Conditions: Healthy Volunteers
Allocation: RANDOMIZED | Intervention model: CROSSOVER
Enrollment: 57
Healthy volunteers: yes

Brief summary:
This was a single-center, multiple-dose, randomized, double-blind, placebo-controlled, positive-controlled, twelve sequence, 3-period cross-over study to investigate the effect of balovaptan on the QTc interval in healthy subjects.

Detailed description:

Arms:
arm 1: None arm 2: None arm 3: None arm 4: None arm 5: None arm 6: None arm 7: None arm 8: None arm 9: None arm 10: None arm 11: None arm 12: None

Interventions: intervention 1: Balovaptan therapeutic dose Treatment A intervention 2: Balovaptan supra-therapeutic dose Treatment B intervention 3: Active control [moxifloxacin] on Day 2 Treatment C intervention 4: Active control [Moxifloxacin] on Day 15 Treatment D intervention 5: Placebo for Balovaptan Treatment C intervention 6: Placebo for Balovaptan Treatment D intervention 7: Placebo for Moxifloxacin Treatment A intervention 8: Placebo for Moxifloxacin Treatment B intervention 9: Moxifloxacin Treatment C intervention 10: Placebo for Moxifloxacin Treatment C intervention 11: Placebo for Moxifloxacin Treatment D intervention 12: Moxifloxacin Treatment D
Intervention descriptions: intervention 1: Days 1-14: A single once daily dose at dose level A of balovaptan for 14 days intervention 2: Days 1-14: A single once daily oral dose at dose level B of balovaptan for 14 days. intervention 3: Day 2: A single oral dose of 400 mg moxifloxacin capsule. intervention 4: Day 15: A single oral dose of 400 mg moxifloxacin capsule. intervention 5: Days 1-14: Matching placebo for balovaptan for 14 days. intervention 6: Days 1-14: Matching placebo for balovaptan for 14 days. intervention 7: Day 2 and 15: A single oral dose of a matching placebo capsule for moxifloxacin. intervention 8: Days 2 and 15: Single oral dose of a matching placebo capsule of moxifloxacin. intervention 9: Day 2: A single oral dose of 400 mg moxifloxacin capsule. intervention 10: Day 15: A single oral dose of a matching placebo capsule for moxifloxacin. intervention 11: Day 2: A single oral dose of a matching placebo capsule for moxifloxacin. intervention 12: Day 15: A single oral dose of 400 mg moxifloxacin capsule.

Protocol document:
Prot_000.pdf: 
Official Title: 
A Single-Center, Multiple-Dose, Randomized, Double-Blind, Placebo-
Controlled, Positive-Controlled, Crossover Study to Investigate the 
Effect of Balovapton on the Qtc Interval in Healthy Subjects 
NCT Number: 
NCT03808298 
Document Date: 
Protocol Version 3: 23-April-2019 


FINAL PROTOCOL APPROVAL
CONFIDENTIAL
This clinical study is being sponsored globally by F. Hoffmann-La Roche Ltd of Basel, 
Switzerland. However, it may be implemented in individual countries by Roche’s local affiliates, 
including Genentech, Inc. in the United States.  The information contained in this document, especially 
any unpublished data, is the property of F. Hoffmann-La Roche Ltd (or under its control) and therefore 
is provided to you in confidence as an investigator, potential investigator, or consultant, for review by 
you, your staff, and an applicable Ethics Committee or Institutional Review Board.  It is understood that 
this information will not be disclosed to others without written authorization from Roche except to the 
extent necessary to obtain informed consent from persons to whom the drug may be administered.
Balovaptan —F. Hoffmann-La Roche Ltd
Protocol WP40734, Version 3.0 
PROTOCOL
TITLE:
A SINGLE-CENTER, MULTIPLE-DOSE, 
RANDOMIZED, DOUBLE-BLIND, PLACEBO-
CONTROLLED, POSITIVE-CONTROLLED, CROSS-
OVER STUDY TO INVESTIGATE THE EFFECT OF 
BALOVAPTAN ON THE QTC INTERVAL IN 
HEALTHY SUBJECTS
PROTOCOL NUMBER:
WP40734
VERSION NUMBER:
V3.0
EUDRACT NUMBER:
NA
IND NUMBER:
116483
TEST PRODUCT:
Balovaptan (RO5285119)
MEDICAL MONITOR:
, MD
SPONSOR:
F. Hoffmann-La Roche Ltd
DATE FINAL:
See electronic date stamp below
 
23-Apr-2019 06:19:00
Title
Approver's Name
Company Signatory
Date and Time (UTC)


Balovaptan —F. Hoffmann-La Roche Ltd
2/Protocol WP40734, Version 3.0
Document
Date of Issue
Approver(s)
Summary of Changes
Original 
Protocol
Oct-23-2018

Not Applicable
Protocol v2.0 
Amendment 1
and 
Administrative 
letter 1, Nov-
13-2018
See date 
stamp on the 
cover page.

Changes in wording regarding intensive PK sampling for 
balovaptan and moxifloxacin outlined in Administrative 
letter from Nov-13-2018 incorporated.

Reference to balovaptan sampling on Day 16 have been 
removed.

The 1.25 h time point for PK sampling and ECG readings 
has been changed to 1.0 h post-dose. This information has 
been updated in Appendices and protocol text.

Section 4.4 has been edited to clarify that concomitant 
therapy should be captured from 30 days prior to initiation 
of study drug.

The text in Section 4.4.2 regarding prohibited therapies 
has been edited for clarity.

Text in Section 4.4.4 has been edited to remove the 
statement “Subjects should have fasted for at least 8 hours 
prior to the screening visit, each admission to the CRU, 
and the Follow up visit” as it redundant with the collection 
description in Section 4.5.5.1.

Text has been added to Section 4.4 to clarify that the 
clinical site is not required to track subject’s fluid intake.

In Section 4.5.3 Physical Examinations the phrase “Any 
abnormality identified” has been updated to read “Any 
clinically significant abnormalities identified”

Instructions for collection and handling of PK samples in 
Section 4.5.5.2 has been updated. 

Text reading safety ECG readings in Section 4.5.6.1 has 
been edited for clarity.

Section 4.6.1 has been edited for clarity regarding 
assessments for subjects who withdraw or are terminated 
from the study early.

Text regarding ECG extraction on consecutive days has 
been edited throughout the document to clarify that when 
a reading is taken on consecutive days (Day 2 and 
Day 15) the 24 h time point can be taken from the 3 pre-
dose Holter extractions.

Text regarding analysis of samples for cardiac biomarkers 
has been edited to indicate that samples will be taken but 
only analyzed for specific subjects on request.

Text in Section 5.3.1 has been modified to make it 
consistent with Section 5.4.2.1 and Appendix 2 clarifying 
that only SAE’s will be recorded prior to study drug 
administration.

Section 6.7.2 has been edited to make text consistent with 
ECG extraction time points throughout protocol.

TQT exclusion criteria #3 and #7 have been edited to 
clarify that criteria should be assessed at both Screening 




Balovaptan —F. Hoffmann-La Roche Ltd
4/Protocol WP40734, Version 3.0
TABLE OF CONTENTS
PROTOCOL ACCEPTANCE FORM .................................................................. 10
PROTOCOL SYNOPSIS .................................................................................... 11
1.
BACKGROUND .......................................................................................... 23
1.1
Autism Spectrum Disorder..................................................... 23
1.2
Pre-Clinical Background on Balovaptan ................................ 23
1.2.1
Previous and Ongoing Clinical Studies.................................. 25
1.2.1.1
Clinical Pharmacology ........................................................... 25
1.2.1.2
Safety .................................................................................... 26
1.3
Study Rationale and Benefit-Risk Assessment...................... 28
2.
OBJECTIVES AND ENDPOINTS ............................................................... 28
3.
STUDY DESIGN......................................................................................... 31
3.1
Description of the Study......................................................... 31
3.2
End of Study and Length of Study......................................... 35
3.3
Rationale for Study Design.................................................... 35
3.3.1
Rationale for Balovaptan and Moxifloxacin Dose 
and Schedule......................................................................... 35
3.3.1.1
Balovaptan............................................................................. 35
3.3.1.2
Moxifloxacin........................................................................... 36
3.3.2
Rationale for Study Population .............................................. 36
3.3.3
Rationale for Transthoracic Doppler-
Echocardiogram and Ambulatory Blood Pressure 
Monitoring.............................................................................. 37
4.
MATERIALS AND METHODS .................................................................... 37
4.1
Subjects................................................................................. 37
4.1.1
Inclusion Criteria.................................................................... 37
4.1.2
Exclusion Criteria................................................................... 38
4.2
Method of Treatment Assignment and Blinding..................... 41
4.2.1
Treatment Assignment........................................................... 41
4.2.2
Blinding.................................................................................. 42


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5/Protocol WP40734, Version 3.0
4.3
Study Treatment and Other Treatments Relevant 
to the Study Design ............................................................... 42
4.3.1
Study Treatment Formulation, Packaging, and 
Handling ................................................................................ 43
4.3.1.1
Balovaptan and Placebo........................................................ 43
4.3.1.2
Moxifloxacin Active Control and Placebo............................... 43
4.3.2
Study Treatment Dosage, Administration, and 
Compliance............................................................................ 43
4.3.2.1
Balovaptan and Placebo........................................................ 43
4.3.2.2
Moxifloxacin (Active Control) and Placebo ............................ 44
4.3.3
Investigational Medicinal Product Accountability ................... 44
4.4
Concomitant Therapy, Prohibited Food, and 
Additional Restrictions ........................................................... 45
4.4.1
Permitted Therapy................................................................. 45
4.4.2
Prohibited Therapy ................................................................ 45
4.4.3
Prohibited Food ..................................................................... 46
4.4.4
Additional Restrictions/Considerations .................................. 46
4.4.4.1
Herbal Therapies................................................................... 47
4.5
Study Assessments ............................................................... 47
4.5.1
Informed Consent Forms and Screening Log........................ 47
4.5.2
Medical History, Concomitant Medication, and 
Demographic Data................................................................. 47
4.5.3
Physical Examinations........................................................... 47
4.5.4
Vital Signs.............................................................................. 48
4.5.5
Laboratory, Genotyping, and Pharmacokinetic 
Samples................................................................................. 48
4.5.5.1
Laboratory Samples .............................................................. 48
4.5.5.2
Pharmacokinetic Samples ..................................................... 49
4.5.5.3
Optional Samples for Genetic Analyses ................................ 49
4.5.6
Electrocardiograms................................................................ 50
4.5.6.1
12-lead Safety Electrocardiograms........................................ 50
4.5.6.2
Cardiodynamic ECG Evaluation ............................................ 51
4.5.7
Transthoracic-Doppler Echocardiography ............................. 53
4.5.8
Ambulatory Blood Pressure Monitoring ................................. 54
4.5.9
Columbia-Suicide Severity Rating Scale ............................... 55




Balovaptan —F. Hoffmann-La Roche Ltd
7/Protocol WP40734, Version 3.0
5.2.3
Adverse Events of Special Interest (Immediately 
Reportable to the Sponsor).................................................... 65
5.3
Methods and Timing for Capturing and 
Assessing Safety Parameters................................................ 66
5.3.1
Adverse Event Reporting Period ........................................... 66
5.3.2
Eliciting Adverse Event Information....................................... 67
5.3.3
Assessment of Severity of Adverse Events ........................... 67
5.3.4
Assessment of Causality of Adverse Events ......................... 67
5.3.5
Procedures for Recording Adverse Events............................ 68
5.3.5.1
Diagnosis versus Signs and Symptoms................................. 68
5.3.5.2
Adverse Events That Are Secondary to Other 
Events.................................................................................... 69
5.3.5.3
Persistent or Recurrent Adverse Events................................ 69
5.3.5.4
Abnormal Laboratory Values ................................................. 69
5.3.5.5
Abnormal Vital Sign Values ................................................... 70
5.3.5.6
Abnormal Liver Function Tests.............................................. 71
5.3.5.7
Deaths ................................................................................... 71
5.3.5.8
Preexisting Medical Conditions.............................................. 71
5.3.5.9
Hospitalization or Prolonged Hospitalization.......................... 72
5.4
Immediate Reporting Requirements from 
Investigator to Sponsor.......................................................... 72
5.4.1
Emergency Medical Contacts................................................ 73
5.4.2
Reporting Requirements for Serious Adverse 
Events and Adverse Events of Special Interest..................... 73
5.4.2.1
Events That Occur prior to Study Drug Initiation.................... 73
5.4.2.2
Events That Occur after Study Drug Initiation........................ 74
5.4.3
Reporting Requirements for Pregnancies.............................. 74
5.4.3.1
Pregnancies in Female Subjects ........................................... 74
5.4.3.2
Pregnancies in Female Partners of Male 
Subjects................................................................................. 74
5.4.3.3
Abortions ............................................................................... 75
5.4.3.4
Congenital Anomalies/Birth Defects ...................................... 75
5.4.4
Reporting Requirements for Cases of Accidental 
Overdose or Medication Error................................................ 75
5.5
Follow-Up of Subjects after Adverse Events.......................... 76


Balovaptan —F. Hoffmann-La Roche Ltd
8/Protocol WP40734, Version 3.0
5.5.1
Investigator Follow-Up........................................................... 76
5.5.2
Sponsor Follow-Up ................................................................ 76
5.6
Adverse Events That Occur after the Adverse 
Event Reporting Period.......................................................... 77
5.7
Expedited Reporting to Health Authorities, 
Investigators, Institutional Review Boards, and 
Ethics Committees................................................................. 77
6.
STATISTICAL CONSIDERATIONS AND ANALYSIS PLAN....................... 77
6.1
Determination of Sample Size ............................................... 78
6.2
Summaries of Conduct of Study............................................ 78
6.3
Summaries of Demographic and Baseline 
Characteristics....................................................................... 78
6.4
Efficacy Analyses .................................................................. 78
6.5
Safety Analyses..................................................................... 78
6.6
Pharmacokinetic Analyses..................................................... 79
6.7
Cardiodynamic ECG Evalutaion ............................................ 80
6.7.1
Analysis Populations ............................................................. 80
6.7.2
Baseline................................................................................. 81
6.7.3
By-Time Point Analysis.......................................................... 81
6.7.4
Secondary Analysis ............................................................... 82
6.7.5
Assay Sensitivity.................................................................... 82
6.7.6
Categorical Analyses............................................................. 82
6.7.7
Concentration-QTc Analysis (Secondary Analysis)
.............................................................................................. 83
6.7.8
Investigation of Hysteresis..................................................... 84
6.8
TT-Doppler Echocardiogram ................................................. 85
6.9
Blood Pressure and Heart Rate Monitoring ........................... 85
7.
DATA COLLECTION AND MANAGEMENT ............................................... 86
7.1
Data Quality Assurance......................................................... 86
7.2
Electronic Case Report Forms............................................... 87
7.3
Source Data Documentation.................................................. 87
7.4
Use of Computerized Systems .............................................. 87
7.5
Retention of Records............................................................. 88
8.
ETHICAL CONSIDERATIONS.................................................................... 88


Balovaptan —F. Hoffmann-La Roche Ltd
9/Protocol WP40734, Version 3.0
8.1
Compliance with Laws and Regulations ................................ 88
8.2
Informed Consent.................................................................. 88
8.3
Institutional Review Board or Ethics Committee.................... 89
8.4
Confidentiality........................................................................ 90
8.5
Financial Disclosure .............................................................. 90
9.
STUDY DOCUMENTATION, MONITORING, AND 
ADMINISTRATION ..................................................................................... 91
9.1
Study Documentation ............................................................ 91
9.2
Protocol Deviations................................................................ 91
9.3
Site Inspections ..................................................................... 91
9.4
Administrative Structure......................................................... 91
9.5
Publication of Data and Protection of Trade 
Secrets .................................................................................. 92
9.6
Protocol Amendments ........................................................... 92
10. REFERENCES ........................................................................................... 93
LIST OF TABLES
Table 1
Objectives and Corresponding Endpoints................................... 29
Table 2
T-wave Morphology Categories (Assessed Manually) ................ 53
Table 3
Adverse Event Severity Grading Scale ....................................... 67
Table 4
Causal Attribution Guidance ....................................................... 68
Table 5
Threshold for Hypertension Diagnosis Based on Ambulatory 
Blood Pressure Monitoring.......................................................... 85
LIST OF FIGURES
Figure 1
Study Schema............................................................................. 34
LIST OF APPENDICES
Appendix 1
Schedule of Activities.................................................................. 95
Appendix 2
Schedule of Pharmacokinetic, Vital Signs, and ECG.................. 97


Balovaptan —F. Hoffmann-La Roche Ltd
10/Protocol WP40734, Version 3.0
PROTOCOL ACCEPTANCE FORM
TITLE:
A SINGLE-CENTER, MULTIPLE-DOSE, 
RANDOMIZED, DOUBLE-BLIND, PLACEBO-
CONTROLLED, POSITIVE-CONTROLLED, CROSS-
OVER STUDY TO INVESTIGATE THE EFFECT OF 
BALOVAPTAN ON THE QTC INTERVAL IN 
HEALTHY SUBJECTS
PROTOCOL NUMBER:
WP40734
VERSION NUMBER:
v3.0
EUDRACT NUMBER:
NA
IND NUMBER:
116483
TEST PRODUCT:
Balovaptan (RO5285119)
MEDICAL MONITOR:
, MD
SPONSOR:
F. Hoffmann-La Roche Ltd
I agree to conduct the study in accordance with the current protocol.
Principal Investigator’s Name (print)
Principal Investigator’s Signature
Date
Please retain the signed original of this form for your study files.  Please return a copy of 
the signed form by your local study monitor to the contact provided below.
Roche Products Limited
6 Falcon Way, Shire Park
Welwyn Garden City,
AL7 1TW, United Kingdom


Balovaptan —F. Hoffmann-La Roche Ltd
11/Protocol WP40734, Version 3.0
PROTOCOL SYNOPSIS
TITLE:
A SINGLE-CENTER, MULTIPLE-DOSE, RANDOMIZED, DOUBLE-
BLIND, PLACEBO-CONTROLLED, POSITIVE-CONTROLLED,
CROSS-OVER STUDY TO INVESTIGATE THE EFFECT OF 
BALOVAPTAN ON THE QTC INTERVAL IN HEALTHY SUBJECTS
PROTOCOL NUMBER:
WP40734
VERSION NUMBER:
v3.0
EUDRACT NUMBER:
NA
IND NUMBER:
116483
TEST PRODUCT:
Balovaptan (RO5285119)
PHASE:
Phase 1
INDICATION:
Not applicable
SPONSOR:
F. Hoffmann-La Roche Ltd
Objectives and Endpoints
This study will investigate the effect of balovaptan (RO5285119) on cardiac repolarization by 
measuring ECGs in healthy subjects following multiple doses of the targeted therapeutic and 
supra-therapeutic doses.
Primary Objective
Corresponding Endpoint
1.
To investigate the effect of balovaptan 
50 mg QD on the QTcF interval after 14 
days of dosing.
1.
Placebo-corrected, change-from-baseline QTcF
(QTcF) measured on 12-lead ECGs extracted 
from continuous recordings at the specified time 
points on Day 14
Secondary Objectives
Corresponding Endpoints
1.
To investigate the effect of a single dose 
of balovaptan 50 mg on the QTcF 
interval at Day 1.
1.
Placebo-corrected, change-from-baseline QTcF 
(QTcF) measured on 12-lead ECGs extracted 
from continuous recordings at the specified time 
points on Day 1.
2.
To investigate the effect of balovaptan 
10 mg QD on the QTcF interval at Day 1 
and Day 14.
2.
Placebo-corrected, change-from-baseline QTcF 
(QTcF) measured on 12-lead ECGs extracted 
from continuous recordings at the specified time 
points on Day 1 and Day 14.


Balovaptan —F. Hoffmann-La Roche Ltd
12/Protocol WP40734, Version 3.0
3.
To investigate the effect of balovaptan
on the following ECG parameters: PR, 
RR, QRS, QT, T-wave and U-wave 
morphology.
3.
Placebo-corrected, change-from-baseline heart 
rate (HR), PR and QRS interval (HR, PR 
and QRS) measured on 12-lead ECGs 
extracted from continuous recordings at the 
specified time points on Day 1 and Day 14.

Categorical outliers for QTcF, HR, PR, and 
QRS.

Frequency of treatment emergent changes of 
T-wave morphology and U-wave presence.
4.
To investigate the pharmacokinetics of 
balovaptan, M2 (as appropriate) and 
M3.
4.
Pharmacokinetic parameters estimation such as 
Tmax, Cmax, and AUC0-24h for plasma 
concentrations of balovaptan, M2 (as 
appropriate) and M3. Other pharmacokinetic 
parameters may also be added.
5.
To investigate the pharmacokinetics of 
moxifloxacin.
5.
Pharmacokinetic parameters estimation such as 
Tmax, Cmax, and AUC0-24h for plasma 
concentrations of moxifloxacin. Other 
pharmacokinetic parameters may also be added.
6.
To investigate the relationship between 
plasma concentrations of balovaptan, 
M2 (as appropriate), M3 and QTcF 
interval length.
6.
Pharmacokinetic-pharmacodynamic model of 
balovaptan, M2 (as appropriate) and M3 
concentrations vs. QTcF changes.
7.
To investigate the relationship between 
plasma concentrations of moxifloxacin 
and QTcF interval length (if warranted).
7.
Pharmacokinetic-pharmacodynamic model of 
moxifloxacin concentrations vs. QTcF changes (if 
warranted).
8.
To investigate the ECG assay sensitivity 
to detect a change in the QTcF interval, 
using moxifloxacin 400 mg as an active 
control
8.
Placebo-corrected, change-from-baseline QTcF 
(QTcF) measured on 12-lead ECGs extracted 
from continuous recordings at the specified time 
points on Day 2 and Day 15.
9.
To investigate the safety and tolerability 
of balovaptan and moxifloxacin at the 
dosing regimens scheduled.
9.
Safety assessments: adverse events, physical 
examination, vital signs, laboratory tests, 
C-SSRS and HR monitoring.
Exploratory Objectives
Corresponding Endpoints
1.
To investigate any effect of balovaptan
at 50 mg QD and 10 mg QD doses on 
pulmonary blood pressure as assessed 
by the use of TT-doppler 
echocardiography.
1.
TT-Doppler echocardiography: pulmonary artery 
acceleration time (PAAT), right ventricular 
outflow tract - velocity time integral (RVOT-VTI) 
and derived mean pulmonary arterial pressure as 
change to baseline relative to placebo treatment.


Balovaptan —F. Hoffmann-La Roche Ltd
13/Protocol WP40734, Version 3.0
2.
To investigate any effect of 50 mg qd 
and 10 mg qd on right and left heart as 
assessed by TT-Doppler echo.
2.
TT-Doppler-echocardiography: LVEF and 
additional secondary TT-Doppler-
echocardiography parameters of left and right 
heart, as described in Section 4.5.7 (also for 
objective 1)
3.
To investigate the effect 10 mg and 50 
mg QD balovaptan on blood pressure 
and heart rate as measured by the use 
of Holter-ECG and ABPM.
3.
Ambulatory blood pressure monitoring (ABPM): 
mean diastolic and systolic blood pressure 
(daytime and nighttime mean).

Heart rate will also be analyzed as 
derived from Holter ECG.
-
Heart rate as derived from ABPM 
and/or Holter ECG may be 
explored against fluctuations in 
blood pressure measured by 
ABPM (if warranted).
4.
To investigate whether genetic variants 
in genes are associated with natural or 
potential drug induced neutropenia; with 
differences in pharmacokinetics of 
balovaptan, M2 or M3, or with any 
potential effect of balovaptan on ECG 
parameters.
5.
The genetic variants in genes that are associated 
with natural or potential drug induced 
neutropenia; genetic variants in genes that are 
associated with pharmacogenetics of 
metabolizing enzymes, transferases, 
transporters, etc, (eg,CYP3A4 and P-
glycoprotein); genetic variants in genes that may 
be associated with a potential effect of 
balovaptan on ECG parameters.
Study Design
This will be a single-center, multiple-dose, randomized, double-blind, placebo-controlled, positive-
controlled, twelve sequence, 3-period cross-over study to investigate the effect of balovaptan on 
the QTc interval in healthy subjects.
Screening
Screening will be conducted up to 28 and 3 days (inclusive) before each subject’s first 
dosing day (Day 1 of Treatment Period 1). 
In-Clinic Confinement
Day -2
Subjects will be confined to the CRU starting on Day -2 until the morning of Day 16 during 
each Treatment Period. Subjects are entered into CRU on Day -2 so that subjects are 
rested and prepared for the baseline ABPM and baseline TT-Doppler echocardiogram on 
Day -1.  Eligibility assessments will be performed on Day -2 as outlined in the Schedule of 
Activities (SoA), Appendix 1.
Day -1
On Day -1 of each Treatment Period, ABPM and TT-Doppler echocardiogram assessments 
will be performed.  These results will serve as the baseline for the subjects.  On days when 
TT-Doppler echocardiogram and ABPM assessments are performed subjects will be given 
low salt standardized meals.


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14/Protocol WP40734, Version 3.0
Randomization (Day -1)
At the start of the study in Treatment Period 1, at the end of the day of Day -1, subjects will 
be randomized to one of the 12 treatment sequences comprised of the 4 treatments 
described:
Treatments
Treatment A (balovaptan therapeutic dose)
Days 1-14:
A single once daily oral dose of 10 mg (1 × 10 mg tablet) of balovaptan and 
4 tablets of matching placebo for balovaptan for 14 days.
Day 2 and 15: A single oral dose of a matching placebo capsule for moxifloxacin.
Treatment B (balovaptan supra-therapeutic dose)
Days 1-14:
A single once daily oral dose of 50 mg (5 x 10 mg tablets) of balovaptan for 
14 days.
Day 2 and 15: A single oral dose of a matching placebo capsule for moxifloxacin.
Treatment C (active control [400 mg moxifloxacin] on Day 2)
Days 1-14:
A single once daily oral dose of 5 tablets of matching placebo for balovaptan 
for 14 days.
Day 2:
A single oral dose of 400 mg moxifloxacin capsule
Day 15: 
A single oral dose of a matching placebo capsule for moxifloxacin 
Treatment D (active control [400 mg moxifloxacin] on Day 15)
Days 1-14:
A single once daily oral dose of 5 tablets of matching placebo for balovaptan 
for 14 days.
Day 2:
A single oral dose of a matching placebo capsule for moxifloxacin
Day 15: 
A single oral dose of 400 mg moxifloxacin capsule.
Treatment Sequences
1.
A, B, C
2.
A, C, B
3.
B, A, C
4.
B, C, A
5.
C, A, B
6.
C, B, A
7.
A, B, D
8.
A, D, B
9.
B, A, D
10. B, D, A
11. D, A, B
12. D, B, A
Dosing (Days 1-15)
Each Treatment Period will involve 15 days of dosing (Day 1 – Day 15).  Subjects must fast 
for at least 8 hours prior to and 4 hours after dosing on study days when 24-hour Holter ECG 
assessments will be performed (study Days 1, 2, 14 and 15).  Dosing on study days with 
24-hour Holter ECG assessment should be at approximately the same time of day ± 1h.
Prior to dosing on Day 1 subjects will undergo pre-dose assessments as outlined in the SoA 
(Appendix 1).  Following pre-dose assessments subjects will receive a single dose of study 
medication preceded by a ≥8 hour fast the night before.  Study drugs will be taken with 
240 mL of water and be followed by a 4-hour post-dose fast. If subjects require more liquid 
to take all study drugs, additional water may be provided in aliquots of 50 mL.  If additional 


Balovaptan —F. Hoffmann-La Roche Ltd
15/Protocol WP40734, Version 3.0
water is provided the amount should be captured. Water provided for study drug 
administration will be identical on all dosing days.  Identical fasting procedures from Day 1 
will be followed on study Days 2, 14 and 15.  No fasting will be required on other dosing 
days.  Subjects will receive standardized low salt meals on study days when 24-hour Holter 
ECGs (Days 1, 2, 14 and 15), ABPM (Days -1 and 12) or TT-Doppler echocardiogram
(Days -1 and 13) assessments will be performed. Meals will be given at approximately the 
same time on each of these study days.
Days 1 and 2: 24-hour Holter ECG Assessments
On study Days 1 and 2 of each Treatment Period, 24-hour ECG assessments will be 
performed.  Subjects will be administered study drug following a ≥ 8 hour fast and continue 
to be fasted for 4 hours post-dose on these days.  Subjects will be provided with 
standardized low salt meals on these days.  Subjects will undergo pre and post-dose 
assessments as outlined in the SoA (Appendix 1).
Day 12
On study Day 12 of each Treatment Period, ABPM assessments will be performed for each 
subject. On study days when ABPM assessments are performed subjects will be provided 
with standardized low salt meals.  Subjects will undergo pre and post-dose assessments as 
outlined in the SoA (Appendix 1).
Day 13
On study Day 13 of each Treatment Period, at approximately the same time of day as on 
Day -1, TT-Doppler echocardiogram will be performed to assess any effects of steady state 
balovaptan levels on cardiac parameters.  Subjects will receive standardized low salt meals 
on this study day.  In addition to TT-Doppler echocardiogram subjects will undergo pre and 
post-dose assessments as outlined in the SoA (Appendix 1).
Days 14 and 15: 24-hour Holter ECG Assessments
On study Days 14 and 15 of each study Treatment Period, 24-hour ECG assessments will 
be performed. Subjects will be administered study drug following a ≥8 hour fast and 
continue to be fasted for 4 hours post-dose.  Subjects will be provided with standardized low 
salt meals on these days.  Subjects will undergo pre and post-dose assessments as outlined 
in the SoA (Appendix 1).
Day 16
Subjects will leave the clinical research unit (CRU) on the morning of Day 16 after final 
assessments as described in the SoA (Appendix 1).
Washout Period:
Following each Treatment Period subjects will undergo a washout period lasting a minimum 
of 13 and a maximum of 20 days from the last dose of study drug.
Follow-up:
Subjects will return to the unit for a follow-up visit between 14 and 21 days after the final 
dose of study drug.  This will include a general physical examination, 12-lead-ECG, vital 
signs, C-SSRS, adverse event review, body weight and laboratory safety tests.
Number of Subjects
56 subjects will be enrolled with the goal of obtaining 48 evaluable subjects.
Target Population
The target population will be healthy male or female volunteers aged 18 to 60 years inclusive.  An 
attempt will be made to have each Treatment population contain at least 1/3 male subjects by 
ensuring that a minimum of 40% of enrolled population are male.


