Conditions: Healthy Volunteers
Allocation: NON_RANDOMIZED | Intervention model: SINGLE_GROUP
Enrollment: 16
Healthy volunteers: yes

Brief summary:
This study was a single-center, non-randomized, open-label, one-sequence, two-period, within-subject study to investigate the effects of multiple doses of rifampicin on the PK and safety of multiple doses of balovaptan in healthy subjects. The study was conducted at 1 site in the Netherlands.

Detailed description:

Arms:
arm 1: Participants received the study drugs in 2 periods. There was a minimum of a 14-day to a maximum of a 21-day washout between the last dose in Period 1 and the first dose in Period 2.

Interventions: intervention 1: Balovaptan intervention 2: Rifampicin
Intervention descriptions: intervention 1: In Period 1, balovaptan was administered alone as a once daily (qd) dose on Days 1 to 10.

In Period 2, balovaptan was administered as a qd dose on Days 7 to 16. intervention 2: In Period 2, 600 mg of rifampicin will be administered alone as a qd dose from Day 1 to Day 6, and as a qd dose on Days 7 to 16.

Protocol document:
Prot_000.pdf: 
 
Official Title: 
A SINGLE-CENTER, NON-RANDOMIZED, OPEN-LABEL, 
ONE-SEQUENCE, TWO-PERIOD WITHIN-SUBJECT 
STUDY TO INVESTIGATE THE EFFECT OF RIFAMPICIN 
ON THE PHARMACOKINETICS OF MULTIPLE DOSES 
OF BALOVAPTAN IN HEALTHY VOLUNTEERS 
NCT Number: 
NCT03586726  
Document Date: 
Protocol version 1: 03-Jun-2018 
 
 


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  The information contained in this document, especially any unpublished data, 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 WP40608, Version 1.0
PROTOCOL
TITLE:
A SINGLE-CENTER, NON-RANDOMIZED, OPEN-LABEL, 
ONE-SEQUENCE, TWO-PERIOD WITHIN-SUBJECT 
STUDY TO INVESTIGATE THE EFFECT OF RIFAMPICIN 
ON THE PHARMACOKINETICS OF MULTIPLE DOSES 
OF BALOVAPTAN IN HEALTHY VOLUNTEERS
PROTOCOL NUMBER:
WP40608
VERSION NUMBER:
1.0
EUDRACT NUMBER:
2018-001783-40
IND NUMBER:
NA
TEST PRODUCT:
Balovaptan (RO5285119)
MEDICAL MONITOR:
, MD
SPONSOR:
F.  Hoffmann-La Roche Ltd
DATE FINAL:
See electronic date stamp below
 
03-Jun-2018 13:34:40
Title
Approver's Name
Company Signatory
Date and Time (UTC)


Balovaptan—F.  Hoffmann-La Roche Ltd
2/Protocol WP40608, Version 1.0
TABLE OF CONTENTS
PROTOCOL ACCEPTANCE FORM .................................................................... 8
PROTOCOL SYNOPSIS ...................................................................................... 9
1.
BACKGROUND .......................................................................................... 17
1.1
Autism Spectrum Disorder..................................................... 17
1.2
Background on Balovaptan.................................................... 18
1.2.1
Previous and Ongoing Clinical Studies.................................. 20
1.2.1.1
Clinical Pharmacology ........................................................... 20
1.2.1.2
Safety .................................................................................... 21
1.3
Study Rationale and Benefit-Risk Assessment...................... 22
2.
OBJECTIVES AND ENDPOINTS ............................................................... 22
3.
STUDY DESIGN......................................................................................... 23
3.1
Description of the Study......................................................... 23
3.2
End of Study and Length of Study......................................... 25
3.3
Rationale for Study Design.................................................... 25
3.3.1
Rationale for Open-Label Design .......................................... 25
3.3.2
Rationale for Balovaptan and Rifampicin Dose 
and Schedule......................................................................... 25
3.3.2.1
Balovaptan............................................................................. 25
3.3.2.2
Rifampicin.............................................................................. 25
3.3.3
Rationale for Study Population .............................................. 26
4.
MATERIALS AND METHODS .................................................................... 26
4.1
Subjects................................................................................. 26
4.1.1
Inclusion Criteria.................................................................... 26
4.1.2
Exclusion Criteria................................................................... 27
4.1.2.1
Rifampicin-related Exclusion Criteria..................................... 29
4.2
Method of Treatment Assignment and Blinding..................... 30
4.3
Study Treatment and Other Treatments Relevant 
to the Study Design ............................................................... 30
4.3.1
Study Treatment Formulation, Packaging, and 
Handling ................................................................................ 30
4.3.1.1
Balovaptan............................................................................. 30


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3/Protocol WP40608, Version 1.0
4.3.1.2
Rifampicin.............................................................................. 30
4.3.2
Study Treatment Dosage, Administration, and 
Compliance............................................................................ 31
4.3.2.1
Balovaptan............................................................................. 31
4.3.2.2
Rifampicin.............................................................................. 31
4.3.3
Investigational Medicinal Product Accountability ................... 31
4.4
Concomitant Therapy, Prohibited Food, and 
Additional Restrictions ........................................................... 32
4.4.1
Permitted Therapy................................................................. 32
4.4.2
Prohibited Therapy ................................................................ 32
4.4.3
Prohibited Food ..................................................................... 33
4.4.4
Additional Restrictions/Considerations .................................. 33
4.5
Study Assessments ............................................................... 34
4.5.1
Informed Consent Forms and Screening Log........................ 34
4.5.2
Medical History, Concomitant Medication, and 
Demographic Data................................................................. 34
4.5.3
Physical Examinations........................................................... 34
4.5.4
Vital Signs.............................................................................. 35
4.5.5
Laboratory, Pharmacokinetic, and Genotyping 
Samples................................................................................. 35
4.5.5.1
Laboratory Samples .............................................................. 35
4.5.5.2
Pharmacokinetic Samples ..................................................... 36
4.5.5.3
Genotyping Samples ............................................................. 36
4.5.6
Electrocardiograms................................................................ 37
4.5.7
Columbia-Suicide Severity Rating Scale ............................... 37
4.5.8
Drug and Metabolite Concentration 
Measurements....................................................................... 37
4.5.9
Timing of Study Assessments ............................................... 37
4.6
Treatment, Subject, Study, and Site 
Discontinuation...................................................................... 37
4.6.1
Study Treatment Discontinuation........................................... 37
4.6.2
Subject Discontinuation from Study....................................... 38
4.6.3
Study Discontinuation............................................................ 38
4.6.4
Site Discontinuation............................................................... 38


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4/Protocol WP40608, Version 1.0
5.
ASSESSMENT OF SAFETY....................................................................... 39
5.1
Safety Plan ............................................................................ 39
5.1.1
Potential Important Risks Associated with 
Balovaptan............................................................................. 39
5.1.1.1
Hematotoxicity....................................................................... 39
5.1.1.2
Skeletal Muscle Toxicity ........................................................ 39
5.1.1.3
Cardiac Toxicity ..................................................................... 40
5.1.1.4
Neurologic Toxicity ................................................................ 40
5.1.1.5
Orthostasis ............................................................................ 41
5.1.2
Management of Toxicities and of Subjects Who 
Experience Adverse Events................................................... 41
5.1.2.1
Management Guidelines........................................................ 41
5.1.2.2
Management of Hematological Toxicity................................. 42
5.1.2.3
Management of Skeletal Muscle Toxicity............................... 42
5.1.2.4
Management of Cardiac Toxicity ........................................... 43
5.1.2.5
Management of Neurologic Toxicity ...................................... 44
5.1.2.6
Management of Orthostatic Hypotension............................... 45
5.2
Safety Parameters and Definitions ........................................ 45
5.2.1
Adverse Events ..................................................................... 45
5.2.2
Serious Adverse Events (Immediately Reportable 
to the Sponsor)...................................................................... 45
5.2.3
Adverse Events of Special Interest (Immediately 
Reportable to the Sponsor).................................................... 46
5.3
Methods and Timing for Capturing and 
Assessing Safety Parameters................................................ 47
5.3.1
Adverse Event Reporting Period ........................................... 47
5.3.2
Eliciting Adverse Event Information....................................... 47
5.3.3
Assessment of Severity of Adverse Events ........................... 48
5.3.4
Assessment of Causality of Adverse Events ......................... 48
5.3.5
Procedures for Recording Adverse Events............................ 49
5.3.5.1
Diagnosis versus Signs and Symptoms................................. 49
5.3.5.2
Adverse Events That Are Secondary to Other 
Events.................................................................................... 49
5.3.5.3
Persistent or Recurrent Adverse Events................................ 50
5.3.5.4
Abnormal Laboratory Values ................................................. 50


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5.3.5.5
Abnormal Vital Sign Values ................................................... 51
5.3.5.6
Abnormal Liver Function Tests.............................................. 51
5.3.5.7
Deaths ................................................................................... 52
5.3.5.8
Pre-existing Medical Conditions ............................................ 52
5.3.5.9
Hospitalization or Prolonged Hospitalization.......................... 52
5.4
Immediate Reporting Requirements from 
Investigator to Sponsor.......................................................... 53
5.4.1
Emergency Medical Contacts................................................ 54
5.4.2
Reporting Requirements for Serious Adverse 
Events and Adverse Events of Special Interest..................... 54
5.4.2.1
Events that Occur Prior to Study Drug Initiation .................... 54
5.4.2.2
Events that Occur after Study Drug Initiation......................... 54
5.4.3
Reporting Requirements for Pregnancies.............................. 55
5.4.3.1
Pregnancies in Female Subjects ........................................... 55
5.4.3.2
Pregnancies in Female Partners of Male 
Subjects................................................................................. 55
5.4.3.3
Abortions ............................................................................... 55
5.4.3.4
Congenital Anomalies/Birth Defects ...................................... 56
5.4.4
Reporting Requirements for Cases of Accidental 
Overdose or Medication Error................................................ 56
5.4.5
Reporting Requirements for Medical Device 
Complaints............................................................................. 57
5.5
Follow-Up of Subjects after Adverse Events.......................... 57
5.5.1
Investigator Follow-Up........................................................... 57
5.5.2
Sponsor Follow-Up ................................................................ 57
5.6
Adverse Events That Occur after the Adverse 
Event Reporting Period.......................................................... 58
5.7
Expedited Reporting to Health Authorities, 
Investigators, Institutional Review Boards, and 
Ethics Committees................................................................. 58
6.
STATISTICAL CONSIDERATIONS AND ANALYSIS PLAN....................... 58
6.1
Determination of Sample Size ............................................... 58
6.2
Summaries of Conduct of Study............................................ 59
6.3
Summaries of Demographic and Baseline 
Characteristics....................................................................... 59


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6.4
Efficacy Analyses .................................................................. 59
6.5
Safety Analyses..................................................................... 59
6.6
Pharmacokinetic Analyses..................................................... 59
6.7
Other Analyses...................................................................... 60
6.7.1
Clinical Genotyping................................................................ 60
7.
DATA COLLECTION AND MANAGEMENT ............................................... 60
7.1
Data Quality Assurance......................................................... 60
7.2
Electronic Case Report Forms............................................... 61
7.3
Source Data Documentation.................................................. 61
7.4
Use of Computerized Systems .............................................. 62
7.5
Retention of Records............................................................. 62
8.
ETHICAL CONSIDERATIONS.................................................................... 62
8.1
Compliance with Laws and Regulations ................................ 62
8.2
Informed Consent.................................................................. 63
8.3
Institutional Review Board or Ethics Committee.................... 63
8.4
Confidentiality........................................................................ 64
8.5
Financial Disclosure .............................................................. 64
9.
STUDY DOCUMENTATION, MONITORING, AND 
ADMINISTRATION ..................................................................................... 65
9.1
Study Documentation ............................................................ 65
9.2
Protocol Deviations................................................................ 65
9.3
Site Inspections ..................................................................... 65
9.4
Administrative Structure......................................................... 65
9.5
Publication of Data and Protection of Trade 
Secrets .................................................................................. 65
9.6
Protocol Amendments ........................................................... 66
10. REFERENCES ........................................................................................... 67
LIST OF TABLES
Table 1
Objectives and Corresponding Endpoints................................... 22
Table 2
Adverse Event Severity Grading Scale ....................................... 48
Table 3
Causal Attribution Guidance ....................................................... 48


Balovaptan—F.  Hoffmann-La Roche Ltd
7/Protocol WP40608, Version 1.0
LIST OF FIGURES
Figure 1
Study Schema............................................................................ 24
LIST OF APPENDICES
Appendix 1
Schedule of Activities................................................................. 68
Appendix 2
Schedule of Pharmacokinetic Samples and 
Electrocardiogram, and Vital Signs Measurements .................... 70




Balovaptan—F.  Hoffmann-La Roche Ltd
9/Protocol WP40608, Version 1.0
PROTOCOL SYNOPSIS
TITLE:
A SINGLE-CENTER, NON-RANDOMIZED, OPEN-LABEL, 
ONE-SEQUENCE, TWO-PERIOD WITHIN-SUBJECT STUDY TO 
INVESTIGATE THE EFFECT OF RIFAMPICIN ON THE 
PHARMACOKINETICS OF MULTIPLE DOSES OF BALOVAPTAN 
IN HEALTHY VOLUNTEERS
PROTOCOL NUMBER:
WP40608
VERSION NUMBER:
1.0
EUDRACT NUMBER:
2018-001783-40
IND NUMBER:
NA
TEST PRODUCT:
Balovaptan (RO5285119)
PHASE:
I
INDICATION:
Not applicable
SPONSOR:
F.  Hoffmann-La Roche Ltd
Objectives and Endpoints
This study will investigate the effect of rifampicin treatment on the PK of balovaptan and its 
major metabolites M2 and M3.
Objectives
Endpoints
Primary
To investigate the effect of multiple doses of 
rifampicin, a potent cytochrome P450 
(CYP)3A4 inducer, on the pharmacokinetics 
of balovaptan 10 mg once daily (qd) and its 
major metabolites M2 and M3 at steady state 
in healthy subjects
Primary PK endpoints: maximum observed 
plasma concentration (Cmax) and area under 
the plasma concentration curve from time 0 to 
24 hours (AUC0-24h) for balovaptan, M2, and 
M3 following 10 days of qd dosing of 
balovaptan 10 mg
Secondary
To evaluate the safety and tolerability of 
balovaptan 10 mg qd administered alone and 
in combination with rifampicin in healthy 
subjects
Adverse events, clinical laboratory values, 
vital signs, electrocardiogram (ECG), 
Columbia-Suicide Severity Scale and 
physical examination
To investigate the effect of multiple doses of 
rifampicin on secondary PK parameters of 
multiple doses of balovaptan 10 mg qd and 
its major metabolites M2 and M3 at steady 
state in healthy subjects
Balovaptan and metabolite M2 and M3: time 
to maximum observed plasma concentration 
(tmax); metabolite to parent ratio; any other 
parameter as appropriate


