Clinical trial NCT02287896
Conditions: Rh Disease
Keywords: RhD-negative pregnant woman anti-RhD antibody
Sponsor: Laboratoire français de Fractionnement et de Biotechnologies
Start date: 2014-04-01; Completion date: 2017-09-13
Enrollment: 62 participants; Allocation: NON_RANDOMIZED; Intervention model: PARALLEL

Brief summary:
The aim of this study is to assess the pharmacokinetic profile of Roledumab 300μg IM / IV in RhD-negative pregnant women carrying an RhD-positive foetus.

To assess the safety of Roledumab in RhD-negative pregnant women and in RhD-positive fetus and newborns.

In addition the efficacy of Roledumab 300μg IM and IV to prevent RhD alloimmunisation in RhD-negative pregnant women carrying an RhD-positive fetus and the immunogenicity of Roledumab will be assessed.

Detailed description:

Arms: arm 1: \- Planned antenatal prophylaxis: A single dose of 300 µg IM of Roledumab at 28 or 29 weeks of gestation.

\- Antenatal prophylaxis following sensitizing events: One or more dose(s) of 300μg IM anti-RhD antibodies (Rhophylac or Roledumab) based on the Kleihauer-test as soon as possible and no later than 72 hours after event occurrence.

\- Postnatal prophylaxis: Roledumab should be administered to the mother as soon as possible within 72 hours of delivery of an RhD positive infant.

The postnatal dose must still be given even when antenatal prophylaxis has been administered.

Before Roledumab 300μg IM postnatal administration, a Kleihauer-Betke test will be performed on maternal blood sample taken no earlier than 30 min after delivery in order to determine the volume of foetomaternal hemorrhage (FMH). arm 2: \- Planned antenatal prophylaxis: A single dose of 300 µg IV of Roledumab at 28 or 29 weeks of gestation.

\- Antenatal prophylaxis following sensitizing events: One or more dose(s) of 300μg IV anti-RhD antibodies (Rhophylac or Roledumab) based on the Kleihauer-test as soon as possible and no later than 72 hours after event occurrence.

\- Postnatal prophylaxis: Roledumab should be administered to the mother as soon as possible within 72 hours of delivery of an RhD positive infant.

The postnatal dose must still be given even when antenatal prophylaxis has been administered.

Before Roledumab 300μg IV postnatal administration, a Kleihauer-Betke test will be performed on maternal blood sample taken no earlier than 30 min after delivery in order to determine the volume of foetomaternal hemorrhage (FMH).
Interventions: intervention 1: ROLEDUMAB
Intervention descriptions: intervention 1: See Arm description

Protocol documents:
Prot_002.pdf: 
 
COVER PAGE 
 
 
TITLE: Pharmacokinetics and Safety of Roledumab, a fully human recombinant monoclonal 
anti-RhD antibody, in RhD-negative pregnant women carrying an RhD-positive foetus: a 
phase IIb, multicenter, open-label study. 
 
NCT Number: NCT02287896 
 
DOCUMENT: Protocole (v12.0)  
 
 
VERSION & DATE OF DOCUMENT: Version 12.0; October 27, 2017 




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TABLE OF CONTENTS
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS..................................................... 6
SYNOPSIS ............................................................................................................................................ 8
1.
INTRODUCTION AND BACKGROUND INFORMATION............................................... 18
Disease and Context .............................................................................................................. 18
1.1.
1.1.1.
Disease ........................................................................................................................... 18
1.1.2.
Context........................................................................................................................... 18
Target..................................................................................................................................... 19
1.2.
IMP, Non-Clinical and Clinical Information ........................................................................ 19
1.3.
1.3.1.
IMP and non-clinical information.................................................................................. 19
1.3.2.
Clinical information ....................................................................................................... 20
Rationale of the Study........................................................................................................... 21
1.4.
1.4.1.
Rationale of the dose...................................................................................................... 22
1.4.2.
Appropriateness of this study......................................................................................... 23
2.
STUDY OBJECTIVES.............................................................................................................. 23
Primary Objective.................................................................................................................. 23
2.1.
Secondary Objective(s) ......................................................................................................... 23
2.2.
3.
STUDY DESIGN........................................................................................................................ 23
Design.................................................................................................................................... 26
3.1.
Endpoints............................................................................................................................... 29
3.2.
3.2.1.
Primary endpoints .......................................................................................................... 29
3.2.2.
Secondary endpoints ...................................................................................................... 30
Study Investigational Site(s) ................................................................................................. 30
3.3.
Data and Safety Monitoring Board........................................................................................ 30
3.4.
4.
DURATION AND TIMELINES OF THE STUDY................................................................ 31
5.
STUDY POPULATION ............................................................................................................ 32
Number of Subjects............................................................................................................... 32
5.1.
Eligibility Criteria.................................................................................................................. 32
5.2.
5.2.1.
Inclusion criteria............................................................................................................. 32
5.2.2.
Exclusion criteria ........................................................................................................... 32
Duration of Subject Participation.......................................................................................... 33
5.3.
Early Discontinuation Criteria / Stopping Rule(s) ................................................................ 33
5.4.
5.4.1.
Handling of early terminations....................................................................................... 34
5.4.2.
Sponsor’s study termination........................................................................................... 34
5.4.3.
Lost to follow-up............................................................................................................ 34
6.
INVESTIGATIONAL MEDICINAL PRODUCT(S) (IMP).................................................. 35
Description of IMP(s)............................................................................................................ 35
6.1.
IMP Packaging and Labelling ............................................................................................... 35
6.2.
6.2.1.
Roledumab packaging.................................................................................................... 35


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6.2.2.
Labelling ........................................................................................................................ 35
Management of IMP.............................................................................................................. 36
6.3.
6.3.1.
Shipment and receipt...................................................................................................... 36
6.3.2.
Storage requirements...................................................................................................... 36
6.3.3.
IMP re-supplying ........................................................................................................... 36
6.3.4.
IMP accountability......................................................................................................... 37
6.3.5.
IMP return, destruction and recall.................................................................................. 37
Treatment of Subjects............................................................................................................ 38
6.4.
6.4.1.
Methods for assigning subjects to treatment groups...................................................... 38
6.4.2.
Dispensing...................................................................................................................... 38
6.4.3.
Compliance .................................................................................................................... 43
Randomization Codes and Procedures for Unblinding ......................................................... 43
6.5.
7.
PRIOR AND CONCOMITANT MEDICATION................................................................... 43
Prior Medication.................................................................................................................... 43
7.1.
Concomitant Medication ....................................................................................................... 43
7.2.
8.
STUDY PLAN ............................................................................................................................ 44
Subject Recruitment .............................................................................................................. 44
8.1.
8.1.1.
Informed consent............................................................................................................ 44
8.1.2.
Subject enrolment........................................................................................................... 45
8.1.3.
Subject allocation........................................................................................................... 45
8.1.4.
Replacement of early withdrawals................................................................................. 45
Schedule of Visits.................................................................................................................. 45
8.2.
8.2.1.
Screening period: visit 1 (12-27 GA weeks).................................................................. 45
8.2.2.
Antenatal period (from visit V2 to visit V7).................................................................. 46
8.2.3.
Antenatal period - in case of a sensitizing event............................................................ 49
8.2.4.
Postnatal period - visit 8 to visit 9.................................................................................. 50
8.2.5.
Follow-up period (from visit 10 to visit 13, additional visits AV1 and AV2)............... 51
8.2.6.
Follow-up in case of alloimmunization ......................................................................... 53
8.2.7.
Follow-up in case of ADA production after IMP administration .................................. 54
Assessments........................................................................................................................... 55
8.3.
8.3.1.
Assessments performed on site...................................................................................... 55
8.3.2.
Centralized assessments................................................................................................. 59
Compliance with the Study Plan ........................................................................................... 62
8.4.
9.
SAFETY...................................................................................................................................... 62
Safety Reference Document.................................................................................................. 62
9.1.
Benefit / Risk Information..................................................................................................... 63
9.2.
9.2.1.
Potential risk(s) related to the IMP(s)............................................................................ 64
9.2.2.
Benefit/risk balance........................................................................................................ 69
Risk Minimization Actions/Monitoring Throughout the Protocol........................................ 69
9.3.
Alternative Therapeutic Management - Emergencies Handling ........................................... 70
9.4.
Definition and Reporting of (Serious) Adverse Events......................................................... 70
9.5.
9.5.1.
Definition of adverse event and serious adverse event (see also Section 9.5.2: 
Definition of specific events in the study) ................................................................................... 70
9.5.2.
Definition of specific events in the study....................................................................... 71
9.5.3.
Adverse events recording and reporting ........................................................................ 71


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9.5.4.
Procedures for reporting serious adverse events............................................................ 72
9.5.5.
Follow-up of adverse events .......................................................................................... 73
Reporting of Pregnancies ...................................................................................................... 73
9.6.
10.
PARAMETERS AND ASSESSMENT CRITERIA............................................................. 74
Assessment of Pharmacokinetics....................................................................................... 74
10.1.
Assessment of Safety......................................................................................................... 74
10.2.
10.2.1.
Adverse events ........................................................................................................... 74
10.2.2.
Physical examination.................................................................................................. 75
10.2.3.
Laboratory data........................................................................................................... 75
Assessment of Efficacy: Occurrence of RhD-Alloimmunization...................................... 75
10.3.
Other Assessments............................................................................................................. 76
10.4.
11.
DATA MANAGEMENT ........................................................................................................ 76
CRF Completion Guidelines.............................................................................................. 76
11.1.
11.1.1.
Introduction ................................................................................................................ 76
11.1.2.
General instructions.................................................................................................... 76
11.1.3.
Completion of the e-CRF ........................................................................................... 78
11.1.4.
Specific case report form instructions........................................................................ 78
E-CRF and Data Handling................................................................................................. 78
11.2.
12.
STATISTICS ........................................................................................................................... 78
Study Objectives and Design............................................................................................. 78
12.1.
Statistical Analysis Plan..................................................................................................... 79
12.2.
Sample Size Determination ............................................................................................... 79
12.3.
Randomization................................................................................................................... 79
12.4.
Protocol Deviations and Analysis Sets.............................................................................. 79
12.5.
General Rules for Handling of Missing or Inconsistent Data............................................ 80
12.6.
Demographic and Baseline Characteristics ....................................................................... 80
12.7.
12.7.1.
Demographic characteristics, medical history, diagnoses.......................................... 80
12.7.2.
Previous treatments .................................................................................................... 81
Investigational Product (IMP) and Concomitant Treatments............................................ 81
12.8.
12.8.1.
Extent of exposure...................................................................................................... 81
12.8.2.
Treatment compliance ................................................................................................ 81
12.8.3.
Concomitant treatment ............................................................................................... 81
Efficacy Analysis............................................................................................................... 81
12.9.
12.9.1.
Primary efficacy variable(s) ....................................................................................... 81
12.9.2.
Secondary efficacy variables...................................................................................... 81
Safety Analysis .................................................................................................................. 82
12.10.
12.10.1.
Extent of exposure...................................................................................................... 82
12.10.2.
Adverse events ........................................................................................................... 82
12.10.3.
Fetal anemia ............................................................................................................... 82
12.10.4.
Vitals signs and physical findings.............................................................................. 82
12.10.5.
Laboratory data........................................................................................................... 83
12.10.6.
Other endpoints .......................................................................................................... 83
Pharmacokinetics, PK/PD Analysis................................................................................... 83
12.11.
12.11.1.
Primary analysis ......................................................................................................... 83
12.11.2.
Secondary analysis ..................................................................................................... 84


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13.
STUDY REPORT ................................................................................................................... 84
14.
CONFIDENTIALITY AND PUBLICATION...................................................................... 84
Subject Confidentiality...................................................................................................... 84
14.1.
Use of Information............................................................................................................. 84
14.2.
15.
ARCHIVING........................................................................................................................... 85
16.
RESPONSIBILITIES OF PARTICIPANTS........................................................................ 86
Responsibilities of the Investigator(s) ............................................................................... 86
16.1.
Responsibilities of the Monitor.......................................................................................... 88
16.2.
Responsibilities of the Data Manager................................................................................ 88
16.3.
17.
ETHICS AND REGULATORY CONSIDERATIONS....................................................... 88
18.
AUDIT AND INSPECTION .................................................................................................. 90
19.
REFERENCES........................................................................................................................ 91
20.
APPENDICES ......................................................................................................................... 93
LIST OF TABLES
Table 6–1: Dosing guidelines for administration................................................................................39
Table 6–2: Adapted dose according to the Kleihauer-Betke test CNGOF table.................................41
LIST OF FIGURES
Figure 3–1: Study Plan for IM arm.....................................................................................................27
Figure 3–2: Study Plan for IV arm......................................................................................................28
Figure 6–1: Flow diagram for FMH testing and subsequent actions after delivery or sensitizing event
.............................................................................................................................................................42


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LIST OF ABBREVIATIONS AND DEFINITION OF TERMS
ADA
Anti-drug antibody
ADCC
Antibody-dependent cell mediated cytotoxicity
AE
Adverse Event
ALT
Alanine aminotransferase
aPTT
Activated partial thromboplastin time
AST
Aspartate aminotransferase
ATC
Anatomical Therapeutic Chemical
AUC
Area Under the Curve
AV
Additional Visit
Cmax
Maximum Concentration
CNGOF
Collège National des Gynécologues et Obstétriciens Français
CRF
Case Report Form
CRO
Contract Research Organization
CRP
C-Reactive Protein
CSP
Code de la Santé Publique
CV
With respect to staff: Curriculum Vitae
With respect to PK: Coefficient of Variation
DAT
Direct Antiglobulin Test
DSMB
Data and Safety Monitoring Board
EC
Ethics Committee
e-CRF
Electronic Case Report Form
ENT
Ear-Nose-Throat
FAS
Full Analysis Set
FMH
Fetomaternal hemorrhage
FOCE
First Order Conditional Estimation
GA
Gestational Age
GCP
Good Clinical Practice
HDF
Hemolytic Disease of the Fetus
HDFN
Hemolytic Disease of the Fetus and Newborn
HDN
Hemolytic Disease of the Newborn
HIV
Human Immunodeficiency Virus
IA
Indirect Antibodies
IAT
Indirect Antiglobulin Test
ICF
Informed Consent Form


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ICH
International Conference on Harmonisation
IEC/IRB
Independent Ethics Committee (IEC) or International Review Board (IRB)
Ig
Immunoglobulin
IM
Intramuscular
IMP
Investigational Medicinal Product
IV
Intravenous
KB
Kleihauer-Betke test
MAA
Marketing Authorization Application
mAb
Monoclonal Antibody
MCA-PSV
Middle Cerebral Artery - Peak Systolic Velocity
MCH
Mean corpuscular hemoglobin
MCHC
Mean corpuscular hemoglobin concentration
MCV
Mean corpuscular volume
MedDRA
Medicinal Dictionary for Regulatory Activities
MoM
Multiple of Medians
NAB
Neutralizing Antibodies
NCA
Non Compartmental Analysis
NCR
None Carbon Required
NOAEL
No Observed Adverse Effect Limit
PK
Pharmacokinetic
PT
Preferred Terms
RBC
Red Blood Cells
RhD
Rhesus D
SAE
Serious Adverse Event
SAF
Safety set
SAP
Statistical Analysis Plan
SOC
System Organ Class
Tmax
Time to reach maximum concentration
UCHP
Unité Centre d’Hémobiologie Périnatale (250 UCHP corresponds to 1 µg anti-D)
WBC
White blood cells




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STUDY No.
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Postnatal prophylaxis:
Roledumab should be administered to the mother as soon as 
possible within 72 hours of delivery of an RhD-positive infant.
The postnatal dose must still be given even when antenatal 
prophylaxis has been administered.
Before roledumab 300 µg IM/IV postnatal administration, a KB
test will be performed on a maternal blood sample taken no
earlier than 30 min after delivery in order to determine the 
volume of fetomaternal hemorrhage (FMH).
If the result of the KB test is 4 mL, additional dose(s) of
roledumab will need to be administered.
If the KB test before IMP administration is positive, a 
subsequent test should be done 24 hours after IMP 
administration. 
If the second KB test is positive, additional 300 µg dose(s) of 
roledumab will be administered according to the volume of 
FMH.
STUDY DESIGN
Phase 2b, interventional, multicenter, open-label, non-
randomized, sequential
OBJECTIVES
Primary:

To assess the pharmacokinetic profile of roledumab 300 µg
IM/IV in RhD-negative pregnant women carrying an RhD-
positive fetus.
Secondary:

To assess the safety of roledumab administered IM and IV in 
RhD-negative pregnant women and in RhD-positive fetus 
and newborns

To assess the efficacy of roledumab IM and IV to prevent 
RhD-alloimmunization in RhD-negative pregnant women 
carrying an RhD-positive fetus

To assess the immunogenicity of roledumab

To measure roledumab concentration in first milk and breast 
milk

To measure roledumab concentration in cord blood after 
delivery
NUMBER OF SUBJECTS
35 planned to reach 30 evaluable subjects in the IM arm
25 planned to reach 20 evaluable subjects in the IV arm
STUDY POPULATION
Pregnant RhD-negative women carrying an RhD-positive fetus


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STUDY No.
ADNC-1301
MAIN INCLUSION/EXCLUSION 
CRITERIA
A subject is eligible when all inclusion and no exclusion criteria 
are met. 
Inclusion criteria

Signed and dated informed consent form provided by the 
subject prior to proceeding with any study-related procedure

At least 18 years old

Pregnancy between 12 and 27 weeks gestational age as 
confirmed by early ultrasound

Pregnant RhD-negative woman carrying an RhD-positive 
fetus confirmed by a non-invasive fetal RhD genotyping test

Negative serology: hepatitis B and C, HIV (1 and 2), except 
for positive results due to vaccinations
Covered by healthcare insurance in accordance with local 
requirements.
MAIN INCLUSION/EXCLUSION 
CRITERIA
Exclusion criteria

RhD-alloimmunized subject

Positive for ADA Test

Multiple fetuses

Occurrence of a documented potential sensitizing event in 
this pregnancy before the antenatal IMP administration

Prior administration of anti-RhD immunoglobulin during the 
current pregnancy

Known clinically relevant maternal or fetal abnormality (e.g. 
as determined by ultrasound or genetic testing), such as 
placenta previa

History of anaphylactic or severe systemic reaction to 
immunoglobulin of any origin

Current diagnosis of an immune disease which by itself or its 
treatment could impair the safety and/or efficacy evaluation 
of roledumab in this study. These diseases are:

All immune deficiencies, particularly those requiring IV-Ig 
supplementation or other systemic treatment

Connective tissue and autoimmune diseases (e.g. systemic 
lupus erythematosus, antiphospholipid syndrome, Sjögren’s 
syndrome, rheumatoid arthritis, ankylosing spondylarthritis) 
requiring systemic immunosuppressive treatment

Allergic and inflammatory diseases requiring systemic 
immunosuppressive treatment.

Clinically significant medical history contraindicating the 
participation in the study according to the judgment of the 
Investigator or Sponsor

Clinically 
significant 
laboratory 
(hematology, 
blood 
chemistry, or urinalysis) parameters

Transfusion of RhD-positive blood or blood derived products 
within the 6 months prior to enrolment

Anticipated poor compliance with the study procedures


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STUDY No.
ADNC-1301

Subject within exclusion period further to her participation in 
a clinical study
For the IM arm only, subject with coagulation disorders 
contraindicating intramuscular injection (subject will still be 
considered for the IV arm)
DURATION OF SUBJECT 
PARTICIPATION
Minimal duration: approximately 9 months
Maximum duration: 18 months approximately
STUDY PLANNING
Expected start date: Q3 2013
Expected end date: Q4 2017
STUDY DESCRIPTION
Selection of country/site: France / multicenter
Inclusion of subjects: subject information and written informed 
consent form must be obtained before any study procedure. 
Subjects will be included when they signed their informed 
consent form. The first 35 subjects will be enrolled in the IM 
arm; the subsequent 25 subjects will be enrolled in the IV arm.
For the IM arm, the subject will attend 13 visits (9 visits will be 
performed at the hospital unit and possibly 4 at home). If further 
follow-up is needed to document alloimmunization, the subject 
may attend 1 or 2 additional visits.
For the IV arm, the subject will attend 14 visits (9 visits will be 
performed at the hospital unit and possibly 5 at home). If further 
follow-up is needed to document alloimmunization, the subject 
may attend 1 or 2 additional visits.
The study consists of a screening period, an antenatal period, a 
postnatal period and a follow-up period:

The screening visit will be performed as close as possible to 
the antenatal IMP administration, as long as all screening 
results are available prior to IMP administration. However,
the screening period can occur between 12 and 27 weeks GA

The antenatal period comprises 6 visits for the IM route and 
7 for the IV route and lasts approximately 12 weeks

The postnatal period comprises 2 visits and lasts 3 1 days

The follow-up period comprises at least 4 visits (6 months 
2 weeks) and possibly 2 additional visits.
Clinical examinations and non-clinical tests will be performed 
as detailed in the flow chart.
ENDPOINTS AND EVALUATION 
PARAMETERS
Primary endpoint
Serum concentrations of roledumab will be measured at the 
defined time points. The PK profile will be described by:
1)
A compartmental population PK model with 3 primary 
parameters: Volume of distribution (Vd), Clearance (CL) 
and for the IM route absorption rate constant (Ka) and 
derived parameters: Cmax, Tmax, Area under the curve 
(AUCinf), terminal half-life (t1/2) and Elimination rate 
constant.