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Inclusion Criteria
Subjects must meet the following criteria for study entry:
1.
Healthy male or female volunteers aged 18 to 60 years inclusive, at screening.  Healthy 
status is defined by absence of evidence of any active or chronic disease following a 
detailed medical and surgical history, a complete physical examination including vital 
signs, 12-lead ECG, hematology, blood chemistry, urinalysis, and serology.
2.
Body Mass Index (BMI) between 18 and 32 kg/m2 inclusive, at Screening.
3.
For women of childbearing potential: agreement to use at least 1 acceptable form of 
contraception during the entire study and for 90 days following last dose of study drug. 
a.
A woman is considered to be of childbearing potential if she is postmenarcheal, 
has not reached a postmenopausal state (> 12 continuous months of 
amenorrhea with no identified cause other than menopause), and has not 
undergone surgical sterilization (removal of ovaries and/or uterus). 
b.
The following are acceptable contraceptive methods: abstinence from 
heterosexual activity; bilateral tubal ligation; sterilization of male partner; 
hormonal contraceptives; hormone-releasing intrauterine devices; copper 
intrauterine devices; Essure.
c.
Barrier methods alone are not considered an acceptable form of contraception 
for this study and if utilized must be supplemented by one of the acceptable 
methods described above. For this study, barrier contraception methods are 
defined as: male or female condom with or without spermicide (male and female 
condom must not be used simultaneously); and cap, diaphragm, or sponge with 
spermicide.
4.
For men: Vasectomized, agreement to remain abstinent (refrain from heterosexual 
intercourse) or use of a condom during intercourse. Must also agree to refrain from 
donating sperm, as defined below:
a.
With female partners of childbearing potential or pregnant female partners, men 
must remain abstinent or use a condom during the entire study and for 90 days 
after the last dose of study drug to avoid exposing the embryo.  Men must refrain 
from donating sperm during this same period.
5.
Able to participate, and willing to give written informed consent and to comply with the 
study restrictions.
6.
Fluent in English.
Exclusion Criteria
Subjects who meet any of the following criteria will be excluded from study entry:
Past Medical History
1.
History of any clinically significant gastrointestinal, renal, hepatic, broncho-pulmonary, 
neurological, psychiatric, cardiovascular, endocrinological, hematological, lymphatic, 
musculoskeletal, genitourinary, immunological, dermatological or connective tissue or 
allergic disease, metabolic disorder, or cancer.
2.
Signs and symptoms potentially indicative of peripheral neuropathy.
3.
Unexplained orthostatic reaction with syncope within last 6 months.
4.
A history of clinically significant hypersensitivity (e.g., to drugs, including moxifloxacin or any 
quinolone antibiotics and balovaptan, and excipients) or allergic reactions.
5.
History of alcohol and/or drug abuse or addiction within the last 2 years before study 
enrollment.


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6.
Subjects on hormone replacement therapy if post-menopausal status cannot be documented 
from medical history.
7.
Previous treatments with iron for iron deficiency anemia within three months of screening.
8.
History of coagulopathies, bleeding disorders, or blood dyscrasias.
9.
History of porphyria, active liver disease or permanent or unexplained elevation of serum 
transaminases.
10. In the opinion of the Investigator or designee, any major illness within one month before the 
screening examination or any febrile illness within one week prior to Screening and up to first 
study drug administration.
Current Status
1.
Clinically relevant deviation from normal in the physical examination including vital signs.
2.
Positive result on human immunodeficiency virus (HIV) 1, HIV 2, hepatitis C virus (HCV) 
antibody, or hepatitis B virus surface antigen (HBsAg).
3.
Clinically significant abnormalities in laboratory test results (including complete blood count, 
chemistry panel, and urinalysis). In the case of uncertain or questionable results, tests 
performed during Screening may be repeated on Day -1 or Day -2 to confirm eligibility.  
Repeat of abnormal test results are allowed during Screening and eligibility assessment at 
the discretion of the Investigator or designee provided results will be available prior to 
Randomization on Day -1.
4.
If female, a positive pregnancy test at screening or prior to Day 1 of any Treatment Period.
5.
Lactating women.
6.
Any condition or disease detected during the medical interview / physical examination that 
would render the subject unsuitable for the study, place the subject at undue risk or interfere 
with the ability of the subject to complete the study in the opinion of the Investigator or 
designee. 
7.
Subjects with CPK values > 2x upper limit of normal (ULN), confirmed by repeat sample.
8.
Positive urine alcohol test or urine drug test at Screening or Day -2 of each Treatment Period 
(amphetamines, barbiturates, benzodiazepines, cannabinoids, cocaine, opiates, methadone, 
methamphetamines, and 3,4-methylenedioxymethamphetamine [MDMA]).
9.
Subject has been a regular smoker (> 5 cigarettes, > 1 pipeful, > 1 cigar per day or 
equivalent as determined by Investigator or designee) within 6 months of study enrollment.
10. Coffee (or tea) consumption > 5 cups per day or methylxanthine-containing drinks >1.5 
liter/day or more than 250 g/day of chocolate within 3 months of first dose of study drug. 


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11. Alcohol consumption of > 18 units per week for males and females. One unit of alcohol 
equals 12 ounces of regular beer, 5 ounces of wine or 1.5 ounces of 80 proof spirits within 
3 months of first dose of study drug.
12. Subjects who have lost or donated in excess of ~450 mL blood in the 3 months prior to 
Day 1 of Treatment Period 1.
13. Subjects under judicial supervision, guardianship, or curatorship.
14. Dietary restrictions that would prohibit the consumption of standardized meals.
15. Poor venous access for blood sampling.
Concomitant Medication
1.
Taken greater than 2 g/day paracetamol within 48 hours of study drug administration or 
greater than 4 g/day paracetamol within 7 days of study drug administration. 
2.
Any nonprescribed systemic or topical medication or herbal remedies are prohibited for 
7 days prior to the first study drug administration through Follow-up.
3.
Participation in an investigational drug study within 60 days or 5 times the elimination half-life, 
whichever is longer, prior to first dosing, or within 5 months prior to first dosing in case of a 
study with a biological, as calculated from the last day of follow-up from the previous study.
4.
Participation in an investigational device study within 60 days of first dosing.
5.
Subjects who have received any prescribed systemic or topical medication within 14 days of 
the first dose administration are excluded, unless in the opinion of the Investigator (or 
designee) the medication will not interfere with the study procedures or compromise safety. 
6.
Any slow release medicinal formulations considered to still be active within 4 weeks (or within 
5 times the elimination half-life of the medication, whichever is longer) prior to the first study 
drug administration are prohibited, unless in the opinion of the Investigator or designee the 
medication will not interfere with the study procedures or compromise safety.
7.
Taking any inhibitor of CYP3A taken within 4 weeks (or within 5 half-lives, whichever is longer) 
prior to study drug administration or taking any inducer of CYP3A within 4 weeks prior to the 
first dosing.
8.
Use of any prohibited medications and food before study start and subjects who do not agree 
to refrain from consuming prohibited medications or food during the study.
9.
Subjects who have previously enrolled in, or withdrawn after enrollment, from this study.
TQT-related Exclusion Criteria
1.
Personal history of unexplained black-outs or faints.
2.
Personal history of reactions or faints at having their blood taken, unexplained black-outs or 
faints or any condition which could relapse during or immediately after the study which in the 
opinion of the Investigator or designee makes the subject unsuitable to participate in this 
study.
3.
Subjects experiencing fainting or pre-syncopal events during orthostatic challenge testing, 
performed according to site SOP, at Screening or Day -2 of Treatment Period 1.
4.
Subjects with Screening or Treatment Period 1 (Day -2) baseline mean QT interval corrected 
using Fridericia’s formula (QTcF) > 450 ms or < 300 ms (using the same upper QTcF limit in 
both males and females) or > 30 ms difference between the highest and the lowest of the 
triplicate readings.


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5.
Screening or baseline (Day -2, Treatment Period 1) ECG evidence of atrial fibrillation, atrial 
flutter, second or third-degree AV block, complete right or left bundle branch block, 
Wolff-Parkinson-White-syndrome, or cardiac pacemaker.
6.
Screening or baseline ECGs (Day -2 of Treatment Period 1) with QRS and/or T-wave judged 
to be unfavorable for a consistently accurate QT measurement (i.e., neuromuscular artifact 
that cannot be readily eliminated, arrhythmias, indistinct QRS onset, low amplitude T-wave, 
merged T- and U-waves, prominent U-waves, etc.).
7.
Confirmed (based on the average of ≥3 consecutive measurements) systolic blood pressure 
greater than 139 or less than 90 mmHg, and diastolic blood pressure greater than 89 or less 
than 45 mmHg at Screening or Day -2 of Treatment Period 1.
8.
Confirmed (based on the average of ≥3 consecutive measurements) resting HR > 100 bpm 
or < 40 bpm at Screening or Day -2 of Treatment Period 1.
9.
Notable resting bradycardia (HR < 40 bpm) on ECG at Screening or Day -2 of Treatment 
Period 1. Notable resting tachycardia (mean HR > 100 bpm) on ECG at Screening or Day -2
of Treatment Period 1.
10. Family history of congenital long QT syndrome or sudden death.
11. History or presence of clinically significant ECG abnormalities, as determined by study 
Investigator or designee, before study drug administration.
End of Study
The end of this study is defined as the date when the last subject, last visit occurs, or safety 
follow-up is received from the last subject, whichever occurs later.
Length of Study
The total length of the study, from screening of the first subject to the end of the study, is 
expected to be approximately 14-20 weeks.  For each enrolled subject the duration of the study 
from Screening to the end of Follow-up will be a minimum of 122 days or a maximum of 143 days.

Screening: up to 28 days

Dosing periods including washout: 80 – 94 days
o
Treatment Period 1: 18 Days (Day -2 – Day 16)
o
Washout: 13-20 Days
o
Treatment Period 2: 18 Days (Day -2 – Day 16)
o
Washout: 13-20 Days
o
Treatment Period 3: 18 Days (Day -2 – Day 16)

Follow up: 14 - 21 days after last dosing
Investigational Medicinal Products
Test Product (Investigational Drug)
Balovaptan 10 mg (Therapeutic dose) orally, once-daily for 14 days
Balovaptan 50 mg (Supratherapeutic dose) orally, once-daily for 14 days
Moxifloxacin 400 mg, single, oral dose
Comparator
Placebo controlled


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Statistical Methods
For cardiodynamic ECG evaluation, the primary analysis will be based on by-time point analysis 
to evaluate the effect of balovaptan on the placebo-corrected change-from-baseline QTcF 
(ΔΔQTcF) at each post-dosing time point using the Intersection Union Test. The effect of 
balovaptan on placebo-corrected change-from-baseline in HR, PR, and QRS (ΔΔHR, ΔΔPR, and 
ΔΔQRS) will also be evaluated using the Intersection Union Test. In addition, the relationship 
between plasma concentrations of balovaptan and its metabolites (M2 [as appropriate] and M3) 
and Δ∆QTcF will be evaluated using a linear mixed-effects modeling approach. An analysis of 
categorical outliers will be performed for changes in HR, PR, QRS, QTcF, T-wave morphology 
and U-wave presence. Assay sensitivity will also be evaluated using by-time point analysis of the 
effect on Δ∆QTcF of moxifloxacin using a similar model as for the primary analysis.
Determination of Sample Size:
A sample size of 56 subjects was chosen to obtain 48 evaluable subjects.  Subjects are evaluable 
if ECGs are available for all three periods to allow (1) the derivation of the primary endpoint (i.e., 
change from baseline on Day 14), and (2) the assessment of the effect of moxifloxacin.  
Assuming a 1-sided 5% significance level and a within-subject standard deviation (SD) of 8 ms 
for ΔQTcF and a true mean difference of 3 ms in ΔQTcF between balovaptan and placebo, a 
sample size of 48 evaluable subjects will provide a power of 96.6% to demonstrate that the upper 
bound of all the 2-sided 90% CIs will fall below 10 ms for up to 7 post-dose time points on Day 14.  
Sample Size Determination for Assay Sensitivity: Assuming a 1-sided 5% significance level and a 
within-subject SD of 8 ms for ΔQTcF, a sample size of 48 evaluable subjects will provide a power 
of 99% to exclude a mean difference of 5 ms in ΔQTcF between moxifloxacin and placebo groups 
from the lower bound of the 2-sided 90% CI at least one of the three pre-specified time points.


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LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS
Abbreviation
Definition
ABPM
                       
ambulatory blood pressure monitoring
AE
adverse event
ALT
alanine transaminase 
ANC
absolute neutrophil count
ASD
autism spectrum disorder
AST
aspartate transaminase
AUC
area under the curve
BMI
body mass index
BP
blood pressure
BPM
beats per minute
Cl
confidence interval
Cmax
mean maximum observed plasma concentration
cTnT
cardiac troponin
CNS
central nervous system
CPK
creatine phosphokinase
CRO
contract research organization
CRU
clinical research unit
C-SSRS
Columbia-suicide severity rating scale (C-SSRS)
CTCAE
common terminology criteria for adverse events
Δ
change-from-baseline
ΔΔ
placebo-corrected change-from-baseline
DMC
data monitoring committee
EC
ethics committee
ECG
electrocardiogram
eCRF
electronic Case Report Form
EPQT
expert precision QT technique
EDC
electronic data capture
FDA
Food and Drug Administration
FSH
follicle-stimulating hormone
GLP
good laboratory practice
HBsAg
hepatitis B virus surface antigen
HCV
hepatitis C virus
HIPAA
Health Insurance Portability and Accountability Act
HIV
human immunodeficiency virus
hERG
human ether-à-go-go related gene


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HR
heart rate
ICH
International Council for Harmonisation
IMP
investigational medicinal product
IND
Investigational New Drug (Application)
IRB
Institutional Review Board
LDH
lactate dehydrogenase
LPLV
last patient, last visit
LS
least squares
LVEF
left ventricular ejection fraction
MAD
multiple ascending dose
MDMA
3,4 Methylenedioxymethamphetamine
mEq/L
milliequivalents per liter
mmHg
millimeters of Mercury
NCI
National Cancer Institute 
NT-proBNP
N-terminal pro B-type natriuretic peptide
NOAEL
no observed adverse effect level
NCI
National Cancer Institute
PAAT
pulmonary artery acceleration time
P-gp
P-Glycoprotein
PK
pharmacokinetic
PND
postnatal day
PR
PR interval of the ECG
PRO
patient-reported outcome
QRS
QRS interval of the ECG
Q
QT interval of the ECG
QTc
corrected QT interval
QTcF
QT interval corrected through use of Fridericia’s 
formula
RR
RR interval of the ECG
RVOT
right ventricular outflow tract
SAD
single ascending dose
SAE
serious adverse event
SD
standard deviation
SE
standard error
SOA
schedule of activities
Tmax
time to reach Cmax
TT
trans thoracic
TVI
transvalvular impedance
ULN
upper limit of normal


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1.
BACKGROUND
1.1
AUTISM SPECTRUM DISORDER
Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental disorder
characterized by two core domains: impairments in social interaction and communication
and the presence of repetitive or restricted behaviors, interests, or activities (American
Psychiatric Association, Diagnostic and statistical manual of mental disorders (DSM-5®) 
2013). The Autism and Developmental Disabilities Monitoring Network recently reported, 
based on children records, that the estimated prevalence of ASD in the United States for 
the year 2010 was 14.7 per 1000 (one in 68) children aged 8 years (Autism and 
Developmental Disabilities Monitoring Network Surveillance Year 2010 Principal 
Investigators, 2014). Core symptoms of ASD affecting domains of socialization, 
communication, and repetitive behavior are usually observed by 3 years of age, although 
typical language development might delay identification of symptoms.
Evidence from both human and animal studies strongly implicates the V1a receptor in 
mediating and modulating key social behaviors that are deficient in ASD. Together with 
the known negative effects of intranasal administration of vasopressin on emotional 
cognition, a V1a receptor antagonist may provide a novel and first approach to treat the 
deficits at the core of these disorders.
Balovaptan (RO5285119) is a potent and selective human V1a receptor antagonist that 
blocks the activation of the V1a G protein-coupled receptor. More detailed information 
around ASD and Balovaptan are presented in the Investigator’s Brochure.
1.2
PRE-CLINICAL BACKGROUND ON BALOVAPTAN
The in vitro metabolism of balovaptan was studied in hepatic preparations from human, 
rat, dog, minipig, cynomolgus monkey, mouse, and rabbit. In microsomes from all 
species tested, major metabolism pathways were 
 (formation of M3) 
and 
 (formation of M1), whereas further secondary metabolites including 
oxidations and desaturations were found at low amounts.
The in vivo pharmacokinetic (PK) studies with balovaptan were conducted in rats, dogs, 
and cynomolgus monkeys. In animals, balovaptan showed a high (rats) to moderate or 
low (dogs and cynomolgus monkeys) plasma clearance and moderate extravascular
distribution. High to moderate bioavailability, depending on the species, was observed 
after oral application. In human, 2 major metabolites, M2 
and M3 
, have been identified, which are present in rats and dogs. 
 
 
Balovaptan and M3 have been shown in vitro to be potentially strong, 
substrate dependent P-glycoprotein (P-gp) inhibitors.
 


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In vitro cardiovascular safety was assessed in a Good Laboratory Practice (GLP) human 
ether-à-go-go related gene (hERG) assay, and the balovaptan concentration required for 
20% inhibition (IC20) was 4.4 μM, which is orders of magnitude above the free 
concentrations expected to be achieved in man at a 10 mg daily dose. Mild effects on
cardiovascular functions observed in the pivotal dog telemetry study consisted of a mild 
sustained increase in systolic and mean blood pressures, paralleled by a left ventricular 
peak systolic increase. An additional mild sustained increase in positive and negative 
dP/dt and a mild transient heart rate (HR) increase was noted. The no observed effect 
level (NOEL) was 10 mg/kg. In the GLP safety pharmacology studies conducted with 
rats assessing CNS, respiratory, and renal function, 100 mg/kg was established as the 
NOEL/NOAEL.
Balovaptan was not irritating to the skin or eye in the rabbit, nor did the compound prove 
to be a skin sensitizer.
1.2.1
Previous and Ongoing Clinical Studies
To date, balovaptan has been investigated in 6 completed Phase I studies, including a 
proof-of-mechanism study in healthy volunteers (BP29412). A Phase II study in adult 
male ASD patients has also been completed (BP28420, also referred to as VANILLA).
Balovaptan is currently being evaluated in 4 ongoing studies.  Two of these are Phase I 
studies in healthy subjects examining pharmacokinetic CYP3A related drug-drug 
interactions (DDI), (WP40608 [rifampicin DDI] and WP40609 [itraconazole DDI]).
Balovaptan is also currently being evaluated in a Phase II study (BP30153, also referred 
to as AV1ATION) in pediatric ASD patients and a Phase III study (WN39434 also 
referred to V1aduct) in adult ASD patients.
1.2.1.1
Clinical Pharmacology
Exposure of balovaptan increased in a greater than dose-proportional manner following 
single doses of 0.5 to 76 mg, whereas an approximately linear increase in exposure was 
observed after repeated dosing with 12 to 52 mg once daily (qd) for 14 days.
Balovaptan was rapidly absorbed with a median time to maximum observed plasma 
concentration (Tmax) between 1 and 4.5 hours after administration of single doses in 
fasted state and between 3 and 4 hours in fed state following multiple dosing. Steady 
state was achieved after approximately 7 days; the mean apparent elimination half-life 
after the last dose (Day 14) was approximately 46 hours. Food had no relevant effect on 
the overall exposure of balovaptan.
In the human mass balance study (Study BP29279), an almost complete absorption of 
the study drug was observed within the first 72 hours following administration.
In plasma, steady state exposure to 
 M3 was similar to or 
slightly higher than that of balovaptan and exposure to 
 M2 
was lower than that of parent. M3 and M2 qualify as major metabolites in plasma.


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Mean cerebral spinal fluid (CSF)-to-plasma concentration ratios of balovaptan and M3 
following 7 days of dosing of 10 and 52 mg qd balovaptan were independent of dose. 
The ratio was approximately 12% for balovaptan, which is comparable to the balovaptan 
plasma free fraction (approximately 13%), and 6% for M3, which is lower than the M3 
plasma free fraction (15 to 17%) reflecting M3 being a P-gp substrate. Data regarding 
M2 exposure in CSF is not available at this time.
Hepatic metabolism was found to be the major pathway of elimination with the primary 
route of excretion via urine (approximately 53% of the material recovered, mostly 
composed of metabolites). A further 30% of the administered dose was recovered in 
feces. The majority of excretion occurred within the first 7 days.
There was no clinically/statistically relevant PK interaction with the P-glycoprotein 
substrate risperidone, one of only 2 approved medications for ASD symptoms in the US, 
and fluoxetine, a weak CYP3A4 inhibitor often used to treat associated symptoms of 
ASD. Administration of a single dose of balovaptan 12 mg in combination with 
itraconazole, a strong CYP3A4 inhibitor, resulted in an increase in the exposure of 
balovaptan by 36% for maximum observed plasma concentration (Cmax) and an 
approximately 3.1-fold increase for area under the concentration curve from time 0 to 
120 hours (AUC0-120h).
1.2.1.2
Safety
To date, balovaptan has been dosed to 217 healthy subjects in 6 completed Phase 1 
studies (Studies BP25694, BP28318, BP28977, BP29279, BP29412, and WP40038).
Additionally, balovaptan was investigated in a completed Phase II study in 148 adult 
male patients with ASD (BP28420, “VANILLA”), about 30 of whom received 10 mg qd for 
12 weeks.
Balovaptan has been found to be safe and well tolerated in clinical trials. No particular 
safety concerns have emerged, and no adverse drug reactions have been identified per 
reference safety information (Section 6.4 of the IB). The maximum tolerated dose was 
not reached in either the single ascending dose (SAD) study with single doses (up to 76 
mg) or the multiple ascending dose (MAD) study with multiple doses (up to 52 mg qd for 
14 days) in healthy subjects. In order to keep an approximately 10-fold safety margin to
observed in dogs, dose escalation was stopped so as not to exceed a Cmax (mean) of 
810 ng/mL in the SAD and MAD studies which compares to Cmax of about 100 and 
500 ng/mL after 10 and 50 mg qd dosing.
 
 
 
 
  


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Balovaptan has also been found to be safe and well tolerated in male patients with ASD 
at doses up to 10 mg/day (highest dose administered) for a period of 12 weeks (Study 
BP28420). No particular safety patterns attributable to the administration of balovaptan 
were identified. A total of 8 serious adverse events (SAEs) (all but one [rhabdomyolysis] 
considered unrelated to study drug) in 4 patients and 1 adverse event (AE) of special 
interest (neutropenia) were reported in the study. No relevant safety concern has 
emerged from the ongoing study in pediatric ASD patients (BP30153) and the recently 
started Phase 3 study in adults ASD patients (WN39434).
 
 
Cardio-vascular SAEs in the clinical program refer to clinically asymptomatic sinus node 
dysfunction / arrhythmia observed in a subject diagnosed with ASD dosed with 1.5 mg 
od in the Phase 2 study BP28420 in whom sinus arrhythmia was interpreted to be pre-
existing because follow-up examinations with 24-hour Holter ECG monitoring several 
weeks after the SAE showed the same pattern as seen when the SAE was reported. 
Second, another subject with ASD in Phase 2 study BP28420 suffered from syncope 
reported as SAE about a week after having voluntarily stopped treatment after 6 weeks 
of 4 mg qd. The same subject showed another syncope 4 weeks after the first one and 
died due to suspect heart failure about 16 weeks after last dosing. ECG monitoring and 
troponin measurements, which were part of the safety monitoring in this study, had not 
shown abnormalities at any time point. The event was considered “not related” to study 
treatment by the Investigator or designee. Of note, the subject had been taking multiple 
co-medications for several years including methylphenidate and aripiprazole, a 
combination that has been reported to be associated with a higher cardiovascular risk 
potentially.1
ECGs recorded in Phase I studies, including the MAD part of study BP25694 were 
analyzed by balovaptan concentration vs QTcF modelling. The preliminary data from 
these studies showed an increase in QTcF of < 5 ms for concentrations up to 550 ng/mL 
with an upper CI of < 13 ms at 550 ng/mL. In none of the subjects treated with 
balovaptan in the completed Phase I studies were QTcF values observed as 
QTcF > 450 ms or increases of QTcF from baseline of > 30 ms, except for a single 
observation but which was < 50 ms.
Refer to the Investigator's Brochure for more details on non-clinical and clinical studies.


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1.3
STUDY RATIONALE AND BENEFIT-RISK ASSESSMENT
This study investigates the effect of balovaptan on cardiac repolarization by measuring 
ECGs in healthy subjects following multiple doses of the targeted therapeutic (10 mg) 
and supra-therapeutic (50 mg) doses.
The study will be performed in healthy subjects who will not receive any health benefit 
from participating in this study. The study is necessary for the further development of
balovaptan and may be of benefit for future subjects.
The doses for the treatment periods planned in this study have been explored in the 
MAD part of study BP25694 and found to be safe and well tolerated. In proof-of-
mechanism study BP29412 a dose of 52 mg, once daily (qd), was administered for 
6 days to healthy volunteers (N=35) not revealing a balovaptan associated safety signal.
QTc analyses based on previous ECG recordings do not indicate relevant ECG changes.
The safety monitoring across all clinical studies has not identified a balovaptan linked 
adverse drug reaction.
Moxifloxacin, a fluoroquinolone antibiotic, was approved for use in the United States in 
1999 and is currently on the World Health Organization's list of essential medicines. The 
dose of moxifloxacin planned for this study 400 mg is the recommended dosage for 
adults in the package insert. Moxifloxacin has been shown to increase HR and is
associated with mild QT prolongation but only seldom with torsade de pointes.2 The 
single dose of moxifloxacin planned for this study will be used as a positive control to 
evaluate the sensitivity of the study to detect increases in QT interval measurements.
The FDA has proposed the use of moxifloxacin in this manner.3
While planned dosing of subjects in this study are expected to be well tolerated, diligent 
safety monitoring and in-house stays are included in this study during dosing. Subjects 
will be asked to report any events of feeling “not well” to study staff immediately.
In conclusion, any safety risk for the healthy volunteers participating in this study is 
considered minimal.
2.
OBJECTIVES AND ENDPOINTS
This study will investigate the effect of balovaptan (RO5285119) on cardiac 
repolarization by measuring ECGs in healthy subjects following multiple doses of the 
targeted therapeutic and supra-therapeutic doses.


