatment group and overall. 
Each subject will be counted only once in each SOC or SOC/PT summary. 
7.4. 
PRIOR AND CONCOMITANT MEDICATIONS 
Prior and concomitant medications will be coded using the latest WHO Drug Dictionary version 
available. Medication will be presented for the safety population by ATC level 2 (therapeutic main 
group) and ATC level 5 (standardized medication name) with counts and percentages for each 
treatment group and overall.  A subject who took more than one medication will be counted only 
once if these medications belong to the same extended ATC classification.   
In addition, a summary of prior emollients (excluding study emollients) and concomitant 
emollients will be presented for Cohorts 2 and 3 with counts and percentages for each treatment 
group. Prior emollients will exclude study emollients which will be identified as emollients started 
prior to dosing but on or after the screening visit. The study emollients used were 
 for Cohort 
2 and 
 or Epaderm for Cohort 3. A clinical review of all unique medications will be done prior 
to database lock to identify emollients.  
Prior medications will be defined as those medications started prior to the administration of study 
drug on Day 1. Concomitant medications will be defined as those medications taken following the 
first administration of study drug on Day 1. Hence medications started before study dosing, but 
continuing after are considered as both prior and concomitant medications.  The listing of 
medications will identify prior and concomitant medications. 
If either the start or stop date of medication is missing, the worst or most conservative case will 
be considered when assigning medications to categories. So for a missing start date (where stop 
date is after date of first dose) the date will be imputed as the date of first dose; for a missing 
stop date the date will be imputed as the subject’s last study day or start date if start date is after 
last dose.  If a partial date is recorded, the following convention will be used to assign the 
medication: 
 
If a partial date is missing a start day and the month/year is the same as first dose date 
then use first dose date, else ‘01’ will be used for the day; if a start date is missing a 
month and the year is the same as first dose date then use first dose date, else January 
will be used for the start month. 
 
If a partial date is missing a stop day and month/year is same as last study date then use 
last study date, else last day of the given month will be used for the stop day; if a stop 
date is missing a month and the year is the same as last study date then use last study 
date, else December will be used for the stop month. 


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8. 
EFFICACY 
As Cohort 1 includes healthy volunteers no efficacy data will be collected.  Though some efficacy 
assessments are included for Cohort 2 the data will only be listed and no formal comparisons will 
be made between the treatment groups.  Consequently efficacy will only be assessed for Cohort 
3. Hence the following analyses apply to Cohort 3 data. 
The observed data summaries for all efficacy endpoints, except PGIC, will only use ‘on-treatment’ 
records (i.e. on or before the date of last study medication plus 1 day) for all visits, except 
baseline and follow-up. 
For all ANCOVA analyses the treatment least square means (LS means) and LS mean standard 
errors, treatment difference estimate, standard error, 1-sided p-value, and 90% CI will be 
presented.  The corresponding SAS ANCOVA output will also be outputted for internal statistical 
review. 
8.1. 
PRIMARY EFFICACY ENDPOINT AND ANALYSIS 
The primary endpoint is the percentage change from baseline to Week 2 in the EASI score.  
Baseline will be taken as the last non-missing measurement prior to administration of the first 
dose of IMP, this should be Day 1 for the primary endpoint.   
The observed percentage change from baseline to Week 2 in the EASI score will be analysed for 
the FAS with an Analysis of Covariance (ANCOVA) model that includes baseline score as a 
covariate and treatment (ZPL521 or placebo). The treatment effect will be estimated with the 
difference between the least square means (LS means), defined as ZPL521 - placebo.  
The ANCOVA analysis assumptions will be checked by evaluating the model residuals. A Shapiro-
Wilks test will be performed on the residuals; a p-value > 0.1 will be considered proof of 
normality. If the Shapiro-Wilks p-value is ≤ 0.1, the residual plot will be evaluated for symmetry. If 
the evaluation of residuals indicates a problem with the model assumptions, a transformation of 
the data will be considered, including a rank-transformation of the data.  
The EASI score will be summarised by treatment and visit for the FAS with observed data.  
Summaries will present the descriptive statistics for baseline, change from baseline and 
percentage change from baseline data.  
Patient profile plots by treatment group, of the percentage change from baseline in EASI score 
will also be presented.  Note that only observed data will be included in these plots; i.e. no 
imputations will be used. 
8.1.1. 
Sensitivity Analyses of the Primary Endpoint 
The robustness of the primary analysis will be evaluated by repeating the primary analysis with 


Ziarco Pharma Ltd 
 
Date: 16th March 2017 
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the per-protocol analysis set.   
In addition, the following sensitivity analyses will be conducted using the FAS: 
 
an analysis where missing values will be imputed using MCMC multiple imputation 
methods (see Section 6.2.1).  These imputations will be performed separately within each 
treatment group. 
 