Balovaptan—F.  Hoffmann-La Roche Ltd
10/Protocol WP40608, Version 1.0
Tertiary/Exploratory
To evaluate the plasma exposure of 
rifampicin 600 mg qd 
Rifampicin Cmax, AUC0-24h, tmax, area under 
the plasma concentration-time curve from 
time zero to the last measurable plasma 
concentration time point (AUC0-last) following 6 
and 16 days of qd dosing of rifampicin, and 
any other parameter as appropriate
To investigate the relationship between the 
CYP3A4 genotype, among othersa, and 
steady state balovaptan exposure, and the 
influence of the CYP3A4 genotype, among 
others, on the effect of rifampicin on 
balovaptan PK
The pharmacogeneticsb of metabolizing 
enzymes, tranferases, transporters, etc, 
possibly involved in the absorption, 
distribution, metabolism and excretion of 
balovaptan and its major metabolites (e.g., 
CYP3A and P-glycoprotein)
aThe relationship between other genotypes and PK of balovaptan may also be explored.
bResults may be pooled with data from other studies of balovaptan.
Study Design
This study will be a single-center, non-randomized, open-label, one-sequence, two-period, 
within-subject study to investigate the effects of multiple doses of rifampicin on the PK and 
safety of multiple doses of balovaptan in healthy subjects.  The study will be conducted at 1 site 
in the Netherlands.  
Screening will be conducted up to 3 weeks prior to admission in the clinical research unit (CRU).  
Subjects will be in the CRU for 2 study periods. In both periods, subjects will be admitted on 
Day -1 (the day before dosing begins); Day 1 will be the first day of dosing. In Period 1, subjects 
will be discharged on Day 11 after the last assessment is completed.  There will be a minimum 
of a 14-day to a maximum of a 21-day washout between the last dose in Period 1 and the first 
dose in Period 2.  Subjects will therefore return to the CRU on Day 24 or up to Day 30 of Period 
1 (Day -1 of Period 2) to begin Period 2.  In Period 2, subjects will be discharged on Day 17 
after all assessments have been performed.  Subjects will return for a follow-up visit between 14 
to 21 days after the last dose.  
Subjects will receive the following study drugs in the 2 study periods, after an overnight fast of 
at least 8 hours:
Period 1:

Days 1 to 10: oral balovaptan 10 mg qd

Days 11 to 23 (or up to Day 30): no study drug
Period 2:

Days 1 to 6: rifampicin 600 mg qd

Days 7 to 16: balovaptan 10 mg qd + rifampicin 600 mg qd 
Number of Subjects
A total of 16 subjects will be enrolled to ensure 12 evaluable subjects.  


Balovaptan—F.  Hoffmann-La Roche Ltd
11/Protocol WP40608, Version 1.0
Target Population
Healthy male (at least 50% of enrolled subjects) and female subjects.
Inclusion Criteria
Subjects must meet the following criteria for study entry:
1.
Healthy male and female subjects, aged 18 to 65 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 of 18 to 30 kg/m2, inclusive, at Screening.
3.
Women of childbearing potential must agree use one highly effective non-hormonal 
contraceptive method combined with a barrier method from screening until 90 days 
after the last dose of study drug.  

Highly effective methods include:
o
A non-hormonal intrauterine device implanted at least 2 months prior to 
screening.

Acceptable barrier methods include:
o
Condom, diaphragm, cervical/vault cap, or vaginal sponge.
Women are considered to be of non-child bearing potential if postmenopausal at least 
12 months of amenorrhea and confirmed by follicle stimulating hormone, or at least 
24 months if on hormone replacement therapy), or surgically sterile (absence of ovaries 
and/or uterus; or has undergone bilateral tubal ligation). Female subjects who have 
undergone bilateral tubal ligation must also use a barrier method.
4.
For men:  agreement to use contraceptive measures, and agreement to refrain from 
donating sperm, as defined below:
o
With female partners of childbearing potential or pregnant female partners, men 
must use a condom during the treatment period 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 or Dutch.
Exclusion Criteria
Subjects who meet any of the following criteria will be excluded from study entry:
1.
Female subjects who are pregnant or lactating.
2.
If female of childbearing potential, a positive serum pregnancy test at Screening or at 
admission (Day -1 of either period).  
3.
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.  
4.
History of any clinically significant gastrointestinal, renal, hepatic, broncho-pulmonary, 
neurological, psychiatric, cardiovascular, endocrinological, hematological, lymphatic, 
musculoskeletal, genitourinary, immunological, dermatological, connective tissue or allergic 
disease, metabolic disorder, or cancer.  
5.
Signs or symptoms potentially indicative of peripheral neuropathy.
6.
History or evidence of any medical condition potentially altering the absorption, metabolism 
or elimination of drugs.  Surgical history of the gastrointestinal tract affecting gastric motility 
or altering the gastrointestinal tract, with the exception of uncomplicated appendectomy, 
which is allowed.  


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12/Protocol WP40608, Version 1.0
7.
A history of clinically significant hypersensitivity (e.g., to drugs, including rifampicin and 
balovaptan, and excipients) or allergic reactions.  
8.
In the opinion of the Investigator, 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.
9.
Known personal or family history of congenital long QT syndrome or sudden death.
10. Subjects with screening or predose baseline mean QT interval corrected using Fridericia’s 
formula (QTcF) >450 msec or <300 msec (using the same upper QTcF limit in both males 
and females).
11.
Notable resting bradycardia (mean HR <40 bpm) on screening or predose baseline ECG. 
Notable resting tachycardia (mean HR > 90 bpm) on screening or predose baseline ECG.
12.
Screening or baseline ECGs 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).
13. Screening or baseline ECG with evidence of clinically relevant abnormalities, e.g. atrial 
fibrillation, atrial flutter, complete bundle branch block, Wolf-Parkinson-White Syndrome, or 
cardiac pacemaker.
14. History or presence of clinically significant ECG abnormalities before study drug 
administration.  
15. Clinically significant abnormalities in laboratory test results (including hepatic and renal 
panels, complete blood count, chemistry panel, and urinalysis).  In the case of uncertain or 
questionable results, tests performed during screening may be repeated before Day -1 of 
Period 1 to confirm eligibility.  
16. Confirmed (based on the average of ≥ 3 consecutive measurements) resting pulse rate 
greater than 100 or less than 40 bpm.  
17. Any suspicion or history of alcohol abuse and/or suspicion or history of regular consumption 
of drugs of abuse.  Past alcohol and/or other substance abuse or addiction is also not 
allowed.
18. Alcohol consumption of >24 units per week for males and females. One unit of alcohol 
equals ½ pint (285 mL) of beer or lager, 1 glass (125 mL) of wine, or 1/6 gill (25 mL) of 
spirits
19. Positive urine alcohol test or urine drug screen at Screening or Day -1 of each study period 
(barbiturates, benzodiazepines, methadone, amphetamines [including ecstasy], 
methamphetamines, opiates, cocaine, and cannabinoids).
20. Smokers who regularly smoke more than 5 cigarettes daily or equivalent and are unable or 
unwilling not to smoke or use tobacco for 48 hours prior to dosing and during both in-house 
periods.  
21. A screen positive for hepatitis B surface antigen (HBsAg), hepatitis C virus (HCV) antibody, 
or antibodies against human immunodeficiency viruses 1 and 2 (HIV-1/HIV-2 antibodies).
22. Participation in an investigational drug or device study within 90 days 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.  
23. Any donation of blood or plasma or significant blood loss within 3 months prior to Screening.  
24. 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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25. History of coagulopathies, bleeding disorders or blood dyscrasias.
26. Confirmed (based on the average of ≥3 consecutive measurements) systolic blood pressure 
greater than 139 or less than 90 mm Hg, and diastolic blood pressure greater than 89 or less 
than 45 mm Hg.
27. Dietary restrictions that would prohibit the consumption of standardized meals.
28. Subjects likely to need concomitant medication during the study period (including for dental 
conditions).  
29. Subjects who are unwilling to practice safe sex (use of male or female condom) for the 
duration of the study.
30.
Subjects who have received any prescribed systemic or topical medication within 4 weeks 
(or within 5 times the elimination half-life of the medication, whichever is longer)the first dose 
administration, unless in the opinion of the Investigator the medication will not interfere with 
the study procedures or compromise safety, and subjects who have received 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)of the first dose administration will 
also be excluded unless in the opinion of the Investigator the medication will not interfere 
with the study procedures or compromise safety.
31.
Used the following within 7 days before the first study drug administration, unless in the 
opinion of the Investigator the medication will not interfere with the study procedures or 
compromise safety: 
- any non-prescribed systemic or topical medication
- any herbal remedies.
Paracetamol (up to 4 g/day within 7 days before study drug administration and 2 g/day 
within 48 hours before study drug administration), dietary supplements, vitamins and 
minerals, hormonal contraceptives, and hormone replacement therapy are allowed.
32.
Received any medications known to chronically alter drug absorption or elimination 
processes within 4 weeks before the first study drug administration, unless in the opinion of 
the Investigator the medication will not interfere with the study procedures or compromise 
safety.
33. Subjects under judicial supervision, guardianship, or curatorship.
Rifampicin-related Exclusion Criteria:
Contraindications
1.
Subjects diagnosed, or suspected of having porphyria, and subjects with first-degree 
relatives diagnosed, or suspected of having porphyria.
2.
Subjects unwilling not to use contact lenses for the duration of rifampicin dosing (Day 1 to 
Day 16 of Period 2) until 7 days after the last dose of rifampicin (subjects may resume 
wearing contacts on Day 23 or Period 2).
3.
Known hypersensitivity to rifampicin
Additional Exclusion Criteria:
Taking any drug or herbal CYP3A modulator (e.g., erythromycin; St. John’s wort) within 
4 weeks (or 5 half-lives, whichever is longer) or any other nutrients known to modulate CYP3A 
activity (e.g., grapefruit juice; Seville orange) within 2 weeks prior to the first administration of 
study drugs.


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14/Protocol WP40608, Version 1.0
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 up to approximately 12 weeks.
Investigational Medicinal Products
Balovaptan
RO5285119/F17 10 mg dispersible tablet will be administered alone as a qd dose on Days 1 to
10 in Period 1 and as a qd dose in combination with rifampicin 600 mg qd on Days 7 to 16 in 
Period 2.
Rifampicin
Rifampicin 600 mg (two 300-mg capsules) will be administered alone as a qd dose from Day 1 
to Day 6 in Period 2, and as a qd dose in combination with balovaptan 10 mg on Days 7 to 16.
Statistical Methods
Primary Analysis
The primary PK parameters are Cmax and AUC0-24h for balovaptan, M2, and M3 following 
10 days of qd dosing of balovaptan 10 mg.
A linear mixed model analysis will be applied to analyze the log-transformed primary PK 
parameters.  The model assumes fixed effect for treatment and a random effect for subject.  
Estimates of geometric mean ratios together with the corresponding 90% confidence intervals
(CIs) will be derived for the comparisons of the PK parameters for balovaptan plus rifampicin 
(test) vs balovaptan alone (reference).
Determination of Sample Size 
This is an exploratory study for which no formal statistical hypothesis will be tested.  Therefore, 
the sample size is chosen to estimate with sufficient precision the effect of CYP3A4 induction on 
balovaptan.  
The sample size of 12 evaluable subjects was chosen based on balovaptan within-subject 
variability (CV%) of around 39% for AUC0-24h and 36% for Cmax as obtained from the 12 mg 
repeat dose in study BP25694.  Based on the higher value of 39%, it was estimated that with 
12 subjects the half-width of the 90% CI for the ratio of treatment geometric means of the 
combination (balovaptan + rifampicin) versus balovaptan alone would be obtained by 
dividing/multiplying the ratio estimate by a factor of 1.30.  
Interim Analyses
Not applicable


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LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS
Abbreviation
Definition
AE
adverse event
ALT
alanine aminotransferase
ANC
absolute neutrophil count
ASD
Autism Spectrum Disorder
AST
aspartate amino transferase
AUC0-24h
area under the plasma concentration vs time curve 
from time zero to 24 hours
AUC0-120h
area under the concentration curve from time 0 to 120
hours
AUC0-last
area under the plasma concentration-time curve from 
time zero to the last measurable plasma concentration 
time point
CI
confidence interval
Cmax
maximum observed plasma concentration
CNS
central nervous system
CPK
creatine phosphokinase
CRO
contract research organization
CRU
clinical research unit
C-SSRS
Columbia-Suicide Severity Rating Scale
CYP
cytochrome P450
EC
Ethics Committee
ECG
electrocardiogram
eCRF
electronic Case Report Form
EDC
electronic data capture
EU
European Union
FSH
follicle-stimulating hormone
GLP
Good Laboratory Practice
HBsAg
hepatitis B virus surface antigen
HCV
hepatitis C virus
HIV
human immunodeficiency virus
ICH
International Council for Harmonisation
IMP
investigational medicinal product
IND
Investigational New Drug (Application)
IRB
Institutional Review Board
IUD
intrauterine device
LC/MS/MS
liquid chromatography-tandem mass spectrometry
MAD
multiple ascending dose