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Table 1
Objectives and Corresponding Endpoints
Primary Objective
Corresponding Endpoint
1.
To investigate the effect of balovaptan 
50 mg qd on the QTcF interval after 
14 days of dosing.
1.
Placebo-corrected, change-from-baseline QTcF 
(QTcF) measured on 12-lead ECGs extracted 
from continuous recordings at the specified time 
points on Day 14
Secondary Objectives
Corresponding Endpoints
1.
To investigate the effect of a single dose 
of balovaptan 50 mg on the QTcF 
interval at Day 1.
1.
Placebo-corrected, change-from-baseline QTcF 
(QTcF) measured on 12-lead ECGs extracted 
from continuous recordings at the specified time 
points on Day 1.
2.
To investigate the effect of balovaptan 
10 mg qd on the QTcF interval at Day 1 
and Day 14.
2.
Placebo-corrected, change-from-baseline QTcF 
(QTcF) measured on 12-lead ECGs extracted 
from continuous recordings at the specified time 
points on Day 1 and Day 14.
3.
To investigate the effect of balovaptan 
on the following ECG parameters: PR, 
RR, QRS, QT, T-wave and U-wave 
morphology.
3.
Placebo-corrected, change-from-baseline heart 
rate, PR and QRS interval (HR, PR and 
QRS) measured on 12-lead ECGs extracted 
from continuous recordings at the specified time 
points on Day 1 and Day 14.

Categorical outliers for QTcF, HR, PR, and 
QRS.

Frequency of treatment emergent changes of 
T-wave morphology and U-wave presence.
4.
To investigate the pharmacokinetics of 
balovaptan, M2 (as appropriate) and 
M3.
4.
Pharmacokinetic parameters estimation such as 
Tmax, Cmax, and AUC0-24h for plasma 
concentrations of balovaptan, M2 (as 
appropriate) and M3. Other pharmacokinetic 
parameters may also be added.
5.
To investigate the pharmacokinetics of 
moxifloxacin.
5.
Pharmacokinetic parameters estimation such as 
Tmax, Cmax, and AUC0-24h for plasma 
concentrations of moxifloxacin. Other 
pharmacokinetic parameters may also be added.
6.
To investigate the relationship between 
plasma concentrations of balovaptan, 
M2 (as appropriate), M3 and QTcF 
interval length.
6.
Pharmacokinetic-pharmacodynamic model of 
balovaptan, M2 (as appropriate) and M3 
concentrations vs. QTcF changes.


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Table 1
Objectives and Corresponding Endpoints (cont.)
7.
To investigate the relationship between 
plasma concentrations of moxifloxacin 
and QTcF interval length (if warranted).
7.
Pharmacokinetic-pharmacodynamic model of 
moxifloxacin concentrations vs. QTcF changes
(if warranted)
8.
To investigate the ECG assay sensitivity 
to detect a change in the QTcF interval, 
using moxifloxacin 400 mg as an active 
control.
8.
Placebo-corrected, change-from-baseline QTcF 
(QTcF) measured on 12 lead ECGs extracted 
from continuous recordings at the specified time 
points on Day 2 and Day 15
9.
To investigate the safety and tolerability 
of balovaptan and moxifloxacin at the 
dosing regimens scheduled.
9.
Safety assessments: adverse events, physical 
examination, vital signs, laboratory tests, 
C-SSRS, and heart rate (HR) monitoring.
Exploratory Objectives
Corresponding Endpoints
1.
To investigate any effect of balovaptan 
at 50 mg QD and 10 mg QD doses on 
pulmonary blood pressure as assessed 
by the use of TT-doppler 
echocardiography.
2.
To investigate any effect of 50 mg qd 
and 10 mg qd on right and left heart as 
assessed by TT-Doppler echo.
1.
TT-Doppler echocardiography: pulmonary artery 
acceleration time (PAAT), right ventricular 
outflow tract - velocity time integral (RVOT-VTI) 
and derived mean pulmonary arterial pressure as 
change to baseline relative to placebo 
treatment.(objectives 1 & 2)
2.
TT-Doppler-echocardiography: LVEF and 
additional secondary TT-Doppler-
echocardiography parameters of left and right 
heart, as described in Section 4.5.7 (objectives 1 
& 2)
3.
To investigate the effect 10 mg and 50 
mg QD balovaptan on blood pressure 
and heart rate as measured by the use 
of Holter-ECG and ABPM.
3.
Ambulatory blood pressure monitoring (ABPM): 
mean diastolic and systolic blood pressure 
(daytime and nighttime mean).

Heart rate will also be analyzed as 
derived from Holter ECG.
-
Heart rate as derived from ABPM 
and/or Holter ECG may be 
explored against fluctuations in 
blood pressure measured by 
ABPM (if warranted).
4.
To investigate whether genetic variants 
in genes are associated with natural or 
potential drug induced neutropenia; with 
differences in pharmacokinetics of 
balovaptan, M2 or M3, or with any 
potential effect of balovaptan on ECG 
parameters. 
4.
The genetic variants in genes that are associated 
with natural or potential drug induced 
neutropenia; genetic variants in genes that are 
associated with pharmacogenetics of 
metabolizing enzymes, transferases, 
transporters, etc, (eg,CYP3A4 and P-
glycoprotein); genetic variants in genes that may 
be associated with a potential effect of 
balovaptan on ECG parameters.


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3.
STUDY DESIGN
3.1
DESCRIPTION OF THE STUDY
This will be a single-center, multiple-dose, randomized, double-blind, placebo-controlled, 
positive-controlled, twelve sequence, 3-period cross-over study to investigate the effect 
of balovaptan on the QTc interval in healthy subjects.
Screening
Screening will be conducted up to 28 and 3 days (inclusive) before each subject’s first 
dosing day (Day 1 of Treatment Period 1).
In-Clinic Confinement
Day -2
Subjects will be confined to the CRU starting on Day -2 until the morning of Day 16 
during each Treatment Period. Subjects are entered into CRU on Day -2 so that 
subjects are rested and prepared for the baseline ABPM and baseline TT-Doppler
echocardiogram on Day -1. Eligibility assessments will be performed on Day -2 as 
outlined in the Schedule of Activities (SoA), Appendix 1.
Day -1
On Day -1 of each Treatment Period, ABPM and TT-Doppler echocardiogram
assessments will be performed. These results will serve as the baseline for the subjects.
On days when TT-Doppler echocardiogram and ABPM assessments are performed 
subjects will be given low salt standardized meals.
Randomization (Day -1)
At the start of the study Treatment Period 1, at the end of the day of Day -1, subjects will 
be randomized to one of the 12 treatment sequences comprised of the 4 treatments 
described:
Treatments
Treatment A (balovaptan therapeutic dose)
Days 1-14:
A single once daily oral dose of 10 mg (1 × 10 mg tablet) of balovaptan and 4 
tablets of matching placebo for balovaptan for 14 days.
Day 2 and 15: A single oral dose of a matching placebo capsule for moxifloxacin.


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Treatment B (balovaptan supra-therapeutic dose)
Days 1-14:
A single once daily oral dose of 50 mg (5 x 10 mg tablets) of balovaptan for 
14 days.
Day 2 and 15: A single oral dose of a matching placebo capsule for moxifloxacin.
Treatment C (active control [400 mg moxifloxacin] on Day 2)
Days 1-14:
A single once daily oral dose of 5 tablets of matching placebo for balovaptan for
14 days.
Day 2:
A single oral dose of 400 mg moxifloxacin capsule
Day 15: 
A single oral dose of a matching placebo capsule for moxifloxacin 
Treatment D (active control [400 mg moxifloxacin] on Day 15)
Days 1-14:
A single once daily oral dose of 5 tablets of matching placebo for balovaptan for
14 days.
Day 2:
A single oral dose of a matching placebo capsule for moxifloxacin
Day 15: 
A single oral dose of 400 mg moxifloxacin capsule.
Treatment Sequences
1. A, B, C
2. A, C, B
3. B, A, C
4. B, C, A
5. C, A, B
6. C, B, A
7. A, B, D
8. A, D, B
9. B, A, D
10. B, D, A
11. D, A, B
12. D, B, A
Dosing (Days 1-15)
Each Treatment Period will involve 15 days of dosing (Day 1 – Day 15). Subjects must 
fast for at least 8 hours prior to and 4 hours after dosing on study days when 24-hour 
Holter ECG assessments will be performed (study Days 1, 2, 14 and 15). Dosing on 


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study days with 24-hour Holter ECG assessment should be at approximately the same 
time of day ± 1h.
Prior to dosing on Day 1 subjects will undergo pre-dose assessments as outlined in the 
SoA (Appendix 1). Following pre-dose assessments subjects will receive a single dose 
of study medication preceded by a ≥8 hour fast the night before.  Study drugs will be 
taken with 240 mL of water and be followed by a 4-hour post-dose fast. If subjects 
require more liquid to take all study drugs, additional water may be provided in aliquots 
of 50 mL. If additional water is provided the amount should be captured. Water 
provided for study drug administration will be identical on all dosing days. Identical 
fasting procedures from Day 1 will be followed on study Days 2, 14 and 15. No fasting 
will be required on other dosing days. Subjects will receive standardized low salt meals 
on study days when 24-hour Holter ECGs (Days 1, 2, 14 and 15), ABPM (Days -1 
and 12) or TT-Doppler echocardiogram (Days -1 and 13) assessments will be performed.  
Meals will be given at approximately the same time on each of these study days.
Days 1 and 2: 24-hour Holter ECG Assessments
On study Days 1 and 2, of each Treatment Period, 24-hour ECG assessments will be 
performed. Subjects will be administered study drug following a ≥8 hour fast and 
continue to be fasted for 4 hours post-dose on these days. Subject will be provided with
standardized low-salt meals on these days. Subjects will undergo pre and post-dose 
assessments as outlined in the SoA (Appendix 1).
Day 12
On study Day 12 of each Treatment Period, ABPM assessments will be performed for 
each subject. On study days when ABPM assessments are performed subjects will be 
provided with standardized low salt meals. Subjects will undergo pre and post-dose 
assessments as outlined in the SoA (Appendix 1).
Day 13
On study Day 13, of each Treatment Period, at approximately the same time of day as 
on Day -1, TT-Doppler echocardiogram will be performed to assess any effects of steady 
state balovaptan levels on cardiac parameters. Subjects will receive standardized low 
salt meals on this study day. In addition to TT-Doppler echocardiogram subjects will 
undergo pre and post-dose assessments as outlined in the SoA (Appendix 1).
Days 14 and 15- 24-hour Holter ECG Assessments
On study Days 14 and 15 of each Treatment Period, 24-hour ECG assessments will be 
performed. Subjects will be administered study drug following a ≥8 hour fast and 
continue to be fasted for 4 hours post-dose. Subjects will be provided with standardized 


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low salt meals on these days. Subjects will undergo pre and post-dose assessments as 
outlined in the SoA (Appendix 1).
Day 16
Subjects will be discharged from the CRU on the morning of Day 16 after final 
assessments as described in the SoA (Appendix 1).
Washout Period:
Following each Treatment Period subjects will undergo a washout period lasting a 
minimum of 13 and a maximum of 20 days from the last dose of study drug.
Follow-up:
Subjects will return to the unit for a follow-up visit between 14 and 21 days after the final 
dose of study drug. This will include a general physical examination, 12-lead ECG, vital 
signs, C-SSRS, adverse event review, body weight and laboratory safety tests.
Figure 1 presents an overview of the study design.  A SoA is provided in Appendix 1.
Figure 1
Study Schema


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3.2
END OF STUDY AND LENGTH OF STUDY
The end of this study is defined as the date when the last subject, last visit occurs, or 
safety follow-up is received from the last subject, whichever occurs later.
The total length of the study, from screening of the first subject to the end of the study, is
expected to be approximately 14 - 20 weeks. For each enrolled subject the duration of 
the study from Screening to the end of Follow-up will be a minimum of 122 days or a
maximum of 143 days.

Screening: up to 28 days

Dosing periods including washout: 80 - 94 days
o
Treatment Period 1: 18 Days (Day -2 – Day 16)
o
Washout: 13 - 20 Days
o
Treatment Period 2: 18 Days (Day -2 – Day 16)
o
Washout: 13 - 20 Days
o
Treatment Period 3: 18 Days (Day -2 – Day 16)

Follow up: 14 - 21 days after last dosing
3.3
RATIONALE FOR STUDY DESIGN
The study will utilize a positive-control, 400 mg moxifloxacin, to assure study design and 
measurements will be able to detect potential ECG effects of balovaptan. Moxifloxacin, 
a fluoroquinolone antibiotic, when utilized at therapeutic doses (400 mg) is associated 
with QT prolongation and regulatory authorities have recommended it as a positive 
control to evaluate the sensitivity of clinical studies designed to detect small but 
significant increases in QT interval measurements.4
Across clinical trials, no balovaptan associated changes in blood pressure or HR are
apparent. Therefore, this study uses QTcF as primary parameter rather than an 
individualized corrected QTc. Since a cross-over design corrects for diurnal fluctuations 
of QTc, a separate Day -1 reading is not needed, 3 time points (replicates at each time 
point) pre-dose are used as baseline.
3.3.1
Rationale for Balovaptan and Moxifloxacin Dose and Schedule
3.3.1.1
Balovaptan
A dose of 10 mg balovaptan has shown statistically significant changes in efficacy 
parameters in adult male patients with ASD (Phase 2) and is the current recommended 
Phase 3 dose.
Balovaptan is being developed for use in ASD, so it is expected that patients will need to 
take it for prolonged periods of time, likely for years or for their lifetime. Hence multiple 
dose testing is relevant from a clinical point of view. The multiple dose PK of balovaptan 
with 14 days of dosing was previously characterized in humans and was found to be 
time-dependent. Accumulation was observed with multiple dosing, and steady state of 


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balovaptan was observed after approximately 7 days of dosing and after 10 days for M3.
The plasma PK time course of M2 has not been closely studied but steady state has 
been demonstrated after 2 weeks of once daily dosing with 10 mg balovaptan.
The primary objective of this study is to investigate the effect of balovaptan on cardiac 
repolarization by measuring ECGs in healthy subjects at both therapeutic and supra-
therapeutic doses. To this end subjects will be dosed once daily with balovaptan for 
14 days at either the projected therapeutic dose of 10 mg or a supra-therapeutic dose of 
50 mg and ECG parameters monitored.
In the MAD part of study BP25694 multiple doses of 12 mg (just above the clinical dose 
of 10 mg) resulted in a Cmax of 99 ng/mL. In Study BP28977 co-administration of 
itraconazole led to a 36% increase in Cmax. In the MAD part of study BP25694 multiple 
doses of 52 mg qd in healthy subjects were well tolerated and resulted in a Cmax of 
484 ng/mL. In light of convulsions observed in dogs, dose escalation in study BP25694 
was stopped so as not to exceed a Cmax of 810 ng/mL. In conclusion, based on 
previous clinical experience a once-daily dose of 50 mg balovaptan for 14 days is 
expected to be well tolerated considering the short duration of dosing in this study. 
3.3.1.2
Moxifloxacin
The study will utilize moxifloxacin (400 mg) as a positive-control to assure the study as 
designed will be able to detect potential ECG effects of balovaptan. Moxifloxacin, a 
fluoroquinolone antibiotic, when utilized at its recommended therapeutic dose of 400 mg
qd is associated with QT prolongation. Regulatory authorities have recommended 
moxifloxacin as a positive control to evaluate the sensitivity of clinical studies designed 
to detect small but significant increases in QT interval measurements. Subjects will 
receive 400 mg oral moxifloxacin in a nested cross-over fashion using a joint 
placebo/positive control period. In this period, half of the subjects will be dosed with 
moxifloxacin on Day 2 and the other half dosed on Day 15. To keep the blind between 
balovaptan and placebo, this nested design has 4 Holter extraction days and 
moxifloxacin is blinded to avoid unblinding of the placebo period. The moxifloxacin 
dosing day is the same number apart from the baseline as in the balovaptan periods to 
maintain the blind.
3.3.2
Rationale for Study Population
Healthy male or female volunteers aged 18 to 60 years inclusive have been chosen as 
the study population due to the low risk of clinically significant toxicity at anticipated 
exposure levels. Moreover, use of healthy subjects, as opposed to patients, will allow a 
clearer interpretation of the study results, as there will be no confounding factors that 
result from changes in disease state and/or concomitant medication use.


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3.3.3
Rationale for Transthoracic Doppler-echocardiogram and 
Ambulatory Blood Pressure Monitoring
As an exploratory marker transthoracic Doppler echo-cardiography is included to 
estimate pulmonary blood pressure with the a priori expectation that no changes in 
pulmonary circulatory dynamics will be seen.
In animals as well as preparations of pulmonary vasculature, vasopressin has been 
shown to decrease pulmonary blood pressure and to relax arterial constriction through 
V1a receptors and in animals these changes were accompanied by systemic blood 
pressure increases.8,9 However, GLP toxicology animal studies with balovaptan have 
not pointed to a right heart challenge strain nor to pulmonary fibrosis. No systemic blood 
pressure changes with balovaptan were noted in clinical studies.
In clinical trials, blood pressure monitoring has not revealed apparent blood pressure 
changes. However, blood pressure monitoring focused on few time points around 
pharmacokinetic Tmax and no night-time blood pressure data are available. Hence, 
ABPM is included in this study to supplement the existing blood pressure profiling, 
exploring in a more intense way any effects of balovaptan on daytime blood pressure
(9:00 am to 9:00 pm) and to collect data for nighttime blood pressure (1:00 am to 
5:30 am).
This study provides an opportunity for these exploratory measures considering a 
homogeneous population of healthy volunteers and the inclusion of a multiple dose 
higher than the dose of 10 mg qd tested in the ongoing Phase 3 study WN39434.
4.
MATERIALS AND METHODS
4.1
SUBJECTS
The target population will be healthy male or female volunteers aged 18 to 60 years 
inclusive.  An attempt will be made to have each Treatment population contain at least 
1/3 male subjects by ensuring that a minimum of 40% of enrolled population are male.
Initially 56 subjects will be enrolled with the goal of obtaining 48 evaluable subjects.
Enrolled subjects who withdraw from the study may be replaced at the discretion of the 
Investigator (or designee) and Sponsor to ensure adequate numbers of evaluable 
subjects.
4.1.1
Inclusion Criteria
Subjects must meet the following criteria for study entry:
1. Healthy male or female volunteers aged 18 to 60 years inclusive, at Screening. 
Healthy status is defined by absence of evidence of any active or chronic disease 
following a detailed medical and surgical history, a complete physical 


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examination including vital signs, 12-lead ECG, hematology, blood chemistry, 
urinalysis, and serology.
2. Body Mass Index (BMI) between 18 and 32 kg/m2 inclusive, at Screening.
3. For women of childbearing potential: agreement to use at least 1 acceptable form 
of contraception during the entire study and for 90 days following last dose of 
study drug.
a. A woman is considered to be of childbearing potential if she is 
postmenarcheal, has not reached a postmenopausal state (> 12 
continuous months of amenorrhea with no identified cause other than 
menopause), and has not undergone surgical sterilization (removal of 
ovaries and/or uterus).
b. The following are acceptable contraceptive methods: abstinence from 
heterosexual activity; bilateral tubal ligation; sterilization of male ‘partner; 
hormonal contraceptives; hormone-releasing intrauterine devices; copper 
intrauterine devices; Essure.
c.
Barrier methods alone are not considered an acceptable form of 
contraception for this study and if utilized must be supplemented by one 
of the acceptable methods described above. For this study barrier 
contraception methods are defined as: male or female condom with or 
without spermicide (male and female condom must not be used 
simultaneously); and cap, diaphragm, or sponge with spermicide.
4. For men: Vasectomized, agreement to remain abstinent (refrain from 
heterosexual intercourse) or use of a condom during intercourse. Must also 
agree to refrain from donating sperm, as defined below:
a. With female partners of childbearing potential or pregnant female partners, 
men must remain abstinent or use a condom during the entire study and 
for 90 days after the last dose of study drug to avoid exposing the embryo. 
Men must refrain from donating sperm during this same period.
5. Able to participate, and willing to give written informed consent and to comply 
with the study restrictions.
6. Fluent in English.
4.1.2
Exclusion Criteria
Subjects who meet any of the following criteria will be excluded from study entry:
Past Medical History
1.
History of any clinically significant gastrointestinal, renal, hepatic, broncho-
pulmonary, neurological, psychiatric, cardiovascular, endocrinological, 
hematological, lymphatic, musculoskeletal, genitourinary, immunological, 
dermatological or connective tissue or allergic disease, metabolic disorder, or 
cancer.
2.
Signs and symptoms potentially indicative of peripheral neuropathy.


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3.
Unexplained orthostatic reaction with syncope within last 6 months.
4.
A history of clinically significant hypersensitivity (e.g., to drugs, including 
moxifloxacin or any quinolone antibiotics and balovaptan, and excipients) or allergic 
reactions.
5.
History of alcohol and/or drug abuse or addiction within the last 2 years before study 
enrollment.
6.
Subjects on hormone replacement therapy if post-menopausal status cannot be 
documented from medical history.
7.
Previous treatments with iron for iron deficiency anemia within three months of 
screening.
8.
History of coagulopathies, bleeding disorders, or blood dyscrasias.
9.
History of porphyria, active liver disease or permanent or unexplained elevation of 
serum transaminases.
10. In the opinion of the Investigator or designee, any major illness within one month 
before the Screening examination or any febrile illness within one week prior to 
Screening and up to first study drug administration.
Current Status
1.
Clinically relevant deviation from normal in the physical examination including 
vital signs.
2.
Positive result on human immunodeficiency virus (HIV) 1, HIV 2, hepatitis C virus 
(HCV) antibody, or hepatitis B virus surface antigen (HBsAg).
3.
Clinically significant abnormalities in laboratory test results (including complete 
blood count, chemistry panel, and urinalysis). In the case of uncertain or 
questionable results, tests performed during Screening may be repeated on Day -1 
or Day -2 to confirm eligibility. Repeat of abnormal test results are allowed during 
Screening and eligibility assessment at the discretion of the Investigator or designee
provided results will be available prior to Randomization on Day -1.
4.
If female, a positive pregnancy test at screening or prior to Day 1 of any Treatment 
Period.
5.
Lactating women.
6.
Any condition or disease detected during the medical interview / physical 
examination that would render the subject unsuitable for the study, place the subject 
at undue risk or interfere with the ability of the subject to complete the study in the 
opinion of the Investigator or designee.
7.
Subjects with CPK values > 2x upper limit of normal (ULN), confirmed by repeat 
sample.
8.
Positive urine alcohol test or urine drug test at screening or Day -2 of each 
treatment period (amphetamines, barbiturates, benzodiazepines, cannabinoids,
cocaine, opiates, methadone, methamphetamines, and
3,4-methylenedioxymethamphetamine [MDMA]).


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9.
Subject has been a regular smoker (> 5 cigarettes, > 1 pipeful, > 1 cigar per day or 
equivalent as determined by Investigator or designee) within 6 months of study 
enrollment.
10. Coffee (or tea) consumption > 5 cups per day or methylxanthine-containing 
drinks > 1.5 liter/day or more than 250 g/day of chocolate within 3 months of first 
dose of study drug. 
11. Alcohol consumption of > 18 units per week for males and females. One unit of 
alcohol equals 12 ounces of regular beer, 5 ounces of wine or 1.5 ounces of 
80 proof spirits within 3 months of first dose of study drug.
12. Subjects who have lost or donated in excess of ~450 mL blood in the 3 months prior 
to Day 1 of Treatment Period 1.
13. Subjects under judicial supervision, guardianship, or curatorship.
14. Dietary restrictions that would prohibit the consumption of standardized meals.
15. Poor venous access for blood sampling.
Concomitant Medication
1. Taken greater than 2 g/day paracetamol within 48 hours of study drug administration
or greater than 4 g/day paracetamol within 7 days of study drug administration.
2. Any nonprescribed systemic or topical medication or herbal remedies are prohibited 
for 7 days prior to the first study drug administration through Follow-up visit.
3. Participation in an investigational drug study within 60 days or 5 times the elimination 
half-life, whichever is longer, prior to first dosing, or within 5 months prior to first 
dosing in case of a study with a biological, as calculated from the day of Follow-up 
visit from the previous study.
4. Participation in an investigational device study within 60 days of first dosing of study 
drug.
5. Subjects who have received any prescribed systemic or topical medication within 
14 days of the first dose administration are excluded, unless in the opinion of the 
Investigator (or designee) the medication will not interfere with the study procedures 
or compromise safety. 
6. Any slow release medicinal formulations considered to still be active within 4 weeks 
(or within 5 times the elimination half-life of the medication, whichever is longer) prior 
to the first study drug administration are prohibited, unless in the opinion of the 
Investigator or designee the medication will not interfere with the study procedures or 
compromise safety.
7. Taking any inhibitor of CYP3A taken within 4 weeks (or within 5 half-lives, whichever 
is longer) prior to study drug administration or taking any inducer of CYP3A within 
4 weeks of the first dose of study drug.
8. Use of any prohibited medications and food before study start and subjects who do 
not agree to refrain from consuming prohibited medications or food during the study.


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9. Subjects who have previously enrolled in, or withdrawn after enrollment, from this 
study.
TQT-related Exclusion Criteria:
1.
Personal history of unexplained black-outs or faints.
2.
Personal history of reactions or faints at having their blood taken, unexplained 
black-outs or faints or any condition which could relapse during or immediately after 
the study which in the opinion of the Investigator or designee makes the subject 
unsuitable to participate in this study.
3.
Subjects experiencing fainting or pre-syncopal events during orthostatic challenge 
testing, performed according to site SOP, at Screening and Day -2 of Treatment 
Period 1.
4.
Subjects with Screening or Treatment Period 1 (Day -2) baseline mean QT interval 
corrected using Fridericia’s formula (QTcF) > 450 ms or < 300 ms (using the same 
upper QTcF limit in both males and females) or > 30 ms difference between the 
highest and the lowest readings of the triplicate ECGs recorded.
5.
Screening or baseline (Day -2 Treatment Period 1) ECG evidence of atrial fibrillation, 
atrial flutter, second or third-degree AV block, complete right or left bundle branch 
block, Wolff-Parkinson-White-syndrome, or cardiac pacemaker.
6.
Screening or baseline ECGs (Day -2 of Treatment Period 1) with QRS and/or 
T-wave judged to be unfavorable for a consistently accurate QT measurement (i.e., 
neuromuscular artifact that cannot be readily eliminated, arrhythmias, indistinct QRS 
onset, low amplitude T-wave, merged T- and U-waves, prominent U-waves, etc.).
7.
Confirmed (based on the average of ≥3 consecutive measurements) systolic blood 
pressure greater than 139 or less than 90 mmHg, and diastolic blood pressure 
greater than 89 or less than 45 mmHg at Screening and Day -2 of Treatment Period 
1.
8.
Confirmed (based on the average of ≥3 consecutive measurements) resting 
HR > 100 bpm or < 40 bpm at Screening or Day -2 of Treatment Period 1.
9.
Notable resting bradycardia (HR < 40 bpm) on ECG at Screening or Day -2 of 
Treatment Period 1. Notable resting tachycardia (mean HR > 100 bpm) on ECG at 
Screening or Day -2 of Treatment Period 1.
10. Family history of congenital long QT syndrome or sudden death.
11. History or presence of clinically significant ECG abnormalities, as determined by 
study Investigator or designee, before study drug administration.
4.2
METHOD OF TREATMENT ASSIGNMENT AND BLINDING 
This is to be a randomized, double-blind study.
4.2.1
Treatment Assignment
After obtaining informed consent, subjects will be screened according to the inclusion
and exclusion criteria. Subjects who have met all eligibility criteria will receive a 


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randomization number on Day -1 of the first treatment period and according to the 
randomization code generated by the Biostatistics Department of PRA. The 
randomization number will ensure identification throughout the study. Replacement 
subjects will receive the randomization number of the subject to be replaced, increased 
by 1000 (e.g., 1101 replacement number for randomization number 101), and will be 
administered the same treatment.
Subjects who drop out or withdraw for any reason without completing all screening
evaluations successfully, will be considered “screening failures”. Such subjects will not
receive a randomization number, and only applicable data will be entered in the 
electronic case report forms (eCRFs).
Subjects will be randomly assigned to one of the 12 treatment sequences according to 
the randomization code.
4.2.2
Blinding
The following controls will be employed to maintain the double-blind status of the study.
A double dummy design will be used, all treatments are blinded (balovaptan and 
matching placebo, moxifloxacin and matching placebo). Groups will receive placebo
and study drugs as described in Section 3.1. The oral tablets/capsules will be 
indistinguishable in appearance and taste to balovaptan and moxifloxacin, respectively.
The number of tablets and capsules will be identical for each subject regardless of 
treatment and dose.
Study site personnel (with the exception of the unblinded pharmacy staff) and subjects
will be blinded to treatment assignment during the study.  The Sponsor and its agents 
will also be blinded to treatment assignment, with the exception of individuals who 
require access to subject treatment assignments to fulfill their job roles during a clinical 
trial.
As per health authority reporting requirements, the Sponsor's Drug Safety representative
will break the treatment code for all serious, unexpected suspected adverse reactions
that are considered by the Investigator or designee or Sponsor to be related to study 
drug.  The subject may continue to receive treatment, and the Investigator or designee, 
subject, and Sponsor personnel, with the exception of the Drug Safety representative 
and personnel who must have access to subject treatment assignments to fulfill their 
roles (as defined above), will remain blinded to treatment assignment.
4.3
STUDY TREATMENT AND OTHER TREATMENTS RELEVANT 
TO THE STUDY DESIGN
The investigational medicinal products (IMPs) for this study are balovaptan, moxifloxacin
and placebo.