an analysis using a worst case imputation for missing data considered as ‘informative’. 
Missing data will be reviewed by the study team to identify situations where the 
missingness is considered informative; e.g. subject withdrew for lack of efficacy. In these 
situations the Week 2 score will be taken as equal to the worst score recorded for either 
treatment group.  For the ‘non-informative’ values the MCMC imputed values will be 
used.  Note, the worst case imputation i.e. worst score, will occur after the MCMC values 
have been generated. 
 
an analysis using an LOCF method for handling missing data  
In addition, the EASI score will be summarised by treatment and visit for the FAS with MCMC 
imputation data. For this summary, the mean of the imputed data for a subject is used in the 
calculation. Hence when multiple imputations are performed for the analysis, a missing data point 
will be estimated with the mean of the multiple different imputations; this individual mean will be 
used to calculate the overall treatment summary statistics. 
8.1.2. 
Additional Analyses of the Primary Endpoint 
The percentage change from baseline to Day 5, Day 8 (Week 1) and Day 10 will also be analysed 
for the FAS using the same approach as for the primary analysis. 
Additional summaries and analyses of the Week 2 data will be performed for the subgroups 
detailed in Section 6.5. These analyses will be based on the FAS and will be the same as the 
primary analysis. 
8.2. 
SECONDARY ENDPOINTS AND ANALYSIS 
8.2.1. 
EASI-50 and EASI-75 
The proportion of subjects who achieve an EASI-50 response will be summarised in the FAS with 
counts and percentages by treatment group at each visit. The number of missing values will also 
be presented.  
An exact logistic regression analysis will be performed at Week 2. The model will include 
treatment and baseline EASI score. The odds ratio, p-value and associated exact 90% CI will be 
presented. Subjects with missing responses will be counted as non-responders. 
The proportion of subjects who achieve an EASI-75 response will be analysed in the same way as 
described for the EASI-50. 


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The cumulative proportion of subjects achieving a ≥x% reduction in EASI at Week 2 will be plotted 
against x, with different lines for each treatment group. Subjects with missing data will be 
regarded as having the worst change from baseline and labelled as such on the plot. Horizontal 
reference lines will be drawn that relate to the 0, 25, 50 and 75% reductions in EASI scores.  
8.2.2. 
EASI Components 
The 4 EASI component scores (erythema, oedema/papulation, excoriation and lichenification) will 
be summarised by treatment and visit for the FAS. Summaries will present the descriptive 
statistics for baseline, change from baseline and percentage change from baseline data.  
Percent change from baseline to Week 2 for each EASI component will be analysed for the FAS 
with an ANCOVA model.  The model will include baseline value and treatment. The treatment 
effect will be estimated with the difference between least square means (LS means), defined as 
ZPL521 - placebo. The estimate, standard error, 1-sided p-value, and 90% CI will be presented. 
The analysis will be repeated for all other time points (Day 5, Day 8 [Week 1] and Day 10). 
8.2.3. 
Investigators Global Assessment (IGA) 
IGA will be summarised for the FAS with counts and percentages by treatment at each visit. The 
number of missing values will also be presented. Shift tables of change from baseline will also be 
presented with marginal totals. 
IGA response will be summarised at each visit with counts and percentages; missing responses 
will also be tabulated.  An exact logistic regression analysis will be performed for Week 2. Baseline 
IGA, stratification variable (baseline EASI; ≤20 or >20) and treatment will be included in the 
models as factors; subjects with missing responses will be counted as non-responders. The odds 
ratio, p-value and associated exact 90% CI will be presented.  This analysis will be repeated for 
IGA success. 
8.2.4. 
Numerical Rating Scale (NRS) 
NRS for pruritus will be summarised by treatment and visit for the FAS. Summaries will present 
the descriptive statistics for baseline, change from baseline and percentage change from baseline 
data.  
Change from baseline to Week 2 of the NRS for pruritus will be analysed for the FAS with an 
ANCOVA model.  The model will include baseline NRS, stratification variable (baseline EASI; ≤20 or 
>20) and treatment. The treatment effect will be estimated with the difference between least 
square means (LS means), defined as ZPL521 - placebo. The estimate, standard error, 1-sided p-
value, and 90% CI will be presented. 
This analysis will be repeated for the average change from baseline in NRS pruritus over Days 8, 
10 and 15. 
Summaries over time will be presented for the FAS for the following NRS endpoints: 