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MedDRA
Medical Dictionary for Regulatory Activities
NOAEL
no observed adverse effect level
NOEL
no observed effect level
PK
pharmacokinetic(s)
PND
postnatal day
qd
once daily
SAD
single ascending dose
SAE
serious adverse event
tmax
time to maximum observed plasma concentration
ULN
upper limit of normal
US
United States
V1a
vasopressin receptor 1A
WBC
white blood cell


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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 presence of repetitive or restricted behaviors, interests, or activities (American 
Psychiatric Association, 2013).  The Autism and Developmental Disabilities Monitoring 
Network recently reported, based on children records, that the estimated prevalence of 
ASD in the United States (US) 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.  
Deficits in socialization manifest themselves as impaired use of non-verbal 
communication, delayed and reduced interactions with peers, absent sharing of 
enjoyable experiences and interest with peers, and lack of social judgment.  
Abnormalities in communication include a delay in verbal language development, 
impaired expressive language, deficient language pragmatics, as well as stereotyped, 
repetitive, or idiosyncratic use of language.  Stereotyped and repetitive behavior 
manifests as a preoccupation with stereotyped or restricted interests, adherence to 
routines, rigidity, and perseverative and stereotyped behavior, motor mannerisms, and 
preoccupation or fascination with parts of items, and unusual visual exploration (Levy et 
al., 2009).  In addition to these core deficits, patients with ASD suffer from a range of 
comorbid conditions, including irritability, depression or anxiety, attention deficits, 
obsessive compulsive symptoms, seizures, and sleep disruption.  Functional and 
structural deficits in the brain network subserving the processing of social cues (such as 
faces and speech) and social affiliative behavior have been implicated in the core 
symptom of abnormal socialization associated with ASD.  Abnormalities have been 
found in key areas including structures in the superior temporal lobe, fusiform gyrus, 
amygdala, and prefrontal cortex.  At present, no pharmacological treatment exists for the 
core deficits of ASD, and currently available treatments address only associated 
behavioral problems.  Non-pharmacological treatments have been developed to address 
the core symptoms; however, efficacy has not been proven in large clinical trials
(Meyer-Lindenberg et al., 2009).  Accordingly, there is a big unmet medical need for 
pharmacological treatments that target and modify these key deficits.
The etiology of ASD is highly genetic, although environmental factors also contribute to 
the overall risk.  Heritability estimates from family and twin studies suggest that about 90% 
of variance can be attributed to genetic factors, making ASD the neuropsychiatric 
disorder most affected by genetic factors.  The vasopressin system has been implicated 
in autism by studies of the arginine vasopressin receptor 1A (V1a) gene, which encodes 
V1a and is located on chromosome 12q (Meyer-Lindenberg et al., 2009).  Both over-
and under-transmission of specific but different alleles of this gene have been reported 


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in autistic individuals and are associated with specific traits.  One of these risk alleles 
has been found to increase activation of the amygdala during face processing and is 
associated with increased V1a receptor messenger RNA levels in the human 
hippocampus post-mortem.  In particular, both nonclinical and clinical research support a 
specific role for vasopressin in affiliative behavior.
In healthy subjects, intranasal vasopressin administration has been shown to increase 
the perception of ambiguous social stimuli as threatening and to impair emotion 
recognition in males.  In non-human mammals, vasopressin receptors are distributed in 
various brain regions associated with central nervous system (CNS) control of stress 
and anxiety and, more importantly in this context, with social behavior, including parental 
care, pair-bonding, social memory, and social aggression.  Specifically, vasopressin has 
been implicated more in male-typical social behaviors, including aggression, pair-bond 
formation, scent marking, and courtship.  Recently, vasopressin was found to affect 
social recognition and interaction in rodents via its modulatory effect on olfaction (Tobin 
et al., 2010) – a sensory modality that may also be impaired in autism (May et al., 2011).
In summary, evidence from both human and animal studies strongly implicates the V1a 
receptor in mediating and modulating key social behaviors that are deficient in ASD.  
Given 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.
1.2
BACKGROUND ON BALOVAPTAN
Chronic administration of balovaptan reversed social behavior and cognitive impairments 
induced by prenatal valproate treatment in a rat model that reflects some of the 
behavioral changes seen in patients with ASD.  
 
 
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.  Balovaptan, M2, and M3 are mainly metabolized through 
cytochrome P450 (CYP)3A4.  Balovaptan and M3 have been shown in vitro to be strong, 
substrate-dependent P-gp inhibitors.  In human, 2 major metabolites, M2 (RO7045402) 
and M3 (RO5273004), have been identified, which are present in rats and dogs.


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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 been completed (BP28420, also
referred to as VANILLA) and a Phase II study in pediatric ASD patients (male and 
female) is ongoing (BP30153, also referred to as AV1ATION).
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 and 
between 3 and 4 hours 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 PK 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 the 
 M3 was similar to or 
slightly higher than that of balovaptan and exposure to the 
 M2 
was lower than that of parent.  M3 and M2 qualify as major metabolites in plasma.
 
 
Renal excretion was found to be the major pathway of elimination (approximately 53% of 
the material recovered, mostly composed of metabolites) with the primary route of 


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excretion via metabolism to M1 into urine, accounting for approximately 23% of the dose.  
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 found to be safe and well tolerated in clinical trials.  No 
particular safety concerns have emerged, and no adverse drug reactions have been 
identified. More specifically, creatine phosphokinase (CPK) was monitored in previous 
trials based on observations of muscle toxicity in rats; no changes of any importance 
were noticed.  Similarly, no hematology or neurotoxicity findings have been observed in 
clinical trials.  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 light of convulsions 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.  In healthy volunteer 
studies, one serious adverse event (SAE) of bacterial infection (suspected Lyme disease) 
was reported in Study BP29412; this SAE was considered unrelated to study drug.
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 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 recently started study in ASD pediatric patients (BP30153).
Based on the review of all available data, it is currently recommended not to exceed a 
plasma exposure equivalent to 20 mg qd in adult subjects in clinical trials with a 
continuous treatment duration longer than 4 weeks.
Refer to the Investigator's Brochure for details on nonclinical and clinical studies.


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1.3
STUDY RATIONALE AND BENEFIT-RISK ASSESSMENT
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 patients.  
To date, balovaptan has been investigated in 209 healthy subjects in 6 completed Phase 
1 studies (Studies BP25694, BP28318, BP28977, BP29279, BP29412, and WP40038).
Additionally, balovaptan was investigated in a Phase II study in 148 adult male patients 
with ASD (BP28420, “VANILLA”), about 30 of whom received 10 mg qd for 12 weeks.  
Adverse event reporting in healthy subjects did not reveal any particular safety alert for 
balovaptan.  The most frequently reported AE was headache, without an apparent 
dose-response relationship for balovaptan.  For the vast majority of AEs, the intensity 
was rated as mild or moderate.  The dose range in these studies was 0.5 mg to 76 mg 
(single dose) and 12 mg to 52 mg (multiple doses for 14 days).
As balovaptan exposure is expected to be decreased in the presence of rifampicin, the 
exposure of some of its metabolites may increase.  Given that multiple doses of
balovaptan from 12 mg to 52 mg qd showed linear M3 metabolite and parent PK 
exposures, i.e., a 5-fold higher exposure than the selected dose of 10 mg, and were well 
tolerated, the risk of AEs due to elevated metabolite levels is considered low. The 
metabolite M2 plasma concentrations have only been measured after daily doses of 
balovaptan 10 mg, but as M2 itself is metabolized by CYP3A4, the risk of AEs due to 
elevated M2 metabolite levels is also considered to be low. However, 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.
2.
OBJECTIVES AND ENDPOINTS
This study will investigate the effect of rifampicin treatment on the PK of balovaptan and 
its major metabolites M2 and M3.  Table 1 presents the objectives and endpoints.
Table 1 Objectives and Corresponding Endpoints
Objectives
Endpoints
Primary
To investigate the effect of multiple doses 
of rifampicin, a potent CYP3A4 inducer, on 
the pharmacokinetics of balovaptan 10 mg 
qd and its major metabolites M2 and M3 at 
steady state in healthy subjects
Primary PK endpoints: Cmax and area 
under the concentration curve from time 0 
to 24 hours (AUC0-24h) for balovaptan, M2, 
and M3 following 10 days of qd dosing of 
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Secondary
To evaluate the safety and tolerability of 
balovaptan 10 mg qd administered alone 
and in combination with rifampicin in 
healthy subjects
AEs, clinical laboratory values, vital signs, 
electrocardiogram (ECG), Columbia-
Suicide Severity Rating Scale, and 
physical examination
To investigate the effect of multiple doses 
of rifampicin on secondary PK parameters 
of multiple doses of balovaptan 10 mg qd 
and its major metabolites M2 and M3 at 
steady state in healthy subjects
Balovaptan and metabolite M2 and M3: 
tmax; metabolite to parent ratio; any other 
parameter as appropriate
Tertiary/Exploratory
To evaluate the plasma exposure of 
rifampicin 600 mg qd
Rifampicin Cmax, AUC0-24h, tmax, area under 
the plasma concentration-time curve from 
time zero to the last measurable plasma 
concentration time point (AUC0-last)
following 6 and 16 days of qd dosing of 
rifampicin, and any other parameter as 
appropriate
To investigate the relationship between the 
CYP3A4 genotype, among othersa, and 
steady state balovaptan exposure, and the 
influence of the CYP3A4 genotype, among 
others, on the effect of rifampicin on 
balovaptan PK
The pharmacogeneticsb of metabolizing 
enzymes, tranferases, transporters, etc, 
possibly involved in the absorption, 
distribution, metabolism and excretion of 
balovaptan and its major metabolites (e.g., 
CYP3A and P-glycoprotein)
aThe relationship between other genotypes and PK of balovaptan may also be explored.
bResults may be pooled with data from other studies of balovaptan.
3.
STUDY DESIGN
3.1
DESCRIPTION OF THE STUDY
This study will be a single-center, non-randomized, open-label, one-sequence, two-
period within-subject study to investigate the effects of multiple doses of rifampicin on 
the PK and safety of multiple doses of balovaptan in healthy subjects.  The study will be 
conducted at 1 site in the Netherlands.
Screening will be conducted up to 3 weeks prior to admission to the clinical research unit
(CRU).  Subjects will be in the CRU for 2 study periods. In both periods, subjects will be 
admitted on Day -1 (the day before dosing begins); Day 1 will be the first day of dosing. 
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completed.  There will be a minimum of a 14-day to a maximum of a 21-day washout 
between the last dose in Period 1 and the first dose in Period 2.  Subjects will therefore 
return to the CRU on Day 24 or up to Day 30 of Period 1 (Day -1 of Period 2) to begin 
Period 2. Day 1 will be the first day of dosing. In Period 2, subjects will be discharged on 
Day 17 after all assessments have been performed.  Subjects will return for a follow-up 
visit between 14 to 21 days after the last dose.  
Subjects will receive the following study drugs in the 2 study periods after an overnight 
fast of at least 8 hours:
Period 1:

Days 1 to 10: oral balovaptan 10 mg qd

Days 11 to 23 (or up to Day 30): no study drug
Period 2:

Days 1 to 6: rifampicin 600 mg qd

Days 7 to 16: balovaptan 10 mg qd + rifampicin 600 mg qd 
Figure 1 presents an overview of the study design.  A schedule of activities is provided in 
Appendix 1.
Abbreviations: PK = pharmacokinetic; qd = once daily; 
aFull balovaptan PK profile on Day 10/11 of Period 1 and Day 16/17 of Period 2; trough concentrations on 
other days. Full rifampicin PK profile on Days 6/7 and 16/17 of Period 2
Subjects will undergo study assessments and procedures as detailed in the schedule of 
assessments (Appendix 1), including assessments of AEs and concomitant medications, 
Screening
(Day -21 to 
Day -2)
Days 1 to 10:
balovaptan qd
Days 11 to 23 (or up to 
Day 30): washout
Days 1 to 6: Rifampicin 600 mg qd
Days 7 to 16: balovaptan 10 mg qd
+ rifampicin 600 mg qd
Balovaptan, M2 and M3 
PK samplinga on 
Days 2, 4, 8, 9, 10/11
Balovaptan M2 and M3 PK samplinga on 
Days 8, 10, 14, 15, 16/17
Rifampicin PK samplinga on Days 6/7 and 
16/17
Follow-up
(14 to 21 days 
after last dose)
Period 1
Period 2
Figure 1
Study Schema


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vital signs, 12-lead ECGs, clinical laboratory evaluations and blood sampling for PK
assessments.
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 end of the 
study is expected to occur 12 weeks after the last subject is enrolled.
The total length of the study, from screening of the first subject to the end of the study, is 
expected to be approximately 12 weeks.
3.3
RATIONALE FOR STUDY DESIGN
3.3.1
Rationale for Open-Label Design
The primary objective in this study is to assess whether and to what extent exposure to 
balovaptan may change in the presence of the CYP3A4 inducer rifampicin.  Drug
concentrations in plasma are assumed to be objective measurements, which cannot be 
affected by volunteer or staff behavior.  This is different for tolerability reporting, for 
which it is conceivable that knowing what treatment is being administered may affect 
reporting or causality interpretation.  Therefore, it is generally accepted to conduct
drug-drug interaction studies with an open-label design, without placebo treatments, and 
since evaluation of safety and tolerability is only a secondary objective, an open-label 
design was deemed appropriate.  
3.3.2
Rationale for Balovaptan and Rifampicin Dose and Schedule
3.3.2.1
Balovaptan
A dose of 10 mg balovaptan has shown statistically significant changes in efficacy 
parameters in patients with ASD and is the current recommended Phase 2 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 life.  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 
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 qd dosing with balovaptan 10 mg.  Based on these 
observations, steady state plasma levels of balovaptan and its major metabolites M2 and 
M3 under induction of CYP3A4 can be expected to be observed with 10 days of dosing 
with balovaptan.
3.3.2.2
Rifampicin
Multiple doses of rifampicin lead to induction of CYP3A4 (Yamashita et al., 2013); the 
dose of rifampicin to be used in this study is an approved and therapeutically relevant 
dose and has an acceptable safety profile; common AEs following daily or intermittent 


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rifampicin therapy include thrombocytopenia with or without purpura, headache, 
dizziness, nausea, vomiting, and increases in liver function enzymes (Sanofi Aventis, 
2018).
3.3.3
Rationale for Study Population
Healthy subjects 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.
Female and male subjects may be included.  Female subjects of childbearing potential 
must use one highly effective non-hormonal contraception combined with a barrier 
method. Female subjects of childbearing potential must therefore use a non-hormonal 
intra-uterine device (IUD) combined with a barrier method. Female subjects who have 
undergone tubal ligation must also use a barrier method of contraception.  Male subjects 
must agree to use contraceptive measures and agree to refrain from donating sperm.  
The study will enroll at least 50% male subjects, as ASD is more prevalent in males. 
4.
MATERIALS AND METHODS
4.1
SUBJECTS
The target population will be healthy male (at least 50% of enrolled subjects) and female 
subjects.  Sixteen subjects will be enrolled to obtain 12 evaluable subjects.  Enrolled 
subjects who withdraw from the study may be replaced at the discretion of the 
Investigator 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 and female subjects, aged 18 to 65 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 of 18 to 30 kg/m2, inclusive, at Screening.
3.
Women of childbearing potential must agree to use one highly effective 
non-hormonal contraceptive method combined with a barrier method from screening 
until 90 days after the last dose of study drug.  