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STUDY No.
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2)
A secondary PK analysis (NCA), which will provide the 
following parameters Vd, CL, Cmax, Tmax, AUCt, AUCinf, 
terminal half-life and elimination rate constant.
All samples (ante- and postnatal samples) will be used for 
population PK modeling whereas only samples post-first 
administration (ante-natal sample) will be analyzed by NCA for 
each study arm. 
Secondary endpoints
Safety of the mother and the fetus/newborn will be 
assessed throughout the study by the incidence, nature, 
severity, seriousness, relationship to the IMP of AEs and 
by changes in physical examination findings, vital signs,
clinical laboratory parameters, and Doppler ultrasound.
Safety of newborns/infants up to 6 months age will be 
assessed by recording and reporting any AE and related 
concomitant medications.
RhD-alloimmunization rate will be assessed at 6 months 
and up to 12 months if applicable and described. 
Roledumab first milk/breast milk concentration/maternal 
serum concentration ratio will be described.
Roledumab cord blood concentration/maternal serum 
concentration ratio after delivery will be described.
STATISTICS
The statistical analysis will be descriptive on all endpoints. No 
statistical hypothesis will be tested.
There will be two PK analyses using two analysis populations
for both the IM and IV arm:
-
The primary PK analysis (population PK modeling) will 
be performed on PK set 1 (PKS1).
-
The secondary pharmacokinetic analysis will be a non-
compartmental analysis that will be performed on PK set 
2 (PKS2) and PKS3


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STUDY FLOW CHART FOR THE IM arm
Visit (V)
Day (D)
Week (W)
Month (M)
Screening
visit
(V1)
12-27 GA
Antenatal period 
Postnatal period 
Follow-up period
V2
Week 28 or 29 GA 
(D1)
V310
48h24
V410
120h24
V5
9d3
V6
29d4
V7
59d4
V8
Postnatal Treatment 
72 hours of 
delivery
V 9
Discharge
48h24
V1010
72h24 
after post-
natal
injection*
V1110
9d2 after 
postnatal
injection*
V12
6W2W
V13
6M2W
Additional
AV1
9M
2W
Additional 
AV2
12M
2W
Inclusion
Injection
Before
Injection*
Informed Consent
x
Eligibility Criteria
x
x
Demographics/ 
Medical History/
Life habits
x
Serology: hepatitis 
B and C, HIV
x
RhD status
x
ABO status
x
Non-inv. fetal RhD
genotyping
x
Vital Signs1
x
x
x 30 min 
and 6h 
after 
injection
x
x
x
x
x
x
x 30 min 
after 
injection
x
x
x
x
x
x
x
Physical 
Examination
x
x2
x2
x2
x2
x2
Laboratory Tests
x3
x4
x4
x4
x12
x4
x4
Cytokines5
x
x
6h after 
injection
x
x
6h after 
injection
CRP
x
x
x
x
Obstetric and fetal
Doppler ultrasound
x
x
x
x
IAT in mothers 
x
x
x6
x 6
x 6
x 6
Anti-D 
Quantification
x
x
x
x
x
x
x
IM IMP 
administration
x
x
PK sample
x
x
x
x
x
x
x
x
x
x


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Visit (V)
Day (D)
Week (W)
Month (M)
Screening
visit
(V1)
12-27 GA
Antenatal period 
Postnatal period 
Follow-up period
V2
Week 28 or 29 GA 
(D1)
V310
48h24
V410
120h24
V5
9d3
V6
29d4
V7
59d4
V8
Postnatal Treatment 
72 hours of 
delivery
V 9
Discharge
48h24
V1010
72h24 
after post-
natal
injection*
V1110
9d2 after 
postnatal
injection*
V12
6W2W
V13
6M2W
Additional
AV1
9M
2W
Additional 
AV2
12M
2W
Inclusion
Injection
Before
Injection*
KB test 
x
x11
Breast milk sample
x
x
ADA tests
x13
x
x
x
x
x
x
x
x
x
Adverse events 
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
Concomitant 
treatment 
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
Delivery7
x
Cord blood 
sample8
x
Newborn RhD 
status9
x
Newborn ABO 
status14
x
Newborn general 
status
x
x
x
1
Include blood pressure, heart rate and body temperature
2
Partial examination (Weight, General Appearance, Skin, Heart, Lungs, Abdomen)
3
Hematology, blood chemistry, aPTT, urinalysis
4
Hematology, blood chemistry
5
Cytokines: IFN- , IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13 and TNF-
6
If IAT positive, IA characterization and Anti-D antibodies quantification 
7
Collection of APGAR score, AEs, CMs
8
Cord blood sample: DAT and hematocrit, hemoglobin, reticulocytes, total bilirubin including routine tests (RhD status), detection of roledumab
9
Only in case of a confirmed alloimmunization: RhD genotyping in cord blood will be performed to search for potential D-variants
10
Visit performed at home
11
Kleihauer-Betke test to be performed within 24h after IMP administration only if the one prior to the IMP administration was positive.
12
Hematology, blood chemistry, urinalysis
13
Evaluation of pre-existing antibody presence
14
If performed in routine practice during the delivery hospitalization of the mother
* Postnatal injection should be performed within 72h after delivery. All subsequent visits are based from this time point.
GA: gestational age refers to the duration of the pregnancy since the 1st day of the last menstrual period and will be expressed in weeks


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STUDY FLOW CHART FOR THE IV ARM
Visit (V)
Hour (H)
Day (D)
Week (W)
Month (M)
Screening
visit
(V1)
12-27 GA
Antenatal period 
Postnatal period 
Follow-up period
V2
Week 28 or 29 GA (D1)
V2b10
24h
2h
V310
48h
6 h
V410
96h-
120h
V5
9d
3
V6
29d4
V7
59d4
V8
Postnatal 
Treatment <72 
hours of delivery
V 9
Discharge
48h24
V1010
72h24
after
post-
natal
injection*
V1110
9d2 
after 
post-
natal 
injection*
V12
6W2W
V13
6M2W
Additional
AV1
9M
2W
Additional
AV2
12 M
2W
Inc. Inj. 
(H0)
30
5min
H1 
5min
H6 
15min
Before Injection*
Informed Consent
x
Eligibility Criteria
x
x
Demographics/ 
Medical History/ 
Life habits
x
Serology: hepatitis 
B and C, HIV
x
RhD status 
x
ABO status
x
Non-inv. fetal RhD
genotyping
x
Vital Signs1
x
x
x
x
x
x
x
x
x
x
x
x
x
x 30 min 
after inj.
x
x
x
x
x
x
x
Physical 
Examination
x
x2
x2
x2
x2
x2
Laboratory Tests
x3
x4
x4
x4
x12
x4
x4
Cytokines5
x
x
x
x
6h after 
inj.
CRP
x
x
x
x
x
x
Obstetric and fetal
Doppler ultrasound
x
x
x
x
IAT in mothers 
x
x
x 6
x 6
x 6
x 6
Anti-D 
Quantification
x
x
x
x
x
x
x
IV IMP 
administration
x
x
PK sample
x
x
x
x
x
x
x
x
x
x
x
KB test 
x
x11


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Visit (V)
Hour (H)
Day (D)
Week (W)
Month (M)
Screening
visit
(V1)
12-27 GA
Antenatal period 
Postnatal period 
Follow-up period
V2
Week 28 or 29 GA (D1)
V2b10
24h
2h
V310
48h
6 h
V410
96h-
120h
V5
9d
3
V6
29d4
V7
59d4
V8
Postnatal 
Treatment <72 
hours of delivery
V 9
Discharge
48h24
V1010
72h24
after
post-
natal
injection*
V1110
9d2 
after 
post-
natal 
injection*
V12
6W2W
V13
6M2W
Additional
AV1
9M
2W
Additional
AV2
12 M
2W
Inc. Inj. 
(H0)
30
5min
H1 
5min
H6 
15min
Before Injection*
Breast milk 
sample
x
x
ADA tests
x13
x
x
x
x
x
x
x
x
x
Adverse events 
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
Concomitant 
treatment 
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
Delivery7
x
Cord blood 
sample8
x
Newborn RhD 
status9
x
Newborn ABO 
status14
Newborn general 
status
x
x
x
1
Include blood pressure, heart rate and body temperature
2
Partial examination (Weight, General Appearance, Skin, Heart, Lungs, Abdomen)
3
Hematology, blood chemistry, aPTT, urinalysis
4
Hematology, blood chemistry
5
Cytokines: IFN- , IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13 and TNF-
6
If IAT positive, IA characterization and Anti-D antibodies quantification 
7
Collection of APGAR score, AEs, CMs 
8
Cord blood sample: DAT and hematocrit, hemoglobin, reticulocytes, total bilirubin including routine tests (RhD status), detection of roledumab
9
Only in case of a confirmed alloimmunization: RhD genotyping in cord blood will be performed to search for potential D-variants
10
Visit performed at home 
11
Kleihauer-Betke test to be performed within 24h after IMP administration only if the one prior to the IMP administration was positive
12
Hematology, blood chemistry, urinalysis
13
Evaluation of pre-existing antibody presence
14
If performed in routine practice during the delivery hospitalization of the mother
* Postnatal injection should be performed within 72h after delivery. All subsequent visits are based from this time point.
Inc: Inclusion / Inj: Injection / h: Hours / Min: Minutes; GA: gestational age refers to the duration of the pregnancy since the 1st day of the last menstrual period and will be expressed 
in weeks


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STUDY FLOWCHART – ADDITIONAL VISITS IN CASE OF SENSITIZING EVENT FOR 
IM/IV ARMS
Visit
Before injection
6 hours after IMP 
administration
24 hours after IMP 
administration
Sensitizing event
(description)
x
Vital signs
x
x
(6h after injection)
x
Partial physical 
examination
x
IAT
x*
Kleihauer-Betke test
x**
x***
Cytokines
x
(6h after injection)
CRP
x
Adverse Events
x
x
x
Concomitant treatment
x
x
x
*
Perform a ponderal dosage analysis if IAT is positive, to confirm the anti-D concentration
** Before and after (if the first one is positive) anti-D administration
*** The dose will be adapted to the KB results
The PK sampling schedule after the antenatal injection should stay the same in case of a sensitizing 
event treated with roledumab.


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1. INTRODUCTION AND BACKGROUND INFORMATION
Disease and Context
1.1.
1.1.1. Disease
The RhD (Rh) factor is a red blood cell (RBC) surface antigen that was named according to the 
monkeys in which it was first discovered. Rh incompatibility, also known as Rh disease, is a 
condition that occurs when a woman with Rh-negative blood type is exposed to Rh-positive blood 
cells, leading to the development of antibodies. Although the Rh blood group systems consist of 
several antigens (D, C, c, E, e), the D antigen is the most immunogenic and most commonly 
involved in Rh incompatibility.
Sensitization or alloimmunization can happen at any time during pregnancy, but is more common in 
the third trimester and during childbirth. The first exposure from an RhD-negative mother bearing 
an RhD-positive fetus does not generally affect the pregnancy during which it occurs, even if
fetomaternal hemorrhage sensitizes the mother against the D antigen. Subsequent hemolytic disease, 
leading to morbidity and mortality, can affect the fetus or the newborn in future pregnancies [1], [2]. 
Hemolytic disease of the fetus and newborn (HDFN) can result in complications before birth, or 
anemia and hyperbilirubinemia after birth or both including intrauterine fetal death.
The risk of sensitization depends on the volume of transplacental hemorrhage, the level of the 
maternal immune response and the concomitant presence of ABO incompatibility. The risk and the 
severity of the sensitization response increase with each subsequent pregnancy involving a fetus
with RhD-positive blood.
Routine antenatal care was first proposed in the 1970s to all RhD-negative pregnant women i.e. an 
anti-RhD IgG immunoglobulin injection at about 28 weeks of GA (with or without a booster at 
34 weeks gestation). This reduces the effect of the vast majority of sensitizing events which mostly 
occur after 28 weeks of gestation. 
Anti-RhD immunoglobulin is also given to non-sensitized Rh-negative women immediately (within 
72 hours) after potentially sensitizing events occurring earlier in pregnancy.
Therefore the incidence of RhD-alloimmunization has significantly decreased from 13% to 0.35% of 
pregnancies, as a consequence of the preventive use of polyclonal human anti-RhD 
immunoglobulins [3].
1.1.2. Context
These anti-RhD immunoglobulins derived from human plasma of immunized RhD-negative donors 
exposed to RhD-positive RBC raise several concerns regarding i) the ethics of deliberately exposing 
healthy volunteers to human RBC, ii) the potential appearance in blood derived products concerning 
unknown transmissible agents, iii) the potential future shortage of donors and subsequent RhD 
immunoglobulin supply.
These concerns can be circumvented by the development of a human monoclonal recombinant anti-
D antibody such as roledumab, which is manufactured with consistent quality, high purity as well as 


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with large-scale capacities, ensuring unlimited supplies. Thus, this process could enhance 
substantially the benefit-risk ratio of an anti-D treatment as not extracted from human plasma.
Roledumab is being developed to prevent RhD-alloimmunization in RhD-negative women carrying 
an RhD-positive fetus as well as treatment of RhD-negative individuals after incompatible 
transfusions with blood components containing RhD-positive RBCs.
Target
1.2.
Roledumab is a human recombinant monoclonal antibody that specifically recognizes the RhD 
antigen on human RBC with high affinity. 
Anti-D IgGs prevent alloimmunization by first binding to the RhD antigen of the RhD-positive 
RBC, resulting in the elimination of these RBCs from the circulation by mononuclear effector cells, 
mainly in the spleen. This clearance is associated with a dynamic suppression of the primary 
immune response, thus preventing immunization. The most serious clinical consequence of 
alloimmunization to fetal RhD-positive RBCs in pregnant RhD-negative females is HDFN during 
subsequent pregnancies. 
IMP, Non-Clinical and Clinical Information
1.3.
1.3.1. IMP and non-clinical information
Roledumab is a recombinant IgG1 deriving from a natural anti-RhD antibody isolated from an 
immunized donor. This IgG1 belongs to a new generation of monoclonal antibodies based on the 
proprietary EMABlingtechnology which confers to roledumab enhanced FcRIIIa interaction. As 
a consequence, this anti-RhD antibody triggers in vitro Antibody Dependent Cell-mediated 
Cytotoxicity (ADCC) against RhD-positive RBC. In the absence of relevant in vivo efficacy animal 
models caused by a high level of species specificity for RhD antigen in mammals [4], extensive in 
vitro pharmacology studies comparative to the reference polyD antibodies have been performed. 
Roledumab affinity for the RhD antigen is similar to that reported for polyclonal anti-D antibody
[5], [6].
Roledumab complies with global quality and safety criteria required at this stage of product 
development, considering that: 
The original clone, from which roledumab is produced, is genetically stable over time
The global structure of the recombinant roledumab, especially of the N-terminal chains, is 
identical to the original human IgG1, with a voluntarily introduced mutation of a free cystein in 
phenylalanine at position 68 of the heavy chains
The manufacturing of roledumab is compliant with cGMP
Although the mode of action of anti-RhD products is not completely known, yet binding of the 
mAbs to the RhD-antigen is important for the efficacy. Due to the monoclonal nature of mAbs, each 
mAb recognizes one specific epitope of the RhD-antigen and binds to it solely. RhD-variants which 
do not express this specific epitope will not be recognized by the individual mAb.
Roledumab is assumed to recognize epitope 13.1 that is not expressed by certain variants i.e. DIVb, 
DVI, DBT, DHAR and DHMi. Roledumab failed to bind to certain variants: Weak types: 10, 11, 18, 


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38 and 58; partial: DIV type 4 (DIVb) DV type 5 (also called DHK or DYO), DVI (type 1, 2 and 4), 
DBT (type 1 and 2), DHAR (ce-d (5)-ce (former ROHAR)), DAU4, DAR-E (also called DAR2) and 
DAR-A (DAR-E+silent mutation); not tested but probably not recognized on the basis of epitope 
13.1 recognition profile: DHMi (epitope 13.1).
Roledumab did not induce any sign of toxicity, nor local intolerability at the injections site. No 
mutagenic activity was detected in the Ames test. Furthermore, non-specific cross reactivity was not 
observed. No differences were showed between roledumab and polyclonal D in in vitro and ex vivo
experiments designed to compare the transplacental transfer of roledumab and poly-D and to assess 
the capacity of roledumab to cross react with fetal tissues.
In conclusion, no evidence of roledumab toxicity has been observed in the safety assessment 
experiment performed.
1.3.2. Clinical information
Further to pre-clinical evidence for activity, LFB BIOTECHNOLOGIES decided to start a clinical 
development program for a marketing authorization application (MAA) with these indications: 
prevention of RhD-alloimmunization in RhD-negative women with an Rh-incompatible pregnancy 
and treatment of RhD-negative individuals after incompatible transfusions with blood components 
containing RhD-positive RBCs.
The current guideline on the clinical investigation of human anti-D immunoglobulin for intravenous 
and/or intramuscular use (CPMP/BPWG/575/99 Rev.1) for the clinical investigation of plasma-
derived polyclonal human anti-D IgG does not apply to a new product such as a recombinant 
monoclonal anti-D antibody. Consequently, a stepwise approach for the development of roledumab 
comprising two clinical studies in healthy male and female RhD-negative volunteers was adopted. 
Results from these clinical studies are summarized below.
1.3.2. 1. Phase 1 pharmacokinetic study (study ADNC-0701)
A phase 1, double-blind escalating single-dose design, randomized, placebo-controlled study in
healthy RhD-negative volunteers was conducted to assess the safety and pharmacokinetic (PK) 
profile of roledumab after IM and IV administration (up to 3000 µg) in 46 subjects including 36 
with roledumab.
The PK characteristics of roledumab (non-compartmental analysis) can be summarized as follows:
The maximum concentration Cmax (geometric mean = 24.5 ng/mL) was reached around 10 days 
after IM administration (300 µg dose). For IV administration the Cmax (geometric mean) was 
144.5 ng/mL reached theoretically just after administration
The terminal elimination half-life was approximately 18 days with 29.9% CV
The absolute bioavailability of a dose of 300 µg IM administration is estimated between 73% and
80%
Exposure after IV and IM route was similar from 18 days after administration (300 µg dose)
Inter-individual variability was relatively low (25%)
The PK of roledumab was not significantly affected by the FCGRT polymorphisms.


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Roledumab was well tolerated up to 3000 µg IV and 300 μg IM in healthy RhD-negative volunteers. 
No concern of site injection reactions was raised. And no immune responses were reported 
throughout the study (internal report [7]).
1.3.2. 2. Phase 2 dose-finding study (ADNC-0726)
This dose-finding study was designed to assess the ability of roledumab IV or IM to effectively 
eliminate 15 mL of exogenously-administered RhD-positive RBCs from the circulation of RhD-
negative individuals (male and non-child bearing female), thereby preventing RhD-
alloimmunization when compared to Rhophylac300 µg IM and IV.
The secondary objectives were to assess potential RhD-alloimmunization following RhD-positive 
RBCs administration, to evaluate the pharmacokinetic profile of roledumab and Rhophylacin the 
presence of RhD-positive RBCs following IV and IM administrations and to assess the safety of 
roledumab in the presence of RhD-positive RBCs.
Roledumab 100, 200 and 300 µg IV were concluded to be similarly effective in terms of clearance 
of RhD-positive RBCs when compared to Rhophylac300 µg IV. Roledumab 300 µg IM was 
similarly effective in the clearance of RhD-positive RBCs when compared to Rhophylac300 µg
IM.
The exposure to roledumab after IV administration of doses 100, 200 and 300 µg increased more 
than proportionally to the dose administered. 
In healthy volunteers, the PK of roledumab was comparable to Rhophylac.
The PK parameters determined from phase 2 study are summarized as follows:
After IM injection median tmax of roledumab was 117 hours (5 days) while that of Rhophylac
IM was 141 hours (6 days)
The absolute bioavailability of a dose of 300 µg IM administration was 76%
This study did not raise any significant safety concerns: no subject withdrawal for Adverse Event 
occurred and no immunization event was reported with roledumab. No difference in the nature and 
the incidence of AEs between roledumab and Rhophylacwas observed regardless of the route of 
administration (internal report [8]).
Rationale of the Study
1.4.
The PK profile of roledumab has been studied, with doses ranging from 30 to 3000 µg (IV) and 
300 µg IM in healthy volunteers without (phase 1 ADNC-0701) and with (phase 2 ADNC-0726) the 
target cells (RhD-positive RBCs). Conclusions are in favor of a comparable PK profile of roledumab 
with both IM and IV compared to a reference plasma derived polyclonal anti-D treatment: 
RhophylacIM and IV [9].
Roledumab has demonstrated its activity in non-clinical studies and in clinical experience in clearing 
RhD-positive RBCs in healthy volunteers. No relevant or significant safety issues were observed (up 
to 3000 µg IV) in subjects treated in the absence of target cells (RhD-positive RBCs) and up to 
300 µg IV in the presence of the target cells.