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4.3.1
Study Treatment Formulation, Packaging, and Handling
4.3.1.1
Balovaptan and Placebo
RO5285119/F17 10 mg film-coated dispersible tablets and matching placebo tablets will 
be supplied by the Sponsor in high-density polyethylene bottles. The balovaptan and 
matching placebo tablets must be stored and handled according to the details specified 
on the product label. For additional details, refer to the balovaptan Investigator's 
Brochure.
4.3.1.2
Moxifloxacin Active Control and Placebo
Moxifloxacin (400 mg qd, administered as 400 mg over-encapsulated tablets) will be 
supplied by the Sponsor. The over-encapsulated moxifloxacin and matching placebo
must be stored and handled according to the details specified on the product label.
4.3.2
Study Treatment Dosage, Administration, and Compliance
The treatment regimens are summarized in Section 3.1
Any overdose or incorrect administration of any of the study treatments should be noted 
on the Study Drug Administration electronic Case Report Form (eCRF). In this study an 
overdose is defined as any dose of balovaptan greater than 50 mg per day and any dose 
of moxifloxacin greater than 400 mg per day. Adverse events associated with an 
overdose or incorrect administration of any of the study treatments should be recorded 
on the Adverse Event Report Form.
Guidelines for treatment interruption or discontinuation for subjects who experience 
adverse events are provided in Section 5.1.
4.3.2.1
Balovaptan and Placebo
Treatment A: A single once daily oral dose of 10 mg (1 × 10 mg tablet) of balovaptan 
and 4 tablets of matching placebo for balovaptan for 14 days.
Treatment B: A single once daily oral dose of 50 mg (5 x 10 mg tablets) of balovaptan 
for 14 days.
Treatment C: A single once daily oral dose of 5 tablets of matching placebo for
balovaptan for 14 days.
Treatment D: A single once daily oral dose of 5 tablets of matching placebo for 
balovaptan for 14 days
Doses of balovaptan, and matching placebos on Days 1, 2, 14 or 15 will be administered 
with approximately 240 mL of water (at room temperature) after at least 8 hours of 
fasting. Dosing on these days will be at approximately the same time of day ± 1h. 
Post-dose subjects will maintain a 4-hour fast during which no food will be allowed.


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While in the clinical research unit, subjects will be allowed to consume water ad libitum, 
except for 2 hours before and 2 hours after oral dosing. However, excessive 
consumption of liquids should be avoided (Section 4.4.4). Liquids are permitted with the 
breakfast meal on non-fasting days.
4.3.2.2
Moxifloxacin (Active Control) and Placebo
Treatment A: A single oral dose of a matching placebo capsule for moxifloxacin on 
Day 2 and 15.
Treatment B: A single oral dose of a matching placebo capsule for moxifloxacin on 
Day 2 and 15.
Treatment C: A single oral dose of 400 mg moxifloxacin capsule on Day 2. A single oral 
dose of a matching placebo capsule for moxifloxacin on Day 15.
Treatment D: A single oral dose of a matching placebo capsule for moxifloxacin on 
Day 2. A single oral dose of 400 mg moxifloxacin capsule on Day 15.
The moxifloxacin and matching placebos on Day 2 and 15 will be administered with 
approximately 240 mL of water (at room temperature) after at least 8 hours of fasting.
On Day 2, moxifloxacin or matching placebo will be administered immediately after the 
balovaptan or placebo dose with the same 240 mL of water. Dosing on these days will 
be at approximately the same time of day ± 1h. Post-dose subjects will maintain a
4-hour fast during which no food will be allowed.
4.3.3
Investigational Medicinal Product Accountability
Balovaptan and placebo will be provided by the Sponsor. The study site will 
acknowledge receipt of IMPs supplied by the Sponsor by returning the appropriate 
documentation form to confirm the shipment condition and content. Any damaged 
shipments will be replaced. Moxifloxacin and matching placebo will be provided by the 
Sponsor.
The IMPs will either be disposed of at the study site according to the study site’s 
institutional standard operating procedure or be returned to the Sponsor with the 
appropriate documentation. The site’s method of destroying Sponsor-supplied IMPs 
must be agreed to by the Sponsor. The site must obtain written authorization from the 
Sponsor before any Sponsor-supplied IMP is destroyed, and IMP destruction must be 
documented on the appropriate form.
Accurate records of all IMPs received at, dispensed from, returned to, and disposed of by 
the study site should be recorded on the Drug Inventory Log.


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4.4
CONCOMITANT THERAPY, PROHIBITED FOOD, AND 
ADDITIONAL RESTRICTIONS
Concomitant therapy consists of any medication (e.g., prescription drugs, 
over-the-counter drugs, vaccines, or homeopathic remedies, nutritional supplements) 
used by a subject in addition to protocol-mandated treatment from 30 days prior to 
initiation of study drug to the study completion/ discontinuation visit. All such 
medications should be reported to the Investigator or designee and recorded on the 
Concomitant Medications eCRF.
4.4.1
Permitted Therapy 
Existing hormonal contraceptives and hormone replacement therapy regimens may 
remain unchanged during the conduct of the study. Women of childbearing potential: 
agreement to use at least 1 acceptable form of contraception during the entire study and 
for 90 days following last dose of study drug.
Acetaminophen/paracetamol is allowed up to a maximum dose of 4 g/day within 7 days 
before study drug administration and 2 g/day within 48 hours before study drug 
administration, but total should not exceed 4 g during the week prior to dosing. Subjects 
will be instructed not to exceed these limits.
4.4.2
Prohibited Therapy
Participation in an investigational drug study within 60 days or 5 times the elimination 
half-life, whichever is longer, prior to first dosing, or within 5 months prior to first dosing 
in case of a study with a biological, as calculated from the day of follow-up from the 
previous study. Participation in an investigational device study within 60 days prior to 
first dosing is prohibited.
Any non-prescribed systemic, topical medication or herbal remedies are prohibited for 
7 days prior to the first study drug administration through follow-up.
Any slow release medicinal formulations considered to still be active within 4 weeks (or 
within 5 times the elimination half-life of the medication, whichever is longer) prior to the 
first study drug administration are prohibited, unless in the opinion of the Investigator or 
designee the medication will not interfere with the study procedures or compromise 
safety.
Any prescribed systemic or topical medication taken or applied within 14 days prior to 
the first dose of study drug through the follow-up visit (with the exception of permitted 
therapies per Section 4.4.1).
Additional exceptions maybe made for medications to treat AEs, if the rationale for the 
exception is discussed and approved by the Investigator (or designee) as well as the 
Roche medical monitors.
All medication administered to manage AEs should be 
recorded on the Adverse Event Report Form.


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The following concomitant medications in particular are prohibited, unless an exception 
has been agreed to by the Investigator (or designee), and the Roche medical monitors.

Any inhibitor of CYP3A taken within 4 weeks (or within 5 half-lives, whichever is 
longer) prior to study drug administration, including but not limited to the following 
drugs: ketoconazole, miconazole, itraconazole, fluconazole, erythromycin, 
clarithromycin, ranitidine, and cimetidine, until follow-up.

Any inducer of CYP3A taken within 4 weeks prior to study drug administration, 
including but not limited to the following drugs: rifampicin, rifabutin, glucocorticoids, 
carbamazepine, phenytoin, phenobarbital and St. John's wort, until follow-up.
4.4.3
Prohibited Food

Consumption of methylxanthine-containing products (e.g., coffee, tea, cola, 
chocolate) will be forbidden from 48 hours prior to Day -2 check-in to CRU and 
during the stay.

It is not permitted to take any nutrients known to modulate cytochrome CYP3A 
activity (e.g., grapefruit juice; Seville orange) within 2 weeks prior to first dosing until 
discharge from the CRU at the end of the study.

Subjects should refrain from consumption of any foods containing poppy seeds 
within 48 hours (2 days) prior to screening and each admission to the clinical 
research center to avoid false positive drug screen results.

Alcohol consumption will not be allowed 48 hours before Day -2 admission to CRU 
and while staying in the CRU
4.4.4
Additional Restrictions/Considerations

While in the clinical research unit, subjects will be allowed to consume water ad 
libitum, except for 2 hours before and 2 hours after oral dosing. Subjects will be 
provided guidance at CRU Admission, for each Treatment Period, that they should 
avoid excessive fluid intake during the study. Excessive fluid intake for this study is 
defined as 3 L (100 fl oz.) per day.  Clinic staff will provide guidance as to 
approximate number of drinks in 3 liters as part of the subject orientation process at 
Admission. However, the clinical site will not be required to measure, or record fluid 
intake.

Subjects must fast for at least 8 hours prior to and 4 hours after dosing on Days 1, 2, 
14, and 15. Dosing on these days will be at approximately the same time of day (±
1h). Standardized low salt meals should be provided at approximately the same 
time of day on Days -1, 1, 2, 12, 13, 14 and 15.

Between the screening visit and follow-up visit, alcohol must be restricted to no 
more than 2 drinks per day (one drink is equal to 12 ounces of beer, 5 ounces of 
wine or 1.5 ounces of 80 proof spirits). Alcohol consumption will not be allowed 
48 hours before Day -2 admission to the CRU and while staying in the CRU. 
Additional alcohol/drugs of abuse testing may be employed throughout the study to 
verify compliance.


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
Use of tobacco will not be permitted 48 hours prior to admission and until discharge 
from the CRU in each study period.

Subjects should refrain from strenuous exercise for at least 96 hours prior to dosing
until discharge from the CRU at the end of the study.
4.4.4.1
Herbal Therapies
Any herbal remedies are prohibited for 7 days prior to the first study drug administration 
through follow-up.
4.5
STUDY ASSESSMENTS 
The schedule of activities to be performed during the study is provided in Appendix 1.  
All activities must be performed and documented for each subject.
Subjects will be closely monitored for safety and tolerability throughout the study.
4.5.1
Informed Consent Forms and Screening Log
Written informed consent for participation in the study must be obtained before 
performing any study-related procedures (including screening evaluations).  Informed 
Consent Forms for enrolled subjects and for subjects who are not subsequently enrolled 
will be maintained at the study site.
All screening evaluations must be completed and reviewed to confirm that subjects meet 
all eligibility criteria before enrollment.  The Investigator or designee will maintain a 
screening log to record details of all subjects screened and to confirm eligibility or record 
reasons for screening failure, as applicable.
4.5.2
Medical History, Concomitant Medication, and Demographic 
Data
Medical history, including clinically significant diseases, surgeries, cancer history 
(including prior cancer therapies and procedures), reproductive status, smoking 
history, and use of alcohol and drugs of abuse, will be recorded at baseline.  In addition, 
all medications (e.g., prescription drugs, over-the-counter drugs, vaccines, or 
homeopathic remedies, nutritional supplements, or herbal) used by the subject within 
30 days prior to initiation of study treatment will be recorded. Any nonprescribed 
systemic or topical medication or herbal remedies are prohibited for 7 days prior to the 
first study drug administration through Follow-up. At the time of each Follow-up visit 
physical examination, an interval medical history should be obtained and any changes in 
medications and allergies should be recorded.
Demographic data will include age, sex, and self-reported race/ethnicity.
4.5.3
Physical Examinations
A complete physical examination, performed at Screening and other specified visits
(Appendix 1), should include an evaluation of the head, eyes, ears, nose, and throat, 


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and the cardiovascular, dermatologic, musculoskeletal, respiratory, gastrointestinal, 
genitourinary (at Investigator or designee’s discretion).  Any clinically significant 
abnormality identified at baseline should be recorded on the General Medical History 
and Baseline Conditions eCRF.  Changes from baseline abnormalities should be 
recorded in subject notes.  New or worsened clinically significant abnormalities should 
be recorded as adverse events on the Adverse Event eCRF and source form.
4.5.4
Vital Signs
Oral body temperature, pulse rate, and supine blood pressure will be collected as 
described in the SoA (Appendix 1). ABPM assessment will be performed on study 
Days -1 and 12.
4.5.5
Laboratory, Genotyping, and Pharmacokinetic Samples
For sampling and sample processing procedures, storage conditions, and shipment 
instructions, see the Laboratory Manuall.
Biological samples will be destroyed when the final Clinical Study Report has been 
issued, except where specified otherwise.
When a subject withdraws from the study, samples collected prior to the date of 
withdrawal may still be analyzed, unless the subject specifically requests that the 
samples be destroyed, or local laws require destruction of the samples. However, if 
samples have been tested prior to withdrawal, results from those tests will remain as 
part of the overall research data.
Data arising from sample analysis will be subject to the confidentiality standards 
described in Section 8.4.
4.5.5.1
Laboratory Samples
Laboratory samples will be collected at the time points indicated in the SoA (Appendix 1). 
Samples will be collected at Screening, Day -2, Day 3, Day 7, Day 15 and Follow-up visit
for each Treatment Period. Additional blood or urine samples may be taken at the 
discretion of the Investigator or designee if the results of any test falls outside the 
reference ranges, or clinical symptoms necessitate additional testing to monitor subject 
safety. Subjects must fast for at least 8 hours prior to laboratory safety assessments
(except for urine assessments).
Samples for the following laboratory tests will be sent to the study site’s local laboratory 
for analysis:

Hematology: leukocytes, erythrocytes, hemoglobin, hematocrit, platelets, absolute 
and percent differential WBC count (neutrophils, eosinophils, basophils, monocytes, 
lymphocytes).

Urinalysis: A midstream, clean-catch urine specimen will be collected for dipstick 
analysis of pH, glucose, leukocytes, nitrites, protein and blood. Microscopy will be 
performed if abnormalities are observed in initial analysis.


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
Coagulation: prothrombin time (international normalized ratio) and activated 
thromboplastin time.

Blood chemistry panel: sodium, potassium, glucose (fasting), creatinine, total and 
conjugated bilirubin, alkaline phosphatase, alanine aminotransferase (ALT), 
aspartate aminotransferase (AST), gamma-glutamyl transferase, creatine 
phosphokinase (CPK) and lactate dehydrogenase (LDH).

Biomarkers for cardiac damage: samples will be taken for troponin I, cardiac 
troponin T (cTnT) and N-terminal pro b-type natriuretic peptide (NT-proBNP) but 
these samples will be stored, and assays only run if requested for specific subject(s)
due to concerns regarding cardiac safety.  

Drugs of abuse (including amphetamines, barbiturates, benzodiazepines 
cannabinoids, cocaine, opiates, methamphetamines, methadone and 
3,4-methylenedioxymethamphetamine (MDMA), will be measured in urine at 
Screening and on Day -2 of each Treatment Period.  Further testing may be 
performed throughout the study to confirm compliance at the discretion of the 
Investigator or designee.

Urine alcohol test at Screening and on Day -2 of each study period.  Further testing 
may be performed throughout the study to confirm compliance at the discretion of 
the Investigator or designee.

Viral serology: human immunodeficiency virus (HIV) -1 and -2 antibodies, hepatitis 
C antibody, hepatitis B surface antigen and hepatitis C virus (HCV) antibody will be 
assessed at Screening.

Pregnancy test: beta-human chorionic gonadotropin (serum pregnancy test) will be 
performed at Screening and Day -2 for all female subjects at Treatment Period 1 
and Day -2 for subsequent Treatment Periods.

Follicle-stimulating hormone (FSH) will be assessed at Screening for all females
4.5.5.2
Pharmacokinetic Samples
Pharmacokinetic samples will be taken via an indwelling catheter or by direct 
venipuncture at the time points in Appendix 1 and Appendix 2.
Three (3) aliquoted plasma samples will be prepared for each time point. Two (2) of the
aliquoted PK plasma samples will be shipped to the designated bioanalytical laboratory 
for analysis. The third aliquot will be frozen and stored at PRA (-80°C) for up to 2 years 
after issue of the CSR to allow for repeat analysis if required.
For additional sampling procedures, storage conditions, and shipment instructions, see 
the Laboratory Manual.
4.5.5.3
Optional Samples for genetic analyses
An optional whole blood sample will be taken for DNA extraction and clinical genotyping 
from subjects consenting to this additional sample. See Appendix 1 for collection 
timepoint and associated footnote.


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The DNA may be used for:
 
o
to determine if genetic variants of cytochrome P450s (e.g., CYP3A4), 
transporters, or receptors may affect the metabolism, pharmacokinetics of 
balovaptan, or be associated with any potential effect on ECG parameters.
This sample may be stored for up to two years following the issue of the CSR.
Data arising from all biosamples including samples for analyses of inherited DNA will be 
subject to the confidentiality standards described in Section 8.4.
4.5.6
Electrocardiograms
Safety and diagnostic ECG assessments will be performed in this study.
4.5.6.1
12-lead Safety Electrocardiograms
Safety ECGs will be performed at defined time points (Appendix 1). Safety ECG 
performed at Screening and Day -2 of each treatment period will be done in triplicate. All 
other safety ECG readings will be single recordings. These ECG recordings will only be
used to assess/define a safety baseline and determine the immediate safety of each 
subject during each Treatment Period. Safety ECG recordings may be obtained at 
unscheduled timepoints as required by the Investigator, or designee, to ensure the 
safety of study subjects. For safety monitoring purposes, the Investigator, or designee,
must review, sign, and date all ECG tracings.
All safety ECG recordings must be performed using a standard high-quality, high-fidelity 
digital electrocardiograph machine equipped with computer-based interval 
measurements.  Lead placement should be as consistent as possible.  ECG recordings 
must be performed after the subject has been resting in a supine position for at least 10 
minutes.  All ECGs are to be obtained prior to other procedures scheduled at that same 
time (e.g., vital sign measurements, blood draws) and should not be obtained within 
3 hours after any meal, when possible.  Circumstances that may induce changes in HR, 
including environmental distractions (e.g., television, radio, conversation), should be 
avoided during the pre-ECG resting period and during ECG recording.
Paper copies of ECG tracings will be kept as part of the subjects’ permanent study file at 
the site. The following should be recorded in the appropriate eCRF:  HR, QRS interval, 
PR duration, uncorrected QT interval, and QTcF based on the machine readings of the 
individual ECG tracings.  Any morphologic waveform changes or other ECG 


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abnormalities must be documented on the eCRF.  If considered appropriate by the 
Sponsor, ECGs may be analyzed retrospectively at a central laboratory.
If at a post-dose timepoint the mean QTcF is 500 ms and/or 60 ms longer than the 
baseline value, another ECG must be recorded, ideally within the next 5 minutes, and 
ECG monitoring should continue until QTcF has stabilized on two successive ECGs.  
The Medical Monitor should be notified.  Standard-of-care treatment may be instituted 
per the discretion of the Investigator or designee.  If a PK sample is not scheduled for 
that timepoint, an unscheduled PK sample should be obtained.  A decision on study drug 
discontinuation should be made, as described in Section 4.6.1.  The Investigator or 
designee should also evaluate the subject for potential concurrent risk factors 
(e.g., electrolyte abnormalities, co-medications known to prolong the QT interval, severe 
bradycardia).
4.5.6.2
Cardiodynamic ECG Evaluation
12-lead ECGs will be recorded continuously using a Holter device from at least 1 hour
before to at least 24 hours after drug administration on Day 1, 2, 14 and 15. The 
placement of electrodes for each subject must be marked on the skin with permanent 
marker, to allow electrodes to be positioned as consistently as possible throughout the 
study (Appendix 1 and Appendix 2).
ECGs will be extracted from Holter devices at 3 time points pre-dose (-45, -30 and -15 
minutes) on Days 1, 2, 14, and 15. Post-dose ECGs will be extracted at 0.5, 1.0, 2.5, 4, 
8, 12 and 24 hours post-dose on Days 1, 2, 14 and 15 (Appendix 1 and Appendix 2).
The 24 h Holter extraction time point for Days 1 and 14 can be taken from the 3 pre-
dose Holter time points on Days 2 and 15. 12-lead ECGs will be extracted from the 
continuous recording in up to 10 replicates from a 5-minute window during a 15-minute 
supine period pre-dose and at the nominal Holter timepoints post-dose. Whenever ECG 
extraction and PK sample timepoints are close together the ECG extraction will be 
performed prior to PK sample collection.
During days with Holter ECG monitoring subjects (Appendix 1), will be fed standardized 
low-salt meals.
To reduce variability, Holter recording and supine periods have to be 
performed according to PRA Work Instruction EDSEXE 009 W 131.
The 12-lead Holter will be supplied and supported by ERT.
All ECG data will be
collected using a 
 USA) M12R ECG continuous 
12-lead digital recorder. The continuous 12-lead digital ECG data will be stored onto SD 
memory cards.
ECGs to be used in the analyses will be selected by pre-determined 
time points as defined in the SoA (Appendix 1) and will be read centrally by ERT.
The following principals will be followed in ERT’s core laboratory:

ECG analysts will be blinded to the subject, visit and treatment allocation.


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
Baseline and on-treatment ECGs for a particular subject will be over-read on 
the same lead and will be analyzed by the same reader.

The primary analysis lead is lead II. If lead II is not analyzable, then primary 
lead of analysis will be changed to another lead for the entire subject data 
set.
TQT Plus ECG Extraction Technique
Ten 14-second digital 12-lead ECG tracings will be extracted from the continuous Holter 
recordings using the ‘TQT Plus method’, a computer-assisted and statistical process 
utilized by ERT. The method enables extraction of ECGs with the lowest HR variability 
and noise within the protocol-specified extraction time window (e.g., the HR and QT 
changes from beat-to-beat in the range of < 10%). At each protocol-specified timepoint, 
10 ECG replicates will be extracted from a 5-minute “ECG window” (the last 5 minutes of 
the 15-minute period when the subject is maintained in a supine quiet position).
Expert-Precision QT Analysis
High-precision QT analysis will be performed on all analyzable (non-artifact) beats in the 
10 ECG replicates. Statistical quality control procedures are used to review and assess 
all beats and identify “high” and “low” confidence beats using several criteria, including:

QT or QTc values exceeding or below certain thresholds (biologically unlikely).

HR values exceeding or below certain thresholds (biologically unlikely).

Rapid changes in QT, QTc or HR from beat to beat.
Measurements of all primary ECG parameters (QT, QTc, HR) in all recorded beats of all 
replicates that are deemed “high confidence” is performed using COMPAS software.  
All low confidence beats are reviewed manually and adjudicated using pass-fail criteria.
The final QC assessment is performed by a cardiologist. The beats found acceptable by 
manual review are included in the analysis. The median QT, QTc, and HR value from 
each extracted replicate is calculated, and then the mean of all available medians from a 
nominal timepoint is used as the subject’s reportable value at that timepoint.
Categorical T-wave morphology analysis and the measurement of PR and QRS intervals 
will be performed manually in 3 of the 10 ECG replicates at each timepoint.
Each 
fiducial point (onset of P-wave, onset of Q-wave, offset of S-wave, and offset of T-wave) 
is electronically marked.  




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o
pulmonary valve regurgitation- absent or present. If present, used to 
estimate diastolic PAP
o
Right ventricular wall thickness
o
Right ventricular size; diameter

Doppler-echo parameters for evaluation of left heart:
o
left ventricular ejection fraction (LVEF)
o
left ventricular wall thickness
o
Ratio of transmitral Doppler early filling velocity (E) to tissue Doppler early 
diastolic mitral annular velocity (E’)
o
aortic valve insufficiency: if present, categorization as mild, moderate, 
severe
o
mitral valve regurgitation: if present, categorization as mild, moderate, 
severe per definition in the procedure manual.
4.5.8
Ambulatory Blood Pressure Monitoring
Ambulatory Blood Pressure Monitoring (ABPM) will be performed on Day -1 and Day 12 
and mean daytime and mean nighttime blood pressure will be determined to assess the 
effect of balovaptan on blood pressure.  For this study the daytime period is defined as 
9:00 am to 9:00 pm and the nighttime period is defined as 1:00 am to 5:30 am.  The 
ABPM device needs to be active during these periods.  
The blood pressure cuff for the ABPM device will be placed on the non-dominant arm of 
the subjects. This means the majority of subjects will likely have the cuff fastened to 
their left arm. Since the TT-Doppler echocardiogram requires access to the left side of 
the subject and manipulation of the left arm the APBM device might be turned off during 
this procedure. On completion of the TT-Doppler echocardiogram the ABPM device will 
be reactivated, and assessments collected as usual for the remainder of the testing 
period. All deactivations and or removals of the APBM device during Day -1 and during 
Day 12 should be captured for each subject.  
During the period of time that no assessments are planned on Day -1 and Day 12, i.e.,
9:00 pm to 1:00 am, subjects may remove the ABPM device to perform activities that 
would be hindered by the device, e.g., to shower or change clothing.
Daytime period: Daytime is defined from 9 am to 9 pm for this study and blood pressure 
will be measured in 20-minute intervals during this period. Unusual or strenuous 
physical activities are not allowed during assessment periods. Subjects will be 
instructed not to sleep during this assessment period. The ABPM device maybe 
deactivated due to the need to perform other assessments including TT-Doppler 
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Nighttime period: Nighttime for this study is defined as from 1 am to 5:30 am and blood 
pressure will be measured in 20-minute intervals during this period. It is expected that 
subjects will remain asleep during this assessment period. Subjects will be instructed to 
try and remain asleep during this period and not leave the bed.
Pulse rate will be recorded by ABPM device during each assessment period at the times 
of blood pressure measurements.
Further details of ABPM analysis will be provided in a separate manual.
4.5.9
Columbia-Suicide Severity Rating Scale
The Columbia-Suicide Severity Rating Scale (C-SSRS) is a clinical tool used to assess 
the lifetime suicidality of a subject (C-SSRS lifetime version) as well as any new 
instances of suicidality (C-SSRS since last visit).
The structured interview prompts recollection of suicidal ideation, including the intensity 
of the ideation, behavior, and attempts with actual/potential lethality.
C-SSRS assessments will be completed by the Investigator or designee at Screening, 
Day -2, Day 16 and at Follow-up visits (Appendix 1).
4.5.10
Drug and Metabolite Concentration Measurements
For each Treatment Period plasma samples will be collected, as described in
Appendix 1 and Appendix 2, to determine concentrations of balovaptan and its 
metabolites M2 (as appropriate) and M3 utilizing a specific and validated liquid 
chromatography-tandem mass spectrometry (LC-MS/MS) method. Plasma 
concentrations of moxifloxacin will also be measured using a separate, specific and 
validated LC-MS/MS method.  
Moxifloxacin PK samples: will only be analyzed if the moxifloxacin assay sensitivity test 
fails; in such case, moxifloxacin plasma levels matched to the QT data by timepoint 
across subjects will be used for exposure response analysis of the QT effect.  
4.5.11
Timing of Study Assessments
If performed at the same timepoint, assessments should be prioritized as follows, while 
ensuring PK blood sampling is conducted at the scheduled time, when possible:  

12-lead ECGs

Vital signs measurements

PK blood sampling

Laboratory tests


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4.6
TREATMENT, SUBJECT, STUDY, AND SITE 
DISCONTINUATION
4.6.1
Study Treatment Discontinuation
Subjects must permanently discontinue study treatment if they experience any of the 
following:

Any medical condition that the Investigator, designee or Sponsor determines may 
jeopardize the subject’s safety if he or she continues to receive study treatment.
Section 5.1.2 provides additional guidance on the management of toxicities and of 
subjects who experience AEs within the outline of specific stopping rules.

Investigator, designee or Sponsor determines it is in the best interest of the subject

Pregnancy
The primary reason for study treatment discontinuation should be documented on the 
appropriate eCRF.  Subjects who discontinue study treatment may be replaced.
Subjects who terminate the study early while admitted to the CRU will have safety 
assessments performed as indicated for the Follow-up visit as described in SoA 
(Appendix 1) prior to discharge. In addition, subjects will be encouraged to return to the 
clinic for a Follow-up visit 14 to 21 days after the last dose of study drug (Appendix 1).
4.6.2
Subject Discontinuation from Study
Subjects will return to the clinic for a Follow-up visit 14 to 21 days after the last dose of
study drug.
Subjects have the right to voluntarily withdraw from the study at any time for any reason.  
In addition, the Investigator or designee has the right to withdraw a subject from the 
study at any time.  Reasons for withdrawal from the study may include, but are not 
limited to, the following:

Subject withdrawal of consent

Study termination or site closure

Subject non-compliance, defined as failure to comply with protocol requirements 
as determined by the Investigator, designee or Sponsor
Every effort should be made to obtain information on subjects who withdraw from the 
study.  The primary reason for withdrawal from the study should be documented on the 
appropriate eCRF. Subject requests to be withdrawn from the study must be 
documented in the source documents and signed by the Investigator (or designee).
Subjects who withdraw from the study may be replaced at the discretion of the 
Investigator or Sponsor.