Highly effective methods include:
o
A non-hormonal intrauterine device implanted at least 2 months prior to 
Screening.

Acceptable barrier methods include:
o
Condom, diaphragm, cervical/vault cap, or vaginal sponge.


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Women are considered to be of non-child bearing potential if postmenopausal at 
least 12 months of amenorrhea and confirmed by follicle-stimulating hormone (FSH), 
or at least 24 months if on hormone replacement therapy), or surgically sterile 
(absence of ovaries and/or uterus; has undergone bilateral tubal ligation). Female 
subjects who have undergone bilateral tubal ligation must also use a barrier method.
4.
For men: agreement to use contraceptive measures, and agreement to refrain from 
donating sperm, as defined below:
With female partners of childbearing potential or pregnant female partners, men 
must use a condom during the treatment period 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 or Dutch.
4.1.2
Exclusion Criteria
Subjects who meet any of the following criteria will be excluded from study entry:
1.
Female subjects who are pregnant or lactating.
2.
If female of childbearing potential, a positive serum pregnancy test at Screening or 
at admission (Day -1 of either period).  
3.
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.  
4.
History of any clinically significant gastrointestinal, renal, hepatic, 
broncho-pulmonary, neurological, psychiatric, cardiovascular, endocrinological, 
hematological, lymphatic, musculoskeletal, genitourinary, immunological, 
dermatological, connective tissue or allergic disease, metabolic disorder, or cancer.  
5.
Signs or symptoms potentially indicative of peripheral neuropathy.
6.
History or evidence of any medical condition potentially altering the absorption, 
metabolism or elimination of drugs.  Surgical history of the gastrointestinal tract 
affecting gastric motility or altering the gastrointestinal tract, with the exception of 
uncomplicated appendectomy, which is allowed.  
7.
A history of clinically significant hypersensitivity (e.g., to drugs, including rifampicin
and balovaptan, and excipients) or allergic reactions.  
8.
In the opinion of the Investigator, 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.
9.
History or presence of clinically significant ECG abnormalities before study drug 
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10. Known personal or family history of congenital long QT syndrome or sudden death.
11. Subjects with screening or predose baseline mean QT interval corrected using 
Fridericia’s formula (QTcF) >450 msec or <300 msec (using the same upper QTcF 
limit in both males and females).
12. Notable resting bradycardia (mean HR <40 bpm) on screening or predose baseline 
ECG. Notable resting tachycardia (mean HR > 90 bpm) on screening or predose 
baseline ECG.
13. Screening or baseline ECGs 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).
14. Screening or baseline ECG with evidence of clinically relevant abnormalities, e.g. 
atrial fibrillation, atrial flutter, complete bundle branch block, Wolf-Parkinson-White 
Syndrome, or cardiac pacemaker.
15. Clinically significant abnormalities in laboratory test results (including hepatic and 
renal panels, 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 of Period 1 to confirm eligibility. 
16. Confirmed (based on the average of ≥ 3 consecutive measurements) resting pulse 
rate greater than 100 or less than 40 bpm.  
17. Any suspicion or history of alcohol abuse and/or suspicion or history of regular 
consumption of drug of abuse.  Past alcohol and/or other substance abuse or 
addiction is also not allowed.
18. Positive urine alcohol test or urine drug screen at Screening or Day -1 of each 
treatment period (barbiturates, benzodiazepines, methadone, amphetamines 
[including ecstasy], methamphetamines, opiates, cocaine, or cannabinoids).
19. Alcohol consumption of >24 units per week for males and females. One unit of 
alcohol equals ½ pint (285 mL) of beer or lager, 1 glass (125 mL) of wine, or 1/6 gill 
(25 mL) of spirits.
20. Smokers who regularly smoke more than 5 cigarettes daily or equivalent and unable 
or unwilling not to smoke or use tobacco for 48 hours prior to dosing and during both
in-house periods.  
21. A screen positive for hepatitis B surface antigen (HBsAg), hepatitis C virus (HCV) 
antibody, or antibodies against human immunodeficiency viruses 1 and 2 
(HIV-1/HIV-2 antibodies).
22. Participation in an investigational drug or device study within 90 days 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.  
23. Any donation of blood or plasma or significant blood loss within 3 months prior to 
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24. 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
(see Section 4.4).  
25. History of coagulopathies, bleeding disorders or blood dyscrasias.
26. 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.
27. Dietary restrictions that would prohibit the consumption of standardized meals.
28. Subjects likely to need concomitant medication during the study period (including for 
dental conditions).  
29. Subjects who have received any prescribed systemic or topical medication within 4 
weeks (or within 5 times the elimination half-life of the medication, whichever is 
longer) of the first dose administration, unless in the opinion of the Investigator the 
medication will not interfere with the study procedures or compromise safety, and 
subjects who have received 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) of the first dose administration will also be excluded unless in 
the opinion of the Investigator the medication will not interfere with the study 
procedures or compromise safety.
30. Used the following within 7 days before the first study drug administration, unless in 
the opinion of the Investigator the medication will not interfere with the study 
procedures or compromise safety: 
- any non-prescribed systemic or topical medication
- any herbal remedies.
Paracetamol (up to 4 g/day within 7 days before study drug administration and 
2 g/day within 48 hours before study drug administration), dietary supplements, 
vitamins and minerals, hormonal contraceptives, and hormone replacement therapy 
are allowed.
31. Received any medications known to chronically alter drug absorption or elimination 
processes within 4 weeks before the first study drug administration, unless in the 
opinion of the Investigator the medication will not interfere with the study procedures 
or compromise safety.
32. Subjects who are unwilling to practice safe sex (use male or female condom) for the 
duration of the study.
33. Subjects under judicial supervision, guardianship or curatorship.
4.1.2.1
Rifampicin-related Exclusion Criteria
Contraindications
1.
Subjects diagnosed or suspected of having porphyria, and subjects with first-degree 
relatives diagnosed or suspected of having porphyria.


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2.
Subjects unwilling not to use contact lenses for the duration of rifampicin dosing
(Day 1 to Day 16 of Period 2) until 7 days after the last dose of rifampicin (subjects 
may resume wearing contact lenses on Day 23 of Period 2).
3.
Known hypersensitivity to rifampicin.
Additional exclusion criteria:

Taking any drug or herbal CYP3A modulator (e.g., erythromycin; St. John’s wort) 
within 4 weeks (or 5 half-lives, whichever is longer) or any other nutrients known
to modulate CYP3A activity (e.g., grapefruit juice; Seville orange) within 2 weeks 
prior to the first administration of study drugs.
4.2
METHOD OF TREATMENT ASSIGNMENT AND BLINDING 
This study will be a non-randomized, single-center, open-label, one-sequence, 
two-period study.  Each subject will receive a fixed sequence of 2 treatments.
Subjects who have met all eligibility criteria will receive a subject number upon inclusion 
in the study (subject numbers: 01 to 16).  Subject numbers will be allocated sequentially 
in the order in which the subjects are enrolled.  The subject number will ensure 
identification throughout the study.  Replacement subjects (if needed) will receive the 
number of the subject to be replaced, increased by 100 (e.g., 101 replacement number 
for subject number 01), and will be administered the same treatments in the same order.
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 subject number.  The Investigator will keep a screening log of all subjects 
screened in order to assess the numbers and characteristics of the excluded subjects, 
and the reasons for their exclusion.
4.3
STUDY TREATMENT AND OTHER TREATMENTS RELEVANT 
TO THE STUDY DESIGN
The investigational medicinal products (IMPs) for this study are balovaptan and 
rifampicin.
4.3.1
Study Treatment Formulation, Packaging, and Handling
4.3.1.1
Balovaptan
RO5285119/F17 10 mg dispersible tablet will be supplied by the Sponsor in high-density 
polyethylene bottles.  For information on the formulation and handling of balovaptan, see 
the Investigational Medicinal Product Dossier.
4.3.1.2
Rifampicin
Rifampicin (600 mg qd, administered as two 300-mg capsules) will be obtained 
commercially by the site.  For information on the formulation, packaging, and handling of 
rifampicin, see the local prescribing information for rifampicin.


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4.3.2
Study Treatment Dosage, Administration, and Compliance
The treatment regimens are summarized in Section 3.1.
Any overdose or incorrect administration of study drug should be noted on the 
Study Drug Administration electronic Case Report Form (eCRF).  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 discontinuation for subjects who experience AEs are provided 
in Section 5.1.
4.3.2.1
Balovaptan
A single dose of balovaptan 10 mg will be administered alone each morning of Days 1 to 
10 of Period 1 and in combination with rifampicin on Days 7 to 16 of Period 2, after an 
overnight fast of at least 8 hours in each period.  
4.3.2.2
Rifampicin
A single dose of rifampicin 600 mg will be administered alone on each morning of Days 
1 to 6 of Period 2, and in combination with balovaptan on Days 7 to 16 of Period 2.  In 
both periods, the study drugs will be swallowed whole with 240 mL of still water.  
Rifampicin should be taken at least 30 minutes prior to or at least 2 hours after a meal.
Therefore, rifampicin alone or in combination with balovaptan will be administered at 
least 30 minutes prior to breakfast; lunch, dinner, and afternoon/evening snacks will be 
administered 4 or more hours post dose.
In both periods, the study drugs will be swallowed whole with 240 mL of still water.
4.3.3
Investigational Medicinal Product Accountability
Balovaptan will be provided by the Sponsor where required by local health authority 
regulations.  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.  Rifampicin will be obtained 
commercially by the site.
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 (if supplied by 
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 
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Accurate records of all IMPs received at, dispensed from, returned to, and disposed of by 
the study site should be recorded on the Drug Accountability Form and Drug Dispensing 
Form.
4.4
CONCOMITANT THERAPY, PROHIBITED FOOD, AND 
ADDITIONAL RESTRICTIONS
Concomitant therapy consists of any medication (e.g., prescription drugs, 
over-the-counter drugs, vaccines, herbal or homeopathic remedies, nutritional 
supplements) used by a subject in addition to protocol-mandated treatment from 14 days 
prior to initiation of study drug to the study completion/discontinuation visit.  All such 
medications should be reported to the Investigator 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.  However, women of childbearing 
potential must agree to use an alternative highly effective non-hormonal contraception
method (e.g., non-hormonal IUD) combined with a barrier method for 90 days after the 
last dose of study drug.
Acetaminophen/paracetamol is allowed up to a maximum dose of 2 g/day up to 48 hours 
prior to dosing and during the in-house portion of the study, but total should not exceed 
4 g total during the week prior to dosing. Subjects will be instructed not to exceed these 
limits.
4.4.2
Prohibited Therapy
Any prescribed systemic or topical medication within 4 weeks (or within 5 times the 
elimination half-life of the medication, whichever is longer) prior to the first study drug 
administration through the follow-up visit or 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 through the follow-up visit, unless in the opinion of the Investigator the 
medication will not interfere with the study procedures or compromise safety. Any non-
prescribed systemic or topical medication or herbal remedies are prohibited for 7 days 
prior to the first study drug administration through follow-up. This is with the exception of 
medications to treat AEs, unless the rationale for the exception is discussed and clearly 
documented between the Investigator, the medical and safety monitor and the Roche 
clinical pharmacologist, and with the exception of permitted therapies per Section 4.4.1.  
All medication administered to manage AEs should be recorded on the Adverse Event 
Report Form.
The following concomitant medications in particular are prohibited, unless an exception 
has been agreed to as mentioned above.


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
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 (or within 5 half-lives, whichever is 
longer) prior to study drug administration in Period 1, including but not limited to the 
following drugs: rifampicin, rifabutin, glucocorticoids, carbamazepine, phenytoin, 
phenobarbital, and St. John’s wort, until follow-up, except for rifampicin in Period 2.

Investigational drug or medical device therapy (other than protocol-mandated study 
treatment) is prohibited 90 days 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 of study treatment and during study treatment.
4.4.3
Prohibited Food

Consumption of methylxanthine-containing products (e.g., coffee, tea, cola, 
chocolate) will be forbidden from 48 hours prior to dosing and during the stay in the 
CRU

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.  
4.4.4
Additional Restrictions/Considerations

Subjects should have fasted for at least 4 hours prior to the screening visit, each 
admission of the CRU, and the follow-up visit.

Water will be allowed ad libitum until 1 hour prior to dosing and from 1-hour post 
dosing.  The excessive consumption of fluids (greater than 3 liters per day) should 
be avoided.

For maximum absorption, rifampicin should be taken at least 30 minutes prior to or 
at least 2 hours after a meal. 