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These data from previous experience with roledumab support the conduct of this first study in 
pregnant woman to assess the PK profile of roledumab 300 µg IM/IV dose in the targeted 
population (i.e. pregnant women) and to assess the suitability of this dose in ensuring a sufficient 
residual level at delivery. This study will also document the safety of 300 µg IM/IV route in the 
mother, fetus and newborn. Data on the roledumab fetal concentration (cord blood at birth) and first 
milk/breast milk concentration will be also collected in the study.
The route of administration of anti-D has historically been through both IM and IV routes depending 
on different country’s practices. The IM route is usually applied due to its convenience. The IV 
route, though, may be used in specific situations where, e.g. IM administration is contra-indicated 
(coagulation disorders), a larger volume of the drug is needed (large volume of FMH) or when an 
immediate bioavailability is warranted (after a sensitizing event) as well as according to local 
practice. Therefore, both administration routes (IM/IV) are being investigated in this phase 2b 
clinical study to allow further implementation of an international larger scale phase 3 pivotal clinical 
study required for roledumab regulatory registration.
1.4.1. Rationale of the dose
The current recommendations for Rhophylacuse in the prevention of alloimmunization are: a first 
dose of 300 µg around 28 weeks and a second dose of 300 µg within 72h after delivery (CNGOF, 
2010). The objective of these recommendations for planned prevention is to administer a dose 
sufficient to clear potential fetal RBC in the maternal bloodstream during the third trimester of 
pregnancy and to ensure the coverage until delivery.
A phase 2 study (ADNC-0726) assessing the capability of roledumab to eliminate 15 mL of RhD-
positive RBC from the circulation of healthy male and female RhD-negative volunteers showed that:
300 µg IM/IV dose of roledumab dose was as efficient as the recommended dose of Rhophylac
to clear 15 mL of RhD-positive RBC
Roledumab pharmacokinetics IM/IV profile is comparable to that of Rhophylac
These results confirmed the comparability of the PK parameters observed in the phase 1. Based on 
this comparability on the activity in term of RBCs clearance and in PK parameters, the dose of 
300 µg IM/IV of roledumab is supported to ensure RhD-alloimmunization prevention until delivery.
Although a difference in bioavailability between IM and IV exists (76% vs. a presumed 100%, 
respectively), the exposure after 18 days is similar between the two routes of administration, 
indicating that the same dose IM and IV will lead to the same longer term protection against 
alloimmunization. Hence the same dose of roledumab will be used in both the IM and IV arm.
Previous clinical experience with roledumab did not bring any safety concerns with doses up to 
3000 µg in healthy volunteers. 
All these results support the use of a dose of 300 µg IM/IV roledumab in this PK and safety study.
Furthermore, the repeated dose toxicity rat study showed that the NOAEL (No Observed Adverse 
Effect Limit) is equal or higher than 1650 µg/kg. This dose led to an exposure of more than
700 times of the exposure observed in healthy volunteers after IV administration of 300 µg of 
roledumab (AUClast = 715.5 day µg/mL in rat vs. 0.995 day µg/mL in human). The maximum dose 
in this study in case of massive FMH is 1800 µg as per current practice with reference treatment 
(Rhophylac) corresponding to 6 doses of 300 µg IM/IV. The exposure with this maximum dose is 
still approximately 30 fold lower than the exposure at the NOAEL.


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All these data support further clinical investigation in pregnant women of 300 µg IM/IV for 
systematic prevention during third trimester and additional doses in case of sensitizing event as 
explained in Section 6.4.2. 2.2.
1.4.2. Appropriateness of this study
The primary objective of this study is to describe the pharmacokinetic profile of roledumab 300 µg
administered in RhD-negative pregnant women in order to confirm the suitability of this dose to 
ensure adequate anti-D level coverage up to delivery. 
The design of this study will allow the documentation of the PK profile and safety (for the mother, 
fetus and newborn) of roledumab 300 µg administered through both the IM and IV route in the 
target population. It will provide the basis for a phase 3 clinical study in a larger cohort of pregnant 
women.
2. STUDY OBJECTIVES
Primary Objective
2.1.
The primary objective is to assess the pharmacokinetic profile of roledumab 300 µg IM and IV in 
RhD-negative pregnant women carrying an RhD-positive fetus.
Secondary Objective(s)
2.2.
The secondary objectives are:
To assess the safety of roledumab administered IM and IV in RhD-negative pregnant women and 
in RhD-positive fetus and newborn
To assess the efficacy of roledumab 300 µg IM and IV to prevent RhD-alloimmunization in RhD-
negative pregnant women carrying an RhD-positive fetus
To assess the immunogenicity of roledumab
To measure roledumab concentration in first milk and breast milk
To measure roledumab concentration in cord blood after delivery
3. STUDY DESIGN
This is a phase 2b, interventional, multicenter, open-label, non-randomized, sequential study.


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Visits schedule in the IM arm


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Visits schedule in the IV arm


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Design
3.1.
The study will initially enroll 35 subjects to be treated with 300 µg IM (IM arm) and subsequently 
25 subjects to be treated with 300 µg IV (IV arm).
The subject will attend at least 13 visits in the IM arm and 14 in the IV arm (9 will be performed at 
the hospital unit and 4 (IM) and 5 (IV) at home possibly). Additional visits may occur. 
Additional visits will occur in case of:
Sensitizing events requiring study treatment administration
Potential RhD-immunization requiring follow-up up to 12 months after the study treatment 
administration.
A diagram of the study plan for both the IM and IV arm is given below in Figure 3–1 and 
Figure 3–2.






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The study consists of a screening period, an antenatal period, a postnatal period and a follow-up 
period:
The screening visit will be performed as close as possible to the antenatal IMP administration, as 
long as all screening results are available prior to IMP administration. However the screening 
period could start within 12 and 27 weeks GA.
The antenatal period comprises 6 visits (IM) or 7 visits (IV) and lasts about 12 weeks.
The postnatal period comprises 2 visits and lasts 3 1days.
The follow-up period comprises at least 4 visits (6 months 2 weeks) and possibly 2 additional 
visits.
Screening period (from week 12 to week 27 GA)
Suitable subjects will be provided with appropriate information with regards to the study procedures 
and with an informed consent form. They will be given sufficient time to think about their 
participation and to raise questions. They will give signed written informed consent before any 
protocol related procedures.
Only RhD-negative women carrying an RhD-positive fetus are at risk for alloimmunization. 
Therefore the results of routine or not non-invasive fetal RhD genotyping test on maternal blood will 
be collected at screening for the subjects in the selected sites. The RhD fetus status will be 
determined by the genotyping test prior to the 29th week of gestation. 
In addition, subjects will be screened for any previous sensitization (e.g. due to an incompatible 
transfusion or a previous pregnancy) or pre-existing antibodies against roledumab. The presence of 
anti-RhD IgG will be tested by an indirect antiglobulin test (IAT).
All screening results have to be available and verified before the treatment period: subjects who do 
not meet all inclusion criteria or who meet at least one exclusion criterion will not receive IMP 
administration and be excluded from the study.
Treatment period
This part will be described in Section 8.2.2 and following sections.
Follow-up period (from discharge to 6 months up to 12 months)
This period consists of 4 visits (6 months 2 weeks) and possibly 2 additional visits for safety 
assessment in the mother and collection of routine data for the newborn at the 6 month visit.
Endpoints
3.2.
3.2.1. Primary endpoints
Serum concentrations of roledumab will be measured at the defined time points as described in the 
flowchart. The PK profile of roledumab will be described by:
1. A compartmental population PK model with 3 primary parameters: Volume of distribution (Vd), 
Clearance (CL) and for the IM route absorption rate constant (Ka); and derived parameters: Cmax, 
Tmax, Area under the curve (AUCinf), terminal half-life (t1/2) and Elimination rate constant.


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2. A secondary PK analysis (NCA), which will provide the following parameters Vd, CL, Cmax, 
Tmax, AUCt, AUCinf, terminal half-life and elimination rate constant. Performing PK assessment 
after IV administration will further allow estimation of the bioavailability of roledumab and 
better accuracy of the parameters estimation. 
All samples (ante- and postnatal samples) will be used for population PK modeling whereas only 
samples post-first administration (antenatal samples) will be analyzed by NCA for each study arm. 
3.2.2. Secondary endpoints
Safety of the mother and the fetus/newborn will be assessed throughout the study by the incidence, 
nature, severity, seriousness, relationship to the IMP of AEs and by changes in physical examination 
findings, and vital sign measurements, clinical laboratory tests and Doppler ultrasound. 
Safety of newborns/infants up to 6 months age will be assessed by recording and reporting any AE
and related concomitant medications. 
Roledumab breast milk/first milk concentration, maternal serum concentration and cord blood 
concentration will be also measured.
RhD-alloimmunization rate at 6 months and up to 12 months if applicable.
Study Investigational Site(s)
3.3.
This study will be conducted in hospital’s gynecology/obstetrics units. Their laboratory should 
perform non-invasive antenatal RhD genotyping test.
It is planned that more than one site will take part in the study in order to include 35 subjects and get 
30 evaluable subjects in the IM arm and 25 to get 20 evaluable in the IV arm. 
Data and Safety Monitoring Board
3.4.
An external Data Safety Monitoring Board (DSMB) will monitor safety outcomes and study conduct 
and will provide the Sponsor with recommendations regarding continuing or stopping the study for 
all subjects or subgroups of subjects all along the study. 
DSMB sessions could be regular sessions or ad-hoc sessions.
DSMB sessions are planned to take place to review all safety data gathered, as below:
After the first subject receives roledumab, before administering subsequent subjects in the 
study
After 5 subjects administered at least one dose of roledumab IM
After 10 subjects administered at least one dose of roledumab IM, to allow treatment of 
sensitizing events with roledumab in subsequent subjects
After the last subject received the postnatal roledumab IM
After 10 subjects administered at least one dose of roledumab IV
After the last subject received the postnatal roledumab IV
Ad-hoc sessions can be requested by the Sponsor or the DSMB as needed and at any time along 
the study.


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Data summaries for the DSMB will be prepared by the Sponsor. The safety data will include, but not 
be restricted to (serious) adverse events (AEs) and the safety outcomes listed as secondary endpoints 
(laboratory data, Doppler Ultrasound results…). 
DSMB sessions will contain an open part with DSMB members and Sponsor representatives to 
present data and answer any request from DSMB members, followed by second part with only 
DSMB members to analyze data and provide their decision and recommendations to the Sponsor in 
a written report. 
If the DSMB recommends modification or cessation of the study protocol, this will be discussed 
with the Sponsor who will make the decision. 
The DSMB will be composed of different individuals with different and complementary expertise: a 
neonatologist, two gynecologists-obstetricians who have experience with studies in pregnant women 
and some experience on previous DSMBs, an immunologist with extensive knowledge of the use of 
monoclonal antibodies and a toxicologist who have experience in immunotoxicology.
The Sponsor will propose a detailed mandate and review this with the DSMB, from the Outset (a 
detailed DSMB chart and operating rules will be prepared and signed by DSMB members before 
study start). 
For ad-hoc DSMB sessions, a specific procedure will be set up and be used to monitor special 
conditions and acute situations that need the direct and immediate attention of the DSMB members.
The situations when this procedure must be used by the Sponsor are, but not limited to:
1. Any Serious Adverse Event suspected to be related to the IMP or to the study procedures
2. Any antenatal sensitizing event requiring additional doses
3. Suspicion of occurrence of anti-roledumab antibodies
4. Suspicion of any alloimmunization.
4. DURATION AND TIMELINES OF THE STUDY
The duration of this study is expected to be of approximately 39 months, with a First Patient In (FPI)
expected in Q3 2013, and a Last Patient Out (LPO) in Q4 2017 if no positive IAT and 
alloimmunization confirmed by anti-D quantification at V13.


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5. STUDY POPULATION
Number of Subjects
5.1.
In the IM arm, thirty-five (35) subjects who pass the screening procedures will be included in order 
to get 30 evaluable subjects.
In the IV arm, twenty-five (25) subjects who pass the screening procedures will be included in order 
to get 20 evaluable subjects.
Eligibility Criteria
5.2.
A subject is eligible when all inclusion and no exclusion criteria are met. These criteria will be 
checked during the screening/inclusion visit.
5.2.1. Inclusion criteria
a.
Signed and dated informed consent form provided by the subject prior to proceeding with 
any study-related procedure
b. At least 18 years old
c.
Pregnancy between 12 and 27 weeks gestational age as confirmed by early ultrasound
d. Pregnant RhD-negative woman carrying an RhD-positive fetus confirmed by a non-invasive 
fetal RhD genotyping test
e.
Negative serology: HIV (1 and 2), hepatitis B, hepatitis C except for positive results due to
vaccinations
f.
Covered by healthcare insurance in accordance with local requirements.
5.2.2. Exclusion criteria
a.
RhD-alloimmunized subject
b. Positive for ADA test
c.
Multiple fetuses
d. Occurrence of a documented potential sensitizing event in this pregnancy before the 
antenatal IMP administration
e.
Prior administration of anti-RhD immunoglobulin during the current pregnancy
f.
Known clinically relevant maternal or fetal abnormality (e.g. as determined by ultrasound or 
genetic testing), such as placenta previa
g. History of anaphylactic or severe systemic reaction to immunoglobulin of any origin
h. Current diagnosis of an immune disease which by itself or its treatment could impair the 
safety and/or efficacy evaluation of roledumab in this study. These diseases are:
All immune deficiencies, particularly those requiring IV-Ig supplementation or other systemic 
treatment


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Connective tissue and autoimmune diseases (e.g. systemic lupus erythematosus, 
antiphospholipid syndrome, Sjögren’s syndrome, rheumatoid arthritis, ankylosing 
spondylarthritis) requiring systemic immunosuppressive treatment
Allergic and inflammatory diseases requiring systemic immunosuppressive treatment.
i.
Clinically significant medical history contraindicating the participation in the study 
according to the judgment of the Investigator or Sponsor
j.
Clinically significant laboratory (hematology, blood chemistry, or urinalysis) parameters
k. For the IM arm only, subject with coagulation disorders contraindicating intramuscular 
injection (subject will still be considered for the IV arm)
l.
Transfusion of RhD-positive blood or blood derived products within the 6 months prior to 
enrolment
m. Anticipated poor compliance with the study procedures
n. Subject within exclusion period further to her participation in a clinical study.
Duration of Subject Participation
5.3.
For each subject, the participation will last 9 to 12 months if the IAT result is negative at 6-month 
delivery. Otherwise the maximum duration of the participation should be of 18 months
approximately.
The screening visit will be performed as close as possible to the antenatal IMP administration. 
However the screening period can occur between 12 and 27 weeks GA. 
The subject will be given an IMP injection within 72h after delivery and will be followed-up for 
6 months or up to 12 months depending on the IAT results at 6 months or at 9 months.
Early Discontinuation Criteria / Stopping Rule(s)
5.4.
All subjects are free to withdraw from participation in this study at any time, for any reason, and 
without prejudice. The Investigator must withdraw from the study any subject who requests to stop 
participating in the study.
The Investigator must withdraw a subject from the study at the request of the Sponsor or if the 
subject:
Experiences any related serious or intolerable AE
Develops a clinically significant laboratory abnormality that justifies early discontinuation (e.g. 
confirmed anti-roledumab antibodies i.e. occurring after IMP and absent before)
Requires a medication that is prohibited by the protocol or that could impact fetal safety and 
interfere with the analyses (interpretation of the results)
Is not compliant with the protocol
Is lost to follow-up.


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5.4.1. Handling of early terminations
If a subject is withdrawn before completing the study, the reason for withdrawal will be documented 
on the appropriate e-CRF. The specific reason for the withdrawal should be carefully documented 
on the e-CRF. For instance, rather than stating “withdrew informed consent”, the specific reason for 
withdrawing the informed consent should be stated. Whenever possible and reasonable, the 
evaluations that were to be conducted during the "Follow-up visit/End-of-study visit" should be 
performed at the time of premature discontinuation as follows:
During the antenatal period (before delivery): the subject should have the evaluations planned at 
visit 7 (V7) completed with a physical examination and an ADA test. The Investigator should 
attempt to get an ADA test at 6 months 2 weeks after the last injection and collect all data on 
the subject and the newborn.
During the postnatal period (after delivery): the subject should have the evaluations planned at 
visit 13 (V13) and information will be collected for the newborn as per protocol for a non-early 
study termination.
It is mandatory to obtain follow-up data on any subject who terminated prematurely the study 
because of an AE, abnormal laboratory test, or other finding related to the IMP administration.
In any case, every effort must be made to ensure safety follow-up procedures are completed. 
5.4.2. Sponsor’s study termination
The Sponsor reserves the right to terminate the study at any time for clinical or administrative 
reasons. Such a termination must be implemented by the Investigator, if instructed to do so by the 
Sponsor, in a time frame that is compatible with the subject’s well-being.
5.4.3. Lost to follow-up
Investigators should attempt to minimize the number of lost to follow-up. Any attempt will be 
documented in the subject file including all the involved investigations.


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6. INVESTIGATIONAL MEDICINAL PRODUCT(S) (IMP)
Description of IMP(s)
6.1.
The Investigational Medicinal Product (IMP), roledumab is a recombinant fully human monoclonal 
antibody against RhD-positive RBCs produced by biotechnology.
Roledumab is a sterile, clear to slightly opalescent, colorless to slightly yellow, preservative-free 
solution for intra-muscular and intravenous administration. It is supplied at a concentration of 
0.3 mg/mL in 0.9 mg (3.0 mL) single-use glass vials.
Further information such as ingredients is available in the current Investigator’s Brochure.
IMP Packaging and Labelling
6.2.
6.2.1. Roledumab packaging
Each vial of roledumab will be packaged in an individual box and will be supplied along
with administration device including a combo syringe/safety needle:
For IM administration: syringe of 3 mL and 22G 1 ½ needle
For IV administration: syringe of 3 mL, 5 mL (for any administration of roledumab 3 mL), 
21G x 11/2’’ safety needle and catheter Vasofix safety 33 mm 20G (the choice of the 
needle/catheter is at investigator’s discretion and documented in source documents).
6.2.2. Labelling
Each vial of roledumab will be identified by an individual label with two tear-off parts to be used for 
traceability. In addition, these tear-off label parts will be stuck on the NCR subject prescription, one 
will be kept in the subject file and the other at the pharmacy.
The labels will bear the required regulatory texts. Each vial will be labelled with at least the 
following information:
Study no. ADNC-1301
Sponsor details (LFB BIOTECHNOLOGIES): name, address and phone number
Monoclonal Antibody anti-RhD - Roledumab (LFB-R593) - 0.3 mg/mL
Batch number
Re-test date MM/YYYY
Subject number (to be filled in): |__|__|__|
-  |__|__|__|
Site 
- Subject 
Visit 2
8
Other _____________ (to be filled in)
Investigator XXXXXXXX (to be filled in)
For Intramuscular use or Intravenous use only 
Solution for injection
Storage requirement:


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▪
Between +2°C and +8°C
▪
Protect from light, do not freeze
Standard caution statements:
▪
For clinical use only
▪
For use under medical supervision
Management of IMP
6.3.
6.3.1. Shipment and receipt
Roledumab will be supplied to the Hospital Pharmacist of investigational sites by the Sponsor, free 
of charge for both hospital and subject.
Shipment will be organized by the drug distributor (under the Sponsor responsibility), at a 
temperature between +2°C and +8°C and with temperature monitoring device.
The frequency at which the IMP will be supplied to the site will be adapted to the enrolment rate in 
the site and will take into consideration the re-test date of the IMP.
Upon receipt of treatment supplies, the Pharmacist will inventory the study treatment and complete 
the acknowledgement of receipt. Should any abnormality of the supply boxes be observed, the 
Pharmacist must immediately inform the Monitor/Sponsor. The acknowledgement of receipt must 
be returned to the drug distributor as instructed.
6.3.2. Storage requirements
The Hospital Pharmacist will be responsible for the appropriate storage of the IMP at the study site. 
All IMPs must be stored in a safe and locked place with no access by unauthorized personnel.
Roledumab must be stored in a monitored refrigerator between +2°C and +8°C. Vials must be 
protected from light (kept in their boxes) and must not be frozen.
The Hospital Pharmacist must immediately inform the Monitor/Sponsor of non-respect of the 
required storage conditions. Any temperature deviation must be reported within 1 working day to 
the Monitor/Sponsor, using a temperature deviation form and the official written approval for the 
use of treatment must be obtained prior to any dispensation/administration. In the meantime, IMP 
under temperature deviation should be placed under quarantine at refrigerated conditions. In case of 
decision leading to not using the IMP, the vials will be stored under quarantine at ambient 
temperature until shipment back to the drug distributor using the ‘’Return Request form’’.
6.3.3. IMP re-supplying
The IMP re-supplying will be done according to the recruitment rate upon CRA’s or Investigator’s 
request and will take into consideration the re-test date of the IMP.