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4.6.3
Study Discontinuation
The Sponsor has the right to terminate this study at any time.  Reasons for terminating 
the study may include, but are not limited to, the following:

The incidence or severity of AEs in this or other studies indicates a potential health 
hazard to subjects

Subject enrollment is unsatisfactory
The Sponsor will notify the Investigator if the Sponsor decides to discontinue the study.
4.6.4
Site Discontinuation
The Sponsor has the right to close a site at any time.  Reasons for closing a site may 
include, but are not limited to, the following:

Excessively slow recruitment

Poor protocol adherence

Inaccurate or incomplete data recording

Non-compliance with the International Council for Harmonisation (ICH) guideline for 
Good Clinical Practice

No study activity (i.e., all subjects have completed the study and all obligations have 
been fulfilled)
5.
ASSESSMENT OF SAFETY
5.1
SAFETY PLAN
Balovaptan is not approved in any country and clinical development is ongoing. The 
safety plan for subjects in this study is based on clinical experience with balovaptan in 
completed and ongoing studies. Potential safety risks for balovaptan are outlined below.
Please refer to the balovaptan (RO5285119) Investigator’s Brochure for a complete 
summary of the safety information available for the molecule.
Several measures will be taken to ensure the safety of subjects participating in this study.  
Eligibility criteria have been designed to exclude subjects at higher risk for toxicities.  
Subjects will undergo safety monitoring during the study, including assessment of the 
nature, frequency, and severity of adverse events.  In addition, guidelines for managing 
adverse events, including criteria for dosage modification and treatment interruption or 
discontinuation, are provided below.


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5.1.1.4
Pulmonary Blood Pressure
In animals as well as preparations of pulmonary vasculature, vasopressin has been 
shown to decrease pulmonary blood pressure and to relax arterial constriction through 
V1a receptors and in animals these changes were accompanied by systemic blood 
pressure increases.8,9
GLP toxicology animal study have not pointed to a right heart challenge strain nor to 
pulmonary fibrosis. The cardiomyopathy seen in rats refers to the right and left heart 
and mirrors cardiomyopathy with advanced age.
No systemic blood pressure changes with balovaptan were noted in clinical studies.  
Hence, the a priori expectation is to not see changes in pulmonary circulatory dynamics 
and with a N of 48, this study is powered to detect a change in pulmonary blood 
pressure representing half of 1 SD at of 0.05 and with a power of 80 %.
As an accompanied monitoring measure, the current database on systemic blood 
pressure monitoring is supplemented by ABPM monitoring as scheduled within this 
study.
 
 
 
 


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5.1.1.6
Orthostasis
Inhibition of V1a receptors by balovaptan may impair orthostatic regulation with blood 
pressure decreases, particularly when moving rapidly from a supine or sitting position to 
a standing position.  
However, various biological systems besides the vasopressinergic system are involved 
in regulating blood pressure control and the relevance of isolated V1a receptor inhibition 
may be minor.  Safety pharmacology studies in dogs have not pointed to blood pressure 
lowering effect of balovaptan.
Orthostatic challenge testing was included in the Phase 2 study BP28420 and no 
imbalance was noted between placebo- and balovaptan-treated cohorts in the 
percentage of subjects (approximately 8%) showing relevant blood pressure changes.
No respective safety signals emerged from the MAD arm of study BP25694 nor from the 
proof-of-mechanism study (BP29412) when balovaptan doses of 52 mg qd were 
administered for several days.  However, no orthostatic challenge testing was included 
in these trials.
Given that a dose of either 10 mg or 50 mg qd balovaptan is estimated to block the V1a 
receptor fully throughout the dosing interval, together with the clinical trial experience 
outlined above, the risk of balovaptan-related orthostatic impairment in this study is 
considered low.


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As indicated above, this study monitors systemic blood pressure by the means of ABPM.
5.1.2
Management of Subjects Who Experience Adverse Events
5.1.2.1
Management Guidelines
In general, any emerging AEs must diligently be watched, treated as medically indicated 
according to common medical practice and documented in terms of onset-date, intensity, 
off-set date, and any measures taken in order to treat the AE.
No specific treatment guidance to any potentially emerging AEs in balovaptan clinical 
trials exist and treatment should be according to the common medical practice for the 
given AE.
Temporary dosing interruptions must be discussed with the Sponsor’s Medical Monitor in 
the sense that temporary interruptions will have to be very limited in order to nor 
jeopardize the trial’s pharmacokinetic objectives.  Dose modifications because of an AE 
are not allowed in this study.  
 
 
 
 
 
  
 
  
 
  
 
 
  
 
 
 
 


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5.1.2.5
Changes in Pulmonary / Systemic Blood Pressure
While no relevant changes in pulmonary blood pressure are expected, should the 
TT-Doppler echocardiogram examination unexpectedly reveal such a change all 
available cardio-vascular and potential pulmonary data should be reviewed by 
Investigator (or designee) and cardiologist to assess the subject’s safety and to decide 
on next actions which may include treatment interruption and / or permanent treatment 
stop.  
An increase in mean pulmonary blood pressure of 25 mm/Hg will result in termination of 
study drug dosing and referral of the subject to a cardiologist.  If logistically possible 
abnormal TT-Doppler echocardiograms should be repeated as soon as possible to 
confirm results.
The relevance of changes in the parameters obtained by TT-Doppler echocardiogram
will be interpreted by the cardiologist.  A change in PAAT by 50% or more in a subject, is 
a safety signal that will result in a discussion of discontinuing treatment for the affected
subject. 
Given current clinical experience for systemic blood pressure and HR, relevant changes 
in standard and ambulatory blood pressure monitoring (ABPM) monitoring are not 
expected.  However, repeated blood pressure changes exceeding the 
BP systolic > 160 mm Hg and/or BP diastolic > 100 mm Hg and that cannot be explained 
by any circumstances or low blood pressure linked with moderate to severe clinical AEs 
related to low blood pressure may lead to treatment interruption or permanent 
withdrawal from study treatment.  For orthostatic events, please see also Section 5.1.2.6.
5.1.2.6
Management of Orthostatic Hypotension
Subjects with unexplained orthostatic reaction with syncope within the last 6 months are
excluded from this study.
Vital signs will be monitored frequently throughout the study as indicated in the SoA 
(Appendix 1).
Subjects will be instructed to avoid rapid body movements from supine to standing 
positions, and to inform the Investigator or designee as soon as possible should they 
experience dizziness, nausea, sweating, pale face, or even fainting with such body 
movements.


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5.1.2.7
Management of Signs and Symptoms Indicative for Peripheral 
Neuropathy
Subjects with signs and symptoms potentially indicative of peripheral neuropathy are 
excluded from this study.
As outlined above, the risk of peripheral neuropathy is minimal in study.
Should however signs and symptoms potentially indicative of peripheral neuropathy 
occur, an expert in the field of peripheral neuropathy should be consulted to discuss next 
diagnostic steps as well as the need to interrupt study treatment.  Additional diagnostic 
tests may include a nerve conduction study.
5.2
SAFETY PARAMETERS AND DEFINITIONS
Safety assessments will consist of monitoring and recording AEs, including SAEs and 
AEs of special interest (AESI), performing protocol-specified safety laboratory 
assessments, measuring protocol-specified vital signs, and conducting other protocol-
specified tests that are deemed critical to the safety evaluation of the study. The 
schedule for conducting these activities is provided in the SoA (Appendix 1).
Certain types of events require immediate reporting to the Sponsor, as outlined in 
Section 5.4.
5.2.1
Adverse Events
According to the ICH guideline for Good Clinical Practice, an AE is any untoward 
medical occurrence in a clinical investigation subject administered a pharmaceutical 
product, regardless of causal attribution.  An AE can therefore be any of the following:

Any unfavorable and unintended sign (including an abnormal laboratory finding), 
symptom, or disease temporally associated with the use of a medicinal product, 
whether or not considered related to the medicinal product

Any new disease or exacerbation of an existing disease (a worsening in the 
character, frequency, or severity of a known condition) 

Recurrence of an intermittent medical condition (e.g., headache) not present at 
baseline

Any deterioration in a laboratory value or other clinical test (e.g., ECG, X-ray) that is 
associated with symptoms or leads to a change in study treatment or concomitant 
treatment or discontinuation from study drug

Adverse events that are related to a protocol-mandated intervention, including those 
that occur prior to assignment of study treatment (e.g., screening invasive 
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5.2.2
Serious Adverse Events (Immediately Reportable to the 
Sponsor)
A serious adverse event is any adverse event that meets any of the following criteria:

Is fatal (i.e., the AE actually causes or leads to death)

Is life threatening (i.e., the AE, in the view of the Investigator or designee, places the 
subject at immediate risk of death)
o
This does not include any AE that, had it occurred in a more severe form 
or was allowed to continue, might have caused death.

Requires or prolongs subject hospitalization (Section 5.3.5.9)

Results in persistent or significant disability/incapacity (i.e., the AE results in 
substantial disruption of the subject’s ability to conduct normal life functions)

Is a congenital anomaly/birth defect in a neonate/infant born to a mother exposed to 
study drugs

Is a significant medical event in the Investigator or designee's judgment (e.g., may 
jeopardize the subject or may require medical/surgical intervention to prevent one of 
the outcomes listed above)
The terms "severe" and "serious" are not synonymous.  Severity refers to the intensity of 
an adverse event (e.g., rated as mild, moderate, or severe, or according to National 
Cancer Institute Common Terminology Criteria for Adverse Events [NCI CTCAE]; see 
Section 5.3.3); the event itself may be of relatively minor medical significance (such as 
severe headache without any further findings).
Severity and seriousness need to be independently assessed for each AE recorded on 
the eCRF.
Serious adverse events are required to be reported by the Investigator or designee to 
the Sponsor immediately (i.e., no more than 24 hours after learning of the event; see 
Section 5.4.2 for reporting instructions).
5.2.3
Adverse Events of Special Interest (Immediately Reportable to 
the Sponsor)
Adverse events of special interest are required to be reported by the Investigator (or
designee) to the Sponsor immediately (i.e., no more than 24 hours after learning of the 
event; see Section 5.4.2 for reporting instructions).  AESIs for this study are as follows:

Cases of potential drug-induced liver injury that include an elevated ALT or AST in 
combination with either an elevated bilirubin or clinical jaundice, as defined by Hy's 
Law (see Section 5.3.5.6)

Suspected transmission of an infectious agent by the study drug, as defined below
Any organism, virus, or infectious particle (e.g., prion protein transmitting 
transmissible spongiform encephalopathy), pathogenic or non-pathogenic, is 
considered an infectious agent.  A transmission of an infectious agent may be 


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suspected from clinical symptoms or laboratory findings that indicate an 
infection in a subject exposed to a medicinal product.  This term applies only
when a contamination of the study drug is suspected.

Clinically relevant decreases in ANC and for which no alternative explanation has 
been identified (i.e., the relationship to study drug is rated “yes”).

CPK values that exceed 10× ULN for several days without a trend of normalization 
or are associated with relevant dysfunction of kidneys, and if no alternative 
explanation exists.

Adverse events such as: 
o
Arrhythmia
o
Syncope
o
Dyspnea
o
Palpitations
o
Chest pain.
5.3
METHODS AND TIMING FOR CAPTURING AND ASSESSING 
SAFETY PARAMETERS
The Investigator is responsible for ensuring that all AEs are recorded on the Adverse 
Event eCRF and source form and reported to the Sponsor in accordance with 
instructions provided in this Section and in Sections 5.4-5.6.
For each AE recorded on the Adverse Event eCRF and source form, the Investigator or 
designee will make an assessment of seriousness (see Section 5.2.2), severity (see 
Section 5.3.3), and causality (see Section 5.3.4).  
5.3.1
Adverse Event Reporting Period
All AEs, whether reported by the subject or noted by study personnel, will be recorded in 
the subject’s medical record and on the Adverse Event eCRF and source form.
After informed consent has been obtained but prior to initiation of study drug,only SAEs
caused by a protocol-mandated intervention (e.g., invasive procedures such as biopsies, 
discontinuation of medications) should be reported (see Section 5.4.2 for instructions for 
reporting SAEs).
After initiation of study drug, all AEs will be reported until 21 days after the last dose of 
study drug.
Instructions for reporting AEs that occur after the AE reporting period are provided in 
Section 5.6.


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5.3.2
Eliciting Adverse Event Information
A consistent methodology of non-directive questioning should be adopted for eliciting AE
information.  Examples of non-directive questions include the following:
"How have you felt since your last clinic visit?"
"Have you had any new or changed health problems since you were last here?"
5.3.3
Assessment of Severity of Adverse Events
Table 3 provides guidance for assessing AE severity.
Table 3
Adverse Event Severity Grading Scale
Severity
Description
Mild
Discomfort noticed, but no disruption of normal daily activity
Moderate
Discomfort sufficient to reduce or affect normal daily activity
Severe
Incapacitating with inability to work or to perform normal daily activity
Note:  Regardless of severity, some events may also meet seriousness criteria.  
Refer to definition of a serious adverse event (see Section 5.2.2).
5.3.4
Assessment of Causality of Adverse Events
Investigator or designees should use their knowledge of the subject, the circumstances 
surrounding the event, and an evaluation of any potential alternative causes to 
determine whether an AE is considered to be related to the study drug, indicating "yes"
or "no" accordingly.  The following guidance should be taken into consideration (see also 
Table 4)

Temporal relationship of event onset to the initiation of study drug

Course of the event, with special consideration of the effects of dose reduction, 
discontinuation of study drug, or reintroduction of study drug (as applicable)

Known association of the event with the study drug or with similar treatments

Known association of the event with the disease under study

Presence of risk factors in the subject or use of concomitant medications known to 
increase the occurrence of the event

Presence of non-treatment-related factors that are known to be associated with the 
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Table 4
Causal Attribution Guidance
Is the AE suspected to be caused by the study drug on the basis of facts, evidence, 
science-based rationales, and clinical judgment?
YES
There is a plausible temporal relationship between the onset of the AE and 
administration of the study drug, and the AE cannot be readily explained by the 
subject's clinical state, intercurrent illness, or concomitant therapies; and/or the AE
follows a known pattern of response to the study drug; and/or the AE abates or resolves 
upon discontinuation of the study drug or dose reduction and, if applicable, reappears 
upon re-challenge.
NO
An AE will be considered related, unless it fulfills the criteria specified below.  
Evidence exists that the AE has an etiology other than the study drug (e.g., preexisting 
medical condition, underlying disease, intercurrent illness, or concomitant medication); 
and/or the AE has no plausible temporal relationship to administration of the study drug 
(e.g., cancer diagnosed 2 days after first dose of study drug).
For subjects receiving combination therapy, causality will be assessed individually for 
each protocol-mandated therapy following conclusion of the study and unblinding of 
study data.
5.3.5
Procedures for Recording Adverse Events
Investigator or designees should use correct medical terminology/concepts when 
recording AEs on the Adverse Event eCRF and source form.  Avoid colloquialisms and 
abbreviations.
Only one AE term should be recorded in the event field on the Adverse Event eCRF and 
source form.
5.3.5.1
Diagnosis versus Signs and Symptoms
A diagnosis (if known) should be recorded on the Adverse Event eCRF and source form
rather than individual signs and symptoms (e.g., record only liver failure or hepatitis 
rather than jaundice, asterixis, and elevated transaminases).  However, if a constellation 
of signs and/or symptoms cannot be medically characterized as a single diagnosis or 
syndrome at the time of reporting, each individual event should be recorded on the 
Adverse Event eCRF and source form.  If a diagnosis is subsequently established, all 
previously reported AEs based on signs and symptoms should be nullified and replaced 
by one AE report based on the single diagnosis, with a starting date that corresponds to 
the starting date of the first symptom of the eventual diagnosis.


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5.3.5.2
Adverse Events That Are Secondary to Other Events
In general, AEs that are secondary to other events (e.g., cascade events or clinical 
sequelae) should be identified by their primary cause, with the exception of severe or 
serious secondary events.  A medically significant secondary AE that is separated in 
time from the initiating event should be recorded as an independent event on the 
Adverse Event eCRF and source form.  For example:

If vomiting results in mild dehydration with no additional treatment in a healthy adult, 
only vomiting should be reported on the eCRF.

If vomiting results in severe dehydration, both events should be reported separately 
on the eCRF.

If a severe gastrointestinal hemorrhage leads to renal failure, both events should be 
reported separately on the eCRF.

If dizziness leads to a fall and consequent fracture, all three events should be 
reported separately on the eCRF.

If neutropenia is accompanied by an infection, both events should be reported 
separately on the eCRF.
All AEs should be recorded separately on the Adverse Event eCRF and source form if it 
is unclear as to whether the events are associated.
5.3.5.3
Persistent or Recurrent Adverse Events
A persistent AE is one that extends continuously, without resolution, between subject
evaluation timepoints.  Such events should only be recorded once on the Adverse Event 
eCRF and source form.  The initial severity (intensity or grade) of the event will be 
recorded at the time the event is first reported.  If a persistent AE becomes more severe, 
the most extreme severity should also be recorded on the Adverse Event eCRF and 
source form. If the event becomes serious, it should be reported to the Sponsor 
immediately (i.e., no more than 24 hours after learning that the event became serious; 
see Section 5.4.2 for reporting instructions).  The Adverse Event eCRF and source form
should be updated by changing the event from "non-serious" to "serious," providing the 
date that the event became serious, and completing all data fields related to SAEs.
A recurrent adverse event is one that resolves between subject evaluation timepoints 
and subsequently recurs.  Each recurrence of an adverse event should be recorded as a 
separate event on the Adverse Event eCRF and source form.
5.3.5.4
Abnormal Laboratory Values
Not every laboratory abnormality qualifies as an AE.  A laboratory test result must be 
reported as an AE if it meets any of the following criteria:

Is accompanied by clinical symptoms

Results in a change in study treatment (e.g., dosage modification, treatment 
interruption, or treatment discontinuation)


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
Results in a medical intervention (e.g., potassium supplementation for hypokalemia) 
or a change in concomitant therapy

Is clinically significant in the Investigator or designee’s judgment 
It is the Investigator or designee’s responsibility to review all laboratory findings.  Medical 
and scientific judgment should be exercised in deciding whether an isolated laboratory 
abnormality should be classified as an AE.
If a clinically significant laboratory abnormality is a sign of a disease or syndrome 
(e.g., alkaline phosphatase and bilirubin 5ULN associated with cholestasis), only the 
diagnosis (i.e., cholestasis) should be recorded on the Adverse Event eCRF and source 
form.
If a clinically significant laboratory abnormality is not a sign of a disease or syndrome, 
the abnormality itself should be recorded on the Adverse Event eCRF and source form, 
along with a descriptor indicating whether the test result is above or below the normal 
range (e.g., "elevated potassium," as opposed to "abnormal potassium").  If the 
laboratory abnormality can be characterized by a precise clinical term per standard 
definitions, the clinical term should be recorded as the AE.  For example, an elevated 
serum potassium level of 7.0 mEq/L should be recorded as "hyperkalemia."
Observations of the same clinically significant laboratory abnormality from visit to visit 
should only be recorded once on the Adverse Event eCRF and source form (see 
Section 5.3.5.3 for details on recording persistent AEs).
5.3.5.5
Abnormal Vital Sign Values
Not every vital sign abnormality qualifies as an AE.  A vital sign result must be reported 
as an AE if it meets any of the following criteria:

Is accompanied by clinical symptoms

Results in a change in study treatment (e.g., dosage modification, treatment 
interruption, or treatment discontinuation)

Results in a medical intervention or a change in concomitant therapy

Is clinically significant in the Investigator or designee’s judgment 
It is the Investigator or designee’s responsibility to review all vital sign findings.  Medical 
and scientific judgment should be exercised in deciding whether an isolated vital sign 
abnormality should be classified as an AE.
If a clinically significant vital sign abnormality is a sign of a disease or syndrome 
(e.g., high blood pressure), only the diagnosis (i.e., hypertension) should be recorded on 
the Adverse Event eCRF and source form.


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Observations of the same clinically significant vital sign abnormality from visit to visit 
should only be recorded once on the Adverse Event eCRF and source form (see 
Section 5.3.5.3 for details on recording persistent AEs).
5.3.5.6
Abnormal Liver Function Tests
The finding of an elevated ALT or AST (3ULN) in combination with either an elevated 
total bilirubin (2ULN) or clinical jaundice in the absence of cholestasis or other 
causes of hyperbilirubinemia is considered to be an indicator of severe liver injury (as 
defined by Hy's Law).  Therefore, Investigator or designees must report as an AE the 
occurrence of either of the following:

Treatment-emergent ALT or AST 3ULN in combination with total bilirubin 
2ULN

Treatment-emergent ALT or AST 3ULN in combination with clinical jaundice
The most appropriate diagnosis or (if a diagnosis cannot be established) the abnormal 
laboratory values should be recorded on the Adverse Event eCRF and source form (see 
Section 5.3.5.2) and reported to the Sponsor immediately (i.e., no more than 24 hours 
after learning of the event), either as a SAE or an AESI (see Section 5.4.2).
5.3.5.7
Deaths
All deaths that occur during the protocol-specified adverse event reporting period (see 
Section 5.3.1), regardless of relationship to study drug, must be recorded on the 
Adverse Event eCRF and source form and immediately reported to the Sponsor (see 
Section 5.4.2).
Death should be considered an outcome and not a distinct event.  The event or condition 
that caused or contributed to the fatal outcome should be recorded as the single medical 
concept on the Adverse Event eCRF and source form.  Generally, only one such event 
should be reported.  If the cause of death is unknown and cannot be ascertained at the 
time of reporting, "unexplained death" should be recorded on the Adverse Event eCRF 
and source form.  If the cause of death later becomes available (e.g., after autopsy), 
"unexplained death" should be replaced by the established cause of death.  The term 
"sudden death" should not be used unless combined with the presumed cause of death 
(e.g., "sudden cardiac death").
Deaths that occur after the adverse event reporting period should be reported as 
described in Section 5.6.
5.3.5.8
Preexisting Medical Conditions
A preexisting medical condition is one that is present at the screening visit for this study.  
Such conditions should be recorded on the General Medical History and Baseline 
Conditions eCRF.
A preexisting medical condition should be recorded as an adverse event only if the 
frequency, severity, or character of the condition worsens during the study.  When 


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recording such events on the Adverse Event eCRF and source form, it is important to 
convey the concept that the preexisting condition has changed by including applicable 
descriptors (e.g., "more frequent headaches").
5.3.5.9
Hospitalization or Prolonged Hospitalization
Any AE that results in hospitalization (i.e., inpatient admission to a hospital) or prolonged 
hospitalization should be documented and reported as a serious adverse event (per the 
definition of serious adverse event in Section 5.2.2), except as outlined below.
An event that leads to hospitalization under the following circumstances should not be 
reported as an AE or a SAE:

Hospitalization for respite care

Planned hospitalization required by the protocol

Hospitalization for a preexisting condition, provided that all of the following criteria 
are met:
o
The hospitalization was planned prior to the study or was scheduled 
during the study when elective surgery became necessary because of the 
expected normal progression of the disease
o
The subject has not experienced an AE
An event that leads to hospitalization under the following circumstances is not 
considered to be a SAE, but should be reported as an AE instead:

Hospitalization that was necessary because of subject requirement for outpatient 
care outside of normal outpatient clinic operating hours
5.4
IMMEDIATE REPORTING REQUIREMENTS FROM 
INVESTIGATOR TO SPONSOR
Certain events require immediate reporting to allow the Sponsor to take appropriate 
measures to address potential new risks in a clinical trial.  The Investigator or designee
must report such events to the Sponsor immediately; under no circumstances should 
reporting take place more than 24 hours after the Investigator or designee learns of the 
event.  The following is a list of events that the Investigator or designee must report to 
the Sponsor within 24 hours after learning of the event, regardless of relationship to 
study drug:

Serious adverse events (defined in Section 5.2.2; see Section 5.4.2 for details on 
reporting requirements)

Adverse events of special interest (defined in Section 5.2.3; see Section 5.4.2 for 
details on reporting requirements)

Pregnancies (see Section 5.4.3 for details on reporting requirements)


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The Investigator or designee must report new significant follow-up information for these 
events to the Sponsor immediately (i.e., no more than 24 hours after becoming aware of 
the information).  New significant information includes the following:

New signs or symptoms or a change in the diagnosis

Significant new diagnostic test results

Change in causality based on new information

Change in the event’s outcome, including recovery

Additional narrative information on the clinical course of the event
Investigator or designees must also comply with local requirements for reporting serious 
adverse events to the local health authority and IRB/EC.
5.4.1
Emergency Medical Contacts
Medical Monitor Contact Information:
Medical Monitor:
, M.D. (Primary)
Telephone No.:
 
Mobile Telephone No.:
Medical Monitor:
, M.D. (Secondary)
Telephone No.:
Mobile Telephone No.:
To ensure the safety of study subjects, an Emergency Medical Call Center Help Desk 
will access the Roche Medical Emergency List, escalate emergency medical calls, 
provide medical translation service (if necessary), connect the Investigator or designee
with a Roche Medical Responsible (listed above and/or on the Roche Medical 
Emergency List), and track all calls.  The Emergency Medical Call Center Help Desk will 
be available 24 hours per day, 7 days per week.  Toll-free numbers for the Help Desk, as 
well as Medical Monitor and Medical Responsible contact information, will be distributed 
to all Investigators or designees. 
5.4.2
Reporting Requirements for Serious Adverse Events and 
Adverse Events of Special Interest
5.4.2.1
Events That Occur prior to Study Drug Initiation
After informed consent has been obtained but prior to initiation of study drug, only 
serious adverse events caused by a protocol-mandated intervention should be reported.  
The paper Clinical Trial Serious Adverse Event (SAE)/AE of Special Interest Reporting 
Form provided to Investigator or designees should be completed and submitted to the 
Sponsor or its designee immediately (i.e., no more than 24 hours after learning of the 
event), either by faxing or by scanning and emailing the form using the fax number or 
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5.4.2.2
Events That Occur after Study Drug Initiation
After initiation of study drug, SAEs and AEs of special interest will be reported until 
21 days after the last dose of study drug.  Investigator or designees should record all 
case details that can be gathered immediately (i.e., within 24 hours after learning of the 
event) on the paper Clinical Trial Serious Adverse Event/Adverse Event of Special 
Interest Reporting Form provided to investigators and submitting the report, either by 
faxing or by scanning and emailing the form using the fax number or email address 
provided to investigators.
Instructions for reporting SAEs that occur 21 days after the last dose of study treatment 
are provided in Section 5.6.
5.4.3
Reporting Requirements for Pregnancies
5.4.3.1
Pregnancies in Female Subjects
Female subjects of childbearing potential will be instructed to immediately inform the 
Investigator or designee if they become pregnant during the study or within 90 days after 
the last dose of study drug.  A paper Clinical Trial Pregnancy Reporting Form should be 
completed and submitted to the Sponsor or its designee immediately (i.e., no more than 
24 hours after learning of the pregnancy), either by faxing or by scanning and emailing 
the form using the fax number or email address provided to the Investigator.  Pregnancy 
should not be recorded on the Adverse Event eCRF and source form.  The Investigator
or designee should discontinue the study drug and counsel the subject, discussing the 
risks of the pregnancy and the possible effects on the fetus.  Monitoring of the subject
should continue until conclusion of the pregnancy.  Any SAE associated with the 
pregnancy (e.g., an event in the fetus, an event in the mother during or after the 
pregnancy, or a congenital anomaly/birth defect in the child) should be reported on the 
Adverse Event eCRF and source form.  In addition, the Investigator or designee will 
submit a Clinical Trial Pregnancy Reporting Form when updated information on the 
course and outcome of the pregnancy becomes available.  
5.4.3.2
Pregnancies in Female Partners of Male Subjects
Male subjects will be instructed through the Informed Consent Form to immediately 
inform the Investigator or designee if their partner becomes pregnant during the study or 
within 90 days after the last dose of study drug.  A paper Clinical Trial Pregnancy 
Reporting Form should be completed and submitted to the Sponsor or its designee 
immediately (i.e., no more than 24 hours after learning of the pregnancy), either by 
faxing or by scanning and emailing the form using the fax number or email address
provided to investigators.  Attempts should be made to collect and report details of the 
course and outcome of any pregnancy in the partner of a male subject exposed to study 
drug.  When permitted by the site, the pregnant partner would need to sign an 
Authorization for Use and Disclosure of Pregnancy Health Information to allow for 
follow-up on her pregnancy.  If the authorization has been signed, the Investigator or 
designee should submit a Clinical Trial Pregnancy Reporting Form when updated 
information on the course and outcome of the pregnancy becomes available.  An 


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Investigator or designee who is contacted by the male subject or his pregnant partner 
may provide information on the risks of the pregnancy and the possible effects on the 
fetus, to support an informed decision in cooperation with the treating physician and/or 
obstetrician.
5.4.3.3
Abortions
Any abortion should be classified as a SAE (as the Sponsor considers abortions to be 
medically significant), recorded on the Adverse Event eCRF and source form, and 
reported to the Sponsor immediately (i.e., no more than 24 hours after learning of the 
event; see Section 5.4.2).  
5.4.3.4
Congenital Anomalies/Birth Defects
Any congenital anomaly/birth defect in a child born to a female subject exposed to study 
drug or the female partner of a male subject exposed to study drug should be classified 
as a serious adverse event, recorded on the Adverse Event form, and reported to the 
Sponsor immediately (i.e., no more than 24 hours after learning of the event; see 
Section 5.4.2).
5.4.4
Reporting Requirements for Cases of Accidental Overdose or 
Medication Error 
Accidental overdose and medication error (hereafter collectively referred to as "special 
situations"), are defined as follows:

Accidental overdose:  accidental administration of a drug in a quantity that is higher 
than the assigned dose

Medication error:  accidental deviation in the administration of a drug 
In some cases, a medication error may be intercepted prior to administration of 
the drug.
Special situations are not in themselves adverse events but may result in adverse 
events.  Each adverse event associated with a special situation should be recorded 
separately on the Adverse Event eCRF.  If the associated adverse event fulfills 
seriousness criteria, the event should be reported to the Sponsor immediately (i.e., no 
more than 24 hours after learning of the event; see Section 5.4.2)  Special situations 
should be recorded as described below:

Accidental overdose:  Enter the drug name and "accidental overdose" as the event 
term.  Check the "Accidental overdose" and "Medication error" boxes.