In each period, study drugs will be administered at least 30 minutes prior to
breakfast, after an overnight fast of at least 8 hours. Standardized meals will be 
served to all subjects at days and the times noted in Appendix 1.  On these days a 
standardized breakfast, lunch, snack, and dinner will be served at similar times of 
the day (the same meal at each respective meal time), and subjects will be required 
to complete the meal.  The standardized breakfast will be served 30 minutes post 
dose; standardized lunch, 4 hours post dose; and standardized dinner 10 hours post 
dose on Day 10 of Period 1, and Days 6 and 16 of Period 2 (post dose is relative to 
time of dosing of balovaptan alone, rifampicin alone, or balovaptan + rifampicin, as 
applicable). All other meals will conform to PRA standards. Lunch, dinner, and 
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
Between the screening visit and follow-up, alcohol must be restricted to no more 
than 2 units per day (1 unit of alcohol equals ½ pint (285 mL) of beer or lager, 
1 glass (125 mL) of wine, or 1/6 gill (25 mL) of spirits).  Alcohol consumption will not 
be allowed 48 hours before balovaptan dosing and while staying in the CRU.  
Random alcohol/drugs of abuse testing may be employed throughout the study to 
verify compliance.

Use of tobacco will not be permitted 48 hours prior to admission to the clinical unit 
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.
4.5
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 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, herbal or 
homeopathic remedies, nutritional supplements) used by the subject within 14 days prior 
to initiation of study treatment will be recorded.  At follow-up, 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, 
should include an evaluation of the head, eyes, ears, nose, and throat, and the 
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systems; complaint-driven genitourinary examination may be performed.  Any 
abnormality identified at baseline should be recorded on the General Medical History 
and Baseline Conditions eCRF.
Limited, symptom-directed physical examinations should be performed at specified 
postbaseline visits and as clinically indicated.  Changes from baseline abnormalities 
should be recorded in subject notes.  New or worsened clinically significant 
abnormalities should be recorded as AEs on the Adverse Event Report Form.
4.5.4
Vital Signs
Body temperature (tympanic), blood pressure, and pulse rate measurements will be 
collected on the days indicated in Appendix 1.  Blood pressure and pulse rate 
measurements will be performed after the subject has rested in a supine position for at 
least 5 minutes.
4.5.5
Laboratory, Pharmacokinetic, and Genotyping Samples
For sampling and sample processing procedures, storage conditions, and shipment 
instructions, see the laboratory manual.
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 Appendix 1.  
Additional blood or urine samples may be taken at the discretion of the Investigator if the 
results of any test fall outside the reference ranges, or clinical symptoms necessitate 
additional testing to monitor subject safety.  
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 
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performed if abnormalities are observed and if deemed necessary by the 
Investigator or designee, in particular when blood or protein is positive or strong 
positive.  If there is an explanation for the positive dipstick result, it should be 
recorded, and there is no need to perform laboratory for microscopy and culture.

Coagulation:  prothrombin time (international normalized ratio) and activated 
thromboplastin time at Screening and Day -1 of Period 2.

Blood chemistry panel: sodium, potassium, glucose (fasting), creatinine, total and 
conjugated bilirubin, alkaline phosphatase, alanine aminotransferase (ALT), 
aspartate aminotransferase (AST), gamma-glutamyl transferase, CPK

Drugs of abuse (including cannabinoids, amphetamines [including ecstasy], 
methamphetamines, opiates, methadone, cocaine, benzodiazepines, and 
barbiturates) and cotinine will be measured in urine at Screening and on Day -1 of 
each study period.

Urine alcohol test at Screening and on Day -1 of each study period.

Viral serology:  HIV-1 and HIV-2 antibodies, HBsAg, and HCV antibody at Screening.

Pregnancy test: FSH and beta-human chorionic gonadotropin (serum pregnancy 
test) will be performed at Screening in all females.  At other time points, a serum
pregnancy test will be performed in 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.  
Plasma samples for PK analysis will be shipped to designated bioanalytical laboratories 
for analysis.
Plasma samples collected for PK analysis will be destroyed after the final Clinical Study 
Report has been issued.
4.5.5.3
Genotyping Samples
A mandatory 3 mL whole blood sample will be taken for DNA extraction from every 
subject during the study on Day -1 of Period 1.  The DNA may be used to determine 
whether genes relating to drug-metabolizing enzymes and/or drug transporter enzymes 
affect the PK and/or the safety profile of balovaptan.  Genotyping samples will be sent to 
the Sponsor or a designee for analysis.  This specimen will be destroyed no later than 2 
years after the final Clinical Study Report has been issued.
The Sponsor maintains confidentiality standards by coding each subject enrolled in the 
study through assignment of a unique subject identification number. This number is used 
to code the samples. Subject names are not included in samples that are sent for 
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4.5.6
Electrocardiograms
Triplicate ECGs will be collected at the time points indicated in Appendix 1.  
Electrocardiograms will be collected after the subject has rested in a supine position for 
at least 5 minutes.  
4.5.7
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 Research Physician at 
the time points as specified in Appendix 1.
4.5.8
Drug and Metabolite Concentration Measurements
Plasma concentrations of balovaptan and its metabolites M2 and M3 will be measured 
using a specific and validated liquid chromatography-tandem mass spectrometry 
(LC-MS/MS) method.  Plasma concentrations of rifampicin will also be measured using a 
separate, specific and validated LC-MS/MS method.
4.5.9
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:

12-lead ECGs

Vital signs measurements

PK blood sampling

Laboratory safety tests 
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 or Sponsor determines may jeopardize 
the subject’s safety if he or she continues to receive study treatment (see 
Section 5.1.2) for additional guidance on the management of toxicities and of 
subjects who experience AEs)

Investigator or Sponsor determines it is in the best interest of the subject

Pregnancy


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The primary reason for study treatment discontinuation should be documented on the 
appropriate eCRF.  Subjects who discontinue study treatment prematurely may be
replaced.
Subjects will return to the clinic for a follow-up visit 14 to 21 days after the last dose of 
study drug (see Appendix 1 for additional details).
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 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 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.  If a subject requests to be withdrawn from the study, this request 
must be documented in the source documents and signed by the Investigator.  Subjects
who withdraw from the study may be replaced.
4.6.3
Study Discontinuation
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 
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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, 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 RO5285119 (balovaptan) Investigator’s Brochure for a complete summary of 
safety information.  
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 AEs.  Treatment will be discontinued in any subject 
with a medical condition that the Investigator or Sponsor determines may jeopardize the 
subject’s safety if he or she continues to receive study treatment.
 
 
 
 
 
 
  
 
 
 
 
  
 


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5.1.2
Management of Toxicities and 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 
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The Investigator should also consider discontinuation of treatment with balovaptan and 
rifampicin, in particular, if an AE is of severe intensity and considered to be possibly 
related to balovaptan and rifampicin, respectively.  
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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5.2
SAFETY PARAMETERS AND DEFINITIONS
Safety assessments will consist of monitoring and recording AEs, including SAEs and 
AEs of special interest, 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.
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) (see Section 5.3.5.8 for more 
information)

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

AEs that are related to a protocol-mandated intervention, including those that occur 
prior to assignment of study treatment (e.g., screening invasive procedures such as 
biopsies)
5.2.2
Serious Adverse Events (Immediately Reportable to the 
Sponsor)
An SAE is any AE 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, places the subject at 
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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 inpatient hospitalization (see 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 drug

Is a significant medical event in the Investigator’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 AE (e.g., rated as mild, moderate, or severe, or according to National Cancer Institute 
Common Terminology Criteria for Adverse Events; 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 AEs are required to be reported by the Investigator 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 to the 
Sponsor immediately (i.e., no more than 24 hours after learning of the event; see 
Section 5.4.2 for reporting instructions).  Adverse events of special interest 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 
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”).


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•
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.
•
Neurological abnormalities emerging from AE reporting or neurologic examination
that are potentially indicative of dysfunction of peripheral nerves or otherwise
represent a neurological deficit and for which no other explanation is apparent
•
Adverse events such as: arrhythmia, syncope, dyspnea, palpitations, or chest pain
5.3 
METHODS AND TIMING FOR CAPTURING AND ASSESSING 
SAFETY PARAMETERS 
The Investigator is responsible for ensuring that all AEs (see Section 5.2.1 for definition) 
are recorded on the Adverse Event Report Form and reported to the Sponsor in 
accordance with instructions provided in this section and in Sections 5.4−5.6.  The 
Investigator is also responsible for reporting medical device complaints (see 
Section 5.4.5). 
For each AE recorded on the Adverse Event Report Form, the Investigator will make an 
assessment of seriousness (see Section 5.2.2 for seriousness criteria), severity (see 
Section 5.3.3), and causality (see Section 5.3.4).   
5.3.1 
Adverse Event Reporting Period 
Investigators will seek information on AEs at each subject contact.  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 Form, if appropriate). 
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 up to 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. 
5.3.2 
Eliciting Adverse Event Information 
A consistent methodology of non-directive questioning should be adopted for eliciting AE 
information at all subject evaluation timepoints.  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?” 


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5.3.3
Assessment of Severity of Adverse Events
Table 2 provides guidance for assessing AE severity.
Table 2
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 an
SAE (see Section 5.2.2).
5.3.4
Assessment of Causality of Adverse Events
Investigators 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 3):

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 
occurrence of the event
Table 3
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 
(RELATED)
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 (NOT 
RELATED)
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).


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For subjects receiving combination therapy, causality will be assessed individually for 
each protocol-mandated therapy.
5.3.5
Procedures for Recording Adverse Events
Investigators should use correct medical terminology/concepts when recording AEs on 
the Adverse Event eCRF (and Form, if appropriate).  Avoid colloquialisms and 
abbreviations.
Only one AE term should be recorded in the event field on the Adverse Event eCRF
(and Form, if appropriate).
5.3.5.1
Diagnosis versus Signs and Symptoms
A diagnosis (if known) should be recorded on the Adverse Event eCRF (and Form, if 
appropriate) 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 Form, if appropriate).  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.
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 Form, if appropriate).  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 3 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 Form, if 
appropriate) if it is unclear as to whether the events are associated.


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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 Form, if appropriate).  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 Form, if appropriate).  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).  If a nonserious AE becomes 
serious, the Investigator will complete the paper Clinical Trial Serious Adverse 
Event/Adverse Event of Special Interest Reporting Form and send the form to Roche 
drug safety.  The Adverse Event eCRF if already completed should also be updated to 
serious to correspond with the Adverse Event Reporting Form information sent to drug 
safety.
A recurrent AE is one that resolves between subject evaluation timepoints and 
subsequently recurs.  Each recurrence of an AE should be recorded as a separate event 
on the Adverse Event eCRF (and Form if appropriate).
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)

Results in a medical intervention (e.g., potassium supplementation for hypokalemia) 
or a change in concomitant therapy

Is clinically significant in the Investigator’s judgment 
It is the Investigator’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 Form if 
appropriate).
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 Form if 
appropriate), 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 


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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 Form if appropriate)
(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’s judgment 
It is the Investigator’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 Form if appropriate).
Observations of the same clinically significant vital sign abnormality from visit to visit 
should only be recorded once on the Adverse Event eCRF (and Form if appropriate)
(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, investigators 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 Form if 
appropriate) (see Section 5.3.5.1) and reported to the Sponsor immediately (i.e., no 
more than 24 hours after learning of the event), either as an SAE or an AE of special 
interest (see Section 5.4.2).


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5.3.5.7
Deaths
All deaths that occur during the protocol-specified AE reporting period (see 
Section 5.3.1), regardless of relationship to study drug, must be recorded on the 
Adverse Event eCRF (and Form if appropriate) 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 Form if appropriate).  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 Form if appropriate).  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 AE reporting period should be reported as described in 
Section 5.6.
5.3.5.8
Pre-existing Medical Conditions
A pre-existing 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 pre-existing medical condition should be recorded as an AE only if the frequency, 
severity, or character of the condition worsens during the study.  When recording such 
events on the Adverse Event eCRF (and Form if appropriate), it is important to convey 
the concept that the pre-existing 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 an SAE (per the definition of SAE
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 an SAE:

Hospitalization for respite care

Planned hospitalization required by the protocol


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
Hospitalization for a pre-existing 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 an 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 must report 
such events to the Sponsor immediately; under no circumstances should reporting take 
place more than 24 hours after the Investigator learns of the event.  The following is a 
list of events that the Investigator must report to the Sponsor within 24 hours after 
learning of the event, regardless of relationship to study drug:

SAEs (defined in Section 5.2.2; see Section 5.4.2 for details on reporting 
requirements)

AEs 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)

Accidental overdoses or medication errors (see Section 5.4.4 for details on reporting 
requirements)

Medical device complaints (see Section 5.4.5 for details on reporting requirements)
The Investigator 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
Investigators must also comply with local requirements for reporting SAEs to the local 
health authority and Institutional Review Board (IRB)/Ethics Committee (EC).


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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 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.  
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 SAEs 
caused by a protocol-mandated intervention should be reported.  The paper Clinical Trial 
Serious Adverse Event/Adverse Event of Special Interest Reporting Form provided to 
investigators 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 email address provided to 
investigators.
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.  Investigators 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 Adverse Event eCRF (and Form if appropriate) 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 more than 21 days after the last dose of study 
treatment are provided in Section 5.6.


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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 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 investigators.  Pregnancy 
should not be recorded on the Adverse Event eCRF.  The Investigator should 
discontinue 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 SAEs 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 
Form, if appropriate).  In addition, the Investigator 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 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 should submit a Clinical Trial 
Pregnancy Reporting Form when updated information on the course and outcome of the 
pregnancy becomes available.  An investigator 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 an SAE (as the Sponsor considers abortions to be 
medically significant), recorded on the Adverse Event eCRF and Form, and reported to 
the Sponsor immediately (i.e., no more than 24 hours after learning of the event; see 
Section 5.4.2).  


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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 an SAE, recorded on the Adverse Event eCRF and 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 AEs but may result in AEs.  All special situations 
associated with balovaptan, regardless of whether they result in an AE, should be 
recorded on the Adverse Event eCRF and Form and reported to the Sponsor 
immediately (i.e., no more than 24 hours after learning of the event).  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.