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6.3.4. IMP accountability
The Sponsor will provide specific forms for drug accountability to the Hospital Pharmacists. These 
forms will allow following the reception, delivery, dispensation, use and return of the drug. Hospital 
Pharmacists will keep IMP accountability up-to-date throughout the study.
Used IMP vials should be kept at ambient temperature in a secure place at the Hospital Pharmacy 
until the records have been verified by the Monitor. 
A Monitor delegated by the Sponsor will visit the Hospital Pharmacy and verify the drug 
accountability.
6.3.5. IMP return, destruction and recall
All used and unused IMP, including empty vials, must be returned to the drug distributor, preferably 
in their original package, as per instructions. When closing the investigational site, the Monitor 
should ensure all IMP vials (used and unused) have been returned to drug distributor.
The Investigator and/or Pharmacist must never destroy the used/unused treatment. 
6.3.5. 1. Return
The return preparation of used/unused IMP will be performed by the Hospital Pharmacist and the 
Monitor. The return will be organized by the drug distributor. The person in charge of IMP at site 
level should fill in and send (e-mail or fax) to the drug distributor the IMP inventory form and the 
return request form in which the Hospital Pharmacist will set a date and time for pick-up of the vials 
by a courier service. The Hospital Pharmacist or a delegate should be available at the predetermined 
date and time when the boxes are collected from the investigational site.
6.3.5. 2. Destruction
After complete reconciliation, destruction of returned and stored (unshipped) vials will be performed 
by drug distributor upon Sponsor's request.
Destruction of syringes will be carried out on an on-going basis by the site under Investigator's 
responsibilities.
The Investigator and/or Pharmacist must never destroy the used/unused treatment. Unusually, it can 
be allowed destroying IMP after motivated written authorization from the Sponsor. In such case, 
destruction certificate should be issued.
6.3.5. 3. Recall
The Sponsor or its representative and the Investigator/Hospital Pharmacist will inform each other of 
any suspected or identified IMP defect. The concerned units of IMP stored at the investigational site 
must immediately be placed in quarantine and must not be administered to subjects until written 
instructions are given by the Sponsor.


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The Monitor will organize with the Investigator/Hospital Pharmacist the return of the concerned 
batch(es) as per the return procedure. Depending on the study status, new batch(es) could be sent to 
the investigational site.
Treatment of Subjects
6.4.
6.4.1. Methods for assigning subjects to treatment groups
Once the target number of subjects is reached in the IM arm, the subsequent subjects will be 
enrolled into the IV arm as per protocol.
All efforts must be taken for subjects to remain in the respective arms, which they were enrolled in
(same antenatal and postnatal administration route).
6.4.2. Dispensing
Under no circumstances will the Investigator allow the IMP to be used differently than as directed in 
the protocol.
6.4.2. 1. Preparation
Roledumab will be supplied at a concentration of 300 µg/mL in a single use vial.
For administration, it will be prepared either at the Hospital Pharmacy or in the unit by the study 
nurse/midwife. In any case, the person in charge of IMP preparation should follow the different 
steps described below:
Check the product appearance, by inspecting visually for particulate matter or discoloration. If 
the appearance is doubtful or diverts from the product description (roledumab is a clear to slightly 
opalescent and colorless to slightly yellow liquid preparation), the person in charge of IMP 
preparation should immediately alert the Monitor/Sponsor
Retrieve more than 1 mL from the vial
Adjust the volume to be injected to 1 mL (as precisely as possible)
Pull the needle out of the vial
If the preparation is performed by the Pharmacist:
The syringe must be recapped with a stopper and identified with an appropriate label, which 
specifies the time of the preparation.
The last step might be skipped if the IMP is prepared in the unit.
6.4.2. 2. Administration
6.4.2. 2.1. Handling
The materials provided by LFB for withdrawal and administration of the drug must in principal be 
used. The site may use other syringes, provided these are made of polypropylene. In such cases, 
LFB must approve these materials prior to actual use.
Roledumab can be administered up to 1 hour after filling up the syringe and having been at room 
temperature.


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IM route: roledumab should be administered within one minute with a needle into the deltoids, the 
anterolateral upper thighs or in the buttocks. 
IV route: roledumab should be administered as a bolus (within approximately one minute). Two 
ways of IV administration are currently supported by compatibility studies:
a. Directly into the vein with a needle
b. Use of an IV catheter of polyurethane. If this is the case, a flush with NaCl 0.9% should be 
performed to clear the catheter while still connected to the subject.
Roledumab should not be administered through a tube (e.g. butterfly needle, IV line).
The administration will be performed under close medical supervision.
6.4.2. 2.2. Dosage
The dosing guidelines for this protocol summarize the indication, the timing of administration and 
the dose in the table below.
Table 6–1: Dosing guidelines for administration
Indication
Timing of 
administration
Dose
Route of 
Administration
Planned antenatal prophylaxis
At week 28 or 29
300 µg*
IV
or 
IM
The same route of 
administration should be 
used throughout the 
subject’s study 
participation
Sensitizing event

Obstetric complications (e.g. 
miscarriage, abortion, 
threatened abortion, FMH)

Invasive procedures during 
pregnancy (e.g. 
amniocentesis, chorionic 
biopsy)
Obstetric manipulative 
procedures (e.g. external 
version) or abdominal 
trauma
Within 72 hours of 
procedure
300 µg* plus 
additional dose (s) if 
needed**
Postpartum prophylaxis (only if 
RhD-positive)
Within 72 hours of 
birth
300 µg* plus 
additional dose (s) if 
needed**
*A dose of 300 µg roledumab will suppress the immunizing potential of 15 mL of RhD-positive RBC. ** see Table 6–2.
Antenatal prophylaxis
Planned antenatal prophylaxis:
A single dose of 300 g of roledumab at 28 or 29 weeks of gestation.
Antenatal prophylaxis following sensitizing events:


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The first 10 subjects will be administered Rhophylacin case of sensitizing event to enable 
the review of all safety data by the DSMB
The subsequent subjects will be administered roledumab after the DSMB confirm this 
recommendation in a written report
All subjects in the IV arm will receive roledumab and not Rhophylacfor treatment of a 
sensitizing event.
Sensitizing events include but are not limited to:
Obstetric complications (e.g. miscarriage, abortion, threatened abortion, FMH)
Invasive procedures during pregnancy (e.g. amniocentesis, chorionic biopsy)
Obstetric manipulative procedures (e.g. external version) or abdominal trauma.
A Kleihauer-Betke test (KB1) will be performed prior to IMP administration in order to determine 
the volume of fetomaternal hemorrhage (FMH).
One dose of Rhophylac(for the first 10 subjects) or roledumab 300 µg will be administered within 
72 hours after the sensitizing event, usually before the KB results being available.
If the result of the KB1 shows that the volume of FMH is greater than 4 mL, additional doses will be 
administered (Table 6–2 and Figure 6–1). The total dose of roledumab as derived from Table 6–2 is 
inclusive of the first dose administered and should never exceed 1800 µg. 
A second Kleihauer-Betke test (KB2) will be performed 24h after the last IMP administration to 
verify the clearance of fetal erythrocytes. If fetal erythrocytes are still present, additional doses will 
be administered according to Table 6–2 and Figure 6–1. All administered doses should not exceed a 
maximum of 1800 µg of roledumab, if more is required, another treatment should be provided. 
Postnatal prophylaxis
Roledumab should be administered to the mother as soon as possible within 72 hours of delivery of 
an RhD-positive infant. 
The postnatal dose must still be given even when antenatal prophylaxis has been administered and 
even if residual activity from antenatal prophylaxis can be demonstrated in maternal serum.
Before roledumab 300 µg IM/IV postnatal administration, a Kleihauer-Betke test (KB1) will be 
performed on a maternal blood sample taken no earlier than 30 min after delivery, in order to 
determine the volume of FMH.
If the result of the KB1 shows that the volume of FMH is greater than 4 mL, additional doses will be 
administered (Table 6–2 and Figure 6–1). The total dose of roledumab as derived from Table 6–2 is 
inclusive of the first dose administered and should never exceed 1800 µg. 
A second Kleihauer-Betke test (KB2) will be performed 24h after the last IMP administration to 
verify the clearance of fetal erythrocytes. If fetal erythrocytes are still present, additional doses will 
be administered according to the standard reference algorithm used with polyclonal anti-RhD IgG 
referenced in Table 6–2 and Figure 6–1. All administered doses (IM or IV) should not exceed a 
maximum of 1800 µg of roledumab, if more is required, another treatment should be provided. 


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Table 6–2: Adapted dose according to the Kleihauer-Betke test CNGOF table
KLEIHAUER
(FE / 10.000 ME)
Dose of 300 µg of 
Roledumab
Doses
µg
0-4
1
300
5-24
1
300
25-44*
1
300
45-64
2
600
65-84
2
600
85-104
2
600
105-124
3
900
125-144
3
900
145-164
3
900
165-184
4
1200
185-204
4
1200
205-224
4
1200
225-244
5
1500
245-264
5
1500
265-284
5
1500
285-304
6
1800
FE: fetal erythrocytes, ME: maternal erythrocytes
* 44 FE/10 000ME corresponds approximately to 4 mL of FMH
This dose regimen is based on the CNGOF (Collège National des Gynécologues et Obstétriciens 
Français - Recommendations 2010 for alloimmunization prevention with 300 µg dose with current 
polyclonal anti-D (Rhophylac)).
The exact dose, the date and time of administration, as well as the site and route of administration 
should be recorded in the e-CRF and subject’s medical records.


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Figure 6–1: Flow diagram for FMH testing and subsequent actions after delivery or sensitizing event


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6.4.3. Compliance
Since the IMP will be only injected by the site staff trained to this sensitization and to the 
requirements of the study protocol, compliance should not be an issue. However any reason for non-
compliance should be recorded in the e-CRF and the subject file.
Randomization Codes and Procedures for Unblinding
6.5.
Not applicable.
7. PRIOR AND CONCOMITANT MEDICATION
Prior Medication
7.1.
The term ‘prior medication’ refers to any medication given and stopped before study entry, i.e. 
before informed consent was signed.
All prior medications taken within one month before entering the study must be recorded in the 
subject’s medical records and documented on the appropriate pages of the e-CRF.
However, any other relevant prior medication taken within more than 1 month before entering the 
study must be documented in the e-CRF. Careful interview should be done in order to ensure that 
none of the below treatments were administered to the subject before inclusion:
Transfusion of RhD-positive blood or blood derived products within the 6 months prior to 
enrolment
Therapy with an immunoglobulin, immuno-stimulating or -suppressive drug or any other therapy 
influencing the immune system, prior to IMP administration (flu vaccinations are allowed)
And no anti-RhD immunoglobulin was given during the current pregnancy
Concomitant Medication
7.2.
The term ‘concomitant medication’ refers to any medication that the subject receives from the study 
entry and at any time during the study, i.e. from informed consent signature to the last study visit. 
This includes the screening/baseline period, treatment period and follow-up as defined in the 
protocol. Concomitant medication should be kept to a minimum during the study. However, if it is 
considered to be necessary for the subject’s welfare or well-being and is unlikely to interfere with 
the study assessments, it may be given at the discretion of the Investigator. All concomitant 
medication must be recorded in the subject’s medical records and documented on the appropriate 
pages of the e-CRF.


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8. STUDY PLAN
A study plan table outlining the study requirements by visit is presented below. 
V2b, V3, V4, V10 and V11 can be done by a study nurse at the subject’s home in agreement with 
the Investigator. 
Study Period
Type of visit
Visit #
Visit timing
Antenatal
Screening
V1
12-27 weeks GA
Inclusion
Roledumab administration
V2
D1 (IM/IV)
Only for the IV arm
V2b
D2
Post-treatment
V3 to V7
D3, D6, D10, D30, D60
Sensitizing events
Sx
Additional visit(s)
Postnatal
Roledumab administration
V8
within 72h
post delivery
Hospital discharge
V9
D3 (post-delivery)
Sampling first milk/breast milk
V10,V11
D4, D10
Follow-up
V12, V13
1.5M (6 Weeks), 6M
If positive IAT and alloimmunization 
confirmed by anti-D quantification
AV1, 
AV2
9M, 12M
Subject Recruitment
8.1.
8.1.1. Informed consent
The Investigator should provide each subject with relevant, comprehensive, verbal and written 
information regarding the objectives and procedures of the study as well as the possible risks 
involved. 
The subject should have enough time and opportunity to inquire about study details. All his/her 
questions should be answered in a satisfying manner. The subject must be informed about her right 
to withdraw from the study at any time. 
Signed informed consent must be obtained from the subject prior to undertaking any study-related 
procedure. 


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Two copies of the informed consent form will be signed and dated by both the subject and the 
Investigator, one will be given to the subject and the second kept in the subject’s file. The process 
for obtaining consent will be documented in the subject’s file.
The ICF will be in accordance with local regulatory requirements. The ICF signed by the subject 
and/or the designated person as described above will also apply for data collected on the newborn.
8.1.2. Subject enrolment
The subject is enrolled when having signed her consent form.
8.1.3. Subject allocation
All subjects fulfilling all inclusion and exclusion criteria will be considered as eligible. They will 
perform the screening/inclusion visit between 12 and 27 weeks (GA) and visit-related procedures. If 
the inclusion is confirmed, the first 35 subjects will be administered roledumab 300 µg IM during 
the second visit (V2) at 28 or 29 weeks (GA). The following 25 subjects will be allocated to the IV 
arm.
8.1.4. Replacement of early withdrawals
For the PK analysis, subjects who do not provide any blood samples prior to delivery will be 
replaced.
Schedule of Visits
8.2.
8.2.1. Screening period: visit 1 (12-27 GA weeks)
At this visit the Investigator will:
Obtain informed consent prior to any study-related procedures
Check the eligibility criteria
Collect demographics, medical history and life habits data
An interview will identify specific information regarding previous pregnancies, abortion and 
other obstetric procedures, anti-RhD IgG administrations in the context of previous or current 
pregnancy. 
Determine or record the RhD status of the mother
Record ABO status
Collect the vital signs
Perform a complete physical examination
The complete physical examination will include Height, Weight, General Appearance, Skin, 
Head, Eye, Ear-Nose-Throat (ENT), lymph nodes, heart, lungs, abdomen, extremities/joints, 
neurological and mental status. 


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Collect blood samples
For laboratory safety parameters (hematology, blood chemistry and aPTT) and serology 
(hepatitis B, hepatitis C, HIV)
For non-invasive fetal RhD genotyping (if not already available)
For indirect antiglobulin test (IAT)
For ADA test
Collect urine (Section 8.3.1)
Collect the information of the first or second trimester obstetric ultrasound (gestational age 
determination and any maternal or fetal abnormality detection) as well as significant events and 
complications of the pregnancy
Collect prior and concomitant medication
Record any AE which occurred between the ICF signature and inclusion visit
Give a diary to the subject and explain her how to complete it
Complete the e-CRF accordingly.
8.2.2. Antenatal period (from visit V2 to visit V7)
8.2.2. 1. Visit 2/D1 (28 or 29 GA)
This visit will be conducted in two parts: before and after IMP administration.
8.2.2. 1.1. Inclusion: prior to IMP administration
At this visit the Investigator will:
Check inclusion/exclusion criteria in order to confirm the inclusion and particularly the results of 
the non-invasive fetal RhD genotyping and ADA tests
Collect the vital signs
Perform a physical examination (Weight, General Appearance, Skin, Heart, Lungs, Abdomen)
Perform an obstetric and fetal Doppler ultrasound
Collect blood samples for 
PK assessment (baseline) for both arms IM/IV
Cytokines
CRP
Immunogenicity assessment – ADA test (baseline)
IAT 
Anti-D quantification by microtitration or ponderal dosage
Record any AE
Record any concomitant medication
Administer the IMP (IM or IV depending on the arm the subject is in) 
Complete the e-CRF accordingly.




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8.2.2. 3. Visit 3/D3 (48 hours)
IM route
48 hours 24h
IV route
48 hours 6h
This visit can be performed at the subject’s home.
At this visit the Investigator/study nurse will:
Collect vital signs
Collect a blood sample for PK assessment and CRP
Record any AE
Record any concomitant medication
Only the investigator will complete the e-CRF accordingly.
8.2.2. 4. Visit 4/D6 (5 days after IMP administration)
IM route
5 days 1 day (120h 24h)
IV route
At 4 or 5 days after injection (96h-120h)
This visit can be performed at the subject’s house.
At this visit the Investigator/study nurse will:
Collect vital signs
Collect a blood sample for PK assessment
Record any AE
Record any concomitant medication
Only the investigator will complete the e-CRF accordingly.
8.2.2. 5. Visit 5/D10 (9 3 days after IMP administration)
IM route
IV route
9 days 3 days
At this visit the Investigator will:
Collect the vital signs
Perform an obstetric and fetal Doppler ultrasound: in case of abnormality or in the Investigator’s 
opinion the subject will be followed up once a week
Record any AE
Record any concomitant medication
Complete the e-CRF accordingly.
Collect blood samples for:
PK assessment
Immunogenicity assessment – ADA test
Perform an anti-D quantification by 
microtitration or ponderal dosage
Collect blood samples for:
Immunogenicity assessment – ADA test
Perform an anti-D quantification by 
microtitration or ponderal dosage


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8.2.2. 6. Visit 6/D30 (29 4 days after IMP administration)
At this visit the Investigator will:
Collect the vital signs
Perform an obstetric and fetal Doppler ultrasound
In case of abnormality or in the Investigator’s opinion the subject will be followed up once a 
week
Collect blood samples for:
PK assessment (for both IM and IV)
Immunogenicity assessment - ADA test
Perform an anti-D quantification by microtitration or ponderal dosage
Record any AE
Record any concomitant medication
Complete the e-CRF accordingly
8.2.2. 7. Visit 7/D60 (59 4 days after IMP administration, if not yet delivered)
At this visit the Investigator will:
Collect the vital signs
Perform an obstetric and fetal Doppler ultrasound: In case of abnormality or in the Investigator’s 
opinion the subject will be followed up once a week
Collect blood samples for:
PK assessment (for both IM and IV)
Laboratory safety tests (hematology, blood chemistry)
Record any AE
Record any concomitant medication
Complete the e-CRF accordingly
8.2.3. Antenatal period - in case of a sensitizing event
In case of a sensitizing event, the subject should attend an additional visit and be administered an 
injection of Rhophylacor roledumab. The first 10 subjects will be administered Rhophylac, the 
subsequent subjects roledumab upon DSMB approval (for both arms).
8.2.3. 1. Additional visit (prior to IMP administration and 6h later)
The Investigator will:
Collect the vital signs
Perform a partial physical examination
Check anti-roledumab antibodies result if available
Collect blood sample before IMP administration for:
IAT testing, if the result is positive, an anti-D quantification will be performed by 
microtitration or ponderal dosage
Kleihauer-Betke test to determine the volume of the FMH
Administration of roledumab according to Section 6.4.2. 2
Collect blood sample 6 hours after the last IMP administration for:
Cytokines testing


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Collect the vital signs 6 h after the IMP administration
Record any AE
Record any concomitant medication
Complete the e-CRF accordingly
8.2.3. 2. Additional visit (24h after IMP administration)
The Investigator will
Collect vital signs
Collect blood sample for a Kleihauer-Betke test (in case the one prior to roledumab 
administration was positive) to determine the volume of the FMH within 24 hours after IMP 
administration, and CRP test
If needed, administer roledumab according to Section 6.4.2. 2
Record any AE
Record any concomitant medication
Complete the e-CRF accordingly
8.2.4. Postnatal period - visit 8 to visit 9
The postnatal treatment period will consist of:
A postnatal visit: visit 8 - before and after IMP administration
A visit at discharge from hospital: visit 9
8.2.4. 1. Postnatal period: within 72h after delivery - Visit 8
8.2.4. 1.1. Postnatal visit: prior to administration
At this visit the Investigator will record data on the mother and on the newborn.
Information on the mother
Collect vital signs
Record data on date and time of delivery, duration of parturition, type of delivery (spontaneous, 
induced, vaginal or C-section), any complications during delivery, instrumental delivery (spatula, 
forceps…)
Perform a partial physical examination (weight, general appearance, skin, heart, lungs and 
abdomen)
Collect blood samples for:
PK assessment (for both IM and IV arm)
Laboratory safety parameters (hematology, blood chemistry)
IAT
Anti-D quantification by microtitration or ponderal dosage
Immunogenicity assessment – ADA test
Kleihauer-Betke test
Cytokines
CRP
Record any AE
Record any concomitant medication