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 
NRS for sleep disturbance   
 
total duration of itching 
All pruritus data will be listed.  
8.2.5. 
Verbal Rating Score (VRS) 
The VRS will be summarised for the FAS with counts and percentages by treatment at each visit. 
Shift tables of change from baseline will also be presented with marginal totals. 
8.2.6. 
Patient Global Impression of Change (PGIC)   
The PGIC scores will be summarised for the FAS with counts and percentages in each treatment 
group.  All data collected will be included. 
The PGIC will also be dichotomized into responders, defined as responses of ‘Very Much 
Improved’, ‘Much Improved’ or ‘Minimally improved’ (1, 2 or 3) and non-responders (4, 5, 6, or 7 
plus missing data). The PGIC responder variable will be analysed in the FAS with exact logistic 
regression. The stratification variable (Baseline EASI; ≤20 or >20) and treatment will be included in 
the model as factors; the odds ratio, p-value and associated exact 90% CI will be presented. 
8.2.7. 
Body Surface Area (BSA) 
The percentage BSA affected will be summarised at each visit, including change and percentage 
change from baseline, by treatment group, using the FAS. 
Change from baseline to Week 2 in the BSA will be analysed for the FAS with an ANCOVA model 
that includes baseline score, stratification variable (baseline EASI; ≤20 or >20) and treatment. The 
treatment effect will be estimated with the difference between the least square means (LS 
means), defined as ZPL521 - placebo. The estimate, standard error, 1-sided p-value, and 90% CI 
will be presented. 
8.3. 
EXPLORATORY ENDPOINT 
8.3.1. 
Ziarco Ointment Questionnaire 
The categorical responses to the Ziarco ointment questionnaire will be summarised by treatment 
for each of the cohorts using the FAS. All data collected will be included. 
All data will be listed. 
9. 
ANALYSIS OF PHARMACOKINETICS 
9.1. 
CONCENTRATION DATA 
Cohorts 2 and 3 concentration data will be summarised for the PK analysis set at each visit and 
time point using descriptive statistics (n, median, minimum, maximum, missing and frequency of 
values <LOQ). The summary for Cohort 2 will be by treatment group and BSA treated and for 


Ziarco Pharma Ltd 
 
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Cohort 3 by treatment group. For samples after the start of dosing only concentrations collected 
within a 15% window of the protocol specified time point will be included. Concentration values 
reviewed prior to the PK analysis and found to be unexpectedly high due to potential 
contamination will also be excluded. Values below the LLOQ will be taken as 0 for summaries.   
An individual subject profile plot will be presented for the trough concentration data for Cohort 3 
on the linear scale. The BSA treated will be presented along with the Subject ID. The actual 
sampling time will be used on the x-axis. Values below the LLOQ will be taken as 0 for linear plots. 
All plasma concentration data will be listed for all cohorts. The listing will include BSA treated, 
actual sampling times and deviations from nominal sampling times. Values below the LLOQ will be 
listed as <LOQ (with relevant value for LOQ) in listings.   
9.2. 
PHARMACOKINETIC PARAMETER DATA 
There is insufficient data to enable PK parameters to be calculated for Cohort 1 due to the very 
low and sporadic systemic exposure in a small number of subjects who had study ointment 
applied over 40% BSA.   
The PK parameters for Cohort 2 will be summarised by BSA treated and Day for the PK analysis 
set.  Geometric mean and CV will be presented for all parameters except t½. Due to the low 
sporadic exposure Tmax will not be summarized. 
All PK parameters for Cohort 2 will be listed. 
10. 
SAFETY 
10.1. 
DRAIZE SCORE 
The modified Draize score which assesses local toleration will be listed.  This is only collected for 
Cohort 1.  
10.2. 
EXTENT OF EXPOSURE 
For Cohorts 1 and 2, the number of days on treatment,  the average daily volume administered 
and average daily amount of ointment used (based on a density of 1.1089 g/mL) will be 
summarised using descriptive statistics by treatment group for the FAS.  The average daily volume 
administered and average daily amount of ointment used will be split by BSA treated within a 
cohort. 
 
Days on treatment  = date of last dose – date of first dose +1 
 
Daily volume administered = Total volume administered / number of days on treatment 
 
Daily amount of ointment used = daily volume administered * density 


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For Cohort 3 the number of days on treatment and daily amount of ointment used will be 
summarised using descriptive statistics by treatment group for the FAS. 
 
Daily amount of ointment used = (total weight of dispensed jars – total weight of returned 
jars) / days on treatment  
 
If a jar is not returned then it will be assumed that the weight for this jar is missing and no daily 
amount of ointment used will be calculated for the subject.  
Study drug administration, including all the variables specified above, will be listed. 
 
10.3. 
ADVERSE EVENTS 
AEs will be coded using the latest MedDRA version available.  
AEs will be considered treatment-emergent (TEAE) unless there is clear indication that the event 
occurred prior to first dose of study drug.  AEs present prior to the first dose of study drug 
administration that increased in severity or relationship to study drug after the first dose of study 
drug will be classed as TEAE.  Events with missing or partial dates will be handled such that in the 
absence of contradictory information an AE is treatment emergent.  So for a missing start date 
(where stop date is after date of first dose or missing) the date will be imputed as the date of first 
dose; for a missing stop date the date will be imputed as the date of the subject’s last study day.  
If a partial date is recorded, the following convention will be used to assign the AE: 
 
If a start date is missing the day information and month/year is the same as first dose 
date then use first dose date, else ‘01’ will be used for the day; if a start date is missing 
the month and the year is the same as first dose date then use first dose date, else 
January will be used for the start month; if start date is missing (i.e. even the year is 
unknown) then the AE will be considered not treatment emergent and the date will be set 
to the date of informed consent. 
 