Medication error that does not qualify as an overdose:  Enter the name of the drug 
administered and a description of the error (e.g., wrong dose administered, wrong 
dosing schedule, incorrect route of administration, wrong drug, expired drug 
administered) as the event term.  Check the "Medication error" box.

Medication error that qualifies as an overdose:  Enter the drug name and "accidental 
overdose" as the event term.  Check the "Accidental overdose" and "Medication 
error" boxes.  Enter a description of the error in the additional case details.


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
Intercepted medication error:  Enter the drug name and "intercepted medication 
error" as the event term.  Check the "Medication error" box.  Enter a description of 
the error in the additional case details.
For study drug, each AE associated with a special situation should be recorded 
separately on the Adverse Event eCRF and source form.  If the associated AE fulfills 
seriousness criteria, the event should be reported to the Sponsor immediately 
(i.e., no more than 24 hours after learning of the event; see Section 5.4.2).  Study drug 
AEs associated with special situations should be recorded as described below for each 
situation:

Accidental overdose:  Enter the adverse event term.  Check the "Accidental 
overdose" and "Medication error" boxes.

Medication error that does not qualify as an overdose:  Enter the adverse event term.  
Check the "Medication error" box.

Medication error that qualifies as an overdose:  Enter the adverse event term.  
Check the "Accidental overdose" and "Medication error" boxes.
As an example, an accidental overdose that resulted in a headache would require the 
completion of two Adverse Event eCRF and source form, one to report the accidental 
overdose and one to report the headache.  The "Accidental overdose" and "Medication 
error" boxes would need to be checked on both eCRF pages.
5.5
FOLLOW-UP OF SUBJECTS AFTER ADVERSE EVENTS
5.5.1
Investigator Follow-Up
The Investigator or designee should follow each adverse event until the event has resolved 
to baseline grade or better, the event is assessed as stable by the Investigator or designee, 
the subject is lost to follow-up, or the subject withdraws consent.  Every effort should be 
made to follow all SAEs considered to be related to study drug or trial-related procedures 
until a final outcome can be reported.
During the study period, resolution of adverse events (with dates) should be documented 
on the Adverse Event eCRF and source form and in the subject’s medical record to 
facilitate source data verification.
All pregnancies reported during the study should be followed until pregnancy outcome.
5.5.2
Sponsor Follow-Up
For SAEs, AESI, and pregnancies, the Sponsor or a designee may follow up by 
telephone, fax, email, and/or a monitoring visit to obtain additional case details and 
outcome information (e.g., from hospital discharge summaries, consultant reports, 
autopsy reports) in order to perform an independent medical assessment of the reported 
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5.6
ADVERSE EVENTS THAT OCCUR AFTER THE ADVERSE 
EVENT REPORTING PERIOD
The Sponsor should be notified if the Investigator or designee becomes aware of any 
SAE that occurs after the end of the AE reporting period (defined as 21 days after the 
last dose of study drug), if the event is believed to be related to prior study drug 
treatment.  These events should be reported through use of the Adverse Event eCRF 
and source form.  The Investigator or designee should report these events directly to the 
Sponsor or its designee, either by faxing or by scanning and emailing the paper Clinical 
Trial Serious Adverse Event/Adverse Event of Special Interest Reporting Form using the 
fax number or email address provided to investigators.
5.7
EXPEDITED REPORTING TO HEALTH AUTHORITIES, 
INVESTIGATORS, INSTITUTIONAL REVIEW BOARDS, AND 
ETHICS COMMITTEES
The Sponsor will promptly evaluate all SAEs and AESI against cumulative product 
experience to identify and expeditiously communicate possible new safety findings to 
Investigator (or designee), IRBs, ECs, and applicable health authorities based on 
applicable legislation.
Independent from the causality assessment, the following SAEs and non-SAEs of 
special interest will be reported to the FDA on an expedited basis: death, arrhythmia, 
syncope, dyspnea, palpitations, and chest pain.  
To determine reporting requirements for single AE cases, the Sponsor will assess the 
expectedness of these events using the following reference documents:

RO5285119 (Balovaptan) Investigator's Brochure

Moxifloxacin package insert as relevant in the United States of America
The Sponsor will compare the severity of each event and the cumulative event 
frequency reported for the study with the severity and frequency reported in the 
applicable reference document.
Reporting requirements will also be based on the Investigator or designee's assessment 
of causality and seriousness, with allowance for upgrading by the Sponsor as needed.
An aggregate report of any clinically relevant imbalances that do not favor the test 
product will be submitted to health authorities.
6.
STATISTICAL CONSIDERATIONS AND ANALYSIS PLAN
A statistical analysis plan (SAP) will be generated by the biostatistics department of the 
Sponsor and will be finalized prior to database lock.  


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6.1
DETERMINATION OF SAMPLE SIZE
A sample size of 56 subjects was chosen to obtain 48 evaluable subjects.  Subjects are 
evaluable if ECGs are available for all three periods to allow (1) the derivation of the 
primary endpoint (i.e., change from baseline on Day 14), and (2) the assessment of the 
effect of moxifloxacin.
Based on the calculation of the sample size for a TQT study (Zhang and Machado, 
2008)6, assuming a 1-sided 5% significance level and a within-subject SD of 8 ms for 
ΔQTcF and a true mean difference of 3 ms in ΔQTcF between balovaptan and placebo, 
a sample size of 48 evaluable subjects who complete the study will provide a power of 
96.6% to demonstrate that the upper bound of all the 2-sided 90% CIs on ΔΔQTcF will 
fall below 10 ms for up to 7 post-dose time points (0.5, 1.25, 2.5, 4, 8, 12 and 24 hours).
Sample Size Determination for Assay Sensitivity:
Based on the calculation of the sample size for a TQT study from Zhang and Machado, 
20086, as the test is performed at three time points separately (1.25, 2.5,and 4 hours), a 
one-sided 5% significance level (with adjusted one-sided significance levels of 5%, 2.5% 
and 1.67%) is used along with a within-subject SD of 8 ms for ΔQTcF, a sample size of 
48 evaluable subjects who complete the study will provide a power of 99% to prove a 
mean difference of 5 ms in ΔQTcF between moxifloxacin and placebo (i.e., ΔΔQTcF) at 
1 ≥of the 3 specified time points.  
6.2
SUMMARIES OF CONDUCT OF STUDY
The number of subjects who enroll, discontinue, or complete the study will be 
summarized.  Reasons for premature study withdrawal will be listed and summarized.  
Enrollment and major protocol deviations will be listed and evaluated for their potential 
effects on the interpretation of study results.
6.3
SUMMARIES OF DEMOGRAPHIC AND BASELINE 
CHARACTERISTICS
Demographic and baseline characteristics (including age, sex) will be summarized using 
means, standard deviations, medians, and ranges for continuous variables and 
proportions for categorical variables, as appropriate.  Summaries will be presented 
overall and by treatment group.
6.4
EFFICACY ANALYSES
Not Applicable
6.5
SAFETY ANALYSES
All subjects who have received at least one dose of balovaptan, whether prematurely 
withdrawn from the study or not, will be included in the safety analysis. The safety data, 
including AEs, laboratory data, and vital signs, will be listed and summarized by study 
treatment. Reasons for withdrawal from study will be listed and summarized. 


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Concomitant medications, 12-lead ECG data and clinically significant neurological 
examinations will be listed. Adverse events will be listed and summarized by treatment, 
body system and preferred term using the Medical Dictionary for Regulatory Activities 
(MedDRA). All verbatim AE terms will be mapped to MedDRA thesaurus terms, and AE 
severity will be graded according to Table 3. For laboratory and 12-lead ECG data, 
subject listings will be presented with abnormalities flagged.
For statistical analyses of results from TT-Doppler echocardiogram and ABPM please 
see Sections 6.8 and 6.9 below.
6.6
PHARMACOKINETIC ANALYSES
Subjects will be excluded from the PK analysis population if they significantly violate the
inclusion or exclusion criteria, deviate significantly from the protocol, or if data are
unavailable or incomplete. Excluded cases will be documented together with the reason
for exclusion. All decisions on exclusions from the analysis will be made prior to
database closure.
Pharmacokinetics are a secondary objective of this study. The specific objectives are

To investigate the pharmacokinetics of balovaptan, M2 (as appropriate), and M3.

To investigate the pharmacokinetics of moxifloxacin.
Individual plasma concentrations at each sampling timepoint for balovaptan, M2 (as 
appropriate), M3, and moxifloxacin will be presented by listings and descriptive summary 
statistics by treatment, including means, geometric means, ranges, standard deviations 
and coefficients of variation. Individual and mean concentration versus time will be 
plotted on linear and semi-logarithmic scales.
PK parameters will be estimated using standard non-compartmental methods. All PK 
parameters for balovaptan, M2 (as appropriate), and M3 will be presented by individual 
listings and summary statistics by treatment including means, geometric means, 
medians, ranges, standard deviations and coefficients of variation.
The following balovaptan, M2 (as appropriate), M3 PK parameters will be estimated:

Cmax: Day 1 and Day 14 for Treatment A and B

AUC0-24h: Day 1 and Day 14 for Treatment A and B

Tmax: Day 1 and Day 14 for Treatment A and B

Ctrough: Day 11, 12, 13, 14 for Treatment A and B

Time to steady state for Treatment A and B


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The following moxifloxacin PK parameters will be estimated:

Cmax, AUC0-24h, and Tmax will be estimated on Days 2 (Treatment C) or 15
(Treatment D)
Additional PK analyses maybe be conducted as appropriate for balovaptan, M2 (as 
appropriate), M3 and moxifloxacin.
Subjects will be excluded from the PK analysis population if they significantly violate the 
inclusion or exclusion criteria, deviate significantly from the protocol, or if data are 
unavailable or incomplete. Excluded cases will be documented together with the reason 
for exclusion. All decisions on exclusions from the analysis will be made prior to 
database closure.
6.7
CARDIODYNAMIC ECG EVALUTAION
The following ECG parameters will be measured and calculated: HR, PR, QT, QTcF, 
and QRS. T-wave morphology and U-wave presence will be assessed. 
The primary analysis will be based on by-time point analysis to evaluate the effect of 
balovaptan on the placebo-corrected change-from-baseline QTcF (ΔΔQTcF) at each 
post-dosing time point using the Intersection Union Test. The effect of balovaptan on 
placebo-corrected change-from-baseline in HR, PR, and QRS (ΔΔHR, ΔΔPR, and 
ΔΔQRS) will also be evaluated using the Intersection Union Test. In addition, the 
relationship between plasma concentrations of balovaptan and its metabolites (M2 [as 
appropriate] and M3) and Δ∆QTcF will be evaluated using a linear mixed-effects 
modeling approach. An analysis of categorical outliers will be performed for changes in 
HR, PR, QRS, QTcF, T-wave morphology and U-wave presence. Assay sensitivity will 
also be evaluated using by-time point analysis of the effect on Δ∆QTcF of moxifloxacin 
using a similar model as for the primary analysis. 
6.7.1
Analysis Populations
The Safety population will include all subjects who are randomized and received at least 
one dose of the study medication (either balovaptan, moxifloxacin or matching placebo), 
whether prematurely withdrawn from the study or not.
The PK population will include all subjects who receive at least 1 dose of investigational 
product and have at least 1 evaluable PK plasma concentration of balovaptan (and 
moxifloxacin, when applicable).
The QT/QTc population will include all subjects in the Safety population with 
measurements at baseline as well as on-treatment with at least 1 post-dose time point 
with a valid ΔQTcF value. The QT/QTc population will be used for the by-time point and 
categorical analyses of cardiodynamic ECG parameters.


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The PK/QTc population will include all subjects who are in both the QT/QTc and PK 
populations with at least 1 pair of post-dose PK and QTcF data from the same time point. 
The PK/QTc population will be used for the concentration-QTc analysis. PK/QTc 
population will be defined for balovaptan (and for moxifloxacin, when applicable).
6.7.2
Baseline
For the analysis of the ECG effect of balovaptan versus placebo, baseline is defined as 
the average of the measured ECG intervals from the 3 pre-dose time points on Day 1.
For the evaluation of assay sensitivity, the following baselines will be used:

Treatment C (Moxifloxacin / Placebo): For moxifloxacin dosed on Day 2, baseline 
is defined as the average of the measured QTcF intervals from the 3 pre-dose 
time points on Day 15. For placebo–correction in this group, placebo values will 
be derived from Day 14, and baseline is defined as the average of the measured 
QTcF intervals from the 3 pre-dose time points on Day 1.

Treatment D (Placebo / Moxifloxacin): For moxifloxacin dosed on Day 15, 
baseline is defined as the average of the measured QTcF intervals from the 
3 pre-dose time points on Day 2. For placebo–correction in this group, placebo 
values will be derived from Day 1, and baseline is defined as the average of the 
measured QTcF intervals from the 3 pre-dose time points on Day 14.
6.7.3
By-Time Point Analysis
The statistical hypothesis to be tested for the primary assessment of QT prolongation for 
each dose of the balovaptan treatment is:
: ∪	
() −	() ≥10,  = 1,2, … ,7
: ∩	
() −	() < 10,  = 1,2, … ,7
where 	
() and 	()are the least squares (LS) mean of ΔQTcF for each dose of 
balovaptan and placebo at time point  on Day 14, respectively. 
The “by-time point” analysis for QTcF will be based on a linear mixed-effects model with 
change-from-baseline QTcF (ΔQTcF) as the dependent variable; gender, period, 
sequence, time (categorical), treatment (balovaptan, and placebo), and time-by-
treatment interaction as fixed effects; and baseline QTcF as a covariate. An unstructured 
covariance matrix will be specified for the repeated measures at time points for subject 
within treatment period. If the model with an unstructured covariance matrix fails to 
converge, other covariance matrices such as compound symmetry and autoregressive 
will be considered. If the fixed effects for period and/or sequence should prove to be 
nonsignificant (i.e., if the p value > 0.1), these effects may be removed from the model 


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and the analysis will be repeated without those covariates. From this analysis, the LS 
mean and 2-sided 90 % CI will be calculated for the contrast “balovaptan versus placebo” 
at each dose of balovaptan and each post-dose time point on Day 1 and Day 14, 
separately. If the upper bound of the CI lies below 10 ms for the highest dose group of 
balovaptan at all seven post-dose time points (0.5, 1.25, 2.5, 4, 8, 12, and 24 hours) on 
Day 14 (steady state), balovaptan will be concluded not to have a significant effect on 
QT interval prolongation. 
6.7.4
Secondary Analysis
For HR, PR, and QRS intervals, the analysis will be based on the change-from-baseline 
post-dosing (ΔHR, ΔPR, and ΔQRS). The same model will be used as described for 
QTcF in the by-time point analysis. The LS mean, SE, and 90% CI from the statistical 
modeling for both change-from-baseline and placebo-corrected change-from-baseline 
values will be listed in tables and graphically displayed.
6.7.5
Assay Sensitivity 
In order to confirm that the assay is sufficiently sensitivity to detect an effect, the 
following statistical hypothesis is tested for values recorded at the three pre-defined 
post-dose time points (1.25, 2.5, and 4 hours):
: ∩	() −	() ≤5,  = 1,2,3
: ∪	() −	() > 5,  = 1,2,3
where 	() and 	() are the LS mean of ΔQTcF for moxifloxacin and placebo at time 
point , respectively.
The analysis to show assay sensitivity will be based on the change-from-baseline post-
dosing QTcF of moxifloxacin. The same model will be used as described for the primary 
analysis. For the 3 predefined time points (1.25, 2.5, and 4 hours post-dosing), the 
contrast in treatment ΔΔQTcF = “moxifloxacin - placebo” will be tested against the 1-
sided null hypothesis ΔΔQTcF ≤5 ms on the 5% level. Multiplicity will be controlled by 
using a Hochberg procedure.10 If after this procedure, the LS mean of ΔΔQTcF is 
significantly larger than 5 ms for at least 1 time point of these 3 time points, assay 
sensitivity will be considered shown. In addition, 2-sided 90% CIs will be obtained for the 
contrast at all time points and used in the figures.
6.7.6
Categorical Analyses
The analyses results for categorical outliers, T wave morphology, and U-wave presence 
will be summarized in frequency tables with counts and percentages for both number of 
subjects and number of time points. For categorical outliers, the number (percentage) of 
subjects as well as time points who had increases in absolute QTcF values > 450 and 


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≤480 ms, > 480 and ≤500 ms, or > 500 ms, and changes from pre-dose baseline of > 
30 and ≤60 ms, or > 60 ms; increase in PR from pre-dose baseline > 25% to a 
PR > 200 ms; increase in QRS from pre-dose baseline > 25% to a QRS > 120 ms; 
decrease in HR from pre-dose baseline > 25% to a HR < 50 bpm; and increase in HR 
from pre-dose baseline > 25% to a HR > 100 bpm will be determined. For T wave 
morphology and U-wave presence, the analyses will be focused on change from 
baseline (i.e., treatment-emergent changes).
6.7.7
Concentration-QTc Analysis (Secondary Analysis)
The concentration-QTc analysis will be based on placebo-corrected change-from-
baseline QTcF (ΔΔQTcF). That is, for the placebo adjustment, the individual ΔQTcF for 
placebo calculated at a specific time point is subtracted from ΔQTcF for the same 
subject on balovaptan at the same time point to generate ΔΔQTcF. The relationship 
between plasma concentrations of balovaptan and its metabolites (M2 [as appropriate]
and M3) and ΔΔQTcF will be investigated by linear mixed-effects modeling. 
A full model will include ΔΔQTcF as the dependent variable, time-matched plasma 
concentrations of each analyte (balovaptan, M2 [as appropriate] and M3) as the 
covariates, centered baseline QTcF as an additional covariate (i.e., baseline QTcF for 
individual subject at each post-dose time point subtracting the population mean baseline 
QTcF for all subjects), and subject as a random effect for both intercept and slopes, 
when applicable. Assay sensitivity may also be evaluated by a concentration-QTc 
analysis of the effect on ΔΔQTcF of moxifloxacin using a similar model as the above one.
The following model selection procedure among the full model and reduced models from 
possible first order combinations (without quadratic and interaction terms) among the 
three analytes (including models with only one analyte and with any two analytes) will be 
conducted by using Akaike Information Criterion (AIC) and the t-value for the intercept 
estimator.5 The selection will start from the full model and remove the analyte with the 
least significant slope from the model until only one analyte is left to evaluate the most 
appropriate model(s). Note that a significant intercept is not biologically plausible and 
therefore indicates model misfit. The selection of the final models will be performed 
among those models with an absolute t-value for the intercept estimate < 1.95. Nonlinear 
models (Emax model) will be included in this set only if the test for linearity indicates 
non-linearity. Among these models, the model with the smallest AIC will be selected as 
primary. If none of the models qualify according to the condition on the t-value for the 
intercept estimator, non-linear models will be considered.
If the primary model selected from model comparison includes two or more analytes, 
then all analytes will be used to predict QTcF effect. The geometric mean of the 
individual Cmax values of each analyte for subjects at each of balovaptan dose groups 
will be determined together with the geometric mean concentrations of the other two 
analytes at the Tmax of this analyte. The predicted effect and its 2-sided 90% CI for 


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ΔΔQTcF interval at these pairs of concentrations will be obtained for the models with two 
or three analytes.
The plot of the observed median-quantile balovaptan (M2 or M3) concentrations and 
associated mean ΔΔQTcF interval (90% CI) together with the regression line presenting 
the predicted ΔΔQTcF interval (90% CI) (Tornoe et al, 2011) will be used to evaluate the 
adequacy of the model fit of the primary model to the assumption of linearity and the 
impact on quantifying the concentration-QTcF relationship. For models with two or three 
analytes, the analyte not presented at the x-axis will be set to the arithmetic mean 
concentration across all subjects under this analyte at the Tmax of the analyte presented 
at the x-axis. For evaluation of the HR corrected QT interval, scatter and quantile plots 
of QTcF and HR intervals by treatment with regression line and linear mixed effects line 
(90% CI) will be also given, respectively. Additional exploratory analyses (via graphical 
displays and/or model fitting) will include accounting for a delayed effect (hysteresis) and 
the justification for the choice of PD model (linear versus nonlinear) as follows.
6.7.8
Investigation of Hysteresis
Hysteresis will be assessed based on joint graphical displays of the least squares (LS) 
mean difference between ΔQTcF under balovaptan and under placebo (ΔΔQTcF) for 
each post-dose timepoint and the mean concentrations of balovaptan, M2 and M3 at the 
same timepoints. In addition, hysteresis plots will be given for mean ΔΔQTcF and each 
of the mean concentrations of balovaptan, M2, and M3. If a QT effect (∆∆QTcF) > 10 ms
cannot be excluded from the by-timepoint analysis and if a delay between peak ∆∆QTcF 
and peak plasma concentration in any of these three plots (ΔΔQTcF versus balovaptan, 
ΔΔQTcF versus M2, and ΔΔQTcF versus M3) of more than 1 hour is observed, other 
concentration-QTc models, such as a model with an effect compartment, may be 
explored. With the provision stated above, hysteresis will be assumed if any of these 
three curves shows a counterclockwise loop.
Appropriateness of a linear model
To assess the appropriateness of a linear model, normal Q-Q plots for the standardized 
residuals and random effects, and plots of standard residuals versus concentration and 
versus fitted values and versus centered baseline QTcF will be produced. The scatter 
plots of standardized residuals versus concentration and versus centered baseline QTcF 
by LOESS fitting (i.e., locally weighted scatter plot smoothing as described by Cleveland 
1979) will also be produced with optimal smoothing parameters selected by the Akaike 
information criterion with a correction.12 In addition, a model with the original terms, and 
quadratic and interaction terms of the concentrations of balovaptan, M2 (as appropriate)
and M3 will be fitted, and the quadratic and interaction terms will be tested on the 
2-sided 5% level. If there is an indication that a linear model is inappropriate, additional 
models will be fitted, in particular an Emax model. The additional non-linear models 
(Emax) will be added to the candidate set for the primary model if the t-value for the 


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intercept estimate is < 1.95 (see above). The concentration-QTc analysis will then be 
repeated for the model found to best accommodate the nonlinearity detected.
6.8
TT-DOPPLER ECHOCARDIOGRAM
Results from TT-Doppler echocardiogram at each time point will be listed by subject for 
each time point. Descriptive summary statistics for each parameter by time point as well
as its change from baseline and change relative to placebo will be provided and 
illustrated by plots by individual subject and treatment cohorts. A more detail discussion 
of statistics for this parameter will be provided in the SAP to be completed prior to 
database lock.
Categorical analysis will refer N subjects per treatment with mPAP exceeding 25 mm Hg.
Additional exploratory analyses may be made post-hoc.
6.9
BLOOD PRESSURE AND HEART RATE MONITORING
Blood pressure and HR will be analyzed combined and separated for 2 time periods, 
daytime and nighttime. For this study, daytime is defined as 9:00 am to 9:00 pm and 
nighttime is defined as 1:00 am to 5:30 am. Summary statistics will be provided for 
systolic and diastolic blood pressure as well as for HR and compared between 
treatments.
Systolic and diastolic blood pressure will be summarized combined and separate across 
daytime and nighttime assessment periods as mean of all valid measurements with only 
implausible measurement results being considered invalid and removed from analysis. 
Categorical analyses for the mean blood pressure of respective period according to the 
following categories:
Table 5
Threshold for Hypertension Diagnosis Based on Ambulatory 
Blood Pressure Monitoring
Assessments Time Period
Ambulatory Blood Pressure
Daytime mean
≥ 130/85 mm Hg
Nighttime mean
≥ 120/70 mm Hg
In addition, categorical analyses for systolic and diastolic blood pressure as well for 
heart rate will be performed (details will be outlined in the SAP).


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The categorical analyses include a comparison of the number of outliers by treatment 
with outliers defined as: A value for a subject is considered to be an outlier at an on-
treatment time point if the mean daily systolic BP at the follow-up time point was 
≥180 mm Hg and it was at least a 20 mm Hg increase from the subject’s baseline mean 
daily systolic BP.  A value for a subject is considered to be an outlier at an on-treatment 
time point if the mean daily diastolic BP at the follow-up time point was ≥105 mm Hg 
and it was at least a 15 mm Hg increase from the subject's baseline mean daily diastolic 
BP.
Additional exploratory analyses may be made post-hoc.
7.
DATA COLLECTION AND MANAGEMENT
7.1
DATA QUALITY ASSURANCE
PRA will be responsible for data management of this study, including quality checking of 
the data.  Data entered manually will be collected through use of CRFs. In the event of 
discrepant data, the Sponsor will request data clarification from the sites.
PRA will produce a Data Quality Plan that describes the quality checking to be 
performed on the data. Central laboratory data will be sent directly to PRA using PRA’s 
standard procedures to handle and process the electronic transfer of these data. CRFs 
and correction documentation will be maintained in the EDC system’s audit trail.  System
backups for data stored by the Sponsor and records retention for the study data will be 
consistent with the Sponsor’s standard procedures.
The Sponsor will perform oversight of the data management of this study, including 
approval of the PRA’s data management plans and specifications. Data will be 
periodically transferred electronically from PRA to the Sponsor, and the Sponsor’s
standard procedures will be used to handle and process the electronic transfer of these 
data. 
Electronic CRFs and correction documentation will be maintained in the EDC system’s 
audit trail. System backups for data stored at the CRO and records retention for the 
study data will be consistent with the CRO’s standard procedures. eCRFs and 
correction documentation will be maintained in the EDC system’s audit trail.  System 
backups for data stored at the CRO and records retention for the study data will be 
consistent with the CRO’s standard procedures. 