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Complete the e-CRF accordingly
Information on the newborn
Collect APGAR scores, gender, weight, 
Collect cord blood sample for: 
DAT, hematocrit, hemoglobin, reticulocytes, total bilirubin
RhD group, RhD genotyping in case of confirmed alloimmunization
Measurement of roledumab concentration
Record any Adverse Event
Record any concomitant therapy
Complete the e-CRF accordingly
Prior to the postnatal administration (within 72 hours of delivery), a Kleihauer-Betke test will be 
performed in order to determine the volume of FMH. Administration of roledumab will be done 
according to Section 6.4.2. 2.2.
8.2.4. 1.2. Postnatal visit: after IMP administration
At this visit the Investigator will:
Collect the vital signs 30 minutes after IMP administration
Record any AE and concomitant medication
Collect blood samples for cytokines testing 6h after the IMP administration
Collect the vital signs 6h after the IMP administration
Complete the e-CRF accordingly
8.2.4. 2. Postnatal period: discharge from hospital (visit 9 - D3 1 after delivery)
The Investigator will:
Collect vital signs
Record newborn general status
If done in routine practice, collect/record ABO status of the newborn, to assess the ABO 
incompatibility with the mother,
Collect blood samples for CRP testing 24 to 48h after the IMP administration
Record any AE
Record any concomitant therapy
Complete the e-CRF accordingly
8.2.5. Follow-up period (from visit 10 to visit 13, additional visits AV1 and AV2)
8.2.5. 1. Visit 10 - D4 1 after postnatal administration
This visit can be performed at the subject’s home.
The Investigator/study nurse will:
Collect vital signs
Collect a first milk sample (for consenting breastfeeding mother)
Collect blood samples for PK assessment (for both IM and IV)
Record any AE


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Record any concomitant therapy
Only the investigator will complete the e-CRF accordingly
8.2.5. 2. Visit 11 - D9 2 after postnatal administration
This visit can be performed at the subject’s home.
The Investigator/study nurse will:
Collect vital signs
Collect a breast milk sample (for consenting breastfeeding mother)
Collect blood samples for:
PK assessment (for both IM and IV)
Immunogenicity assessment – ADA test
Anti-D quantification by microtitration or ponderal dosage
Record any AE
Record any concomitant therapy
Only the investigator will complete the e-CRF accordingly
8.2.5. 3. Visit 12 - at 6 2 weeks after postnatal administration
The Investigator will:
Collect vital signs
Collect blood samples for:
PK assessment (for both IM and IV)
Laboratory safety parameters (hematology, blood chemistry)
Immunogenicity assessment – ADA test
Anti-D quantification by microtitration or ponderal dosage
Record newborn general status
Record any concomitant therapy
Complete the e-CRF accordingly
8.2.5. 4. End of study visit - visit 13 (or visits AV1 or AV2)
In case of positive IAT and alloimmunization confirmed by anti-D quantification, the Investigator
will ask the subject to attend an additional visit AV1. In case of premature discontinuation, the 
Investigator will perform the procedures described at visit 13.
8.2.5. 4.1. Visit 13 (6 months 2 weeks after postnatal administration)
At this visit, the Investigator will:
Collect vital signs
Perform a partial physical examination (weight, general appearance, skin, heart, lungs, abdomen)
Collect blood samples and urines for:
Laboratory safety parameters (hematology, blood chemistry)
Urinary dipstick (glucose, nitrite, blood and protein) and complete urinalysis in case of 
positive dipstick
Immunogenicity assessment - ADA test


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IAT
Anti-D quantification by microtitration or ponderal dosage
Record any AE
Record any concomitant therapy
Complete the e-CRF accordingly
In case of positive IAT and alloimmunization confirmed by anti-D quantification, the subject will 
attend an additional visit (visit AV1).
8.2.5. 4.2. Additional visit AV1 (9 months 2weeks after postnatal administration)
The Investigator will:
Collect vital signs
Perform a partial physical examination (weight, general appearance, skin, heart, lungs, abdomen)
Collect blood samples for:
Laboratory safety parameters (hematology, blood chemistry)
Immunogenicity assessment – ADA test
IAT
Record any AE
Record any concomitant therapy
Complete the e-CRF accordingly
In case of positive IAT and alloimmunization confirmed by anti-D quantification, the subject will 
attend an additional visit (visit AV2) at 12 months 2 weeks. The procedures will be the same as 
those described for visit AV1.
8.2.6. Follow-up in case of alloimmunization
In case of confirmed alloimmunization, the following clinical, biological and ultrasound follow-up 
surveillance will be applied during pregnancy and after delivery:
Irregular antibodies (IA) characterization and Anti-D antibodies titration and quantitation every 
2 weeks
If anti-D antibodies level is equal to, or higher than 1 µg/mL (or 250 UCHP/mL), weekly 
ultrasound examination will be performed, searching for fetal anemia (any signs of fetal hydrops, 
increased middle cerebral artery peak systolic velocity)
In case of suspected fetal anemia, in utero blood transfusion and/or delivery in a reference center 
should be discussed
Neonatal management will include clinical and biological follow up as well as adequate treatment 
in order to prevent or treat jaundice and anemia in a specialized center
In case of alloimmunization, there will be maternal anti-D generation, consequently the PK results 
will not be analyzed and the PK samples will not be drawn after the alloimmunization confirmation.


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8.2.7. Follow-up in case of ADA production after IMP administration
An ad-hoc DSMB session will be held for these events and additional recommended tests will be 
strictly performed and adhered to.
In case of confirmed ADA, the following decision algorithm will be followed:
Anti-roledumab antibodies occurrence (induced by roledumab, i.e. detected after IMP and absent 
before IMP) will be systematically reported as a serious AE and in case of ADA confirmation in 
maternal plasma, rescue treatment (Rhophylac) will be immediately administered and the same 
biological and ultrasound follow-up as for alloimmunization surveillance will be applied during 
pregnancy and after delivery:
IA characterization and Anti-D antibodies titration and quantitation every 2 weeks
If anti-D antibodies level is equal to, or higher than 1 µg/mL (or 250 UCHP/mL), weekly 
ultrasound examination will be performed, searching for fetal anemia (any signs of fetal hydrops, 
increased middle cerebral artery peak systolic velocity)
In case of suspected fetal anemia, in utero blood transfusion and/or delivery in a reference center 
should be discussed
Neonatal management will include clinical and biological follow up as well as adequate treatment 
in order to prevent or treat jaundice and anemia in a specialized center. 


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Assessments
8.3.
8.3.1. Assessments performed on site
The assessments are described in Section 8.2 and summarized in the table below.
Assessment
Time points
RhD status

At screening (V1) for the mother

At delivery (V8) for the newborn on cord blood
ABO status

At screening (V1) for the mother

No later than at the discharge visit (V9) for the Newborn
Vital signs in mother

At screening

At each visit throughout the study
Physical examination in mother

At screening (complete one)

At Day 1 (V2) prior to antenatal administration

At delivery prior to postnatal administration (V8)

At the End of Study visit (V13) (6 months and up to 12 months 
after postnatal treatment, if applicable)
Laboratory safety parameters (hematology, 
blood chemistry, aPTT, urinalysis*) in mother

At screening* (V1)

At V7 before delivery

At delivery prior to postnatal administration (V8)

At the End of Study visit* (V13) (6 months, and up to 12 months 
after postnatal treatment)
CRP 

At Day 1 prior and 1h, 24h and 48 hours after antenatal 
administration (depending of the administration route)

After delivery before and 24 to 48 hours after postnatal 
administration
Obstetric and fetal Doppler ultrasound

At Day 1 prior to antenatal administration

At Day 10 (V5), D30 (V6) and Day 60 (V7) after antenatal 
treatment

Every week follow-up in the case of an abnormal result
Indirect Antiglobulin Test

At screening (V1)

At Day 1 (V2) prior to antenatal administration

At delivery prior to postnatal administration

At the end of Study visit (V13) (6 months)

At the additional visits in case of a positive IAT
and 
alloimmunization confirmed by anti-D quantification at V13 (9 or 
12 months after postnatal treatment)
Kleihauer-Betke test

Prior to and after IMP administration in case of a sensitizing event 

At delivery prior to postnatal administration (and after when 
required)
Record APGAR score (at delivery) and 
newborn general status

At delivery (+ APGAR score)

At discharge (V9) and 6 weeks 2 weeks (V12) after postnatal 
treatment


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The methods of assessments are detailed below.
Non-invasive fetal RhD genotyping
Non-invasive fetal RhD genotyping based on polymerase-chain-reaction (PCR) is an accurate and 
validated technique. It will be performed at the screening visit (V1) on maternal blood sample.
Vital signs
Systolic and diastolic blood pressures (mmHg), heart rate (beats/minute), and temperature (°C) will 
be recorded in supine position. Any clinically significant abnormal findings will be recorded as AEs 
except for the screening visit.
Physical examination
A complete physical examination will be performed at the screening visit only. This will include the 
following observations/measurements: Height, Weight, General Appearance, Skin, Head, Eyes, 
Ears, Nose, and Throat (ENT), Lymph nodes, Heart, Lungs, Abdomen, Extremities/Joints, 
Neurological and Mental status. Any significant abnormalities observed at screening will be 
recorded as Medical History.
A partial examination will also be performed before the IMP administration at Day 1 (V2), before 
the postnatal administration (at V8), and at the End of Study Visit. In case of additional visit(s) AV1
and/or AV2, a partial examination will also be performed. The following body systems will be 
examined: General Appearance, Skin, Heart, Lungs and Abdomen. Any new clinically significant 
abnormalities will be reported as AEs.
Laboratory safety parameters
Laboratory safety parameters will be performed throughout the study: 
Hematology: RBC count, Hematocrit, Hemoglobin, Mean corpuscular hemoglobin (MCH), Mean 
corpuscular hemoglobin concentration (MCHC), Mean corpuscular volume (MCV), platelet 
count, white blood cells (WBC) count with differential
Blood chemistry: Albumin, Alkaline phosphatase, Alanine aminotransferase (ALT), Aspartate 
aminotransferase (AST), Blood urea nitrogen, Creatinine, Gamma-glutamyl transferase (GGT), 
Glucose, Total bilirubin, Total protein including electrophoresis. C Reactive Protein (CRP) will 
be measured
At screening only:
Activated partial thromboplastin time (aPTT)
Serology: Hepatitis C, Hepatitis B, HIV
At screening (V1) and the end of study (V13) visits: 
Urinalysis: Glucose, Microscopic examination of the sediment only in the case of a positive 
dipstick, Nitrite, Occult Blood, Protein.
Standard methods will be used for the measurement of hematology, blood chemistry and serology. 
Urinalysis will be performed using dipsticks. 


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The Investigator will be in charge to assess whether laboratory values out of normal range has to be 
considered as clinically significant or not. After ICF signature, any clinically significant out of range 
values will be reported as AEs.
Obstetric and fetal Doppler ultrasound
The Obstetric and fetal Doppler ultrasound will be performed prior to roledumab administration 
(V2) and 9 (V5), 29 (V6) and 59 days (V7) after roledumab administration. In case of abnormality 
or in the Investigator’s opinion the subject will be followed up once a week.
Trained Investigator will perform the fetal Doppler ultrasound in order to measure the MCA - PSV 
(Middle Cerebral Artery - Peak Systolic Velocity) at the time points defined by the protocol. 
Frequency should be adapted in accordance with the Mari and Deter curves. The MCA-PSV values 
greater than 1.5 MoM (Multiples of Median) will be considered pathological [10].
Direct and Indirect Antiglobulin Test (DAT and IAT) / Direct and Indirect Coombs test
The Indirect Antiglobulin Test (IAT) will be performed locally using the standard ID-Card 
"LISS/Coombs" (BIORAD or a similar supplier) test at the screening visit (V1), prior to roledumab 
administration at Day 1 (V2), before postnatal administration (V8) and at the end of the study (V13 
and the additional visit(s) AV1 and AV2 if applicable) in order to detect any alloimmunization
during pregnancy.
The test consists of performing an antibody screening with the ready-to-use test cell reagent "ID-
DiaCell". In the case of a positive reaction indicating the presence of irregular antibodies, antibody 
identification will be carried out using the ready-to-use test cell reagent "ID-DiaPanel".
The Indirect Antiglobulin Test (IAT), also known as Indirect Coombs Test, is a qualitative 
evaluation of RhD-positive RBC agglutination in the presence of anti-RhD antibodies. In the 
ADNC-1301 clinical study, IAT is performed right before the first and second IMP administrations 
to assess the alloimmunization status of subjects.
LFB conducted an internal preclinical study (report No. 14ENC005R) to compare monoclonal 
(roledumab) and polyclonal anti-RhD activity values obtained by two methods (A and C) from the 
European Pharmacopeia. Both methods were originally developed using polyclonal antibodies. It 
was observed that the specific activity of roledumab measured using method A (based on analysis of 
RhD-positive RBC agglutination on autoanalyser) was two times lower than the one obtained with 
polyclonal antibodies. The anti-RhD specific activity measured with method C (based on direct 
binding of antibodies to RhD-positive RBC and analysis by flow cytometry) was found to be in the 
same range for both roledumab and polyclonal antibodies. These results were in agreement with 
published data from Thorpe et al [11].
This suggested that not all methods are suitable to precisely measure monoclonal anti-RhD activity. 
In particular, methods based on the agglutination potency of anti-RhD antibodies potentially 
underestimate the specific activity of monoclonal antibodies like roledumab. Thus, a low IAT score 
for samples from roledumab treated subjects cannot be used to draw conclusions about a lack of 
protection by passive anti-RhD antibodies or the absence of IMP efficiency. 


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Consequently, IAT is only to be used to check the alloimmunization status of subjects by evaluating 
the production of polyclonal anti-RhD antibodies. Results from IAT must not be used to assess 
passive monoclonal anti-RhD protection after IMP administration. 
The Direct Antiglobulin Test (DAT) will be performed in cord blood at delivery using the standard 
ID-Card "LISS/Coombs" (BIORAD). 
Kleihauer-Betke (KB) test
The Kleihauer-Betke test is the standard method for quantification of FMH 
Antenatal prophylaxis:
In case of a sensitizing event, and before administration of 300 µg of roledumab, a KB test will 
be performed. If positive, a second KB will be performed 24h after the IMP administration.
Postnatal prophylaxis:
A KB test will be performed no earlier than 30 minutes after delivery and prior to administration 
of roledumab in order to measure the amount of fetal hemoglobin transferred from the fetus to the 
mother's bloodstream. If positive, a second KB test will be performed 24 hours after IMP 
administration.
General status of the newborn
APGAR (Appearance, Pulse, Grimace, Activity, and Respiration) score will be reported. The Apgar 
score is usually given to a baby twice: once at 1 minute after birth, and again at 5 minutes after birth. 
The physical examination of the newborn is the routine clinical examination performed at birth, at 
discharge and at V12. This information will be collected.
Laboratory tests in cord blood
In order to detect any anemia in the newborn, the following laboratory parameters will be measured 
in cord blood according to the standard methods: hematocrit, hemoglobin, reticulocytes and total 
bilirubin and RhD status. If the last two parameters are not available on cord blood, they will be 
collected in the e-CRF if available as part of the standard practices. 
The Investigator will be in charge to assess whether laboratory values out of normal range has to be 
considered as clinically significant or not. Any clinically significant out of range values will be 
reported as AEs.
The Investigator is responsible for reviewing all laboratory reports and evaluating any results that 
are outside the normal range. The Investigator must sign and date all laboratory reports.


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8.3.2. Centralized assessments
Central Laboratories have been identified for the measurements of the following parameters listed in 
the table below.
PARAMETER
TIME POINTS
Roledumab serum concentration 
measurement (PK assessment)

Before delivery

IM arm: prior to IMP administration at Day 1 (baseline), then at V3, 
V4, V5, V6 and V7

IV arm: prior to IMP administration at Day 1 (baseline), then at V2 
1h post-administration, V2b, V3, V4, V6 and V7

IM and IV: at delivery, prior to second, postnatal IMP administration 
(V8), then at V10, V11 and V12
Roledumab measurement in cord blood 
and in first milk / breast milk

At delivery in cord blood

At V10 and V11 in the first milk / breast milk
Anti-roledumab antibody detection
(ADA)

At screening visit (V1)

Prior to the IMP administration (V2)

Day 10 (V5) and Day 30 (V6) after antenatal IMP administration

At delivery, prior to postnatal IMP administration (V8)

Day 10 (V11) and 6 weeks (V12) after postnatal IMP administration

At the End of Study visit (V13), and if applicable at Month 9 (AV1), and 
Month 12 (AV2) after postnatal IMP administration
Cytokines

At V2, V8 and in the case of a sensitizing event: before and 6 hours after 
the IMP administration.
Anti-D Quantification

At V2, V5, V6, V8 (before postnatal administration), V11, V12, V13
Pharmacokinetic assessment: roledumab serum concentration measurement
Blood samplings will be drawn in all subjects once (6 mL) before the IMP administration, then 
(6 mL) five times (IM arm) or six times (IV arm) until delivery and (6 mL) four times (both IV and 
IM) after delivery. Two blood samplings will also be drawn during the postnatal follow-up period. 
The blood sampling will be performed either during a study visit or at the subject’s home by a study 
nurse.
A PK sampling will be performed at study site:
For the IM arm during the visits V2 (D1) (baseline value), V5 (9d 3), V6 (29d 4), V7 
(59d 4), V8 (within 72h of delivery and before the second IMP administration) and V12 
(week 6 2w after the second IMP administration).
For the IV arm during the visits V2 (D1) (baseline value), V2 1h post administration, V6 
(29d 4), V7 (59d 4), V8 (within 72h of delivery and before the second IMP administration)
and V12 (week 6 2w after the second IMP administration).
And at subject's home or at the study site at the subject’s preference:
For the IM arm during the visits V3 (48h 24), V4 (120h 24) after the first IMP 
administration and for V10, V11 (respectively 3d 1, and 9d 2 after the second IMP 
administration).
For the IV arm during the visits V2b (24h post-administration 1), V3 (48h 6), V4 
(96-120h) after the first IMP administration and for V10, V11 (respectively 3d 1, and 9d 2
after the second IMP administration).


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Roledumab serum concentrations will be measured by a validated flow cytometry assay with a lower 
limit of quantification of 1 ng/mL.
The analytical method for the quantification of roledumab in human RhD-negative serum consists of 
measuring the fluorescence intensity of RhD-positive RBCs previously incubated with the clinical 
sample containing roledumab. The complex RhD-positive RBC/roledumab is then revealed by a 
human Fab anti-IgG antibody coupled to a fluorochrome (FITC). Reading is done using flow 
cytometry. The recognition of variable regions of the immunoglobulin (roledumab) by RBCs 
expressing the RhD antigen corresponds to the activity of the compound. The couple human Fab-
FITC anti light and heavy chains that reveals roledumab bound to RBCs corresponds to the structure 
of roledumab as human immunoglobulin.
After drawing biological samples and duplicating serum, the tubes in polypropylene should not 
remain at room temperature for a time-period greater than 12 hours, or for a time-period greater than 
7 days at 4°C. Serum samples will be frozen at -70°C at the investigational site then centralized to 
the Central Laboratory where analyses will be performed.
For each subject, 9 duplicated aliquots of serum will be prepared from the baseline sample because 
each assay by flow cytometry will have to be performed in comparison to the baseline value of the 
volunteer. For the remaining samplings, duplicate aliquots will be prepared.
The logistic provider will organize shipment of the serum samples from the investigational site to 
the Central Laboratory for testing, as instructed by LFB BIOTECHNOLOGIES. To secure 
biological samples, duplicated samples will be centralized at the Central Laboratory at separate 
occasion from the centralization of original samples.
First milk/breast milk passage assessment: roledumab concentration testing in first milk/breast milk
All consenting breastfeeding mothers will be asked to provide a breast milk sample 3 and 9 days 
after postnatal injection since it is predicted that Cmax of roledumab after IM injection will occur 
during this period. For the IV arm, the Cmax will be earlier even though the concentration is expected
to be still higher or at the same level as for IM around these time points. Therefore, the probability 
to detect roledumab in breast milk is maximized. The ratio between the concentration in breast milk 
and blood after postnatal administration will then be calculated to assess the passage of roledumab 
into breast milk. For the determination of roledumab concentration in first milk/breast milk, a 
bridging ELISA method has been developed and will be validated using current bioanalysis 
guidelines. The method is based on a coated anti-idiotypic roledumab antibody using the 
electrochemiluminescent technology (from Meso Scale Discovery). After Quality Control and 
samples deposit, revelation is performed using the same anti-idiotypic antibody that is biotinyled 
and the Streptavidin-Sulfo-TAG as electrochemiluminescent material.
Anti-RhD antibodies quantification by microtitration/ponderal dosage
These tests will be performed centrally. In case of positive IAT, the characterization of the 
specificity of the antibodies will be done and, if anti-D alloimmunization is confirmed, the 
quantification of anti-RhD antibodies will be performed by microtitration or ponderal dosage 
(routine assessment in case of suspicion of alloimmunization).