If a stop date is missing the day information and month/year is same as the subject’s last 
study date then use last study date, else last day of the given month will be used for the 
stop day; if a stop date is missing the month and year is the same as last study date then 
use last study date, else December will be used for the stop month. 
All AEs will be listed however only TEAEs will be included in the summary tables. An overall 
summary will present the number and percentage of subjects by treatment group and BSA 
treated for Cohorts 1 and 2 and by treatment group for Cohort 3. The summary will include: 
 
any TEAE 
 
any TEAE considered as related to study drug (evaluated by the investigator as Related, 
Possibly Related or Probably Related or not reported) 
 
any serious TEAE  
 
Maximum intensity TEAE of none, mild, moderate, severe ; i.e. a subject with TEAEs at 


Ziarco Pharma Ltd 
 
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different intensities will be summarised at the most severe intensity 
 
any TEAE leading to study drug discontinuation  
 
any TEAE leading to death 
The table will also include the total number of TEAEs reported in each treatment group.  The total 
number of unique terms within subjects will also be presented, counting each TEAE only once 
within each subject.  
The number and percentage of subjects with TEAEs will be presented by SOC, PT and treatment 
group and BSA treated for Cohorts 1 and 2 and by SOC, PT and treatment group for Cohort 3. 
Subjects with multiple TEAEs within a SOC or SOC/PT combination will be counted only once for 
that SOC or SOC/PT combination. Similar summaries will be presented for related TEAEs.   
Summaries of TEAEs by maximum intensity (Mild, Moderate, or Severe) will be presented for 
TEAEs.  
All TEAEs will be listed by subject and start date. Separate listings of serious TEAEs and TEAEs 
leading to discontinuation of study drug will also be generated.  
10.4. 
LABORATORY EVALUATIONS 
The change from baseline and percentage change from baseline in haematology, chemistry, and 
continuous urinalysis parameters will be summarised by treatment group using descriptive 
statistics.  Discrete urinalysis parameters will be summarised with counts and percentages by 
treatment group. The number of normal and abnormal (low and high) records will also be 
summarised with counts and percentages by treatment group.  Baseline and all protocol specified 
on treatment (i.e. on or before the date of last study medication plus 1) visit data will be 
summarised.  If a baseline value is missing for a parameter, the screening value will be used as the 
baseline value. 
Subjects who satisfy the criteria for values of potential clinical concern will be summarised and 
listed separately.  The summary will present the number of subjects with at least one on-
treatment visit value (including unscheduled visits) satisfying the criteria. The listing will include 
all data for the subject and parameter which satisfied the criteria. The record satisfying the 
criteria will be identified on the listing. The criteria are: 
 
 


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Table 2: Criteria for Laboratory Safety Values of Potential Clinical Concern 
Category 
Parameter 
Criteria 
Haematology 
Haemoglobin 
<0.8 x baseline 
Haematocrit 
<0.8 x baseline 
WBC 
<2.5 or >17.5 x 103/mm3 
Platelets 
<75 or >700 x 103/mm3 
Chemistry 
GGT 
>3 x ULN 
Creatinine 
>1.3 x ULN 
Urea 
>1.3 x ULN 
Glucose, fasting 
<0.6 x LLN or >1.5 x ULN 
Uric acid 
>1.2 x ULN 
Sodium 
<0.95 x LLN or >1.05 x ULN 
Potassium 
<0.9 x LLN or >1.1 x ULN 
Calcium 
<0.9 x LLN or >1.1 x ULN 
Albumin 
<0.8 x LLN or >1.2 x ULN 
Total protein 
<0.8 x LLN or >1.2 x ULN 
Urinalysis 
Urine WBC 
6/HPF 
Urine RBC 
6/HPF 
LLN=lower Limit of Normal; ULN=Upper Limit of Normal. 
Liver function tests (ALT, AST, alkaline phosphatase and total bilirubin) will also be summarised.  
The number of subjects with any on treatment visit (including unscheduled visits) value either 
>1.5 x ULN or >2 x ULN or >3 x ULN will be summarised for each liver function test parameter. 
All laboratory data will be listed; change from baseline, percentage change from baseline and 
abnormal alerts will be included in the listings. Screening laboratory tests for serology, drug 
screen and breath alcohol test will be listed, but not summarised. 
10.5. 
VITAL SIGNS 
Systolic blood pressure (BP), diastolic BP and pulse will be summarised by visit, time point and 
treatment group using descriptive statistics for the observed and change from baseline values. 
Systolic BP, diastolic BP and pulse will be collected in duplicate at each time point. The mean of 
these 2 measurements will be derived prior to summarizing the data. If a baseline value is missing 
for a parameter, the screening value will be used as the baseline value. All vital sign data, 
including the mean values that are summarised, will be listed.  
The maximum and minimum values will be derived for each subject over all on-treatment visits 
(i.e. on or before the date of last study medication plus 1) from all scheduled and unscheduled 
individual readings and the number (%) of subjects in each of the following categories will be 
tabulated:  
 