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 balovaptan and rifampicin, each AE associated with a special situation should be 
recorded separately on the Adverse Event eCRF and 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).  For 
balovaptan, AEs associated with special situations should be recorded as described 
below for each situation:

Accidental overdose:  Enter the AE term.  Check the “Accidental overdose” and 
“Medication error” boxes.


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
Medication error that does not qualify as an overdose:  Enter the AE term.  Check 
the “Medication error” box.

Medication error that qualifies as an overdose:  Enter the AE 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 2 Adverse Event eCRF pages and forms, 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.
5.4.5
Reporting Requirements for Medical Device Complaints
The Investigator must report all medical device complaints to the Sponsor.  The 
Investigator should document as much information as possible on the IMP Deviation 
Form, including the product batch number, and forward the form to the Sponsor 
immediately (i.e., no more than 24 hours after learning of the event) (refer to the 
pharmacy manual for further details).  If the medical device results in an AE to the study 
subject, the event must be reported on the Adverse Event eCRF and Form and 
submitted through the electronic data capture (EDC) system.  If the event is serious, the 
Adverse Event eCRF and Form must be completed immediately (i.e., no more than 
24 hours after learning of the event), as outlined in Section 5.4.2.
5.5
FOLLOW-UP OF SUBJECTS AFTER ADVERSE EVENTS
5.5.1
Investigator Follow-Up
The Investigator should follow each AE until the event has resolved to baseline grade or 
better, the event is assessed as stable by the Investigator, 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 AEs (with dates) should be documented on the 
Adverse Event eCRF (and Form, if appropriate) 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, AEs of special interest, 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 case.


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5.6
ADVERSE EVENTS THAT OCCUR AFTER THE ADVERSE
EVENT REPORTING PERIOD
The Sponsor should be notified if the Investigator becomes aware of any SAE that 
occurs after the end of the AE reporting period (defined in Section 5.3.1) 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 Report Form.  The 
Investigator 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 AEs of special interest against 
cumulative product experience to identify and expeditiously communicate possible new 
safety findings to investigators, IRBs, ECs, and applicable health authorities based on 
applicable legislation.
To determine reporting requirements for single AE cases, the Sponsor will assess the 
expectedness of these events using the following reference document:

RO5285119 (Balovaptan) Investigator’s Brochure

Rifampicin Summary of Product Characteristics
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’s assessment of causality 
and seriousness, with allowance for upgrading by the Sponsor as needed.
6.
STATISTICAL CONSIDERATIONS AND ANALYSIS PLAN
A statistical analysis plan will be generated by the biostatistics department of PRA
Health Sciences and will be finalized prior to database lock.
6.1
DETERMINATION OF SAMPLE SIZE
This is an exploratory study for which no formal statistical hypothesis will be tested.  
Therefore, the sample size is chosen to estimate with sufficient precision the effect of 
CYP3A4 induction on balovaptan exposure.  
The sample size of 12 evaluable subjects was chosen based on balovaptan 
within-subject variability (CV%) of around 39% for AUC0-24h and 36% for Cmax as 
obtained from the 12 mg repeat dose in study BP25694.  Based on the higher value of 


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39%, it was estimated that with 12 subjects the half-width of the 90% confidence interval 
for the ratio of treatment geometric means of the combination (balovaptan + rifampicin) 
versus balovaptan alone would be obtained by dividing/multiplying the ratio estimate by 
a factor of 1.30.
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 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.
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 treatment.  Reasons for withdrawal from study will be listed and 
summarized.  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 2
For laboratory and 12-lead ECG data, subject listings will be presented with 
abnormalities flagged.
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.


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Individual plasma concentrations at each sampling time point for balovaptan, M2, M3, 
and rifampicin 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 semi-logarithmic scales.
The primary PK parameters are balovaptan, M2, and M3 Cmax, and AUC0-24h following 10 
days of qd dosing of balovaptan 10 mg.  Secondary PK parameters are balovaptan, M2, 
and M3 tmax; the metabolite to parent ratio; and any other parameter as appropriate.  
Rifampicin Cmax, AUC0-24h, tmax and AUC0-last are tertiary/exploratory parameters.  
Additional PK parameters may be derived as appropriate.
All PK parameters for balovaptan, M2, M3, and rifampicin will be presented by individual 
listings and summary statistics by treatment including means, geometric means, 
medians, ranges, standard deviations and coefficients of variation. A linear mixed model 
analysis will be applied to analyze the log-transformed primary PK parameters.  The 
model assumes fixed effect for treatment and a random effect for subject.  Estimates of 
geometric mean ratios together with the corresponding 90% CIs will be derived for the 
comparisons of the PK parameters for balovaptan plus rifampicin (test) vs. balovaptan 
alone (reference).
6.7
OTHER ANALYSES
6.7.1
Clinical Genotyping
The relationship between CYP3A4 genotype and steady state balovaptan exposure and 
the influence of CYP3A4 genotype on the effect of rifampicin on balovaptan PK and 
CYP3A enzyme activity will be evaluated. Other isoforms may also be analysed.
7.
DATA COLLECTION AND MANAGEMENT
7.1
DATA QUALITY ASSURANCE
The Sponsor will supply eCRF specifications for this study.  The contract research 
organization (CRO), PRA, will be responsible for data management of this study, 
including quality checking of the data.  Data entered manually will be collected via EDC 
through use of eCRFs.  Sites will be responsible for data entry into the EDC system.  In 
the event of discrepant data, the PRA will request data clarification from the sites, which 
the sites will resolve electronically in the EDC system.
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.
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 


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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.  
7.2
ELECTRONIC CASE REPORT FORMS 
Electronic CRFs are to be completed through use of a Sponsor-designated 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.


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To facilitate source data verification and review, the investigators 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 
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 patient-reported outcome 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.  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 US or under a US Investigational New 
Drug (IND) Application will comply with US Food and Drug Administration regulations 
and applicable local, state, and federal laws.  Studies conducted in the European Union
(EU) or European Economic Area will comply with the EU Clinical Trial Directive 
(2001/20/EC).


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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.
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 


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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.  Investigators 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 AEs to the Sponsor, investigators must 
comply with requirements for reporting SAEs to the local health authority and IRB/EC.  
Investigators may receive written IND safety reports or other safety-related 
communications from the Sponsor.  Investigators 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 Roche 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.
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 


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course of the study and for 1 year after completion of the study (see definition of end of 
study in Section 3.2).
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 and 
governmental 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
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 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
The Sponsor of the trial is F.  Hoffmann-La Roche Ltd.  The Sponsor has contracted 
PRA, who will be delegated responsibility for various aspects of this clinical trial.
One site will participate to enroll up to 16 subjects.  
Central facilities will be used for certain study assessments throughout the study (e.g., 
specified laboratory tests and PK analyses), as specified in Section 4.5.  Accredited local 
laboratories will be used for routine monitoring; local laboratory ranges will be collected.
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 


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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.
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 PRA.  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
American Psychiatric Association.  Diagnostic and statistical manual of mental disorders.  
5th ed.  Washington, DC: American Psychiatric Association, 2013.
Al-Mosalem OA, El-Ansary A, Attas O, et al. Metabolic biomarkers related to energy 
metabolism in Saudi autistic children. Clin Biochem. 2009; 42:949–957.
Developmental Disabilities Monitoring Network Surveillance Year 2010 Principal
Investigators; Centers for Disease Control and Prevention (CDC).  Prevalence of autism 
spectrum disorder among children aged 8 years – Autism and Developmental
Disabilities Monitoring Network, 11 sites, United States, 2010.  MMWR Surveill Summ.  
2014;63(2):1-21.
Levy SE, Mandell DS, Schultz RT: Autism.  Lancet. 2009;374:1627–1638.
May T, Brewer WJ, Rinehart NJ, et al.  Differential Olfactory Identification in Children
with Autism and Asperger’s Disorder: A Comparative and Longitudinal Study.  J Autism 
Dev Disord.  2011;41(7):837–847.
Meyer-Lindenberg A, Kolachana B, Gold B, et al.  Genetic variants in AVPR1A linked to 
autism predict amygdala activation and personality traits in healthy humans.  Mol
Psychiatry.  2009;14(10):968–975.
Sanofi Aventis.  Rifadin 300 mg Capsules Summary of Product Characteristics. Updated 
20-Feb-2018. Accessed 03 May 2018 at: 
https://www.medicines.org.uk/emc/product/6384/smpc
Tobin VA, Hashimoto H, Wacker DW, et al.  An intrinsic vasopressin system in the
olfactory bulb is involved in social recognition.  Nature.  2010;464(18):413-419.
Yamashita F, Sasa Y, Yoshida S, et al. Modeling of rifampicin-induced CYP3A4 
activation dynamics for the prediction of clinical drug-drug interactions from in vitro data. 
PLoS One. 2013;8(9): e70330


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Appendix 1
Schedule of Activities
Washout of minimally 14 and maximally 21 days
between the study periods
Assessments
Screening
Period 1
Period 2
Follow-up
Day -21 to 
Day -2
Day 
-1
Days
1-9
Day 
10
Day 
11
Washout:
for 14-21 
days after 
last dose
Day 
-1
Days 
1-5
Day 
6
Day 
7
Days 
8-15
Day 
16
Day 
17
Days
30 – 37  
of Period 2
Written informed consent
X
Inclusion and exclusion criteria
X
Demographics
X
Medical history
X
Medication history
X
Study drug administration: balovaptana
X
X
X
X
X
Study drug administration: rifampicina
X
X
X
X
X
Standardized mealsb
X
X
X
Pharmacokinetic sampling balovaptanc 
X
X
X
X
X
X
Clinical genotyping sample
X
Pharmacokinetic sampling rifampicind
X
X
X
X
Physical examination
Xe
X
Xf
X
Xf
Triplicate 12-lead ECGg
X
X
X
X
X
X
X
X
X
X
X
X
X
Vital signsh
Xi
Xi
X
Xi
Xi
Xi
Xi
X
X
Xi
Xi
Xi
Hematologyj
X
X
X
X
X
X
X
X
X
X
Serum chemistryj
X
X
X
X
X
X
X
X
X
X
Urinalysisj
X
X
X
X
X
X
Coagulation
X
X
Urine drug screen 
X
X
X
Urine alcohol test
X
X
X
Serology
X
Pregnancy testk
X
X
X
X
Resident at study centerl
X
X
X
X
X
X
X
X
X
X
X
Adverse event monitoring
X
X
Review concomitant medications
X
X
C-SSRS
X
X
X
Abbreviations: BMI = body mass index; C-SSRS= Columbia Suicide Severity Rating Scale; ECG = electrocardiogram; qd = once daily; 


Appendix 1
Schedule of Activities (cont.)
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a
Study drugs will be administered after an overnight fast of at least 8 hours, 30 minutes prior to breakfast. On days rifampicin and balovaptan are coadministered, they will 
be administered at the same time. Water will be allowed ad libitum up to 1 hour prior to dosing and from 1 hour post dosing, and no food is allowed for at least 4 hours 
post dose—lunch, dinner, and afternoon/ evening snacks will be provided 4 or more hours after dosing.
b
A standardized breakfast will be provided at least 30 minutes after dosing of balovaptan alone, rifampicin alone, or balovaptan + rifampicin together in either period. A 
standardized lunch and standardized dinner will be provided 4 and 10 hours post dose, respectively, in both periods.  Standardized afternoon and evening snacks will be 
provided as well.
c
Samples to measure the concentration of balovaptan, M2, and M3 (one sample per time point to measure all 3 analytes) will be taken at predose, and 0.5, 1, 1.5, 2, 2.5, 3, 
4, 6, 8, 12, 16, 24, hours post dose on Days 10/11 of Period 1 (16- and 24-hour samples are on the morning of Day 11) and 16/17 of Period 2 (16- and 24-hour samples 
are on the morning of Day 17).  A predose sample to measure the concentration of balovaptan, M2, and M3 will also be taken on Days 2, 4, 8, and 9 in Period 1, and on 
Days 8, 10, 14, and 15 in Period 2 (see also Appendix 2).  
d
Samples to measure the concentration of rifampicin will be taken predose and at 1, 1.5, 2, 3, 4, 6, 12, and 24 hours post dose of rifampicin on Days 6/7 and 16/17 of 
Period 2 (24-hour samples are on the morning of Days 7 and 17) (see also Appendix 2).  
e
Includes measurement of height, weight and BMI.
f
Includes measurement of weight.
g
Triplicate ECGs will be collected as follows: in Period 1, on Day -1 and predose and 3 hours after dosing of balovaptan on Days 3, 7, and 10 and in the morning of Day 11; 
and in Period 2, on Day -1, and predose and 3 hours post dose on Days 3, 6, 9, 13, and 16; in the morning of Day 17, and at Follow-up (see also Appendix 2).  
h
Vital signs will be collected as follows: in Period 1, on Day -1 and predose and 3 hours after dosing of balovaptan on Days 3, 7, and 10 and in the morning of Day 11; and 
in Period 2, on Day -1, and predose and 3 hours post dose on Days 3, 6, 9, 13, and 16; in the morning of Day 17, and at Follow-up (see also Appendix 2).  
i
Includes measurement of body temperature.
j
Hematology and chemistry at Screening and on Days -1 and 11 of Period 1 and on Days -1, 3, 6, 9, 13, 17 in Period 2, and follow up.  Urinalysis at Screening and on 
Days -1 and 11 of Period 1 and on Days -1 and 17 in Period 2 and at Follow-up.
k
For females only: FSH and serum pregnancy test at Screening, and serum pregnancy test only at all other time points.  
l
Subjects will be admitted to the clinical research unit on Day -1 of Period 1 and Period 2 and will be discharged on Day 11 and Day 17 of Period 1 and 2, respectively,
after completion of study assessments.