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Quantification of anti-D antibodies in subjects samples will be processed by a microtitration method 
if sample concentration is below 24 ng/mL or a ponderal dosage method if the sample concentration 
is above 24 ng/mL in accordance to the central laboratory SOPs.
Anti-Drug Antibodies (ADA) assessment: Anti-roledumab antibodies detection
Any biotechnology derived protein is potentially immunogenic.
Blood samples will be drawn at screening visit, before the antenatal IMP administration (baseline), 
at 9, 29 days after the first administration, after delivery before the postnatal administration, 9 days, 
6 weeks and 6 months or up to 12 months after the postnatal administration. 
Ten (10) mL of blood will be drawn at each time point, except at V2 and V8 where 15 mL will be 
drawn. The blood samples will be centrifuged and serum will be aliquoted. The aliquots will be 
immediately shipped to the Central Laboratory, at -70°C, for testing. 
In case of symptoms of allergic reaction, an additional blood sample will be drawn.
LFB BIOTECHNOLOGIES has developed a specific assay based on ELISA method for the 
detection of anti-roledumab antibodies. In this assay, normal human serum samples have been used 
to determine a cut point corresponding to 95% of untreated human serum mean responses. This 
assay has been validated in a GCLP laboratory.
If ADA are detected at any time-point after IMP administration, the following assays will be 
implemented:
ADA confirmatory assay
Determination of neutralizing effect
Isotypic characterization and determination of ADA
Determination of ADA concentration
Confirmatory assay: the same bridging ELISA is used to perform the confirmatory assay but in 
presence of roledumab in excess. This assay has been also fully validated in a GCLP laboratory
Determination of neutralizing effect: neutralizing anti-drug antibodies (NAB) will be detected by 
an anti-D potency assay based on method C described in section 2.7.13. (method C) of the 
European pharmacopeia. Briefly the ability of roledumab to bind to RhD-positive RBCs will be 
checked in presence of serum containing ADA by a flow cytometric method. The method has 
been validated using a murine neutralizing anti-idiotype antibody specific from roledumab as 
positive control
Isotypic Characterization: currently, the isotype characterization technique has not been 
developed. Since appropriate human anti-roledumab isotype controls are not easily available. It is 
foreseen to establish the isotype by surface plasmonic resonance (SPR) technology. Briefly 
roledumab will be fixed on a sensor chips then the serum containing ADA will be added and 
finally anti-IgM, IgG, IgE, IgG1, IgG2, IgG3 and anti-IgG4 will be used to detect the isotype of 
the ADA


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Determination of ADA concentration: Currently, a method to quantify anti-roledumab antibodies 
has not been developed. Since appropriate human anti-roledumab Ig reference standard controls 
are not easily available. It is foreseen to quantify ADA titles SPR technology. ADA 
quantification in serum could be achieved by a calibration-free concentration analysis SPR based 
approach. As for Isotypic characterization, roledumab will be fixed on a sensor chip and then the 
serum containing ADA will be added. Concentration will be calculated on the basis of antibody 
molecular mass after having characterized the isotype of the anti-roledumab antibody.
Results of screening and confirmatory tests will be available within 7 working days after blood 
withdrawal.
Cytokines
The following cytokines will be tested: IFN- , IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13 
and TNF-. The specific time points are at V2, after a sensitizing event and V8: before and 6 hours 
after the IMP administration.
RhD genotyping on cord blood
Fetal full RhD genotyping will be performed on cord blood in case of RhD sensitization. The fetal
RhD-variant will be identified on BioArray DNA chip and sequenced. One sample will be taken and 
frozen for later determination in case of alloimmunization.
Compliance with the Study Plan
8.4.
The Investigator should make every effort to comply with the study plan. If the Investigator 
encounters difficulties in complying with the study plan, e.g. with regard to the schedule of visits or 
the required procedures, he/she must alert the Sponsor. The Sponsor may consider it relevant to 
generate an amendment.
The Investigator should make every effort to avoid the occurrence of deviations from the study plan. 
If any deviations occur or if the Investigator knows that a deviation will occur, he/she must promptly 
inform the Sponsor to determine how to handle the deviation.
9. SAFETY
Safety Reference Document
9.1.
In this study, the roledumab Investigator’s Brochure will be the Safety Reference Document for 
evaluation of the AE expectedness. AEs will be assessed for expectedness using the current 
Investigator’s Brochure at the time of their notification. 


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Benefit / Risk Information
9.2.
Alloimmunization of RhD-negative pregnant women carrying RhD-positive fetus can lead to HDFN 
resulting in complications before birth (HDF), and/or anemia and bilirubinemia after birth (HDN).
HDFN can range in severity from being detectable only in laboratory tests, through to stillbirth, birth 
of infants with severe disabilities or death of newborns from anemia and jaundice. In most severe 
form, HDF produces hydrops fetalis (characterized by total body edema, hepatosplenomegaly and 
heart failure) leading to intrauterine death.
Passive immunization with anti-D immunoglobulin can prevent the RhD-negative women from 
being actively alloimmunized, if given early after exposure to RhD-positive fetal RBC (within 72h). 
The introduction of routine postpartum prophylaxis in the early 1970s by administration of anti-D 
immunoglobulin to RhD-negative women delivering an RhD-positive fetus has reduced the 
alloimmunization
of at-risk women from 16% to about 1.2%. A further reduction of 
alloimmunization was achieved by introducing antenatal prophylaxis during the third trimester of 
pregnancy, leading to a residual incidence of 0.35% [3].
Currently marketed human polyclonal anti-D IgG are produced by fractionation of pooled plasma 
obtained from RhD-negative plasma donors, immunized with RhD-positive RBCs. Although these 
products have proven safe and effective, it can be difficult to entirely meet the needs because of 
limitations in obtaining hyperimmune plasma from humans and because of the increasing 
prescriptions due to both ante-partum and postpartum prophylaxis. Indeed, as a result of the success 
of anti-D prophylaxis and the lower incidence of alloimmunization, fewer immunized women can 
donate plasma for anti-D fractionation. 
Moreover, the practice of antigenic restimulation of the sensitized RhD-negative donors to maintain 
their anti-D titers has become ethically questionable due to the precautionary principle towards the 
risk of emerging pathogens (for which current tests are insensitive or inexistent) in RBCs used for 
sensitizing donors, and other complications of deliberate sensitization (e.g. immunogenicity).
Roledumab is being developed as a preventive treatment to avoid RhD-negative mothers to be 
sensitized by fetal RhD-positive RBCs after FMH and therefore to prevent HDFN in subsequent 
pregnancies.
The major benefits of developing a human recombinant monoclonal anti-D IgG such as roledumab 
over polyclonal anti-D IgGs currently on the market may be summarized as follows:
Ability to provide virtually unlimited supplies of a standardized and safe product without the 
barriers associated with harvesting hyperimmune plasma from immunized humans
Possibility to generate a product that minimizes the concerns of transmission of infectious 
diseases.


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9.2.1. Potential risk(s) related to the IMP(s)
Potential risks of roledumab administration in the target population of pregnant women are based on 
pre-clinical studies program and the two clinical studies conducted with the IMP in healthy 
volunteers as well as on the comparison to polyclonal anti-D which is used and documented along 
the preclinical and clinical experiences. 
Potential risks are assessed in target population (fetuses and pregnant women) in the line of this 
protocol ADNC-1301 in pregnant women to provide a benefit-risk assessment. 
It is to note that in the two previous clinical studies with roledumab administration: the phase 1 
which included RhD-negative healthy volunteers and the phase 2 which included RhD-negative 
healthy volunteers challenged with RhD-positive RBCs, no AE reported in both studies was 
associated to one of the potential risks described below for roledumab. 
Potential risks associated to administration of roledumab in the target population could be: 
Potential risks associated to potential maternal effects of roledumab (mother & fetus)
Potential risks associated to the transplacental barrier crossing of roledumab (fetal risks)
Potential risk associated to a non-recognition of some rare RhD-variants.
9.2.1. 1. Potential risks associated to potential maternal effects of roledumab
9.2.1. 1.1. Allergic reactions
Allergic reactions can occur as with any biotechnology derived protein. Roledumab is a fully human 
IgG1 monoclonal antibody for which preclinical studies indicated neither local intolerance nor 
signal for expected high risk of allergic reactions. Humans exposed in the 2 previous clinical studies 
did not experience allergic reactions. 
This rare potential risk will be closely monitored and the early signs of hypersensitivity reactions 
will be monitored after each administration (hypotension, dyspnea, cutaneous eruption...).
9.2.1. 2. Immunogenicity
Anti-roledumab antibodies appearance: as any therapeutic protein, roledumab has the potential to 
induce an undesirable immune response in subjects receiving this kind of product. Consequences of 
anti-roledumab responses could include lack of roledumab efficacy if they are neutralizing, 
alterations of pharmacokinetics or allergic reactions. 
The risk of developing anti-drug antibodies (ADA) upon roledumab administration to pregnant 
women is considered to be low with the planned dose regimen since it is a natural fully human IgG1 
with an amino-acid sequence derived from an antibody isolated form an RhD-positive immunized
donor. Previous clinical experience did not show anti-roledumab antibodies occurrence in RhD-
negative healthy volunteers following roledumab administration up to 3000 µg IV and up to 300 µg
IM during the phase 1 and 2 clinical studies. The Sponsor will closely monitor the risk of roledumab 
antibodies formation and will perform required analysis to characterize them if any case of such 
undesirable immune response. A justified and detailed clinical and biological follow-up in case of 
ADA occurrence is provided in Sections 8.3.2 and 8.2.6 of this study protocol.


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ADA could theoretically appear after the first administration of roledumab (W28 or 29).
When ADA are detected, the biological follow-up protocol will be implemented (refer to 
Section 8.3.2).
In case of ADA confirmation in maternal plasma (after IMP administration), rescue treatment 
(Rhophylac) will be immediately administered and the same biological and ultrasound follow-up as 
for alloimmunization surveillance will be applied during pregnancy and after delivery:
Anti-D antibodies titration and quantitation every 2 weeks
If anti-D antibodies level is equal to, or higher than 1 µg/mL (or 250 UCHP/mL), weekly 
ultrasound examination will be performed, searching for fetal anemia (any signs of fetal hydrops, 
increased middle cerebral artery peak systolic velocity)
In case of suspected fetal anemia, in utero blood transfusion and/or delivery in a reference center 
should be discussed
Neonatal management will include clinical and biological follow up as well as adequate treatment 
in order to prevent or treat jaundice and anemia in a specialized center.
If ADA are not confirmed, no specific action is needed.
Breastfeeding
ADA assessment in newborn circulation is not foreseen since clinical undesirable effects of these 
induced antibodies are not predicted (if present, they are directed against roledumab idiotype, and 
not against any human somatic antigenic structures). Thus, breastfeeding will not be contra-
indicated.
9.2.1. 2.1. Risk of cross-reactivity of anti-roledumab antibodies against marketed polyclonal anti-D
As polyclonal anti-D represent therapeutic alternative to treat pregnant women at risk of 
alloimmunization in case of potential anti-roledumab antibodies occurrence, a risk of cross-
reactivity of potential anti-roledumab antibodies against marketed polyclonal anti-D could be 
evoked even if it does not represent an anticipated or a potential risk. 
The RhD antigen includes at least 37 different epitopes [12]. Polyclonal antibody preparations are 
obtained by the immunization of donors and pooling of their anti-D immunoreactive plasmas. By 
definition, their individual anti-RhD immune responses are polyclonal and therefore diverse in terms 
of epitope recognition as well as VH and VL (paratope) repertoire for each recognized epitope. As a 
result, the different commercially available anti-D polyclonal antibodies have different patterns of 
recognition of the RhD antigen since they come from different pools of donors with different 
immune repertoires and immune responses [13].
To the best of our knowledge, immune responses against polyclonal anti-D antibodies have never 
been reported.
Roledumab is a fully human IgG1 monoclonal antibody recognizing the epitope epD13.1 [14]
derived from a single B cell of one donor who had been immunized to obtain polyclonal anti-D 
plasma. Its paratope/idiotype is one of the multiple possibilities of recognizing RhD, which are 
represented in the polyclonal preparations. Therefore, in case of an immune response against the 
paratope/idiotype of roledumab (which has never been observed until now), the ADA would 
probably neutralize a negligible portion of antibodies constituting the diverse anti-RhD collection of 
antibodies contained in the marketed polyclonal anti-D drug. 


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In the context of the unlikelihood of anti-roledumab antibodies occurrence and the extremely rare 
probability of altering significantly the marketed polyclonal drug efficacy by the presence of this 
ADA, the treatment of a woman presenting an anti-roledumab positive confirmatory test with a 
marketed polyclonal anti-D associated to a careful follow-up of this woman as if she would be at 
risk of alloimmunization detailed in Section 8.2.6 (IAT testing every two weeks until delivery and 
anti-roledumab antibody follow-up and characterization until the response had come back to base 
line) represent a safe and justified therapeutic alternative.
9.2.1. 2.2. Cytokine release
Roledumab is a monoclonal antibody with improved biological activities and, in particular, 
enhanced affinity for FcGRIIIa (CD16). These properties could lead to undesired activation of 
CD16-expressing cells, which could potentially lead to cytokine-release syndromes [15], [16].
Non-clinical studies on the ability of roledumab to induce cytokines did not suggest enhanced 
cytokine secretion in comparison to polyclonal anti-D. It should be emphasized that the 
immunogenicity in animals is not predictive of immunogenicity in human.
Data from clinical studies in healthy volunteers, both in the presence and absence of RhD-positive 
RBC showed absent or very low cytokine secretion. The cytokine secretion profile in roledumab-
treated healthy individuals in the presence of RhD-positive RBC was shown to be very similar 
(almost absent) to that observed in the polyclonal anti-D-treated subjects after an administration of 
300 µg IV or IM dose. Given this similarity, it is not expected that roledumab will trigger significant 
cytokine release in pregnant women when administered for prevention of RhD-alloimmunization. 
Since similar amounts of roledumab are expected to cross the placenta (similarly to polyclonal anti-
D antibodies), roledumab is not expected to trigger significant cytokine release in the fetus also.
The risk of significant cytokine release leading to systemic inflammatory syndrome or more severe 
complication like a preterm birth is very unlikely. However, the Sponsor will continue to closely 
monitor the risk of cytokine release with adequate dosing of relevant cytokines and proinflammatory 
parameters when exposing pregnant women during this study.
9.2.1. 3. Specific potential risks associated to the transplacental barrier crossing of roledumab 
(fetal risks)
9.2.1. 3.1. Risk of hemolytic disease of the fetus/newborn
Polyclonal anti-D products have gained extensive clinical experience with administration of 300 µg 
around week 28, 29 of pregnancy as well as additional antenatal doses in case of a sensitizing event. 
The most common immunizing event, though, is delivery, whether this is vaginal or by Cesarean. 
Treatment with anti-D at that point in time is not a safety issue for the fetus anymore.
The French recommendations (CNOGF) have been revised (2006) in light of current scientific 
evidence, with regards to the immune-hematological and instrumental investigations that should be 
performed in the antenatal and postnatal periods and prophylaxis to prevent the HDFN due to RhD 
incompatibility. Polyclonal anti-D (Rhophylac) is currently recommended to be used by the 
CNOGF.


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The doses of anti-D reported in this recommendation are a standard of 300 µg for antenatal 
prophylaxis around week 28 of gestation. In addition, in case of a sensitizing event, a standard dose 
of 300 µg is (again) to be administered. However, depending on the amount of FMH, the dose needs 
to be adjusted. A dose of 300 µg is recommended for every 4 mL of FMH, with a maximum of 
1800 µg.
In the large majority of cases one dose of 300 µg is considered sufficient to prevent active 
alloimmunization as the FMH usually does not exceed 4.0 mL. It is reported that in 99% of 
deliveries, FMH is below this value [17]. It is extremely rare to have massive FMH during the 
antenatal period. 
Administration of 1 or 2 times 300 µg of polyclonal anti-D has been shown to be safe in the 
antenatal period, without any consequences to the fetus [18]. No reports of safety issues with the 
doses recommended by the CNOGF have been identified, assuming that it can be used safely for the 
mother, the fetus and the newborn.
It is known that in cases of an alloimmunized mother, only a level above 1 g/L there may be an issue 
for the fetus [19]. With the administration of 1800 µg roledumab it is expected that there may be a 
short peak potentially reaching this level, but the exposure over time is far less than a whole 
pregnancy with this level of anti-D present in an alloimmunized mother.
Non-clinical results do not suggest that roledumab could cross the placenta with a higher rate than 
Rhophylacand is therefore not expected to lead to a higher concentration in the fetal circulation.
A tissue reactivity study was conducted with roledumab and RhoGAM(PolyD). No unexpected 
specific reactivity was reported with roledumab or RhoGAM. Roledumab and RhoGAMreact 
similarly regarding Tissue Cross Reactivity (TCR) with fetal tissues.
Based on this information, it can be concluded that there is no additional or increased harm expected 
with roledumab administration IM or IV in pregnant women regarding HDFN. 
The protocol requires close and continuous monitoring of this risk when administering roledumab to 
pregnant women including adequate and regular fetal Doppler ultrasound monitoring to detect any 
hemolysis of the fetus.
9.2.1. 4. Potential risk associated to the non-recognition of some rare RhD-variants by 
roledumab
In RhD-negative Caucasians, the RhD gene is deleted while it is not expressed in certain populations 
such as Africans or Japanese. A large number of RHD alleles known as variants are described and 
characterized by the absence of some RhD epitopes and/or weak expression (weak D, partial D and 
Del phenotypes). 
Roledumab is assumed to recognize epitope 13.1 that is not expressed by certain variants i.e. DIVb, 
DVI, DBT, DHAR and DHMi. 


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Roledumab failed to bind to certain variants: 
Weak: types, 10, 11, 18, 38 and 58
Partial: DIV type 4 (DIVb), DV type 5 (also called DHK or DYO), DVI (type 1 and 2), DBT 
(type 1 and 2), DHAR (ce-d (5)-ce (former ROHAR)), DAU4; DAR-E (also called DAR2) and 
DAR-A (DAR-E+ silent mutation) not tested but probably not recognized on the basis of epitope 
13.1 recognition profile: DHMi.
The most important potential risk of the non-recognition of these very rare variants by roledumab is 
the alloimmunization and its consequences, even if the ability of these rare variants to induce 
maternal alloimmunization is uncertain and the necessity of higher volumes of FMH in these cases 
to immunize the mother is likely. 
Once this risk is evaluated and assessed as to its importance, it is also useful to place it in the context 
of acceptability by the subjects to be included in this study and by the anticipated target population 
considering the disease indication and the extreme rarity of the situations which it could occur. 
This unsolved issue which brings uncertainties regarding potential risk of alloimmunization and the 
relevance of this finding to the use in pregnant women humans are discussed below.
For a quantitative approach to address this risk, estimations were done on the basis on the 
prevalence of weak and partial D-variants in the general population and on the estimated probability 
of carrying an RhD-variant fetus by an RhD-negative pregnant woman (refer to Report – Gabriele 
Dallman provided in the Investigator’s Brochure).
The worst case scenario is taken for these estimations combining the facts that:
We consider in the calculation that roledumab does not recognize all D-variants as a worst 
scenario even it does not recognize only a small part of these variants
We consider that all D-variants can immunize an RhD-negative mother even if the 
immunogenicity of some of these variants such as DVI and DBT are commonly admitted to be 
lower than the wild type RhD.
The probability to have an RhD-negative mother at risk of carrying a fetus with D-variant antigen is 
estimated between 0.004% and 0.26% as a worst case scenario. 
This potential risk related to extremely rare cases cannot be totally prevented as there is no available 
test to perform on mother blood sample before systematic prevention with roledumab to determine 
antenatally if the fetus is a D-variant. Differently, in case of sensitizing event which is at-risk 
situation of alloimmunization, a Kleihauer-Betke test will provide information on fetal RBCs 
clearance from maternal blood and will allow close monitoring of roledumab activity in case of 
presence of D-variant in the fetus. This will optimize management of this risk even it concerns only 
extremely rare at-risk situations. 
According to these quantitative evaluations in the context of the disease indication and expected 
benefits of roledumab, these extremely rare situations in which this risk could occur bring an 
acceptable level of risk for the target population. The issue will continue to be addressed throughout 
the study and clinical development for further documentation, monitoring and minimization. 