 


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Table 3: Criteria for Vital Signs 
Parameter 
Category Criteria 
Systolic BP 
1 
 
< 90 mmHg* 
≥ 90 mmHg 
2 
< 30 mmHg change from baseline  
≥ 30 mmHg change from baseline* 
Diastolic BP 
1 
< 50 mmHg* 
≥ 50 mmHg 
2 
< 20 mmHg change from baseline  
≥ 20 mmHg change from baseline* 
Pulse 
1 
< 40 bpm* 
≥ 40 – ≤120 bpm 
> 120 bpm* 
2 
< 40 bpm and at least one value > 120 bpm 
< 40 bpm and no value > 120 bpm 
≥ 40 bpm 
* Indicates criteria to be identified in listing. 
Vital signs data will be listed.  The listing will include actual sampling times and deviations from 
nominal sampling times. The listing will flag records that satisfy the above criteria. 
10.6. 
ECG 
The mean of the triplicate measurements will be derived prior to summarizing the data. If a 
baseline value is missing for a parameter, the screening value will be used as the baseline value.  
ECG parameters, and their change from baseline, will be summarised at each time point using 
descriptive statistics.  
For Cohorts 1 and 2, subjects who satisfy the criteria for values of potential clinical concern will be 
summarised and listed separately.  The summary will present the number of subjects with at least 
one on-treatment (i.e. on or before the date of last study medication plus 1) visit value (including 
unscheduled visits) satisfying the criteria. The listing will include all data for the subject which 
satisfies the criteria. The record satisfying the criteria will be identified on the listing. The criteria 
are: 
 
 


Ziarco Pharma Ltd 
 
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Table 4: Criteria for ECG Safety Values of Potential Clinical Concern 
Parameter 
Category Criteria 
QT/QTcF 
1 
< 500 msec 
≥ 500 msec* 
2 
Increase from baseline < 30 msec 
Increase from baseline ≥ 30 msec and <60 msec* 
 
Increase from baseline ≥ 60 msec* 
PR 
1 
< 300 msec 
≥ 300 msec* 
2 
< 25% increase from baseline 
≥ 25% and < 50% increase from baseline when baseline > 
200msec* 
≥ 25% and < 50% increase from baseline when baseline ≤ 
200msec 
 
≥ 50% increase from baseline when baseline > 200msec* 
≥ 50% increase from baseline when baseline ≤ 200msec* 
QRS 
1 
< 200 msec 
≥ 200 msec* 
2 
< 25% increase from baseline 
≥ 25% and < 50% increase from baseline when baseline > 
100msec* 
≥ 25% and < 50% increase from baseline when baseline ≤ 
100msec 
 
≥ 50% increase from baseline when baseline > 100msec*  
≥ 50% increase from baseline when baseline ≤ 100msec* 
* Indicates criteria to be identified in listing. 
 
All ECG data will be listed, including the mean values that are summarised.  
10.7. 
PHYSICAL EXAMINATION 
A full physical examination will be performed at screening; post-screening physical exams will only 
include selected body systems (as a minimum, general appearance, heart, lungs and skin), 
determined by clinical findings and any AEs reported. All physical exam data will be listed. 
11. 
INTERIM ANALYSES 
Progression from Cohort 1 to Cohort 3 will be dependent on the demonstration of adequate 
safety within the protocol defined boundaries.  There will be a safety review meeting between 
each cohort to assess safety and PK.  The safety review will be conducted on blinded data, 
however some members of the Safety Review Team may be unblinded to treatment. The Safety 
Review Team will decide whether to proceed to the next cohort. In the event that it is decided not 
to proceed to the next cohort, the reviewers may decide to modify the study within the 