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7.2
ELECTRONIC CASE REPORT FORMS 
eCRFs are to be completed through use of a PRA EDC system.  Sites will receive 
training and have access to a manual for appropriate eCRF completion.  eCRFs will be 
submitted electronically to the Sponsor and should be handled in accordance with 
instructions from the Sponsor.
All eCRFs should be completed by designated, trained site staff.  eCRFs should be 
reviewed and electronically signed and dated by the Investigator or a designee.
At the end of the study, the Investigator will receive subject data for his or her site in a 
readable format on a compact disc that must be kept with the study records.  
Acknowledgement of receipt of the compact disc is required.
7.3
SOURCE DATA DOCUMENTATION
Study monitors will perform ongoing source data verification and review to confirm that 
critical protocol data (i.e., source data) entered into the eCRFs by authorized site 
personnel are accurate, complete, and verifiable from source documents. 
Source documents (paper or electronic) are those in which subject data are recorded 
and documented for the first time.  They include, but are not limited to, hospital records, 
clinical and office charts, laboratory notes, memoranda, subject-reported outcomes, 
evaluation checklists, pharmacy dispensing records, recorded data from automated 
instruments, copies of transcriptions that are certified after verification as being accurate 
and complete, microfiche, photographic negatives, microfilm or magnetic media, X-rays, 
subject files, and records kept at pharmacies, laboratories, and medico-technical 
departments involved in a clinical trial.
Before study initiation, the types of source documents that are to be generated will be 
clearly defined in the Trial Monitoring Plan.  This includes any protocol data to be 
entered directly into the eCRFs (i.e., no prior written or electronic record of the data) and 
considered source data.
Source documents that are required to verify the validity and completeness of data 
entered into the eCRFs must not be obliterated or destroyed and must be retained per 
the policy for retention of records described in Section 7.5.
To facilitate source data verification and review, the Investigator (or designees) and 
institutions must provide the Sponsor direct access to applicable source documents and 
reports for trial-related monitoring, Sponsor audits, and IRB/EC review.  The study site 
must also allow inspection by applicable health authorities.
7.4
USE OF COMPUTERIZED SYSTEMS
When clinical observations are entered directly into a study site’s computerized medical 
record system (i.e., in lieu of original hardcopy records), the electronic record can serve 


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as the source document if the system has been validated in accordance with health 
authority requirements pertaining to computerized systems used in clinical research.  An 
acceptable computerized data collection system allows preservation of the original entry 
of data. If original data are modified, the system should maintain a viewable audit trail 
that shows the original data as well as the reason for the change, name of the person 
making the change, and date of the change.
7.5
RETENTION OF RECORDS
Records and documents pertaining to the conduct of this study and the distribution of 
IMP, including eCRFs, electronic or paper subject reported outcome (PRO) data 
(if applicable), Informed Consent Forms, laboratory test results, and medication 
inventory records, must be retained by the Principal Investigator for at least 15 years 
after completion or discontinuation of the study or for the length of time required by 
relevant national or local health authorities, whichever is longer.  After that period of time, 
the documents may be destroyed, subject to local regulations.  
No records may be disposed of without the written approval of the Sponsor.  Written 
notification should be provided to the Sponsor prior to transferring any records to 
another party or moving them to another location.
8.
ETHICAL CONSIDERATIONS
8.1
COMPLIANCE WITH LAWS AND REGULATIONS
This study will be conducted in full conformance with the ICH E6 guideline for Good 
Clinical Practice and the principles of the Declaration of Helsinki, or the laws and 
regulations of the country in which the research is conducted, whichever affords the 
greater protection to the individual.15  The study will comply with the requirements of the 
ICH E2A guideline (Clinical Safety Data Management:  Definitions and Standards for 
Expedited Reporting).  Studies conducted in the United States or under a U.S. 
Investigational New Drug (IND) Application will comply with U.S. FDA regulations and 
applicable local, state, and federal laws.16,17  Studies conducted in the European Union
or European Economic Area will comply with the E.U. Clinical Trial Directive 
(2001/20/EC).
8.2
INFORMED CONSENT
The Sponsor’s sample Informed Consent Form (and ancillary sample Informed Consent 
Forms such as a Child’s Informed Assent Form or Mobile Nursing Informed Consent 
Form, if applicable) will be provided to each site.  If applicable, it will be provided in a 
certified translation of the local language.  The Sponsor or its designee must review and 
approve any proposed deviations from the Sponsor's sample Informed Consent Forms 
or any alternate consent forms proposed by the site (collectively, the "Consent Forms") 
before IRB/EC submission.  The final IRB/ECapproved Consent Forms must be 
provided to the Sponsor for health authority submission purposes according to local 
requirements.


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If applicable, the Informed Consent Form will contain separate sections for any optional 
procedures.  The Investigator or authorized designee will explain to each subject the 
objectives, methods, and potential risks associated with each optional procedure.  
Subjects will be told that they are free to refuse to participate and may withdraw their 
consent at any time for any reason.  A separate, specific signature will be required to 
document a subject's agreement to participate in optional procedures.  Subjects who 
decline to participate will not provide a separate signature.
The Consent Forms must be signed and dated by the subject or the subject’s legally 
authorized representative before his or her participation in the study.  The case history or 
clinical records for each subject shall document the informed consent process and that 
written informed consent was obtained prior to participation in the study.
The Consent Forms should be revised whenever there are changes to study procedures 
or when new information becomes available that may affect the willingness of the 
subject to participate.  The final revised IRB/EC-approved Consent Forms must be 
provided to the Sponsor for health authority submission purposes.
Subjects must be re-consented to the most current version of the Consent Forms (or to a 
significant new information/findings addendum in accordance with applicable laws and 
IRB/EC policy) during their participation in the study.  For any updated or revised 
Consent Forms, the case history or clinical records for each subject shall document the 
informed consent process and that written informed consent was obtained using the 
updated/revised Consent Forms for continued participation in the study.
A copy of each signed Consent Form must be provided to the subject or the subject’s
legally authorized representative.  All signed and dated Consent Forms must remain in 
each subject’s study file or in the site file and must be available for verification by study 
monitors at any time.
8.3
INSTITUTIONAL REVIEW BOARD OR ETHICS COMMITTEE
This protocol, the Informed Consent Forms, any information to be given to the subject, 
and relevant supporting information must be submitted to the IRB/EC by the Principal 
Investigator and reviewed and approved by the IRB/EC before the study is initiated.  
In addition, any subject recruitment materials must be approved by the IRB/EC. 
The Principal Investigator is responsible for providing written summaries of the status of 
the study to the IRB/EC annually or more frequently in accordance with the requirements, 
policies, and procedures established by the IRB/EC.  Investigator or designees are also 
responsible for promptly informing the IRB/EC of any protocol amendments (see 
Section 9.6).
In addition to the requirements for reporting all adverse events to the Sponsor, 
Investigator or designees must comply with requirements for reporting serious adverse 


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events to the local health authority and IRB/EC.  Investigator or designees may receive 
written IND safety reports or other safety-related communications from the Sponsor.  
Investigator or designees are responsible for ensuring that such reports are reviewed 
and processed in accordance with health authority requirements and the policies and 
procedures established by their IRB/EC and archived in the site’s study file. 
8.4
CONFIDENTIALITY
The Sponsor maintains confidentiality standards by coding each subject enrolled in the 
study through assignment of a unique subject identification number.  This means that 
subject names are not included in data sets that are transmitted to any Sponsor location.
Subject medical information obtained by this study is confidential and may be disclosed 
to third parties only as permitted by the Informed Consent Form (or separate 
authorization for use and disclosure of personal health information) signed by the 
subject, unless permitted or required by law.
Medical information may be given to a subject's personal physician or other appropriate 
medical personnel responsible for the subject's welfare, for treatment purposes.
Given the complexity and exploratory nature of exploratory biomarker analyses, data 
derived from these analyses will generally not be provided to study investigators or 
subjects unless required by law.  The aggregate results of any conducted research will 
be available in accordance with the effective Sponsor policy on study data publication 
(see Section 9.5).
Data generated by this study must be available for inspection upon request by 
representatives of national and local health authorities, Sponsor monitors, 
representatives, and collaborators, and the IRB/EC for each study site, as appropriate.
Study data, which may include data on genomic variants, [applicable if the study is 
collecting mandatory (i.e., not requiring separate patient consent) samples for analysis of 
genomic variants]} may be submitted to government or other health research databases 
or shared with researchers, government agencies, companies, or other groups that are 
not participating in this study.  These data may be combined with or linked to other data 
and used for research purposes, to advance science and public health, or for analysis, 
development, and commercialization of products to treat and diagnose disease.  In 
addition, redacted Clinical Study Reports and other summary reports will be provided 
upon request (see Section 9.5).
8.5
FINANCIAL DISCLOSURE
Investigators will provide the Sponsor with sufficient, accurate financial information in 
accordance with local regulations to allow the Sponsor to submit complete and accurate 
financial certification or disclosure statements to the appropriate health authorities.  
Investigators are responsible for providing information on financial interests during the 
course of the study and for 1 year after completion of the study (see definition of end of 
study in Section 3.2).


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9.
STUDY DOCUMENTATION, MONITORING, AND 
ADMINISTRATION
9.1
STUDY DOCUMENTATION
The Investigator must maintain adequate and accurate records to enable the conduct of 
the study to be fully documented, including, but not limited to, the protocol, protocol 
amendments, Informed Consent Forms, and documentation of IRB/EC approval.  In 
addition, at the end of the study, the Investigator will receive the subject data, including
an audit trail containing a complete record of all changes to data.
9.2
PROTOCOL DEVIATIONS
The Investigator should document and explain any protocol deviations. The Investigator 
or designee should promptly report any deviations that might have an impact on subject
safety and data integrity to the Sponsor and to the IRB/EC in accordance with 
established IRB/EC policies and procedures.  The Sponsor will review all protocol 
deviations and assess whether any represent a serious breach of Good Clinical Practice 
guidelines and require reporting to health authorities.  As per the Sponsor's standard 
operating procedures, prospective requests to deviate from the protocol, including 
requests to waive protocol eligibility criteria, are not allowed.
9.3
SITE INSPECTIONS
Site visits will be conducted by the Sponsor or an authorized representative for 
inspection of study data, subjects' medical records, and eCRFs.  The Investigator or 
designee will permit national and local health authorities; Sponsor monitors, 
representatives, and collaborators; and the IRBs/ECs to inspect facilities and records 
relevant to this study.
9.4
ADMINISTRATIVE STRUCTURE
This trial will be sponsored and managed by F. Hoffmann-La Roche Ltd.  The Sponsor 
will provide clinical operations management, data management, and medical monitoring.
Central facilities will be used for certain study assessments throughout the study (e.g., 
specified laboratory tests, biomarker and PK analyses), as specified in Section 4.5.  
Accredited local laboratories will be used for routine monitoring; local laboratory ranges 
will be collected.


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9.5
PUBLICATION OF DATA AND PROTECTION OF TRADE 
SECRETS
Regardless of the outcome of a trial, the Sponsor is dedicated to openly providing 
information on the trial to healthcare professionals and to the public, both at scientific 
congresses and in peer-reviewed journals. The Sponsor will comply with all 
requirements for publication of study results.  For more information, refer to the Roche 
Global Policy on Sharing of Clinical Trials Data at the following Web site:
www.roche.com/roche_global_policy_on_sharing_of_clinical_study_information.pdf
The results of this study may be published or presented at scientific congresses.  For all 
clinical trials in subjects involving an IMP for which a marketing authorization application 
has been filed or approved in any country, the Sponsor aims to submit a journal 
manuscript reporting primary clinical trial results within 6 months after the availability of 
the respective Clinical Study Report.  In addition, for all clinical trials in subjects involving 
an IMP for which a marketing authorization application has been filed or approved in any 
country, the Sponsor aims to publish results from analyses of additional endpoints and 
exploratory data that are clinically meaningful and statistically sound.
The Investigator must agree to submit all manuscripts or abstracts to the Sponsor prior 
to submission for publication or presentation.  This allows the Sponsor to protect 
proprietary information and to provide comments based on information from other 
studies that may not yet be available to the Investigator.
In accordance with standard editorial and ethical practice, the Sponsor will generally 
support publication of multicenter trials only in their entirety and not as individual center 
data.  In this case, a coordinating investigator will be designated by mutual agreement.
Authorship will be determined by mutual agreement and in line with International 
Committee of Medical Journal Editors authorship requirements.  Any formal publication 
of the study in which contribution of Sponsor personnel exceeded that of conventional 
monitoring will be considered as a joint publication by the Investigator and the 
appropriate Sponsor personnel.
Any inventions and resulting patents, improvements, and/or know-how originating from 
the use of data from this study will become and remain the exclusive and unburdened 
property of the Sponsor, except where agreed otherwise.
9.6
PROTOCOL AMENDMENTS
Any protocol amendments will be prepared by the Sponsor.  Protocol amendments will 
be submitted to the IRB/EC and to regulatory authorities in accordance with local 
regulatory requirements.
Approval must be obtained from the IRB/EC and regulatory authorities (as locally 
required) before implementation of any changes, except for changes necessary to 
eliminate an immediate hazard to subjects or changes that involve logistical or 
administrative aspects only (e.g., change in Medical Monitor or contact information).


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10.
REFERENCES
1. Shin JY, Roughead EE, Park BJ, et al. Cardiovascular safety of methylphenidate 
among children and young people with attention-deficit/hyperactivity disorder 
(ADHD):nationwide self-controlled case series study. BMJ. 2016;353:i2550.
2. Sherazi S, DiSalle M, Daubert JP, Shah AH. Moxifloxacin-induced torsades de 
pointes. Cardiol J. 2008;15(1):71-3
3. US Food and Drug Administration. Guidance for industry: E14 clinical evaluation 
of QT/QTc interval prolongation and proarrhythmic potential for non-
antiarrhythmic drugs. Food and Drug Administration. 2005.
4. Darpo B., Benson C, Dota C, et al. Results from the IQ-CSRC prospective study 
support replacement of the thorough QT study by QT assessment in the early 
clinical phase. Clin. Pharmacol Ther. 2015; 97(4): 326-35.
5. Ferber G, Zhou M, Darpo B. Detection of QTc effects in small studies--
implications for replacing the thorough QT study. Annals of Noninvasive 
Electrocardio. 2015;20(4):368-77.
6. Zhang J. Machado G. Statistical issues including design and sample size 
calculation in thorough QT/QTc Studies. J Biopharm Stat. 2008;18:(3): 451—67.
7. Sherazi S, DiSalle M, Daubert JP, Shah AH. Moxifloxacin-induced torsades de
pointes. Cardiol J. 2008;15(1):71-3.
8. Eichinger, MR, Walker BR. "Enhanced pulmonary arterial dilation to arginine 
vasopressin in chronically hypoxic rats." Am J Physiol. 1994;267(6 Pt 2)
H2413-19.
9. Walker BR, Haynes Jr JO, Wang HL, Voelkel NF. Vasopressin-induced 
pulmonary vasodilation in rats. Am J of Physiol. 1989; 257(2):H415-22.
10. Hochberg Y. A sharper Bonferroni procedure for multiple tests of significance. 
Biometrika. 1988; 75, 800–2.
11. Cleveland WS. Robust Locally Weighted Regression and Smoothing Scatter 
Plots. J Amer Statist Assoc. 1979;74(368):829-36.
12. Hurvich CM, Simonoff JS, Tsai CL. Smoothing parameter selection in 
nonparametric regression using an improved Akaike information criterion.  J Roy 
Statist Soc Ser B. 1998;60(2):271-93.


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13. Tornøe CW, Garnett CE, Wang Y, Florian J, Li M, Gobburu JV.  Creation of a 
knowledge management system for QT analyses. J Clin Pharmacol. 2011; 51 (7): 
1035-42.
14. Al-Mosalem OA, El-Ansary A, Attas O, Al-Ayadhi L. Metabolic biomarkers related 
to energy metabolism in Saudi autistic children. Clinical biochemistry. 2009 Jul 
1;42(10-11):949-57.
15. World Medical Association. World Medical Association Declaration of Helsinki: 
ethical principles for medical research involving human subjects. Jama. 2013 
Nov 27;310(20):2191.
16. International Conference on Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, E3: Structure and Content of Clinical Study Reports 
(CPMP/ICH/137/95), November 1995
17. International Conference on Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite
Guideline, E2A: Clinical Safety Data Management: Definitions and Standards for 
Expedited Reporting, Note for Guidance on Clinical Safety Data Management, 
June 1995


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Appendix 1
Schedule of Activities
Assessments
Screening
Nominal Study Day for Each Treatment Period
Follow
-upa
Days 
-28 to -3
Day 
-2
Day 
-1
Day 
1
Day 
2
Day 
3
Day 
4
Day 
5
Day 
6
Day 
7
Day 
8
Day 
9
Day 
10
Day 
11
Day 
12
Day 
13
Day 
14
Day 
15
Day 
16
Written informed 
consent

Inclusion and exclusion 
criteria

Randomization

Medical history

Conmeds review


Physical examination



Height and weightb


Pre and post-dose 
fastingc




Standardized low salt 
mealsd







Vital signse












TT-Doppler 
echocardiogram


ABPM


Laboratory safety testsf






12-lead safety ECGg












12-lead ECG Holter 
monitoringh




Administration of 
investigational 
medicinal product















Drugs of abuse screeni


Orthostatic challenge 
testing


Urine alcohol screenj


Pregnancy testk


Follicle stimulating 
hormonel



Appendix 1
Schedule of Activities (cont.)
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a.
Follow-up visit should occur between 14 and 21 days after the final dose of study drug
b.
Height and weight to calculate BMI will be collected at Screening. During Follow-up visit only weight will be recorded
c.
Subjects will fast for a minimum of 8 hours pre-dose and 4 hours post-dose
d.
Subjects will be fed standardized low salt meals
e.
Assessment of supine blood pressure, pulse rate and body temperature. All measurements to be recorded after the subject has remained in a supine position for at least 10 minutes. 
(See Appendix 2 for details for vital signs taken on days with study drug treatment will be administered [Days 1, 3, 6, 9,11,13,14, and 15])
f.
Includes hematology, biochemistry, urinalysis, coagulation and serology (coagulation and serology will be performed only during Screening)
g.
Safety ECG assessments: Triplicate ECG measurements at Screening and Day -2. Single ECG measurements at Day 1, Day 3, Day 6, Day 9, Day 11, Day 13, Day 14, Day 15, Day 
16 and follow-up visit.  All measurements to be recorded after the subject has remained in a supine position for at least 10 minutes
h.
ECG Holter monitoring assessments will be recorded on study Days 1, 2, 14 and 15 (See Appendix 2 for additional details). 
All ECG measurements are to be recorded while the subject remains supine for at least 15 minutes.
ECGs will be extracted from Holters at 3 time points pre-dose (-45, -30 and -15 minutes) and 7 time points post-dose (0.5, 1.0, 2.5, 4, 8, 12 and 24 h post-dose). The 24-hour post-
dose extraction time point for Days 1 and 14 can be taken from the 3 pre-dose time points on Days 2 and 15.
12-lead ECGs will be extracted from the continuous recording in up to 10 replicates from a 5-minute window during a 15-minute supine period pre-dose and at the nominal Holter 
timepoints post-dose. Whenever ECG extraction and PK sample timepoints are close together the ECG extraction will be performed prior to PK sample collection.
i.
Drugs of abuse: amphetamines, barbiturates, benzodiazepines cannabinoids, cocaine, opiates, methamphetamines, methadone and 3,4 Methylenedioxymethamphetamine (MDMA), 
will be measured in urine at Screening and Day 2. Additional testing may be performed during the study, at the discretion of the Investigator or designee, to confirm subject 
compliance.
j.
Planned at Screening and Day -2, further testing may be performed during the study, at the discretion of the Investigator or designee, to confirm subject compliance.
k.
Serum pregnancy test
l.
Assessed for all females.
m. See Appendix 2 for PK sampling details
n.
Blood samples for the assessment of early markers of cardiac damage: troponin I, cTnT and NT-proBNP, will be taken and stored but not analyzed unless requested for a specific 
subject.
o.
Optional blood sample for Clinical Genotyping can be collected any time during the study. N.B. one sample only to be collected and only if consented for this sample
PRIORITY OF ASSESSMENTS: where appropriate and feas ble: ECG > Vital Signs > Blood Sampling > Lab Safety Tests
Blood sampling for 
balovaptan PKm





Blood sampling for 
moxifloxacin PKm


Cardiac biomarkersn


C-SSRS




Adverse Events



















Blood sampling for 
Clinical Genotypingo


Balovaptan —F. Hoffmann-La Roche Ltd
97/Protocol WP40734, Version 3.0 
Appendix 2
Schedule of Pharmacokinetic, Vital Signs, and ECG
Assessments
Hours post-dose
Pre-dose
0.5
1.0
2.5
3
4
6
8
12
24a
12-lead ECG Holter 
monitoringb








Vital Signs



Blood collection for 
pharmacokineticsc









a.On days when consecutive 24 hour and pre-dose assessments are being done, only one 
assessment needs to be done prior to dose. This includes Holter extractions where the 24-hour
post-dose extraction time point for Days 1 and 14 can be taken from the 3 pre-dose time points 
on Days 2 and 15.
b.Predetermined 24 h Holter ECG time points on Day 1, Day 2, Day 14 and Day 15.  Holter ECG 
assessments will be performed in the 15 minutes prior to the nominal Holter timepoint but prior 
to the PK sample, if applicable. On Day 2 and 15 of each Treatment Period subjects are 
scheduled to have 24 h post-dose ECG Holter reading, PK sampling and daily dosing of study 
drug. These activities will be performed in the following order the 24 h post-dose Holter readings
will be performed first followed by drawing of the 24 h post-dose PK sample and then dosing of 
study drug. Due to this study drug dosing on these days may be delayed relative to dosing on 
other study days.
c. Blood collection for PK assessments will be identical for Treatment Arms in all Treatment
Periods as this is a blinded study. Intensive PK shown above, in Appendix 2, will take place on 
study Days 1, and 14 for balovaptan and Days 2 and 15 for moxifloxacin. On other study days 
with PK sampling and study drug dosing (Days 11, 12, 13) a single PK sample will be drawn 
prior to dosing with study drugs. This pre-dose sample also serves as the 24 h post-dose 
sample for the previous days dosing. A balovaptan PK sample will be taken on Day 2 and Day 
15 for the Day 1 and Day 14, 24 h post-dose sample, respectively. A moxifloxacin PK sample 
will be taken on Day 16 for the Day 15, 24 h post-dose sample.

SAP_001.pdf: 
Official Title: 
A Single-Center, Multiple-Dose, Randomized, Double-Blind, Placebo-
Controlled, Positive-Controlled, Crossover Study to Investigate the 
Effect of Balovapton on the Qtc Interval in Healthy Subjects 
NCT Number: 
NCT03808298 
Document Date: 
SAP Version 1: 07-May-2019 




Statistical Analysis Plan 
RPU029UF-180292 
Protocol: WP40734 
Version Date: 02-May-2019 
EDSREP 009 T 01 G 
Page 2 of 26 
2.0 Table of Contents 
1.0 Approvals ................................................................................................................................................ 1 
2.0 Table of Contents .................................................................................................................................... 2 
3.0 Introduction ............................................................................................................................................. 4 
4.0 Changes from Previous Version of Approved SAP ................................................................................ 4 
5.0 Study Objectives ..................................................................................................................................... 4 
5.1 Primary ................................................................................................................................................ 4 
5.2 Primary Endpoints ............................................................................................................................... 4 
5.3 Secondary ........................................................................................................................................... 4 
5.3.1 Secondary Endpoints ................................................................................................................... 5 
5.3.2 Exploratory Objectives ................................................................................................................. 5 
5.3.3 Exploratory Endpoints .................................................................................................................. 5 
6.0 Study Design ........................................................................................................................................... 5 
Figure 1: Study Schema ................................................................................................................................ 7 
6.1 Sample Size Considerations ............................................................................................................... 7 
6.2 Randomization .................................................................................................................................... 7 
7.0 Overview of Planned Analysis ................................................................................................................ 8 
7.1 Changes from Protocol ....................................................................................................................... 8 
7.2 Interim Analysis and Key Results ....................................................................................................... 8 
7.3 Final Analysis ...................................................................................................................................... 8 
8.0 Data Review ............................................................................................................................................ 8 
8.1 Data Management ............................................................................................................................... 8 
8.2 Acceptance of Data for Summarization .............................................................................................. 8 
9.0 Definitions and General Analysis Methods ............................................................................................. 8 
9.1 Analysis Data Presentation ................................................................................................................. 8 
9.1.1 Rounding ...................................................................................................................................... 8 
9.1.2 Imputation .................................................................................................................................... 9 
9.1.3 Daylight Savings Time Adjustments ............................................................................................ 9 
9.1.4 Descriptive Statistics .................................................................................................................... 9 
9.1.5 Pooling ......................................................................................................................................... 9 
9.1.6 Unscheduled Measurements ....................................................................................................... 9 
9.2 Analysis Data Definitions .................................................................................................................... 9 
9.2.1 Baseline Definition ....................................................................................................................... 9 
9.2.2 Treatment/Subject Grouping ...................................................................................................... 10 
9.2.3 Common Variable Derivations ................................................................................................... 10 
9.2.4 QC .............................................................................................................................................. 10 
9.2.5 ADaM Datasets and Metadata ................................................................................................... 10 
9.3 Software ............................................................................................................................................ 10 
9.4 Statistical Methods ............................................................................................................................ 11 
9.4.1 Statistical Outlier Determination ................................................................................................. 11 
9.4.2 Predetermined Covariates and Prognostic Factors ................................................................... 11 
9.4.3 Hypothesis Testing ..................................................................................................................... 11 
9.5 TFL Layout ........................................................................................................................................ 11 
10.0 Analysis Sets ....................................................................................................................................... 11 
10.1 Safety Set ........................................................................................................................................ 11 
10.2 Pharmacokinetic Set ....................................................................................................................... 11 
11.0 Subject Disposition .............................................................................................................................. 11 
12.0 Protocol Deviations ............................................................................................................................. 11 
13.0 Demographic and Baseline Characteristics ........................................................................................ 12 
13.1 Demographics ................................................................................................................................. 12 
13.2 Medical History ................................................................................................................................ 12 
14.0 Concomitant Medications .................................................................................................................... 12 
15.0 Treatment Compliance and Exposure ................................................................................................ 12 
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Page 3 of 26 
16.0 Pharmacokinetic Analyses .................................................................................................................. 12 
16.1 Pharmacokinetic Variables.............................................................................................................. 12 
16.2 Plasma Pharmacokinetic Summaries ............................................................................................. 12 
16.2.1 Plasma Concentrations ............................................................................................................ 12 
16.2.2 Plasma Pharmacokinetic Parameters ...................................................................................... 13 
17.0 Safety Analyses .................................................................................................................................. 14 
17.1 Safety Variables .............................................................................................................................. 14 
17.1.1 Adverse Events ........................................................................................................................ 15 
17.1.2 Serious Adverse Events ........................................................................................................... 16 
17.1.3 Laboratory Data ....................................................................................................................... 16 
17.1.4 Vital Signs ................................................................................................................................ 16 
17.1.5 12-Lead Safety Electrocardiograms ......................................................................................... 16 
17.1.6 Echocardiograms ..................................................................................................................... 16 
17.1.7 Other Observations Related to Safety ..................................................................................... 17 
18.0 References .......................................................................................................................................... 17 
Appendix 1: Glossary of Abbreviations ....................................................................................................... 18 
Appendix 2: Protocol Schedule of Assessments ........................................................................................ 20 
Appendix 3: Schedule of Pharmacokinetics, Vital Signs, and ECG ........................................................... 22 
Appendix 4: List of End of Text Outputs ..................................................................................................... 23 
Appendix 5: Shells for Post-Text Tables, Figures and Listings .................................................................. 26 
19.0 Document History ................................................................................................................................ 26 
 