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9.2.1. 5. Related to the study procedures
Blood sampling will be performed during this study in order to monitor the safety and to assess PK 
parameters. Although this procedure represents an inconvenience for the subject, it does not 
constitute an additional significant risk. There is a minor risk of hematoma from intramuscular and 
intravenous infusion of the IMP and from drawing blood. Slight pain at the injection site, feeling 
light-headed, bruising and, exceptionally, infection as well as bleeding from the site of the puncture 
may occur. 
9.2.2. Benefit/risk balance
The assessment of the benefit-risk balance is based on the available relevant non-clinical and clinical 
tests and studies performed. All available data on roledumab were obtained out of the target 
conditions since no pregnant woman has been exposed yet to roledumab. 
However, comparison to marketed polyclonal anti-D was performed in most of the tests and studies 
to provide relevant evaluations and conclusions.
The assessment of all relevant data gathered with roledumab together with all measures to be 
implemented to closely monitor important potential risks, provide objectively confidence that 
roledumab administration will have a favorable benefit/risk balance in pregnant women.
In conclusion, the Sponsor considers that these factors support a favorable benefit/risk assessment 
for the use of roledumab in pregnant women in this study.
Risk Minimization Actions/Monitoring Throughout the Protocol
9.3.
Roledumab has not yet been administered to pregnant women, so the safety in this specific 
population is unknown. 
Target population will be screened and included only if inclusion and exclusion criteria are met. 
Allergic reactions may occur in humans. Symptoms of allergy or early signs of hypersensitivity 
reactions including generalized urticaria, tightness of the chest, wheezing, hypotension or/and 
anaphylaxis may occur. The treatment required depends on the nature and the severity of the AEs. If 
necessary, the current medical standards of care for shock treatment should be implemented.
As any therapeutic protein, roledumab antibody has the potential to induce an undesirable immune 
response in subjects receiving this kind of product. Consequences of anti-roledumab responses 
include lack of efficacy if they are neutralizing, alterations of pharmacokinetics or allergic reactions. 
Cytokines panel will be tested at specific time points.
Close surveillance of fetal signs of anemia will be performed through regular fetal Doppler 
ultrasound (fetal well-being, mean MCA-PSV). This investigation will be repeated according to the 
MoM value.
The newborn will be examined as usual. Cord blood will be taken for DAT and other laboratory 
testing (hemoglobin, hematocrit, reticulocytes and bilirubin).


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First milk / breast milk sampling will be also performed in order to measure roledumab 
concentrations although roledumab concentrations are expected to be extremely low as described for 
other monoclonal IgG1 antibodies [20]. 
Furthermore the DSMB will examine safety data at specific time-points during the study and give 
recommendations regarding any changes that may need to be made to the study, how it should be 
continued and if it should be stopped. 
This summary of risks and benefits is not intended to be all inclusive, but is rather intended to 
highlight possible risks, benefits, and the potential important risks. The Investigator must become 
familiar with all sections of the IMP Clinical Investigator's Brochure provided by the Sponsor before 
the start of the study.
Alternative Therapeutic Management - Emergencies Handling
9.4.
The subject may be administered Rhophylacor any other anti-D immunoglobulin. In this case, the 
subject will be withdrawn from the study and followed as described in Sections 5.4.1, 8.2.6, 8.3.1
and 9.2.1. 2.
Definition and Reporting of (Serious) Adverse Events
9.5.
9.5.1. Definition of adverse event and serious adverse event
(see also Section 9.5.2: Definition of specific events in the study)
An adverse event (AE) is any untoward medical occurrence in a subject or clinical investigation 
subject administered, or intended for administration of, a medicinal product that occurs any time 
after the informed consent has been signed and until the final study visit or early termination and 
which does not necessarily have a causal relationship with this treatment. An AE can therefore be 
any unfavorable and unintended sign (including an abnormal laboratory finding, for example), 
symptom or disease temporally associated with the use of a medicinal product, whether or not 
considered related to the medicinal product. Adverse events include, but are not limited to: (1) a 
worsening or change in nature, severity, or frequency of conditions present at the start of the study; 
(2) subject deterioration due to primary illness; (3) intercurrent illness; (4) drug interaction; and/or 
(5) abnormal clinically significant laboratory values.
Subjects should be questioned in a general way, without asking about the occurrence of any specific 
symptom. The Investigator should attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information. In such cases, the diagnosis should be documented as 
the AE and not the individual signs/symptoms.
Following questioning and evaluation, all AEs, whether believed by the Investigator to be related or 
unrelated to the study medication, must be documented in the subject’s medical records, in 
accordance with the Investigator’s normal clinical practice, and on the AE form in the e-CRF.
Each AE is to be evaluated for duration, intensity, seriousness, temporally association and causal 
relationship to the study medication.


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A Serious Adverse Event (SAE) is an AE that, at any time, fulfils one or more of the following 
criteria:
Results in death
Is life threatening, i.e. the subject was at immediate risk of death at the time of the event; it does 
NOT refer to an event which hypothetically might have caused death if it were more severe
Requires in-subject hospitalization or prolongation of existing hospitalization
Results in persistent or significant disability/incapacity, i.e. substantial disruption of a 
person’s ability to carry out normal life functions
Is a congenital anomaly/birth defect
Is any important medical event that may not be immediately life threatening or result in death 
or hospitalization but, based upon appropriate medical judgment, may endanger the subject or 
may require intervention to prevent one of the above characteristics/consequences. Such events 
(subsequently referred to as ‘important medical events’) should also be considered ‘serious’ in 
accordance with the definition.
Medical and scientific judgement should be exercised in deciding whether an event is ‘serious’ in 
accordance with these criteria.
Surgical intervention is not to be considered as SAE but the medical condition requiring the 
surgery is to be reported as such
Events initially reported as an AE may become serious. For example, diarrhea may become 
debilitating and require hospitalization or prolongation of hospitalization and is then reported 
as SAE
Distinction should be made between serious and severe AEs. Severity is a measure of intensity 
whereas seriousness is based on the seriousness criteria described above.
Occurrence of IMP overdose, abuse/misuse, drug dependency or any medication error, should be 
immediately reported to the Sponsor (via a dedicated email address: ADNC1301@lfb.fr) and 
considered as AEs. These special situations should be reported as SAEs only when fulfilling SAE 
criteria or associated with a SAE.
9.5.2.
Definition of specific events in the study
Hospitalisation for planned delivery should not be reported as a SAE. Occurrence of a FMH will 
not be considered as an SAE as it will be reported as targeted disease treated with roledumab. The 
sensitizing event at the origin of the FMH will be also described
RhD-alloimmunization will be systematically reported as a SAE
Anti-roledumab antibodies occurrence (after IMP and absent before IMP) will be systematically 
reported as a SAE
Preterm delivery (37 weeks of Gestational Age) will be systematically reported as a SAE. 
9.5.3. Adverse events recording and reporting
Each individual AE is to be listed as a separate entry on the AE form in the e-CRF. The Investigator
will provide information on dates of onset and resolution, seriousness, severity, action(s) taken, 
outcome and relationship to study medication.
Any AE collected for the newborn during a routine visit will be reported on a specific AE form in 
the e-CRF.


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Documentation of immediately reportable events will follow procedures described in Section 9.5.4.
The Investigator must report to LFB BIOTECHNOLOGIES, or its designee all AEs that occur 
during the study from the time written informed consent is given until the final study visit or early 
termination, regardless of their relationship to IMP. 
If the Investigator detects an (S)AE after the above-defined period of observation and considers 
the event as possibly related to the IMP, he should contact the Sponsor to determine how it must be 
documented and reported.
Apart from AEs clinically observed by the Investigator, or recorded on self-assessment forms (e.g.
subject diary), the subject will be given the opportunity to report AEs spontaneously. A general 
prompt will also be given to detect AEs, e.g. “Did you notice anything unusual about your health 
since your last visit?”
It is the responsibility of the Investigator to record all the relevant information regarding the event.
The Investigator is requested to assess the relationship between the investigational product and the 
occurrence of each (S)AE. Alternatives causes, such as the underlying diseases, concomitant therapy 
or the temporal relationship of the event to the investigational product will be considered. 
9.5.4. Procedures for reporting serious adverse events
At the occurrence of a subject event that fulfils one or more seriousness criteria, the Investigator 
must immediately forward to the Sponsor or its representative a duly completed “SERIOUS 
ADVERSE EVENT FORM” provided by the Sponsor, even if the data are incomplete, but as soon 
as the following minimum information is available:
Identification of a subject (subject number and/or initials). In the context of this study, the subject 
experiencing the SAE (Mother, Newborn or Both) should be specified in the SAE Form. 
Description of the SAE.
The Investigator will ensure a follow-up of an initial SAE to elucidate the nature, the outcome or the 
causality of the SAE. Supporting documentation (discharge summaries, all examinations carried out, 
etc.) will not be sent systematically, as all relevant information must appear and be summarized in 
the narrative of the SAE report.
However if judged important, the Investigator could send these documents or LFB 
BIOTECHNOLOGIES could request them (all documents must be blinded with respect to the 
subject’s name). Any follow up information must be reported within the same timelines.
TIMEFRAME
“Initial” SAE
Form
“Follow-up” SAE
form
IMMEDIATELY
As of awareness of an SAE 
occurrence
24 hours
As of availability of essential follow-
up information




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Any follow up information must be reported within the same 
timelines.
10.
PARAMETERS AND ASSESSMENT CRITERIA
Assessment of Pharmacokinetics
10.1.
The PK of roledumab will be assessed from serum concentrations of roledumab using all samples 
(ante- and postnatal samples) for the PK population approach and only samples after first infusion 
(antenatal samples) for NCA. 
PK in alloimmunized subjects and in subjects with ADA rescued with Rhophylacwill not be 
analyzed, and no samples following the alloimmunization or ADA confirmation will be drawn.
Assessment of Safety
10.2.
The safety of roledumab will be assessed by recording all AEs occurring during the study, i.e. after 
signature of the informed consent and until the end of the study. AEs in both mother and 
fetus/newborn will be collected and recorded.
Safety assessment during pregnancy will include but is not limited to a fetal Doppler ultrasound 
examination at 9, 29 and 59 days after the antenatal roledumab injection (see Section 8.2 for further 
details on safety assessment during pregnancy). If the Doppler ultrasound examination suggests a 
fetal hemolysis (e.g. a mean cerebral artery peak systolic velocity (MCA-PSV) showing a Multiple 
of Medians (MoM) 1.5) or other abnormalities, the subject will attend a follow-up visit weekly 
until resolution or delivery.
The safety of roledumab will be assessed in the RhD-negative pregnant women and in the RhD-
positive fetus during the third trimester of pregnancy during the follow-up visits.
The safety of roledumab in the breastfed newborns will also include clinical follow-up of the 
newborn.
The safety will be assessed by the following variables:
Incidence and severity of AEs
Anti-roledumab antibodies
Vital signs
Physical examination.
10.2.1.Adverse events
Any abnormalities in vital signs, physical examination or laboratory safety parameters considered as 
clinically significant by the Investigator will be reported as an AE in the CRF. 
The safety of roledumab will be assessed in the RhD-negative pregnant women and in the RhD-
positive fetus during the third trimester of pregnancy and after birth in accordance with the study 
flowchart. Safety assessment during pregnancy will include but is not limited to a fetal Doppler 
ultrasound examination after the antenatal IMP administration. If the Doppler ultrasound 


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examinations suggest a fetal hemolysis (e.g. a mean cerebral artery peak systolic velocity (MCA-
PSV) showing a MoM 1.5) or other abnormalities, subjects will attend a follow-up visit every 
week until resolution or delivery.
The immunogenicity of roledumab will be assessed by the detection of anti-roledumab antibodies 
before the antenatal IMP administration (baseline), at 9 and 29 days after IMP administration, at 
delivery before the postnatal administration, at 9 days, 6 weeks, 6 months or up to 12 months after 
the postnatal administration. 
10.2.2.Physical examination
Vital signs including sitting systolic and diastolic blood pressure, pulse rate will be recorded in the 
e-CRF at each visit.
10.2.3.Laboratory data
Refer to Section 8.3.1.
Assessment of Efficacy: Occurrence of RhD-Alloimmunization
10.3.
As recommended in the European guidelines for investigations on polyclonal anti-RhD antibodies 
[CPMP/BPWG/575/99 Rev.1], the efficacy of RhD-alloimmunization prophylaxis in clinical studies 
is usually assessed by screening the blood sample collected at 6 months after the birth of an RhD-
positive fetus. However, it is well-known that in rare cases the passively administered anti-RhD IgG 
may remain detectable up to 6 months. Therefore, RhD-negative pregnant women will be followed-
up until 6 months after delivery and in the case of a positive indirect antiglobulin test (IAT), an 
additional blood sample will be drawn and tested again at 9 and possibly 12 months post-delivery in 
order to estimate whether RhD-alloimmunization occurred or not. 
The RhD-alloimmunization will be determined by a positive IAT (screening and identification of 
anti-RhD antibodies). In case of a positive IAT, the anti-RhD antibodies will be quantified centrally 
by microtitration or ponderal dosage in order to ascertain true alloimmunization from residual anti-
RhD antibodies, depending on anti-RhD antibodies concentration levels. The serum concentration of 
anti-RhD IgG derived from RhD-alloimmunization is not expected to decline while passively 
acquired anti-RhD IgG should be completely eliminated 3 months later. Passively acquired anti-
RhD IgG is usually weakly reactive while a high level of anti-RhD IgG is likely to be of maternal 
origin.
If the IAT is negative at 6 months post-delivery, the subject will be considered as negative at 9 and 
12 months.
If the IAT is positive (i.e. confirmed by anti-D quantification) at 6 months but negative at 9 months 
post-delivery, the subject will be considered as negative at 12 months.
In the case of a sensitizing event occurring before delivery, additional follow-up visits will be 
performed according to the same scheme i.e. 2, 5, 9, 29 and 59 days after the additional IMP
administration. For PK samplings, the schedule will stay the one initially scheduled after the first 
injection, and the data obtained will be used to perform the population PK model.


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Other Assessments
10.4.
A cord blood sample will be collected at delivery in order to perform a direct antiglobulin test 
(DAT) and to determine the fetal free serum concentrations of roledumab. 
All breastfeeding mothers will be asked to provide a first milk/breast milk sample and a blood 
sample to assess the roledumab first milk or breast milk and maternal serum concentrations. 
11.
DATA MANAGEMENT
CRF Completion Guidelines
11.1.
11.1.1.Introduction
All of the information that must be reported to the Sponsor as per the protocol will be entered in the 
electronic case report form (e-CRF) for each study subject. The e-CRFs must be completed by the 
Investigator or any study center staff designated by the Investigator. Access to the data at the clinical 
site will be restricted and monitored through the system-s software with its required logging, 
security procedures, and audit trail. The software used for this study will be Inform (Oracle). All the 
study data will be entered in the database using Inform and will be stored on an Oracle Server, based 
in the US. The study data will be transferred to the server of the contract research organization 
(CRO) in charge of data management and statistics, based in India. The database will be periodically 
transferred to the LFB server based in Lyon, in France. 
The Investigator is responsible for ensuring the data recorded in the e-CRFs are accurate and 
complete. The e-CRFs should be completed within 5 working days after the subject’s visit and the 
latest before review by the Monitor. 
11.1.2.General instructions
11.1.2. 1.Overview
All data will be recorded in the e-CRF. The e-CRF is a web-based application which is a 
computerized system that allows users to create, modify, store, archive, retrieve, or transmit data. 
Access to the e-CRF will be limited to authorized individuals. In this way, each Investigator and 
each person with authorized access to the e-CRF will receive an individual user name and unique 
password before the study starts. The different access will be limited according to the role of 
persons. Therefore users should only work using their own passwords and must not share these with 
other people. The attributed password should be changed when the user first logs in.
Audit trail
To ensure and protect the authenticity, integrity and confidentiality of electronic records an audit 
trail will be integrated into the e-CRF. All entries and changes will be tracked in an audit trail. The 
audit trail is a secure, computer-generated, time-stamped system which independently records the 
date and time of operator entries and actions that create, modify or delete electronic records.
Individual training


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It will be the Sponsor’s responsibility to organize individual training and certification for all the 
people who will use the computerized system. In order to obtain their log and password each person 
should have been performed this training. Each person who enters or processes data should have the 
necessary education, training and experience to perform the assigned functions. In addition the 
individuals responsible for monitoring the study should have the necessary education, training and 
experience in using the computerized system to enable them adequately monitor the study.
All data in the e-CRF must come from and be consistent with the source documents.
Source documents may be: 
The subject’s medical file
The hospital nurse’s sheet
Laboratory results logs
The subject’s diary
Homecare nurse visit information
Prescription form
Central laboratory transmission form
Any other document issued in the routine practice of healthcare and providing information on 
the subject’s medical status.
Any discrepancy between the data in the e-CRF and those in the source documents should be 
documented by the Investigator.
Abbreviations should be avoided since they are often ambiguous. 
11.1.2. 2.Subject identification
The information reported in the header will be automatically populated on all the e-CRF pages when 
the subject is created in the system.
Header information should include at least:
The protocol number
The name of the visit
The subject’s identification number according to local requirements: subject number is 
recorded with the center number provided by the Monitor at the set-up visit. The subject 
number is a sequential number incremented with each subject included in the order the 
informed consent forms are signed.
11.1.2. 3.Confidentiality
Subjects’ names must be kept confidential and should not appear on any e-CRF page or study-
specific documents.
11.1.2. 4.CRF completion in case of screening failure or premature withdrawal
For screening failure subjects, the Investigator or the study staff designated by the Investigator will 
complete the “End of study” section of the e-CRF.
If a subject has been included and prematurely withdrawn, the Investigator will perform the "End of 
study visit" and complete the "End of Study" Section in the e-CRF.


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11.1.3.Completion of the e-CRF
The Investigator should complete the e-CRF within 5 working days of each subject’s visit. He will 
complete all the corresponding pages and, for blank sections or pages, he will complete at least the 
status of these sections/pages. 
11.1.4.Specific case report form instructions
Specific instructions for completing the e-CRF will be on line.
E-CRF and Data Handling
11.2.
Data management will be carried out by a CRO from the electronic-based CRF in accordance with 
Good Clinical Practices (GCP, see ICH-E6, Section 5).
Data processing, from data capture through to database lock, will be carried out in accordance with 
GCP (see ICH-E6, Section 5).
The data handling documents, e.g. annotated CRFs, database structure, coding rules and 
computerized validation, are defined in a Data Management Plan. The database and data entry 
screens will be created in software specifically designed for clinical data management in compliance 
with ICH-E6 requirements.
Tracking of the e-CRFs will be integrated into the computerized system. As soon as a subject has 
signed their informed consent, the Investigator will create the subject in the e-CRF and complete the 
appropriate page(s).
Medical coding will be done using the MedDRA (Medical Dictionary of Regulatory Activities) for 
all medical terms (medical history, AEs) and the WHO Drug dictionary for all drug names (prior 
and concomitant medications).
The consistency of data will be checked by computerized programs and any related queries will be 
generated for resolution by the Investigator. The database will then be updated accordingly. Coding 
of medical terms and drug names, quality controls to ensure the overall quality and consistency of 
the database and reconciliation of SAE reports with the Pharmacovigilance database will also be 
carried out.
At the end of the data handling process, a data review meeting will be held to prepare the database 
lock.
12.
STATISTICS
Study Objectives and Design
12.1.
The main objective is to assess the PK profile of roledumab 300 µg IM/IV in RhD-negative 
pregnant women carrying an RhD-positive fetus
The design is an interventional, open-label study


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Statistical Analysis Plan
12.2.
The material of this section is the basis for the Statistical Analysis Plan (SAP) of the study. 
The detailed technical aspects of the statistical analyses will be provided in the SAP. The SAP will 
possibly take protocol amendments into account and adapt to unexpected issues raised by the study
running and/or data that affect planned analyses in the protocol. 
Any changes from the protocol will be discussed in the study report.
Prior to locking the database, a data review meeting will be planned in order to review individual 
data and validate the SAP.
Sample Size Determination
12.3.
No statistical hypothesis will be tested. Therefore no formal sample size calculation is done. 
However 30 subjects IM and 20 subjects IV should be sufficient to determine the PK parameters in 
pregnant women taking into account the moderate variability between subjects considering the route 
of administrations and the dose [9], [21].
Randomization
12.4.
Not applicable.
Protocol Deviations and Analysis Sets
12.5.
All the deviations from protocol definitions will be listed and defined as major or minor deviations
in the protocol deviation specification document.
For the PK analysis, if any of the following 2 elements is missing, all of that subject’s data will be 
excluded:
a.
Date/time of IMP administration
b. Dose amount
Missing date and time of sampling for a subject will lead to a deviation for the specific PK time 
point.
The intent-to-treat (ITT) population
The intent-to-treat (ITT) population includes all subjects who were enrolled into the study.
The full analysis set (FAS) 
The full analysis set (FAS) is a subset of ITT population and will be used for the efficacy analysis. It 
consists of all ITT subjects who have a negative IAT test at inclusion and who delivered an RhD-
positive newborn.
The safety set (SAF)
The Safety Set (SAF) will be used for all safety analyses. It consists of all subjects who received at 
least one administration of IMP. 