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boundaries of the adaptive features and continue, to submit a substantial protocol amendment or 
suspend the study. 
Further details on the operation of the safety review meetings will be defined in a separate 
document. 
12. 
CHANGE FROM ANALYSIS PLANNED IN PROTOCOL 
The changes from the analyses specified in the protocol are: 
1. The definition of responders for IGA score in the protocol has been further clarified and 
made consistent with the sponsor’s previous atopic dermatitis study. The definition has 
changed from reduction from baseline of at least 2 points, to those subjects who achieve 
a score of ‘Clear’ or ‘Almost clear’, or a reduction of ≥ 2 from baseline. An additional 
definition of IGA success has also be defined as those subjects who achieve a score of 
‘Clear’ or ‘Almost clear’. 
2. The protocol stated that the changes from baseline at Week 2 in NRS sleep disturbance 
and duration of itch will be analysed using ANCOVA, however it has been decided that it is 
sufficient to only summarize these endpoints by treatment group and visit.  Only the NRS 
endpoint daily pruritus will be analysed. 
3. The protocol stated that the Draize score for Cohort 1 would be summarised by treatment 
group and the change from baseline in Draize score would also be summarised using 
categorical shift tables.  However, the safety review of Cohort 1 showed that there were 
very few changes in Draize score; it was therefore decided that summaries were not 
necessary for this endpoint and the data would only be listed. 
4. The PK analyses described in the protocol have been modified to reflect the very low and 
sporadic systemic exposure observed during the safety review meetings for Cohorts 1 and 
2.   
13. 
REFERENCE LIST 
1. Hanifin JM, Thurston M, Omoto M, Cherill R, Tofte SJ & Graeber M ( 2001). The eczema 
area and severity index (EASI): assessment of reliability in atopic dermatitis. EASI 
Evaluator Group. . Exp Dermatol., Feb;10(1):11-8. 
2. Draize JH, Woodard G & Calvery H (1944) Methods for the study of irritation and toxicity 
of substances applied topically to the skin and mucous membranes. J Pharmacol Exp Ther 
82: 377-390. 
 


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14. 
INDEX OF TABLES 
The following tables are planned to be generated and numbering is indicative only and may be 
updated for final reporting based on CSR requirements. 
Table Number 
Title 
Cohort 
Population 
14.1.1.1 
Screened Subject Disposition by Cohort 
1,2,3 
Screened 
14.1.1.2 
Subject Disposition by Cohort 
1,2,3 
Randomised 
14.1.3.1 
Demography by Cohort 
1,2,3 
FAS 
14.1.3.2.1 
Cohort 2: Baseline Characteristics  
2 
FAS 
14.1.3.2.2 
Cohort 3: Baseline Characteristics  
3 
FAS 
14.1.3.3 
Medical History by Cohort 
1,2,3 
FAS 
14.1.4.1 
Prior Medications by Cohort 
1,2,3 
SAF 
14.1.4.2 
Concomitant Medications by Cohort 
1,2,3 
SAF 
14.1.4.3.1 
Prior Emollient Medications by Cohort 
2,3 
SAF 
14.1.4.3.2 
Concomitant Emollient Medications by Cohort 
2,3 
SAF 
14.1.5.1 
Study Drug Exposure by Cohort 
1,2 
FAS 
14.1.5.2 
Cohort 3: Study Drug Exposure  
3 
FAS 
14.2.1.1 
Cohort 3: EASI by Visit 
3 
FAS 
14.2.1.2 
Cohort 3: EASI by Visit with Multiple Imputation 
3 
FAS 
14.2.1.3 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI at Week 2: Observed Case  
3 
FAS 
14.2.1.4 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI at Week 2: Observed Case  
3 
PP 
14.2.1.5 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI at Week 2: Multiple Imputation  
3 
FAS 
14.2.1.6 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI at Week 2: LOCF  
3 
FAS 
14.2.1.7 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI at Week 2: Worst Case 
Imputation  
3 
FAS 
14.2.1.8 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI Over Time: Observed Case  
3 
FAS 
14.2.1.9 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI at Week 2 by Baseline EASI: 
Observed Case 
3 
FAS 
14.2.2.1.1 
Cohort 3: EASI-50 by Visit 
3 
FAS 
14.2.2.1.2 
Cohort 3: Logistic Regression of EASI-50  
3 
FAS 
14.2.2.2.1 
Cohort 3: EASI-75 by Visit 
3 
FAS 
14.2.2.2.2 
Cohort 3: Logistic Regression of EASI-75 
3 
FAS 
14.2.2.3.1 
Cohort 3: EASI Component Erythema by Visit 
3 
FAS 
14.2.2.3.2 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI Component Erythema Over 
Time: Observed Case  
3 
FAS 