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Appendix 2
Schedule of Pharmacokinetic Samples and Electrocardiogram, 
and Vital Signs Measurements
Procedure
Study Day
Predose
or Prior to 
Admission/Discharge
Hours post dosea
0.5
1
1.5
2
2.5
3
4
6
8
12
16
24
Blood sampling balovaptan, M2, and M3 (one sample for all 3 analytes)
Period 1
Days 2, 4, 8, and 9
Xb
Day 10
Xb
X
X
X
X
X
X
X
X
X
X
Xc
Xc
Period 2
Days 8, 10, 14, and 15
Xb
Day 16
Xb
X
X
X
X
X
X
X
X
X
X
Xd
Xd
Blood sampling rifampicin
Period 2
Day 6
Xb
X
X
X
X
X
X
X
Xe
Day 16
Xb
X
X
X
X
X
X
X
Xd
12-lead electrocardiograms
Period 1
Day -1 and Day 11
X f
Days 3, 7, and 10
Xb
X
Period 2
Day -1 and Day 17
X f
Days 3, 6, 9, 13, and 16
Xb
X
Vital signs
Period 1
Day -1 and Day 11
X f
Days 3, 7, and 10
Xb
X
Period 2
Day -1 and Day 17
X f
Days 3, 6, 9, 13, and 16
Xb
X


Appendix 2
Schedule of Pharmacokinetic Samples and Electrocardiogram, 
and Vital Signs Measurements (cont.)
Balovaptan—F.  Hoffmann-La Roche Ltd
71/Protocol WP40608, Version 1.0
aFor sampling to measure balovaptan, M2, and M3, and for ECG and vital signs measurements on days 
balovaptan or balovaptan together with rifampicin are administered, post dose is relative to time of
balovaptan dosing; for sampling to measure rifampicin and for ECG and vital signs measurements on days 
balovaptan is not administered, post dose is relative to time of rifampicin dosing
b10 minutes predose (or time matched, when not dosed)
cDay 11
dDay 17
eDay 7
fPrior to admission or discharge

SAP_001.pdf: 
 
Official Title: 
A SINGLE-CENTER, NON-RANDOMIZED, OPEN-LABEL, 
ONE-SEQUENCE, TWO-PERIOD WITHIN-SUBJECT 
STUDY TO INVESTIGATE THE EFFECT OF RIFAMPICIN 
ON THE PHARMACOKINETICS OF MULTIPLE DOSES 
OF BALOVAPTAN IN HEALTHY VOLUNTEERS 
NCT Number: 
NCT03586726  
Document Date: 
SAP version 1: 09-October-2018 
 
 




Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
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 Objective ................................................................................................................................ 4 
5.1.1 Primary Endpoints ........................................................................................................................ 4 
5.2 Secondary Objectives ......................................................................................................................... 4 
5.2.1 Secondary Endpoints ................................................................................................................... 4 
5.3 Tertiary/ Exploratory Objectives .......................................................................................................... 5 
5.3.1 Tertiary/Exploratory Endpoints ..................................................................................................... 5 
6.0 Study Design ........................................................................................................................................... 5 
6.1 Sample Size Considerations ............................................................................................................... 6 
6.2 Randomization .................................................................................................................................... 6 
7.0 Overview of Planned Analysis ................................................................................................................. 6 
7.1 Changes from Protocol ....................................................................................................................... 6 
7.2 Interim Analysis and Key Results........................................................................................................ 7 
7.3 Final Analysis ...................................................................................................................................... 7 
8.0 Data Review ............................................................................................................................................ 7 
8.1 Data Management ............................................................................................................................... 7 
8.2 Acceptance of Data for Summarization .............................................................................................. 7 
9.0 Definitions and General Analysis Methods ............................................................................................. 7 
9.1 Analysis Data Presentation ................................................................................................................. 7 
9.1.1 Rounding ...................................................................................................................................... 7 
9.1.2 Imputation ..................................................................................................................................... 7 
9.1.3 Descriptive Statistics .................................................................................................................... 7 
9.1.4 Pooling ......................................................................................................................................... 8 
9.1.5 Unscheduled Measurements ....................................................................................................... 8 
9.2 Analysis Data Definitions .................................................................................................................... 8 
9.2.1 Baseline Definition ....................................................................................................................... 8 
9.2.2 Treatment Grouping ..................................................................................................................... 8 
9.2.3 Common Variable Derivations ..................................................................................................... 9 
9.2.4 QC ................................................................................................................................................ 9 
9.2.5 ADaM Datasets and Metadata ..................................................................................................... 9 
9.3 Software .............................................................................................................................................. 9 
9.4 Statistical Methods ............................................................................................................................ 10 
9.4.1 Statistical Outlier Determination ................................................................................................. 10 
9.4.2 Predetermined Covariates and Prognostic Factors ................................................................... 10 
9.4.3 Hypothesis Testing ..................................................................................................................... 10 
9.5 TFL Layout ........................................................................................................................................ 10 
10.0 Analysis Populations ........................................................................................................................... 11 
10.1 Safety Analysis Population .............................................................................................................. 11 
10.2 Pharmacokinetic Analysis Population ............................................................................................. 11 
11.0 Subject Disposition .............................................................................................................................. 11 
12.0 Protocol Deviations and Violations ..................................................................................................... 11 
13.0 Demographic and Baseline Characteristics ........................................................................................ 11 
13.1 Demographics ................................................................................................................................. 11 
13.2 Medical History ................................................................................................................................ 12 
13.3 Other Baseline Characteristics........................................................................................................ 12 
14.0 Concomitant Medications .................................................................................................................... 12 
15.0 Treatment Compliance and Exposure ................................................................................................ 12 
16.0 Pharmacokinetic Analyses .................................................................................................................. 12 
16.1 Pharmacokinetic Variables. ............................................................................................................. 12 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
16.1.1 Concentrations ......................................................................................................................... 12 
16.1.2 Parameters ............................................................................................................................... 12 
16.2 Pharmacokinetic Summaries .......................................................................................................... 14 
16.2.1 Pharmacokinetic Concentrations ............................................................................................. 14 
16.2.2 Pharmacokinetic Parameters ................................................................................................... 14 
17.0 Safety Analyses .................................................................................................................................. 15 
17.1 Safety Variables .............................................................................................................................. 15 
17.1.1 Adverse Events ........................................................................................................................ 15 
17.1.2 Deaths and Serious Adverse Events ....................................................................................... 16 
17.1.3 Laboratory Data ....................................................................................................................... 16 
17.1.4 Vital Signs ................................................................................................................................ 16 
17.1.5 Electrocardiograms .................................................................................................................. 16 
17.1.6 Columbia Suicide Severity Rating Scale .................................................................................. 16 
18.0 References .......................................................................................................................................... 17 
Appendix 1: Glossary of Abbreviations ....................................................................................................... 17 
Appendix 2: Schedule of Assessments ....................................................................................................... 19 
Appendix 3: Schedule of PK, ECG and VS Measurements ........................................................................ 21 
Appendix 4: List of End of Text Outputs ...................................................................................................... 22 
Document History ........................................................................................................................................ 25 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
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. Hoffmann-La Roche Ltd Protocol WP40608. 
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 03-Jun-2018 (including all 
amendments up to this protocol date) and the final eCRF(s) dated 02-Jun-2018. 
An approved and signed SAP is a requirement for database lock. 
This SAP only covers the results that will be processed by the PRA Early Development Services (EDS) 
Biostatistics Department. 
PRA EDS will perform the pharmacokinetic (PK) and 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 Section 9.8.2 of 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 study will investigate the effect of rifampicin treatment on the PK of balovaptan and its major 
metabolites M2 and M3. 
 
5.1 Primary Objective 
To investigate the effect of multiple doses of rifampicin, a potent cytochrome P450 (CYP)3A4 inducer, on 
the PK of balovaptan 10 mg once daily (qd) and its major metabolites M2 and M3 at steady state in healthy 
subjects. 
 
5.1.1 Primary Endpoints 
PK parameters for balovaptan, M2, and M3 following 10 days of qd dosing of balovaptan 10 mg: 
• 
Maximum observed plasma concentration (Cmax) 
• 
Area under the plasma concentration curve from time 0 to 24 hours (AUC0-24h) 
5.2 Secondary Objectives 
• 
To evaluate the safety and tolerability of balovaptan 10 mg qd administered alone and in 
combination with rifampicin in healthy subjects. 
• 
To investigate the effect of multiple doses of rifampicin on secondary PK parameters of multiple 
doses of balovaptan 10 mg qd and its major metabolites M2 and M3 at steady state in healthy 
subjects. 
 
5.2.1 Secondary Endpoints 
Safety and tolerability: 
• 
Adverse events (AEs) 
• 
Clinical laboratory values 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
• 
Vital signs 
• 
Electrocardiogram (ECG) 
• 
Columbia-Suicide Severity Scale (C-SSRS) 
• 
Physical examination 
 
Secondary PK parameters for balovaptan and metabolite M2 and M3: 
• 
Time to maximum observed plasma concentration (tmax) 
• 
Metabolite to parent ratio 
• 
Any other parameter as appropriate (see Table 1) 
5.3 Tertiary/ Exploratory Objectives 
To evaluate the plasma exposure of rifampicin 600 mg qd. 
To investigate the relationship between the CYP3A4 genotype and steady state balovaptan exposure, and 
the influence of the CYP3A4 genotype on the effect of rifampicin on balovaptan PK. The relationship 
between other genotypes and PK of balovaptan may also be explored. 
 
5.3.1 Tertiary/Exploratory Endpoints 
Exploratory PK parameters for rifampicin following 6 and 16 days of qd dosing: 
• 
Cmax 
• 
AUC0-24h 
• 
tmax 
• 
AUC0-last 
• 
any other parameter as appropriate (see Table 1) 
 
The pharmacogenetics of metabolizing enzymes, tranferases, transporters, etc, possibly involved in the 
absorption, distribution, metabolism and excretion of balovaptan and its major metabolites (e.g., CYP3A 
and P-glycoprotein). Results may be pooled with data from other studies of balovaptan. 
 
6.0 Study Design 
This study will be a single-center, non-randomized, open-label, one-sequence, two-period, within-subject 
study to investigate the effects of multiple doses of rifampicin on the PK and safety of multiple doses of 
balovaptan in healthy subjects. The study will be conducted at 1 site in the Netherlands. 
Screening will be conducted up to 3 weeks prior to admission in the clinical research unit (CRU). Subjects 
will be in the CRU for 2 study periods. In both periods, subjects will be admitted on Day -1 (the day before 
dosing begins); Day 1 will be the first day of dosing. In Period 1, subjects will be discharged on Day 11 after 
the last assessment is completed. There will be a minimum of a 14-day to a maximum of a 21-day washout 
between the last dose in Period 1 and the first dose in Period 2. Subjects will therefore return to the CRU 
on Day 24 or up to Day 30 of Period 1 (Day -1 of Period 2) to begin Period 2. In Period 2, subjects will be 
discharged on Day 17 after all assessments have been performed. Subjects will return for a follow-up visit 
between 14 to 21 days after the last dose. 
Subjects will receive the following study drugs in the 2 study periods, after an overnight fast of at least 
8 hours: 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
Study Period 1: 
• 
Days 1 to 10: oral balovaptan 10 mg qd 
• 
Days 11 to 23 (or up to Day 30): no study drug 
 
Study Period 2: 
• 
Days 1 to 6: rifampicin 600 mg qd 
• 
Days 7 to 16: balovaptan 10 mg qd + rifampicin 600 mg qd 
Figure 1 presents an overview of the study design. 
Figure 1: Study Schema 
 
 
 
A total of 16 subjects will be enrolled to ensure 12 evaluable subjects. 
 
6.1 Sample Size Considerations 
This is an exploratory study for which no formal statistical hypothesis will be tested. Therefore, the sample 
size is chosen to estimate with sufficient precision the effect of CYP3A4 induction on balovaptan. 
 
The sample size of 12 evaluable subjects was chosen based on balovaptan within-subject variability (CV%) 
of around 39% for AUC0-24h and 36% for Cmax as obtained from the 12 mg repeat dose in study BP25694. 
Based on the higher value of 39%, it was estimated that with 12 subjects the half-width of the 90% 
confidence interval (CI) for the ratio of treatment geometric means of the combination (balovaptan + 
rifampicin) versus balovaptan alone would be obtained by dividing/multiplying the ratio estimate by a factor 
of 1.30. 
 
6.2 Randomization 
Not applicable, this is a non-randomized study. 
 
7.0 Overview of Planned Analysis 
7.1 Changes from Protocol 
There are no changes from the protocol. 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
7.2 Interim Analysis and Key Results 
No interim reporting is planned for this study. 
 
7.3 Final Analysis 
Draft TFLs will be provided with the draft CSR. After Sponsor comments have been incorporated, the TFLs 
will be finalized and incorporated in the final CSR. 
 
8.0 Data Review 
8.1 Data Management 
Data handling, transfer and coding 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, all data (except the PK data) will be presented with the same precision as the original data. 
Derived data will be rounded for presentation purposes. 
For descriptive statistics of safety data, range values will be presented with the same precision (number of 
decimals or significant digits) as the data they are calculated from, median, arithmetic mean values with 1 
more decimal digit, standard deviation (SD) with 2 more decimal digits. Percentages will be rounded to 
integers. 
For the derived PK parameters Appendix 2 of the Roche Clinical Pharmacology Guideline will be followed 
[3]. For all PK parameters except tmax; individual subject data, range values, median, arithmetic mean, 
geometric mean and standard deviation (SD) will be presented with a precision of 3 significant digits. The 
coefficients of variation (CV) with 1 decimal digit. 
All individual data and descriptive statistics for tmax will be reported with 2 decimals, except for the CVs 
which will be presented with 1 decimal digit. 
 
9.1.2 Imputation 
Except for the substitution of any PK concentrations below the lower limit of quantification (LLOQ) (see 
Section 16.2) and missing start or end date/times of Adverse Events (AEs) for the calculation of onset and 
duration (see Section 17.1.1), any missing data will not be imputed. 
 
9.1.3 Descriptive Statistics 
Unless otherwise indicated, continuous variables will be summarized with the following descriptive 
statistics: 
• 
n (number of observations), 






Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
9.4 Statistical Methods 
9.4.1 Statistical Outlier Determination 
No statistical outlier analysis is planned. 
 