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Statistical Analysis Plan 
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Protocol: WP40734 
Version Date: 02-May-2019 
EDSREP 009 T 01 G 
Page 4 of 26 
3.0 Introduction 
This Statistical Analysis Plan (SAP) describes the statistical methods that will be used during the analysis 
and reporting of data collected under F. Hoffman-La Roche Ltd. Protocol WP40734. 
This SAP should be read in conjunction with the study protocol and electronic case report form (eCRF). 
This version of the plan has been developed using the protocol dated 23-Apr-2019 (including all 
amendments up to this protocol date) and the final eCRF(s) dated 27-Feb-2019. 
An approved and signed SAP is a requirement for database lock. An approved SAP is also required for 
unblinding of the study treatments. 
This SAP defines the data and analyses that will be processed by the PRA Early Development Services 
(EDS) Biostatistics Department.  
PRA EDS will perform the pharmacokinetic (PK), safety and tolerability evaluation.  
This SAP supersedes the statistical considerations identified in the protocol; where considerations are 
substantially different, they will be so identified. Any post-hoc or unplanned analyses, or significant 
changes from the planned analysis in this SAP performed to provide results for inclusion in the clinical 
study report (CSR) but not included in this SAP, will be clearly identified in the CSR. Changes to planned 
analyses do not require an updated SAP but should be included in the CSR if significant. 
4.0 Changes from Previous Version of Approved SAP 
This is the first version of the SAP. 
5.0 Study Objectives 
This SAP will cover secondary objectives/endpoints 4, 5, and 9 and exploratory objectives/endpoints 1 
and 2. All other objectives and endpoints are noted within this section of this SAP for completeness; 
however  an additional SAP authored by eResearch Technologies (ERT) will define the Cardiodynamic 
ECG analyses in detail . 
5.1 Primary 
1. To investigate the effect of balovaptan 50 mg qd on the QT interval corrected through use of 
Fridericia’s formula (QTcF) after 14 days of dosing 
5.2 Primary Endpoints 
1. Placebo-corrected, change-from-baseline QTcF (∆∆QTcF) measured on 12-lead 
electrocardiograms (ECGs) extracted from continuous recordings at the specified time points on 
Day 14 
5.3 Secondary 
1. To investigate the effect of a single dose of balovaptan 50 mg on the QTcF interval at Day 1. 
2. To investigate the effect of balovaptan 10 mg qd on the QTcF interval at Day 1 and Day 14. 
3. To investigate the effect of balovaptan on the following ECG parameters: PR, RR, QRS, QT, 
T-wave and U-wave morphology. 
4. To investigate the pharmacokinetics of balovaptan, M2 (as appropriate) and M3. 
5. To investigate the pharmacokinetics of moxifloxacin. 
6. To investigate the relationship between plasma concentrations of balovaptan, M2 (as 
appropriate), M3 and QTcF interval length. 
7. To investigate the relationship between plasma concentrations of moxifloxacin and QTcF interval 
length (if warranted). 
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8. To investigate the ECG assay sensitivity to detect a change in the QTcF interval, using 
moxifloxacin 400 mg as an active control. 
9. To investigate the safety and tolerability of balovaptan and moxifloxacin at the dosing regimens 
scheduled. 
5.3.1 Secondary Endpoints 
1. ∆∆QTcF measured on 12-lead ECGs extracted from continuous recordings at the specified time 
points on Day 1. 
2. ∆∆QTcF measured on 12-lead ECGs extracted from continuous recordings at the specified time 
points on Day 1 and Day 14. 
3. Placebo-corrected, change-from-baseline heart rate, PR and QRS interval (∆∆HR, ∆∆PR and 
∆∆QRS) measured on 12-lead ECGs extracted from continuous recordings at the specified time 
points on Day 1 and Day 14. 
a. Categorical outliers for QTcF, HR, PR, and QRS. 
b. Frequency of treatment emergent changes of T-wave morphology and U-wave presence. 
4. Pharmacokinetic parameters estimation such as time to maximum concentration (Tmax), mean 
maximum concentration (Cmax), and area under the plasma concentration curve at 0-24 hours 
(AUC0-24h) for plasma concentrations of balovaptan, M2 (as appropriate) and M3.  Other 
pharmacokinetic parameters may also be added. 
5. Pharmacokinetic parameters estimation such as Tmax, Cmax, and AUC0-24h for plasma 
concentrations of moxifloxacin.  Other pharmacokinetic parameters may also be added. 
6. Pharmacokinetic-pharmacodynamic model of balovaptan, M2 (as appropriate) and M3 
concentrations vs.  QTcF changes. 
7. Pharmacokinetic-pharmacodynamic model of moxifloxacin concentrations vs.  QTcF changes (if 
warranted) 
8. Placebo-corrected ∆∆QTcF measured on 12 lead ECGs extracted from continuous recordings at 
the specified time points on Day 2 and Day 15 
9. Safety assessments: adverse events (AEs), physical examination, vital signs, laboratory tests, 
C-SSRS, and heart rate (HR) monitoring. 
5.3.2 Exploratory Objectives 
1. To investigate any effect of balovaptan at 50 mg once per day (QD) and 10 mg QD doses on 
pulmonary blood pressure as assessed by the use of TT-doppler echocardiography. 
2. To investigate any effect of 50 mg qd and 10 mg qd on right and left heart as assessed by TT-
Doppler echo. 
3. To investigate the effect 10 mg and 50 mg QD balovaptan on blood pressure and heart rate as 
measured by the use of Holter-ECG and ambulatory blood pressure monitoring (ABPM). 
5.3.3 Exploratory Endpoints 
1. TT-Doppler echocardiography: pulmonary artery acceleration time (PAAT), right ventricular 
outflow tract - velocity time integral (RVOT-VTI) and derived mean pulmonary arterial pressure as 
change to baseline relative to placebo treatment. 
2. TT-Doppler-echocardiography: left ventricular ejection fraction (LVEF) and additional secondary 
TT-Doppler-echocardiography parameters of left and right heart. 
3. ABPM: mean diastolic and systolic blood pressure (daytime and nighttime mean). 
a. Heart rate will be analyzed as derived from Holter ECG. 
i. Heart rate as derived from ABPM and/or Holter ECG may be explored against 
fluctuations in blood pressure measured by ABPM (if warranted). 
6.0 Study Design 
This is a single-center, multiple-dose, randomized, double-blind, placebo-controlled, positive-controlled, 
twelve sequence 3-period cross-over study. 
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At the start of treatment period 1, at the end of Day -1, subjects will be randomized to one of the 12 
treatment sequences comprised of 3 of the 4 treatments described in 6.2. All subjects will receive 
treatments A and B, half will receive treatment C, and the other half will receive treatment D. Each 
treatment period consists of 15 dosing days. Following each treatment period, subjects will undergo a 
washout period lasting a minimum of 13 and a maximum of 20 days from the last dose of study drug. 
Treatment A (balovaptan therapeutic dose) 
Days 1-14: A single once daily oral dose of 10 mg (1 × 10 mg tablet) of balovaptan and 4 tablets 
of matching placebo for balovaptan for 14 days. 
Day 2 and 15: A single oral dose of a matching placebo capsule for moxifloxacin. 
Treatment B (balovaptan supra-therapeutic dose) 
Days 1-14: A single once daily oral dose of 50 mg (5 x 10 mg tablets) of balovaptan for 14 days. 
Day 2 and 15: A single oral dose of a matching placebo capsule for moxifloxacin. 
Treatment C (active control [400 mg moxifloxacin] on Day 2) 
Days 1-14: A single once daily oral dose of 5 tablets of matching placebo for balovaptan for 14 
days. 
Day 2: A single oral dose of 400 mg moxifloxacin capsule 
Day 15: A single oral dose of a matching placebo capsule for moxifloxacin 
Treatment D (active control [400 mg moxifloxacin] on Day 15) 
Days 1-14: A single once daily oral dose of 5 tablets of matching placebo for balovaptan for 14 
days. 
Day 2: A single oral dose of a matching placebo capsule for moxifloxacin 
Day 15: A single oral dose of 400 mg moxifloxacin capsule. 
Figure 1 presents an overview of the study design. 
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Figure 1: Study Schema 
 
6.1 Sample Size Considerations 
A sample size of 56 subjects was chosen to obtain 48 evaluable subjects. Based on the calculation of the 
sample size for a TQT study (Zhang and Machado, 2008), assuming a 1-sided 5% significance level and 
a within-subject SD of 8 ms for change from baseline QTcF (ΔQTcF) and a true mean difference of 3 ms 
in ΔQTcF between balovaptan and placebo, a sample size of 48 evaluable subjects who complete the 
study will provide a power of 96.6% to demonstrate that the upper bound of all the 2-sided 90% CIs on 
ΔΔQTcF will fall below 10 ms for up to 7 post-dose time points (0.5, 1.25, 2.5, 4, 8, 12 and 24 hours). 
6.2 Randomization 
Subjects who meet all eligibility criteria will be randomized to 1 of the following 12 sequences using a 
computer-generated randomization scheme produced by a PRA statistician prior to the start of the study.  
A, B, C 
A, C, B 
B, A, C 
B, C, A 
C, A, B 
C, B, A 
A, B, D 
A, D, B 
B, A, D 
B, D, A 
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D, A, B 
D, B, A 
Randomization numbers will range from 
 to 
. Replacement subjects will receive the number of 
the subject to be replaced increased by 1000 (eg, replacement number 
 for subject 
), and will 
be administered the same treatment sequence. The randomization schedule will alternate between 
sequences containing treatment C and sequences containing treatment D to ensure a balance between 
the number of subjects who receive treatment C and D. 
7.0 Overview of Planned Analysis 
7.1 Changes from Protocol 
There are no changes from the protocol. 
7.2 Interim Analysis and Key Results 
There will be no interim analyses or summaries of data provided prior to the delivery of the full set of post-
lock tables, figures and listings (TFLs). 
7.3 Final Analysis 
Draft TFLs will be provided after database lock. After Sponsor comments have been incorporated, the 
TFLs will be finalized and incorporated in the first draft CSR. 
8.0 Data Review 
8.1 Data Management 
Data handling and transfer will take place under the PRA Data Management Plan for the study. 
8.2 Acceptance of Data for Summarization 
Programming of analysis datasets and TFLs may be ongoing during the data management of the study. 
However, programming of analysis datasets and TFLs will be completed and quality controlled (QC’d) 
after database lock. Only quality assured (QA’d) results released by the Safety Laboratory, Bioanalytical 
Laboratory, or other external data source will be used for the programming of analysis datasets and TFLs 
for the final report. Any data values requiring investigation or corrections that are identified while 
programming the analysis datasets and TFLs will be sent to the project Data Manager. If the issue affects 
the TFLs the Programmer or Statistician who identified the issue will follow it to resolution. 
9.0 Definitions and General Analysis Methods 
9.1 Analysis Data Presentation 
9.1.1 Rounding 
In listings, data will be presented with the same precision as the original data. Derived data will be 
rounded for presentation purposes.  
For all summaries, the mean and median will be presented to one decimal place greater than the data, 
standard deviation (SD) to two greater than the data, and the minimum (min) and maximum (max) will be 
presented to the same number of decimal places as the data. Percentages will be presented with one 
decimal. 
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The above rule can be applied directly to collected data. For derived data rounding will occur prior to 
summarization so a specific number of decimal places will have to be assumed to apply the above 
rounding rules for summary statistics. 
PK parameters will be rounded in the derived dataset as determined by the pharmacokineticist. Each 
parameter will have a fixed number of decimals. The pharmacokineticist will follow this following rule 
when deciding the number of decimals for each parameter: no decimal if data for a parameter are 
generally greater than or equal to 100; 1 decimal if data are generally between 10 and 100; 2 decimals if 
data are generally between 1 and 10; 3 decimals if data are generally below 1. 
The Tmax will be reported with 2 decimals. 
Additional derived data will be rounded in the derived dataset as determined by the statistician. 
9.1.2 Imputation 
Unless otherwise noted, data will not be imputed. 
9.1.3 Daylight Savings Time Adjustments 
On March 10, 2019 at 2:00 am the clocks change to 3:00 am for Daylight Savings Time. All clinic 
procedures for the remainder of the treatment period will be moved forward by one hour after daylight 
savings time occurs. All duration calculations for times post-daylight savings time that will be relative to a 
time prior to daylight savings will need to be programmatically adjusted for the hour that was lost on the 
morning of March 10. 
9.1.4 Descriptive Statistics 
Unless otherwise indicated, continuous variables will be summarized with the following descriptive 
statistics: n (number of observations), (arithmetic) mean, SD, min, median, and max. 
Categorical data will be summarized with frequencies and percentages. Percentages by categories will be 
based on the number of subjects exposed within a treatment. 
For categorical data the categories will be presented in the tables exactly as they appear in the CRF.  
9.1.5 Pooling 
Summary statistics will be calculated by treatment (and timepoint, if applicable).  
9.1.6 Unscheduled Measurements 
Unscheduled and early termination measurements will be included in the listings. With the exception of 
unscheduled measurements used for baseline, unscheduled measurements will be excluded from the 
descriptive statistics and statistical analysis. 
9.2 Analysis Data Definitions 
9.2.1 Baseline Definition 
Unless otherwise stated, baseline for post-dose evaluations within each period is defined as the last 
observation recorded before the first study drug administration in each period. The last observation can 
be an unscheduled / repeated measurement. If a pre-treatment observation is missing in a given period 
then the screening value may be used. 
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13.0 Demographic and Baseline Characteristics 
13.1 Demographics 
Subject demographics will be summarized by analysis set. The summary will include the subjects’ age 
(years), sex, race, ethnicity, weight (kg), height (cm), and BMI (kg/m2).  
All demographic data as collected during the screening visit will be listed by subject. 
13.2 Medical History 
Medical history, categorized by preferred term according to MedDRA, will be listed by subject. 
14.0 Concomitant Medications 
Concomitant medications, categorized by medication group and subgroup according to WHO Drug 
Dictionary, will be listed by subject. Medications with an end date prior to the first dose of study drug will 
be considered prior medications and will be noted in the listing. If a partial date allows a medication to be 
considered concomitant it will be categorized as such. 
15.0 Treatment Compliance and Exposure 
The number of subjects receiving each dose of study drug will be summarized by visit. 
Exposure data will be listed by subject. 
16.0 Pharmacokinetic Analyses  
16.1 Pharmacokinetic Variables 
Concentrations of balovaptan, M2, M3, and moxifloxacin will be collected in plasma.  
PK parameters of balovaptan, M2 (as appropriate), M3, and moxifloxacin will be calculated for plasma. 
16.2 Plasma Pharmacokinetic Summaries 
16.2.1 Plasma Concentrations 
Plasma concentrations for balovaptan, M2 (as appropriate), M3, and moxifloxacin below the quantifiable 
limit (BQL) will be set to 0 in the computation of mean concentration values. Descriptive statistics (n, 
arithmetic mean, geometric mean, SD, coefficient of variation, median, min, and max) will be used to 
summarize the plasma concentrations by treatment and visit for balovaptan and by treatment for 
moxifloxacin at each scheduled timepoint. If over ½ the subjects in a given cell have values BQL then the 
descriptive statistics will not be presented and will instead display as BQL for the mean and min. With the 
exception of min and max, all other statistics will be missing. 
Linear and semi-logarithmic plots of the arithmetic mean plasma concentration by scheduled sampling 
time will be provided by treatment for balovaptan on day 1 and day 14 and by treatment for moxifloxacin 
on day 2 for treatment C and day 15 for treatment D. These plots will show time in hours. The plots will 
match the summary table results and will not have an observation at a given timepoint if more than half of 
the subjects have values BQL. 
Linear and semi-logarithmic plots of the individual plasma concentration by actual sampling time will be 
provided by subject (one subject per page). These plots will show time in hours. Individual plots will use 
the BQL handling procedure described below for “Plasma Pharmacokinetic Parameters”. 
All individual subject plasma concentration data will be listed by subject. 
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will not be calculated. 
 
Ctrough 
Predose concentration prior to 
each daily dose from Day 11 to 
Day 14 
Balovaptan, 
M2, M3 
A: 11 to 14 
B: 11 to 14 
Taken directly 
from the 
observed plasma 
concentration-
time data 
 
Descriptive statistics (n, mean, geometric mean, SD, %CV, median, min, and max) will be used to 
summarize the calculated PK parameters by treatment. For Tmax, only median, min and max will be 
presented. 
16.2.2.1 Steady State 
A linear plot of the arithmetic mean trough (predose) plasma concentrations versus time will be provided 
for Balovaptan on days 11 through 14 to visually assess the achievement of steady state. This plot will 
show time in days. 
In addition, a statistical evaluation of steady state will also be made by dose level using the trough plasma 
concentrations. All valid trough concentrations on the natural-log scale will be analyzed using an analysis 
of variance (ANOVA) model with study day as a fixed effect. From this model, orthogonal contrasts with 
95% confidence intervals (CIs) will be formed between the adjusted mean trough concentration at each 
study day and the mean trough concentrations for all the following study days using Helmert contrasts. 
Precisely, Day 11 will be compared to Day 12 through Day 14 combined; Day 12 will be compared to Day 
13 through Day 14 combined; and so on.  
The following SAS PROC MIXED pseudo-code may be used:  
proc mixed data= adpc; 
by treatment;  
class day;  
model ln(trough concentration) = day / solution;  
lsmeans day / cl alpha=.05;  
estimate “Day 11 vs Days 12 - 14” day 3 -1 -1 -1 / e cl alpha=0.05 
divisor=3;  
estimate “Day 12 vs Days 13 - 14” day 0 2 -1 -1 / e cl alpha=0.05 
divisor=2; 
estimate “Day 13 vs 14” day 0 0 1 -1 / e cl alpha=0.05 divisor=2; 
run; 
17.0 Safety Analyses  
17.1 Safety Variables 
• 
AEs 
• 
Clinical Laboratory Evaluations 
o 
Clinical Chemistry  
o 
Hematology 
o 
Urinalysis 
o 
Myocyte Biomarkers 
• 
Vital Signs 
o 
Supine Blood Pressure  
 
Systolic Blood Pressure  
 
Diastolic Blood Pressure  
o 
Pulse rate 
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o 
Oral body temperature 
• 
Electrocardiograms (ECG) 
o 
Heart Rate 
o 
PR Interval 
o 
QRS-Duration 
o 
QT Interval 
o 
QTc (Frederica) Interval 
o 
T-Waves 
o 
U-Waves 
• 
Echocardiograms 
o 
PAAT 
o 
RVOT-VTI 
o 
Inferior Vena Cava Diameter (IVC) 
o 
Right Atrial Size 
o 
Right-Ventricular Index of Myocardial Performance (RIMP) 
o 
Right Ventricle (RV): End-Systolic Pressure 
o 
RV: End-Diastolic Pressure 
o 
Tricuspidal Regurgitation 
o 
Systolic PAP 
o 
Pulmonary Valve Regurgitation 
o 
Diastolic PAP 
o 
Right Ventricular Wall Thickness 
o 
Right Ventricular Size 
o 
LVEF 
o 
Left Ventricular Wall Thickness 
o 
Ratio of Transmitral Doppler Early Filling Velocity (E) To Tissue Doppler Early Diastolic 
Mitral Annular Velocity (E’) 
o 
Aortic Valve Insufficiency 
o 
Mitral Valve Regurgitation 
o 
Number of subjects per treatment with mPAP exceeding 25 mm Hg 
• 
Physical Examination 
• 
Columbia-Suicide Severity Rating Scale (C-SSRS) 
17.1.1 Adverse Events 
Treatment emergence will be evaluated for all AEs. Treatment-emergent adverse events (TEAE) are 
those which occur after the first dose of study drug.  
TEAEs occurring following dosing in a specific period but before dosing in the next period will be 
attributed to the treatment in that period. If the time is missing for an AE on a dosing day then the AE will 
be attributed to the treatment given on that day. 
The following missing data will be imputed as defined (for calculations only / will not be presented): 
• 
Missing AE start and / or end times for the calculation of onset and duration will be assumed to be 
at 00:01 for a start time and 23:59 for end times 
• 
Missing AE severity or relationship will be assumed to be severe or related, respectively 
• 
Missing AE start times for the determination of treatment emergence will be assumed to occur 
after treatment unless partial date documents the AE as happening prior to treatment 
• 
Missing AE start times for the determination of treatment assignment will be assumed to occur 
after treatment on the recorded date one minute after dosing 
• 
Missing AE start date will be assumed to be after treatment for the determination of TEAE but will 
not be attributed to a specific treatment 
A summary of number and percentage of subjects reporting TEAEs, TEAEs by severity and relationship, 
serious AEs (SAEs), and subjects who discontinued study drug due to an AE will be provided. 
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A summary of the number and percentage of subjects reporting each TEAE, categorized by system organ 
class and preferred term coded according to the Medical Dictionary for Regulatory Activities (MedDRA), 
will be presented by treatment and overall. Counting will be done by subject only, not by event; subjects 
will only be counted once within each body system or preferred term. 
A summary of the number and percentage of subjects reporting each TEAE will be presented by 
relationship to study drugs (as recorded on the eCRF) and by treatment and overall. Subjects with 
multiple events within a system organ class or preferred term will be counted under the category of their 
most drug-related event within that system organ class or preferred term.  
A summary of the number and percentage of subjects reporting each TEAE will be presented by severity 
(as recorded on eCRF) and by treatment and overall. Subjects with multiple events within a system organ 
class or preferred term will be counted under the category of their most severe event within that system 
organ class or preferred term. 
A summary presenting the preferred terms for the events in descending order of frequency for the overall 
treatment group will also be presented. 
All AEs (including non-treatment-emergent events) recorded on the eCRF will be listed by subject. 
A separate listing of AEs leading to study drug discontinuation will be provided. 
17.1.2 Serious Adverse Events 
A listing of SAEs will be provided by subject. 
17.1.3 Laboratory Data 
Clinical laboratory data will be presented using units from the study data tabulation model (SDTM) 
Controlled Terminology. 
Descriptive statistics summarizing continuous laboratory results of clinical chemistry, hematology, 
urinalysis, and myocyte by treatment and visit will be provided.  
All laboratory data will be listed by subject, including laboratory variables not listed in the protocol. A 
separate listing of out-of-range values will also be provided. Normal ranges will be used directly from the 
clinical laboratory and will be included in the listings for reference. 
17.1.4 Vital Signs 
Descriptive statistics summarizing vital signs by treatment and scheduled time will be provided. 
All vital signs will be listed by subject, including orthostatic challenge testing taken at supine and standing 
positions.  
Orthostatic change will be calculated for orthostatic challenge testing results for Systolic Blood Pressure, 
Diastolic Blood Pressure, and Pulse Rate as: Orthostatic Vital Sign = Standing Vital Sign – Supine/Sitting 
Vital Sign. 
17.1.5 12-Lead Safety Electrocardiograms 
Descriptive statistics summarizing ECG parameters by treatment and visit will be provided. 
All ECG parameters and the corresponding abnormalities will be listed by subject. 
17.1.6 Echocardiograms 
Descriptive statistics summarizing echocardiogram parameters, changes from baseline, and changes 
from baseline relative to placebo by treatment and visit will be provided. Treatments C and D will be 
pooled for this summary and displayed as “Placebo.” Change from baseline relative to placebo will be 
calculated as change from baseline for the placebo period minus the change from baseline for treatment 
A and B periods. 
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All ECG parameters and the corresponding categorical indicators will be listed by subject. 
17.1.7 Other Observations Related to Safety 
Physical examination results will be listed by subject. Any clinically significant findings will be reported as 
AEs. 
C-SSRS results will be listed by subject. 
18.0 References 
SAS Institute, Inc., SAS® Version 9.4 software, Cary, NC. 
Clinical Study Protocol. A SINGLE-CENTER, MULTIPLE-DOSE, RANDOMIZED, DOUBLE-BLIND, 
PLACEBO-CONTROLLED, POSITIVE-CONTROLLED, CROSS-OVER STUDY TO INVESTIGATE THE 
EFFECT OF BALOVAPTAN ON THE QTC INTERVAL IN HEALTHY SUBJECTS. Version 2.0, Final, 16 
Jan 2018. 
Zhang, Joanne and Machado, Stella G. (2008) 'Statistical Issues Including Design and Sample Size 
Calculation in Thorough QT/QTc Studies', Journal of Biopharmaceutical Statistics,18:3, 451 — 467 
 
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TFL(s) 
Tables, figures and listings 
Tmax 
Time to maximum plasma concentration 
WHO-DDE 
World Health Organization – Drug Dictionary Enhanced 
WNL 
WinNonlin 
 
 
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Appendix 2: Protocol Schedule of Assessments 
 
Assessments 
 
Screening 
 
Nominal Study Day for Each Treatment Period 
Follow-
upa 
 
Days  
-28 to -3 
Day 
-2 
Day 
-1 
Day 
1 
Day 
2 
Day 
3 
Day 
4 
Day 
5 
Day 
6 
Day 
7 
Day 
8 
Day 
9 
Day 
10 
Day 
11 
Day 
12 
Day 
13 
Day 
14 
Day 
15 
Day 
16 
 
Written informed 
consent 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion and exclusion 
criteria 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Randomization 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical history 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Conmeds review 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Physical examination 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Height and weightb 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Pre and post-dose 
fastingc 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Standardized low salt 
mealsd 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Vital signse 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
TT-Doppler 
echocardiogram 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
ABPM 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Laboratory safety testsf 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
12-lead safety ECGg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
12-lead ECG Holter 
monitoringh 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Administration of 
investigational 
medicinal product 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Drugs of abuse screeni 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Orthostatic challenge 
testing 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Urine alcohol screenj 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Pregnancy testk 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Follicle stimulating 
hormonel 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Blood sampling for 
balovaptan PKm 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Blood sampling for 
moxifloxacin PKm 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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a. Follow-up visit should occur between 14 and 21 days after the final dose of study drug 
b. Height and weight to calculate BMI will collected at Screening. During Follow-up visit only weight will be recorded 
c. Subjects will fast for a minimum of 8 hours predose and 4 hours post-dose 
d. Subjects will be fed standardized low salt meals 
e. Assessment of supine blood pressure, pulse rate and body temperature. All measurements to be recorded after the subject has remained in a supine position for at least 10 
minutes. (See Appendix 3 for details) 
f. 
Includes hematology, biochemistry, urinalysis, coagulation and serology (coagulation and serology will be performed only during Screening) 
g. Safety ECG assessments: Triplicate ECG measurements at Screening and Day -2. Single ECG measurements at Day 1, Day 3, Day 6, Day 9, Day 11, Day 13, Day 14, Day 15, 
Day 16 and follow-up visit.  All measurements to be recorded after the subject has remained in a supine position for at least 10 minutes 
h. ECG Holter monitoring assessments will be recorded on study Days 1, 2, 14 and 15 (See Appendix 3 for additional details).  
All ECG measurements are to be recorded while the subject remains supine for at least 15 minutes.  
ECGs will be extracted from Holters at 3 time points predose (-45, -30 and -15 minutes) and 7 time points post-dose (0.5, 1.0, 2.5, 4, 8, 12 and 24 h post-dose). The 24-hour post-
dose extraction time point for Days 1 and 14 can be taken from the 3 predose time points on Days 2 and 15. 
12-lead ECGs will be extracted from the continuous recording in up to 10 replicates from a 5-minute window during a 15-minute supine period pre-dose and at the nominal Holter 
timepoints post-dose. Whenever ECG extraction and PK sample timepoints are close together the ECG extraction will be performed prior to PK sample collection. 
i. 
Drugs of abuse: amphetamines, barbiturates, benzodiazepines cannabinoids, cocaine, opiates, methamphetamines, methadone and 3,4 Methylenedioxymethamphetamine 
(MDMA), will be measured in urine at Screening and Day 2. Additional testing may be performed during the study, at the discretion of the Investigator or designee, to confirm 
subject compliance. 
j. 
Planned at Screening and Day -2, further testing may be performed during the study, at the discretion of the Investigator or designee, to confirm subject compliance. 
k. Serum pregnancy test 
l. 
Assessed for all females. 
m. See Appendix 3 for PK sampling details 
n. Blood samples for the assessment of early markers of myocyte damage: Troponin I, Troponin T and NT-proBNP, Troponin I will be assessed following sample collection while 
Troponin T and NT--proBNP samples will be stored for potential future analysis. 
o. Optional blood sample for Clinical Genotyping can be collected any time during the study. N.B. one sample only to be collected and only if consented for this sample 
 
PRIORITY OF ASSESSMENTS: where appropriate and feasible: ECG > Vital Signs > Blood Sampling > Lab Safety Tests 
 
Myocyte biomarkersn 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
C-SSRS 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Adverse Events 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Blood sampling for 
Clinical Genotypingo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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Appendix 3: Schedule of Pharmacokinetics, Vital Signs, and ECG 
 
 
Assessments 
 
Hours post-dose 
Pre-dose 
0.5 
1.00 
2.5 
3 
4 
6 
8 
12 
24a 
12-lead ECG Holter 
monitoringb 
 
 
 
 
 
 
 
 
 
 
Vital Signs 
 
 
 
 
 
 
 
 
 
 
Blood collection for 
pharmacokineticsc 
 
 
 
 
 
 
 
 
 
 
 
a. On days when consecutive 24 hour and predose assessments are being done, only one assessment needs to be done prior to 
dose. This includes Holter extractions where the 24-hour post-dose extraction time point for Days 1 and 14 can be taken from 
the 3 predose time points on Days 2 and 15. 
b. Predetermined 24 h Holter ECG time points on Day 1, Day 2, Day 14 and Day 15.  Holter ECG assessments will be performed in 
the 15 minutes prior to the nominal Holter timepoint but prior to the PK sample, if applicable. On Day 2 and 15 of each Treatment 
Period subjects are scheduled to have 24 h post-dose ECG Holter reading, PK sampling and daily dosing of study drug. These 
activities will be performed in the following order the 24 h post-dose Holter readings will be performed first followed by drawing of 
the 24 h post-dose PK sample and then dosing of study drug.  Due to this study drug dosing on these days may be delayed 
relative to dosing on other study days. 
c. Blood collection for PK assessments will be identical for Treatment Arms in all Treatment Periods as this is a blinded study. 
Intensive PK shown above, in Appendix 2, will take place on study Days 1, and 14 for balovaptan and Days 2 and 15 for 
moxifloxacin. On other study days with PK sampling and study drug dosing (Days 11, 12, 13) a single PK sample will be drawn 
prior to dosing with study drugs. This predose sample also serves as the 24 h post-dose sample for the previous days dosing. A 
moxifloxacin PK sample will be taken on Day 16 for the Day 15, 24 h post-dose. 
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