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Table Number 
Title 
Cohort 
Population 
14.2.2.4.1 
Cohort 3: EASI Component oedema/papulation 
by Visit 
3 
FAS 
14.2.2.4.2 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI Component 
oedema/papulation  Over Time: Observed Case  
3 
FAS 
14.2.2.5.1 
Cohort 3: EASI Component Excoriation by Visit 
3 
FAS 
14.2.2.5.2 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI Component Excoriation Over 
Time: Observed Case  
3 
FAS 
14.2.2.6.1 
Cohort 3: EASI Component Lichenification by 
Visit 
3 
FAS 
14.2.2.6.2 
Cohort 3: ANCOVA of Percent Change from 
Baseline in EASI Component Lichenification 
Over Time: Observed Case  
3 
FAS 
14.2.3.1 
Cohort 3: Investigators Global Assessment (IGA) 
and Shift from Baseline by Visit  
3 
FAS 
14.2.3.2 
Cohort 3: Logistic Regression of Investigators 
Global Assessment (IGA) - Responder 
3 
FAS 
14.2.3.3 
Cohort 3: Logistic Regression of Investigators 
Global Assessment (IGA) - Success 
3 
FAS 
14.2.4.1.1 
Cohort 3: NRS for Pruritus by Visit 
3 
FAS 
14.2.4.1.2 
Cohort 3: ANCOVA of Change from Baseline in 
NRS for pruritus at Week 2: Observed Case 
3 
FAS 
14.2.4.1.3 
Cohort 3: ANCOVA of Average Change from 
Baseline in NRS for pruritus Over Days 8, 10 and 
15: Observed Case 
3 
FAS 
14.2.4.2 
Cohort 3: Sleep Disturbance for pruritus by Visit 
3 
FAS 
14.2.4.3 
Cohort 3: Total Duration of itch by Visit 
3 
FAS 
14.2.5.1 
Cohort 3: Verbal Rating Score (VRS) and Shift 
from Baseline by Visit  
3 
FAS 
14.2.6.1 
Cohort 3: Patient Global Impression of Change 
(PGIC)  
3 
FAS 
14.2.6.2 
Cohort 3: Logistic Regression of Patient Global 
Impression of Change (PGIC) 
3 
FAS 
14.2.7.1 
Cohort 3: Body Surface Area (BSA) by Visit 
3 
FAS 
14.2.7.2 
Cohort 3: ANCOVA of Change from Baseline 
Body Surface Area (BSA) by Visit at Week 2: 
Observed Case 
3 
FAS 
14.2.8 
Ziarco ointment Questionnaire by Cohort 
1,2,3 
FAS 
14.2.9.1 
Cohort 2: ZPL-5212372 Plasma Concentrations 
2 
PK 
14.2.9.2 
Cohort 3: ZPL-5212372 Plasma Concentrations 
3 
PK 
14.2.10 
Cohort 2: Pharmacokinetic Parameters 
2 
PK 
14.3.1.1.1 
Summary of Adverse Events by Cohort 
1,2 
SAF 


Ziarco Pharma Ltd 
 
Date: 16th March 2017 
ZPL521/101                                                                          
Statistical Analysis Plan 
 
 
SAP Version v2.0     
 
Page 41 of 43 
Table Number 
Title 
Cohort 
Population 
14.3.1.1.2 
Cohort 3: Summary of Adverse Events  
3 
SAF 
14.3.1.2.1 
Treatment Emergent Adverse Events By SOC 
and Preferred Term by Cohort 
1,2 
SAF 
14.3.1.2.2 
Cohort 3: Treatment Emergent Adverse Events 
By SOC and Preferred Term 
3 
SAF 
14.3.1.3.1 
Treatment Related Treatment Emergent 
Adverse Events By SOC and Preferred Term by 
Cohort 
1,2 
SAF 
14.3.1.3.2 
Cohort 3: Treatment Related Treatment 
Emergent Adverse Events By SOC and Preferred 
Term  
3 
SAF 
14.3.1.4.1 
Treatment Emergent Adverse Events By 
Maximum Severity by Cohort 
1,2 
SAF 
14.3.1.4.2 
Cohort 3: Treatment Emergent Adverse Events 
By Maximum Severity 
3 
SAF 
14.3.4.1.1 
Haematology Clinical Laboratory Tests by 
Cohort 
1,2,3 
SAF 
14.3.4.1.2 
Haematology Clinical Laboratory Tests – 
Abnormality Counts by Cohort 
1,2,3 
SAF 
14.3.4.1.3 
Haematology Clinical Laboratory Tests of 
Potential Clinical Concern by Cohort 
1,2,3 
SAF 
14.3.4.2.1 
Chemistry Clinical Laboratory Tests by Cohort 
1,2,3 
SAF 
14.3.4.2.2 
Chemistry Clinical Laboratory Tests – 
Abnormality Counts by Cohort 
1,2,3 
SAF 
14.3.4.2.3 
Chemistry Clinical Laboratory Tests of Potential 
Clinical Concern by Cohort 
1,2,3 
SAF 
14.3.4.2.4 
Liver Function Test of Potential Clinical Concern 
by Cohort 
1,2,3 
SAF 
14.3.4.3.1 
Urinalysis Clinical Laboratory Tests by Cohort 
1,2,3 
SAF 
14.3.4.3.2 
Categorical Urinalysis Clinical Laboratory Tests 
by Cohort 
1,2,3 
SAF 
14.3.4.3.3 
Urinalysis Clinical Laboratory Tests of Potential 
Clinical Concern by Cohort 
1,2,3 
SAF 
14.3.4.4 
Vital Signs by Cohort 
1,2,3 
SAF 
14.3.4.5 
Vital Signs – Categorical by Cohort 
1,2,3 
SAF 
14.3.5.1 
ECGs by Cohort 
1,2,3 
SAF 
14.3.5.2 
ECGs Categorical Summary of Potential Clinical 
Concern by Cohort 
1,2 
SAF 
 