9.4.2 Predetermined Covariates and Prognostic Factors 
CYP3A4 genotype will be investigated as a factor in this study. PK tables will be presented by CYP3A4 
genotype. 
The pharmacogenetics of other isoforms, metabolizing enzymes, transferases, transporters, etc, possibly 
involved in the absorption, distribution, metabolism and excretion of balovaptan and its major metabolites 
(e.g., CYP3A and P-glycoprotein) may be investigated additionally (e.g. CYP3A5 genotype). 
 
9.4.3 Hypothesis Testing 
No formal hypothesis testing will be done. 
 
9.5 TFL Layout 
Report layout will be according to the PRA EDS – ICH E3 compliant – CSR Template. The layout of TFLs 
will be according to the PRA EDS standards. 
No table shells will be provided. The TFLs will be provided in Adobe PDF format. 
Format: 
• 
Page size: A4 
• 
Data in listings will be sorted by subject number and time point. 
• 
Data in tables will be sorted by treatment and time point. 
• 
Column titles will be in title case letters. 
• 
All tables and listings will be in landscape format. 
• 
The treatment labels will be as outlined in Section 9.2.2 Treatment/Subject Grouping Definition 




Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
13.2 Medical History 
Medical history will be listed including the coding according to the Medical Dictionary for Regulatory 
Activities (MedDRA; latest version). 
 
13.3 Other Baseline Characteristics 
• 
Drug and alcohol screen: The results of urine drug screen (barbiturates, benzodiazepines, 
methadone, amphetamines [including ecstasy], methamphetamines, opiates, cocaine, and 
cannabinoids and urine alcohol test will be listed. 
• 
Serology: The results of serology (human immunodeficiency virus (HIV)-1 and HIV-2, hepatitis B 
virus surface antigen (HBsAg), hepatitis C virus (HCV) antibody) at screening will be listed. 
• 
Pregnancy test (females only): Serum pregnancy test results (beta-human chorionic gonadotropin) 
and follicle-stimulating hormone (FSH) will be listed for each female subject at screening, each 
admission to the clinical research center and at follow-up. 
• 
Body weight will be listed at screening and at follow-up. 
• 
Non-compliance to in- or exclusion criteria (if any) will be listed. 
 
14.0 Concomitant Medications 
Concomitant medications, categorized by medication group and subgroup according to GNE Drug 
dictionary, will be summarized. The number and percentage of subjects using each medication will be 
displayed with the number and percentage of subjects using at least one medication within each medication 
group and subgroup, by treatment. 
Concomitant medication will be listed. 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 
Exposure data will be listed by subject. 
The number of subjects receiving each dose of study drug will be summarized. 
 
16.0 Pharmacokinetic Analyses 
PK concentrations will be collected in plasma. 
 
16.1 Pharmacokinetic Variables 
16.1.1 Concentrations 
• 
Plasma concentration of balovaptan 
• 
Plasma concentration of metabolite M2 
• 
Plasma concentration of metabolite M3 
• 
Plasma concentration of rifampicin 
16.1.2 Parameters 
• 
PK Parameters for balovaptan, M2, M3 and rifampicin as defined in Table 1. 




Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
16.2 Pharmacokinetic Summaries 
16.2.1 Pharmacokinetic Concentrations 
Plasma concentrations for balovaptan, M2, M3 and rifampicin below the quantifiable limit (BQL) will be set 
to ½ lower limit of quantification (LLOQ) in the computation of mean concentration values. 
Descriptive statistics (number of subjects, arithmetic mean, geometric mean, SD, coefficient of variation, 
median, min, and max) will be used to summarize the serum concentrations by treatment at each scheduled 
time point. 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 minimum. With the exception of maximum all 
other statistics will be missing. 
Linear and semi-logarithmic plots of the geometric mean plasma concentration by scheduled sampling time 
will be provided by treatment. These plots will show time in hours. The plots will match the summary table 
results and will not have an observation at a given time point 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. Separate plots will be 
created for the total profile including all analysis periods/treatments and for the Analysis Day 10 time-profiles 
(balovaptan versus balovaptan + rifampicin) separately. Individual plots will use the BQL handling 
procedure described below for “Pharmacokinetic Parameters”. 
Individual plasma concentration data will be presented together with descriptive statistics by analyte and 
treatment. 
 
16.2.2 Pharmacokinetic Parameters 
PK parameters for balovaptan, M2, M3 and rifampicin will be estimated using non-compartmental methods 
with WinNonlin®. 
The plasma PK parameters will be estimated from the concentration-time profiles. In estimating the PK 
parameters, BQL values will be set to zero for the pre-dose PK sample, as well as for all other samples 
being BLQ and occurring before tmax. For subsequent time points, the result will be set to missing. Actual 
sampling times, rather than scheduled sampling times, will be used in all computations involving sampling 
times. If the actual time or dose time is missing, the scheduled time may be substituted in order to calculate 
the PK parameter. 
Descriptive statistics (number of subjects, arithmetic mean, geometric mean, SD, geometric and arithmetic 
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. 
The points to be included in the λz range will be determined by the pharmacokineticist after inspection of 
the semi-log concentration-time profiles. At least 3 points will be required to be used and the range should 
ideally be spread over a time interval representing at least 2 half-lives. The Cmax data point will not be 
included. 
Parameters based on R2 below 0.70 or %AUCextra above 20% will be flagged and excluded from descriptive 
statistics. 
 
16.2.2.1 Drug-Drug Interaction 
The effect of rifampicin on the PK of balovaptan will be assessed. The effect of rifampicin on the natural 
log-transformed Cmax and AUC0-24h will be assessed with a linear mixed effects model. Treatment will be 
fitted as fixed effect and subject as a random effect. Point estimates for the means and point estimates and 
corresponding 90% CI for the differences in means between the two treatments (balovaptan plus rifampicin 
as the test treatment, versus balovaptan alone as the reference) will be obtained from the linear mixed 
effects model and exponentiated to obtain geometric means, geometric mean ratios and respective 90% 
CI on the original scale. 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
In addition, as an exploratory analysis CYP3A4 genotype will be added as a factor. The relationship 
between other genotypes (e.g. CYP3A5) may also be explored. 
 
17.0 Safety Analyses 
17.1 Safety Variables 
The following safety variables will be summarized: 
• 
Adverse Events (AEs) 
• 
Vital Signs 
o 
Supine Blood Pressure 
 
Systolic Blood Pressure (SBP) 
 
Diastolic Blood Pressure (DBP) 
o 
Pulse rate 
o 
Body temperature (tympanic) 
o 
Respiratory rate 
• 
Electrocardiograms (ECG) 
o 
Heart Rate 
o 
PR Interval 
o 
QRS-Duration 
o 
QT Interval 
o 
QTc (Fridericia) Interval 
• 
Clinical Laboratory Evaluations 
o 
Serum Chemistry 
o 
Hematology 
o 
Urinalysis 
o 
Coagulation 
• 
Columbia-Suicide Severity Rating Scale (C-SSRS) 
o 
Suicidal ideation 
o 
Suicidal behavior 
17.1.1 Adverse Events 
All AEs will be coded to the current version of Medical Dictionary for Regulatory Activities (MedDRA, the 
latest version) by F. Hoffmann-La Roche, Ltd. 
All AE summaries will include only treatment emergent adverse events (TEAEs). Treatment-emergent 
adverse events are those which occur after the first dose of study drug. AEs starting prior to medication 
dosing in the first period will be regarded as pre-dose AEs (i.e. non-TEAEs). 
TEAEs occurring following dosing in a specific analysis period but before dosing in the next analysis period 
will be attributed to that specific analysis period, thus to the last received treatment. 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. 
All adverse events (including non-TEAEs) recorded on the eCRF will be listed (including analysis period 
and treatment information). Pre-dose AEs will be presented in a separate listing. In addition, a separate 
listing of AEs leading to withdrawal from study will be provided. 
TEAEs will be tabulated by system organ class and preferred term: one table with all TEAEs (number and 
percentage of subjects) overall and by treatment; one table with related TEAEs (number and percentage 
of subjects) by treatment; one table with all TEAEs (number and percentage of subjects) by treatment and 
relationship to study drug; one table with TEAEs (number and percentage of subjects) by treatment and 
severity. 
Subjects are counted once, per preferred term per treatment, for the most severe of multiple occurrences 
(in case of severity) or most drug-related event (in case of relationship) of a specific MedDRA term. AEs 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
whose causal relationship was characterized as ‘Yes’/’No’ will be regarded as being related/not related to 
the study medication. 
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 attributed to the treatment from analysis period 1 unless the AE end 
date occurs before first IMP administration 
17.1.2 Deaths and Serious Adverse Events 
A listing of deaths and other serious adverse events (SAE) 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. 
All laboratory data will be listed, including laboratory variables not listed in the protocol. A separate listing, 
including out-of-range values will also be provided. Normal ranges will be used directly from the clinical 
laboratory. 
Descriptive statistics summarizing continuous laboratory results of clinical chemistry, hematology and 
urinalysis (observed and derived changes from baseline) by treatment and scheduled time will be provided. 
For categorical urinalysis parameters frequency tables, showing the number of subjects in a category (n) 
and the percentage of the total number of subjects per treatment (N), will be provided. Sporadic urinalysis 
tests that were performed when abnormalities were observed (i.e. microscopy/sediment) will only be listed. 
 
17.1.4 Vital Signs 
All vital signs data including derived changes from baseline will be listed. 
Descriptive statistics will be provided to summarize vital signs including changes from baseline by treatment 
and scheduled time. 
 
17.1.5 Electrocardiograms 
All ECG parameters (including changes from baseline) and the corresponding abnormalities and 
physician’s conclusions will be listed by subject. 
The observed measurements for all ECG parameters (including T-wave, U-wave) and the corresponding 
abnormalities will be listed for all time points. The means of triplicate measurements for continuous 
parameters and the change from baseline of the mean triplicate measurements at each scheduled time 
point will be listed by subject. 
Descriptive statistics will be provided to summarize mean continuous ECG parameters (observed and 
changes from baseline) by treatment and scheduled time. A frequency table will be provided as well to 
summarize the ECG parameters by physicians conclusion (normal, abnormal not clinically significant, 
abnormal clinically significant). 
 
17.1.6 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). It captures the occurrence, severity and 




Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
 
HCV 
Hepatitis C virus 
ICH 
The International Conference on Harmonization of Technical Requirements 
for Registration of Pharmaceuticals for Human Use 
LLOQ 
Lower limit of quantification 
MedDRA 
Medical Dictionary for Regulatory Activities 
M2, M3 
Metabolites 
PK 
Pharmacokinetic 
QA’d 
Quality assured 
QC’d 
Quality controlled 
qd 
Once daily 
SAP 
Statistical analysis plan 
SAE 
Serious adverse event 
SBP 
Systolic Blood Pressure 
SD 
Standard deviation 
SDTM 
Study data tabulation model 
TEAE 
Treatment-emergent adverse event 
TFL(s) 
Tables, figures and listings 
WNL 
WinNonlin 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
 
Appendix 2: Schedule of Assessments 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 


Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
 
 
Appendix 3: Schedule of PK, ECG and VS Measurements 




Statistical Analysis Plan 
RPU034EC-180341 
Protocol: WP40608 
Version Date: 04-Oct-2018 
 
Figure 14.2.9.3 
Total Profile of Individual Rifampicin Plasma Concentrations 
versus Time (Linear and Semi-Logarithmic Scale) 
Safety 
Figure 14.2.9.4 
Combined Total profile of Individual Balovaptan Plasma 
Concentrations versus Time (Linear and Semi-Logarithmic 
Scale) 
Safety 
Figure 14.2.9.5 
Combined Total profile of Individual Balovaptan Metabolite (M2, 
M3) Plasma Concentrations versus Time (Linear and Semi- 
Logarithmic Scale) 
Safety 
Figure 14.2.9.6 
Combined Individual Rifampicin Plasma Concentrations versus 
Time (Linear and Semi-Logarithmic Scale) 
Safety 
Figure 14.2.9.7 
Individual Balovaptan Plasma Concentrations versus Time – 
Analysis Day 10 (Linear and Semi-Logarithmic Scale) 
Safety 
Figure 14.2.9.8 
Individual Balovaptan Metabolite (M2, M3) Plasma 
Concentrations versus Time – Analysis Day 10 (Linear and 
Semi-Logarithmic Scale) 
Safety 
Section 14.3 – Safety Data 
Section 14.3.1 Adverse Events 
Table 14.3.1.1 
Summary of TEAEs for Each System Organ Class and 
Preferred Term (Number and Percentage of Subjects) by 
Treatment 
Safety 
Table 14.3.1.2 
Summary of Related TEAEs for Each System Organ Class and 
Preferred Term (Number and Percentage of Subjects) by 
Treatment 
Safety 
Table 14.3.1.3 
Summary of TEAEs for Each System Organ Class and 
Preferred Term (Number and Percentage of Subjects) by 
Treatment and Relationship to Study Drug 
Safety 
Table 14.3.1.4 
Summary of TEAEs for Each System Organ Class and 
Preferred Term (Number and Percentage of Subjects) by 
Treatment and Severity 
Safety 
Section 14.3.2 Lists of Deaths, Other Serious and Significant Adverse Events 
Table 14.3.2.1 
Listing of Deaths and Other Serious Adverse Events 
Safety 
Section 14.3.3 Clinical Laboratory 
Table 14.3.3.1 
Listing of Abnormal Laboratory Values 
Safety 
Table 14.3.3.2 
Descriptive Statistics of Clinical Laboratory Results - 
Hematology 
Safety 
Table 14.3.3.3 
Descriptive Statistics of Clinical Laboratory Results – 
Serum Chemistry 
Safety 
Table 14.3.3.4 
Descriptive Statistics of Clinical Laboratory Results - Urinalysis 
Safety 
Section 14.3.4 Other Safety 
Table 14.3.4.1 
Descriptive Statistics of Vital Signs 
Safety 
Table 14.3.4.2 
Summary of 12-Lead Electrocardiogram 
Safety 
Table 14.3.4.3 
Frequency of 12-Lead Electrocardiogram Physicians 
Conclusion 
Safety