 
 


Ziarco Pharma Ltd 
 
Date: 16th March 2017 
ZPL521/101                                                                          
Statistical Analysis Plan 
 
 
SAP Version v2.0     
 
Page 42 of 43 
15. 
INDEX OF LISTINGS 
The following listings are planned to be generated and numbering is indicative only and may be 
updated for final reporting based on CSR requirements. 
Listing 
Number 
Title 
Cohort 
16.2.1.1 
Subject Disposition by Cohort 
1,2,3 
16.2.1.2 
Screen Failures by Cohort 
1,2,3 
16.2.2.1 
Protocol Deviations by Cohort 
1,2,3 
16.2.3.1.1 
Analysis Sets by Cohort 
1,2 
16.2.3.1.2 
Cohort 3: Analysis Sets 
3 
16.2.4.1.1 
Demography by Cohort 
1,2 
16.2.4.1.2 
Cohort 3: Demography 
3 
16.2.4.2 
Medical History by Cohort 
1,2,3 
16.2.4.3 
Prior and Concomitant Medications by Cohort 
1,2,3 
16.2.5.1.1 
Study Drug Administration by Cohort 
1,2 
16.2.5.1.2 
Cohort 3: Study Drug Administration 
3 
16.2.5.2 
Cohort 3: Study Drug Dispensed and Returned 
3 
16.2.6.1.1 
EASI Total Score by Cohort 
2,3 
16.2.6.1.2.1 
Cohort 3: EASI Components Derived 
3 
16.2.6.1.2.2 
EASI Components by Cohort 
2,3 
16.2.6.1.3 
Investigator Global Assessment (IGA) by Cohort 
2,3 
16.2.6.1.4 
Cohort 3: Pruritus (Itch) Questionnaire 
3 
16.2.6.1.5 
Cohort 3: Patient Global Impression of Change (PGIC) 
3 
16.2.6.2 
Body Surface Area (BSA) by Cohort 
2,3 
16.2.6.3 
Ziarco Ointment Questionnaire by Cohort 
1,2,3 
16.2.6.4.1 
ZPL-5212372 Concentrations by Cohort 
1,2,3 
16.2.6.4.2 
Cohort 2: Pharmacokinetic Parameters 
2 
16.2.7.1 
Adverse Events by Cohort 
1,2,3 
16.2.7.2 
Serious Adverse Events by Cohort 
1,2,3 
16.2.7.3 
Adverse Events Leading to Discontinuation by Cohort 
1,2,3 
16.2.7.4 
Cohort 1: Modified Draize Score 
1 
16.2.8.1.1 
Haematology Clinical Laboratory Data by Cohort 
1,2,3 
16.2.8.1.2 
Haematology Clinical Laboratory Data of Potential Clinical 
Concern by Cohort 
1,2,3 
16.2.8.1.3 
Chemistry Clinical Laboratory Data by Cohort 
1,2,3 
16.2.8.1.4 
Chemistry Clinical Laboratory Data of Potential Clinical Concern 
by Cohort 
1,2,3 
16.2.8.1.5 
Urinalysis Clinical Laboratory Data by Cohort 
1,2,3 
16.2.8.1.6 
Urinalysis Clinical Laboratory Data of Potential Clinical Concern 
by Cohort 
1,2,3 
16.2.8.1.7 
Serology and Screening Laboratory Data by Cohort 
1,2,3 
16.2.8.1.8 
Pregnancy Test Data by Cohort 
1,2,3 


Ziarco Pharma Ltd 
 
Date: 16th March 2017 
ZPL521/101                                                                          
Statistical Analysis Plan 
 
 
SAP Version v2.0     
 
Page 43 of 43 
Listing 
Number 
Title 
Cohort 
16.2.8.2.1 
Vital Signs by Cohort 
1,2,3 
16.2.8.2.2 
ECGs by Cohort 
1,2,3 
16.2.8.2.3 
ECGs of Potential Clinical Concern by Cohort 
1,2 
16.2.8.2.4 
Physical Examination by Cohort 
1,2,3 
 
 
 
16. 
INDEX OF FIGURES 
The following figures are planned to be generated and numbering is indicative only and may be 
updated for final reporting based on CSR requirements. 
Figure Number 
Title 
Cohort 
Population 
14.2.1.11 
Cohort 3: Individual Profiles of Percent Change from 
Baseline in EASI Over Time by Treatment Group 
3 
FAS 
14.2.2.2.3 
Cohort 3: Cumulative Proportion of Subjects 
Achieving Reductions in EASI at Week 2 
3 
FAS 
14.2.9.3 
Cohort 3: Individual ZPL-3893787 Plasma Trough 
Concentration Over Time (linear) 
3 
PK