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PK populations
PKS1: Analysis population 1 for population PK modeling
The PK set 1 consists of all ITT subjects enrolled in study ADNC-1301 (IM or IV arm), treated at 
least once with roledumab, and providing at least one roledumab evaluable concentration after 
administration (including antenatal and postnatal measurements after 1st or 2nd administration).
PKS2: Analysis population 2 for NCA IM arm
The PK set 2 consists of all ITT subjects enrolled in the IM arm of study ADNC-1301, who have at 
least one valid PK assessment after the first IMP administration, where valid PK assessment is 
defined as providing at least one evaluable PK parameter.
PKS3: Analysis population 3 for NCA IV arm
The PK set 3 consists of all ITT subjects enrolled in the IV arm of study ADNC-1301, who have at 
least one valid PK assessment after the first IMP administration, where valid PK assessment is 
defined as providing at least one evaluable PK parameter.
General Rules for Handling of Missing or Inconsistent Data
12.6.
No general rule for the replacement of missing values is used. Any effort should be made to collect 
missing data.
Demographic and Baseline Characteristics
12.7.
12.7.1.Demographic characteristics, medical history, diagnoses
Demographic and baseline characteristics of subjects will be summarized by route of administration 
and overall. This analysis will be performed in the SAF on all available variables including the 
following elements, but not limited to:
Demographic characteristics
Medical and surgical history
Prior medication
Prior pregnancies
Information about the current pregnancy
Physical examination
Laboratory data
The distribution of parameters will be summarized using descriptive statistics depending on the 
characteristics of the variable:
Categorical variables (binary, nominal and ordinal) will be summarized by number of missing 
observations as well as frequencies and percentages for each category. Percentages will be 
calculated on the observed cases
Quantitative variables will be summarized by their number of missing observations, number of 
valid observations, mean, standard deviation, standard error, median, quartiles, minimum and 
maximum values.


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12.7.2.Previous treatments
All previous treatments will be classified using anatomical therapeutic chemical (ATC) codes and 
listed by subject. The standard dictionary for coding treatments will be WHO Drug. Descriptive 
statistics will be produced to analyze the previous treatments by their associated ATC code. 
Investigational Product (IMP) and Concomitant Treatments
12.8.
The analyses described in this section will be performed on the SAF presented by route of 
administration and overall.
12.8.1.Extent of exposure
Subjects are planned to receive two injections of the IMP, one at the 28th week of gestation 
(or 29th week) and one within 72h after delivery. If other drug administration is given (i.e. in case of 
sensitizing event), a description of events and doses will be provided. Duration of exposure will be 
summarized as the time between the date of the first injection until the date of the last injection 
plus 1. The actual administered dose will be presented per injection as well as the number of 
injections and the cumulative dose of the administered IMP.
12.8.2.Treatment compliance
Compliance is defined as compliance to the administration regimen (dosage, schedule and 
administration route) as defined in Table 6–1. 
12.8.3.Concomitant treatment
All concomitant treatment will be classified using ATC codes and listed by subject. The standard 
dictionary for coding treatments will be WHO Drug.
Descriptive statistics will be produced to analyze the concomitant treatments by their associated 
ATC code.
Efficacy Analysis
12.9.
12.9.1.Primary efficacy variable(s)
The primary objective is to assess the PK of roledumab 300 µg IM and IV. Therefore, no primary 
efficacy variable is defined but only secondary efficacy variables.
12.9.2.Secondary efficacy variables
All efficacy analyses will be performed on the FAS set. Secondary efficacy variables are the 
following:
RhD-alloimmunization status at 6, 9 and 12 months after delivery
RhD-alloimmunization will be assessed at 6 months post-delivery. In the case of a positive IAT and 
alloimmunization confirmed by anti-D quantification at 6 months, the subject will be followed until 
9 and eventually 12 months in order to distinguish residual anti-RhD antibodies from anti-RhD 


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alloantibodies. If the IAT is negative at 6 months post-delivery, the subject will be considered as 
negative at 9 and 12 months. If the IAT is positive at 6 months but negative at 9 months post-
delivery, the subject will be considered as negative at 12 months.
The number and percentage of subjects by alloimmunization status will be presented by route of 
administration and overall. A particular attention will be paid of the alloimmunization status in case 
of sensitizing events. No statistical hypothesis is tested.
Safety Analysis
12.10.
All safety analyses will be performed on the SAF presented by route of administration and overall. 
Despite the genotyping on maternal blood in order to include only women carrying an RhD-positive 
child it may happen that at delivery the child is observed as being RhD-negative. Therefore, safety 
on the fetus/infant will be presented by subgroups of their RhD-status (positive / negative).
12.10.1.
Extent of exposure
Please refer to Section 12.8.1.
12.10.2.
Adverse events
With respect to AEs, all AEs of the mother and/or the fetus/newborn/infants occurring during 
treatment and follow-up periods (up to 6 months age for the infant and up to End of Study visit for 
the mother) will be summarized using the Preferred Term (PT) and System Organ Class (SOC). AEs
(occurring during screening period (between V1 and V2) before the first injection of the IMP) will 
only be listed. The duration of the related and specific AEs listed in Section 9.5.2 and the duration 
between the last dose of IMP and onset of these AEs will be calculated and summarized.
The number and percentage of AEs by SOC and PT on all AEs will be tabulated. SAEs, drug-related 
AEs and other significant AEs will be presented in the same way.
The number and percentage of subjects with at least one AE, at least one SAE, at least one drug-
related AE or at least one other significant AE will be tabulated by SOC and PT.
The standard dictionary for coding events will be MedDRA.
12.10.3.
Fetal anemia
The number and percentage of fetal anemia and other abnormalities detected by Doppler ultrasound 
and obstetric ultrasound will be summarized. Details are given in Section 10.2 of the protocol.
12.10.4.
Vitals signs and physical findings
Vital signs and physical examination findings including APGAR score will be summarized using 
appropriate statistics. Any significant abnormalities will be listed.


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12.10.5.
Laboratory data
Laboratory values over time and changes in individual subjects will be summarized using 
appropriate statistics. All clinically significant abnormal values will be listed.
The immunogenicity of roledumab (ADA) will be assessed using descriptive statistics.
12.10.6.
Other endpoints
The analysis of these endpoints will be performed on the SAF. 
Roledumab cord blood concentration
The endpoints will be presented using descriptive statistics. The ratios of roledumab in cord blood 
concentration to maternal serum concentration will be calculated.
Roledumab first milk/breast milk concentration
The endpoints will be presented using descriptive statistics. The ratios of roledumab in first 
milk/breast milk concentration to maternal serum concentration will be calculated.
These endpoints will be only listed. 
Pharmacokinetics, PK/PD Analysis
12.11.
The PK analysis is detailed in the SAP. 
12.11.1.
Primary analysis
A population PK analysis will be performed as the primary PK analysis using nonlinear mixed effect 
modeling with both IM and IV data. 
The model will be created using a two compartments disposition model with first order absorption 
and elimination. The effects of covariate body size will be evaluated on the PK parameters clearance 
and volume of distribution. Bioavailability of roledumab will also be estimated. 
The effect of other covariates such as gestational age, glomerular filtration rate, effect of delivery, 
age and site of injection, will be explored using the likelihood ratio test. Given the small sample size 
of the study, no other covariate will be explored. The first-order conditional estimation (FOCE) 
method will be used throughout analyses.
Various random effects structures and residual errors approaches will be explored as needed. 
Diagnostic plots of observed data versus population and individual predicted values and various 
residuals plots will be used to detect any significant deviation from the model fit. 
A table of population PK parameters with confidence intervals will be produced.
Derived secondary parameters will be obtained from individual posterior estimates, including 
AUCinf, Cmax, Tmax, t1/2, elimination rate constant. Those derived parameters will be summarized by 
appropriate statistics (N, geometric mean, 90% confidence intervals, median, min and max, with the 
exception of Tmax to be summarized by median, min and max only). 


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12.11.2.
Secondary analysis
Two non-compartmental analyses will be used using Phoenixsoftware with IM and IV data.
Volume of distribution (Vd)
Clearance (CL)
Cmax
Tmax
Area under the curve (AUCt, AUCinf)
Terminal half-life (t1/2) and Elimination rate constant
Those derived parameters will be summarized by appropriate statistics (N, geometric mean, 90% 
confidence intervals, median, min and max, with the exception of Tmax to be summarized by median, 
min and max only).
13.
STUDY REPORT
A Clinical Study Report will be prepared in accordance with the ICH-E3 guidelines, by the Sponsor 
or subcontractor in collaboration with the coordinating Investigator and, if any, with the scientific 
committee. 
Within 1 year after the end of the study (Last Visit Last Subject), the Sponsor will provide the 
Health Authorities with the full Clinical Study Report or summary. Only the Sponsor is entitled to 
make the study report available to the Authorities.
Neither the complete report nor any part of the study report may be used without the approval of the 
Sponsor.
14.
CONFIDENTIALITY AND PUBLICATION
Subject Confidentiality
14.1.
Subject data will be kept strictly confidential and subject anonymity will be protected by using 
number codes and /or initials.
The Sponsor or its representative(s) and the Health Authorities are obligated to respect medical 
secrecy and to refrain from divulging any personal subject information they might fortuitously be 
aware of.
Use of Information
14.2.
The Investigator shall not divulge unpublished data or information related to the study provided by 
the Sponsor, including but not limited to the study product characteristics, the Investigator brochure, 
the study protocol, case report forms, assay methods and scientific data, to any third party without 
written approval from the Sponsor.


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In addition, any new information that may become available during the course of the study shall be 
considered as confidential and shall not be used for any purpose other than the performance of the 
clinical study.
The study data are the property of the Sponsor. The Investigator and any of the research staff shall 
obtain written approval from the Sponsor prior to the publication/communication of the results of 
any work carried out during or in relation to the study.
Publication and/or communication of the results of the clinical study will be of a cooperative nature 
involving authors representing the Sponsor, the Study Coordinator, and the scientific committee, if 
any.
The Sponsor reserves the right to request modification of the content and/or timing of any 
publication or presentation if a patent application, an existing patent or other proprietary rights may 
be jeopardized.
Authorship of any publication related to the study and the order of presentation of the authors’ 
names shall be approved by the Sponsor. The Sponsor shall not use an Investigator’s name in any 
publication without his/her written permission and vice versa.
15.
ARCHIVING
The Investigators should retain all essential study-related documents, i.e. documents which permit 
evaluation of the conduct of a study and the quality of the data produced, in accordance with the 
applicable regulatory requirements of his/her country. These essential documents include but are not 
limited to signed protocol, Investigator Brochure, e-CRFs, medical records, laboratory reports, 
informed consent forms, drug disposition records, safety reports, information regarding participants 
who discontinued, and other relevant documents and data.
The study-related documents should be kept together in the Investigator site file provided to the 
Investigator by the Sponsor.
Sufficient information about the identity of all study subjects, e.g. name, medical records number, 
subject number and study number, should be retained by the Investigator so that any Sponsor 
representatives, auditors or inspectors may access this information when required.
The Investigator must retain all records for 15 years or longer if required by specific local 
requirements.
The Investigator will contact the Sponsor for authorization prior to the destruction of any study 
records or in the event of accidental loss or destruction of any of them. 
The Investigator will also notify the Sponsor should he/she relocate or move the study-related files 
to a location other than that specified in the Sponsor’s Study Master File.
All records should be kept in a secure area, however in the cases of audit or inspection, they should 
be rapidly made available.


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16.
RESPONSIBILITIES OF PARTICIPANTS
Responsibilities of the Investigator(s)
16.1.
The Investigators will conduct the study in accordance with ICH-E6, all applicable laws in the 
country where the study is conducted and in accordance with this study protocol.
The responsibilities of the Investigators are summarized below but not limited to:
Subject information and consent
Prior to undertaking any study-related procedure, it is the responsibility of the Investigator, or a 
formal designee, to provide each subject and/or a legal representative/witness, with relevant, 
comprehensive, verbal and written information, including the written information which received 
approval or a favorable opinion from the IEC/IRB and the Health Authorities. Signed informed 
consent must be obtained prior to undertaking any study-related procedure. Obtaining of consent and 
how it was obtained must be described and documented in the subject’s file.
Information on the overall results of the study
Pursuant to the French “Subject’s rights” law adopted on 09 August 2004, the Investigator must 
provide any subject who so requests it with the overall results of the study. The Sponsor will provide 
the Investigator with the overall results beforehand. The Investigator should document in the 
subject’s file the fact that the information has been provided.
Information to other practitioners (if relevant)
In agreement with the subject, the Investigator will formally inform other practitioners of the 
subject’s participation in the study, to avoid any interference or bias in the conduct of the study.
Institutional Review Board / Independent Ethics Committee (IRB/IEC)
In accordance with local regulations, the Investigator may be required to interface with the IRB/IEC.
Adverse events
The Investigator is responsible for ensuring adequate safety monitoring and follow-up of the study 
subjects. The Investigator must report and handle any serious and non-serious AE, whether 
clinically observed or spontaneously reported by the subject, using concise medical terminology in 
accordance with Section 9 of the protocol.
Data recording
It is the Investigator’s responsibility to ensure, on an on-going basis, completion and validation of 
all case report forms as well as study-related supportive data. e-CRFs must be signed by the 
Investigator. If the Investigator formally delegates completion of the e-CRFs, the Investigator 
nevertheless has the final responsibility for signing the e-CRFs to certify the accuracy and reliability 
of the data recorded therein.


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Record retention
To enable inspections and audits from Health Authorities or the Sponsor, the Investigator agrees to 
keep records, including the identity of all participating subjects, i.e. sufficient information to link 
records, all original signed informed consent forms, copies of all CRFs and detailed records of 
treatment disposition. The Investigator should maintain a site file with all essential documents.
See also Section 15.
Use of study-related information
The Investigator must provide the Sponsor with complete test results and all data derived from the 
study. Only the Sponsor may make information available to physicians, Health Authorities and/or 
subjects enrolled in the study (via the Investigators), except as required by local regulations.
IMP
Responsibility for IMP accountability at the study site rests with the Investigator or with the 
institution, depending on local regulations.
Quality control
The Investigator and the relevant personnel should be available during monitoring visits and 
possible audits or inspections and ensure that sufficient time is devoted to the process. The 
Investigator guarantees the Sponsor or its representative and appropriate Health Authorities direct 
access to source documents.
Study discontinuation
Should the Health Authorities or the Sponsor decide to discontinue the study prematurely for any 
reason, the Investigator must promptly contact all participating subjects so they can be appropriately 
followed-up. All study supplies must be collected and all case report forms must be completed as 
fully as possible.
Liability and insurance
Liability and insurance provisions for this study are set out in separate agreements.
Delegation of Investigator duties
The Investigator can delegate tasks to the research team but he/she remains responsible for 
coordinating and informing his/her staff about the protocol and the possible changes made to it.
The Investigator should ensure that all persons assisting with the study are adequately qualified, and 
are informed about the study protocol, any amendments to the protocol, the study treatments, and 
their study-related duties and functions. The Investigator should maintain a list of appropriately 
qualified persons to whom significant study-related duties will be delegated. The list is to be kept 
up-to-date. The Investigator should supply an up-to-date curriculum vitae (CV) in English, dated 
and signed, together with a list of his/her collaborators responsible for the practical conduct of the 
study. These collaborators should also provide a recent English version of their CVs, dated and 
signed.
Study agreement discontinuation
During the study, if events such as retirement, promotion or relocation prevent the Investigator from 
conducting the study as agreed, the Investigator should appropriately transfer his/her 
responsibilities, knowledge and documents to another willing individual, with the agreement of the 
Sponsor. Study specific contracts must be signed between the Sponsor and the newly assigned 
person.


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Responsibilities of the Monitor
16.2.
The responsibilities of the study Monitor are defined in ICH-E6, Chapter 5. The Monitor, who is 
mandated by the Sponsor, must ensure that the study is conducted in accordance with Good Clinical 
Practice guidelines and all applicable local laws, and that the rights, the security and the well-being 
of the subjects are respected.
During the conduct of the study, the Monitor reports any deviations or persistent poor compliance 
with the study requirements and the Sponsor makes decisions about appropriate corrective actions.
Communication
The Monitor is the main line of communication between the Investigator and the Sponsor.
Training
The Monitor must present the protocol and all procedures related to the study during the study set-up 
visit and provide the Investigator with CRF completion guidelines.
Compliance
During periodic monitoring visits at mutually convenient times, the Monitor has the responsibility of 
assessing the progress of the study, of checking that the informed consent forms have been signed, 
of ensuring adhesion to and compliance with the study protocol and other study-related documents, 
and of ensuring the accuracy and completeness of the e-CRFs. Inconsistencies in the study records 
are to be resolved.
Source data verification
The Monitor will perform source document verification and validation and request clarification to 
ensure the accuracy, completeness and reliability of data.
IMP
The Monitor must ensure that IMP handling is properly carried out and documented.
He/she must ensure that the Investigator's file is up-to-date with regard to essential documents.
Responsibilities of the Data Manager
16.3.
Cognizant will be in charge of the management of clinical data from data entry to database lock 
under the supervision of LFB Data Manager.
17.
ETHICS AND REGULATORY CONSIDERATIONS
The current study is to be conducted in accordance with Good Clinical Practice (ICH-E6), European 
Directive 2001/20/EC, and the revised version of the Declaration of Helsinki set out in the European 
Directive, as well as with applicable local requirements.
In France, this study will be conducted in accordance with the CSP. 
The protocol will be submitted to the Health Authorities and a properly constituted Ethics 
Committee (EC) for formal approval of the study conduct in accordance with local regulations. The 
study should not begin until the protocol has received written approval from the EC and Health 
Authorities in accordance with local requirements.


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In accordance with specific local requirements, the Investigator may be responsible for submitting 
the protocol and any amendments to the local EC. A copy of the decision letter, a list and versions of 
documents submitted, the list of EC members and their affiliation should be provided by the 
Investigator to the Sponsor.
During the study, the Sponsor should promptly notify the Investigators, Health Authorities and EC 
of any relevant information that could affect the safety of subjects and could impact on the conduct 
of the study.
Personal Data Protection Committee
For biomedical research in France: the Sponsor attests his conformity regarding the Personal Data 
Protection French requirements ("Méthodologie de Réference MR001" dated 5 January 2006).
Insurance
The Sponsor will contract civil liability insurance to provide subjects with compensation for any 
injury, including the consequences of administration of the investigational product and of the study 
procedures. The insurance company is HDI Gerling and registration number is 01006575-14276.
In case of injury or disability resulting from participation in the study, the subject is requested to 
promptly inform the Investigator responsible for the study.
Indemnity
Subjects will receive compensation for their participation in this study for the following constraints 
imposed by the protocol: 
Participating to up to 16 study visits most of which will require the subject to come to the 
hospital, and some of which (V2 and V8) lasting more than several hours as the subject will 
have to wait 6 hours between the IMP administration and blood sample collection for 
cytokines, CRP analyses
Having urine sample and several blood samples collected during these visits
Having some first milk/breast milk sample collected for the breast feeding subjects.
The subjects will be indemnified based on the visits performed, with a total of 3000 € if completing 
the entire study (providing first milk/breast milk samples at V10 and V11, and participating to the 
additional visits AV1 and AV2). The indemnity per visit will be as follow:
Screening visit V1: 200 €
Antenatal IM roledumab administration V2: 375 € (200 € for the inclusion + 175 € for the IMP 
administration and follow-up sampling), or
Antenatal IV roledumab administration V2: 275 € (100 € the for inclusion + 175 € for the IMP 
administration and follow-up samplings)
V2b (IV only): 100 € per visit
V3 and V4: 100 € (this PK sampling visit can be performed at home) per visit
V5, V6, V7: 200 € per visit
Postnatal IMP administration V8: 375 € (200 € before administration + 175 € for the IMP 
administration and follow-up sampling)
V9: 100 €


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V10 and V11: 250 € (for first milk /breast milk sampling done at home) per visit
V12: 200 €
End of study visit V13: 150 €
Additional visit AV1: 150 € and Additional visit AV2: 150 €
A subject participating to the screening visit but who will not meet all the inclusion criteria will then 
be indemnified 200 €, and a subject who for example will withdraw at V3 will receive 675 €.
Changes to the protocol
The Sponsor will not assume any responsibility or liability resulting from implementation of 
unapproved deviations or changes.
The only circumstance in which an amendment may be initiated prior to approval by the Health 
Authorities is where the change is necessary to eliminate apparent immediate hazards to the 
subjects. In this event, the Investigator must notify the Sponsor and if applicable the Ethics 
Committee, in writing within 5 working days after implementation.
18.
AUDIT AND INSPECTION
An audit/inspection may be carried out by qualified Sponsor staff, by subcontracted auditors or by 
representatives of national or foreign Health Authorities to ensure that the study is conducted as per 
protocol and in accordance with regulatory requirements, and to ensure the validity of the data.
Participation in this study implies acceptance to cooperate in any potential audit/inspection.
The audit/inspection may consist of an inspection of the premises and equipment together with 
verification of the study documents and data. The investigational team must be available for 
inspection or audit.
When the Sponsor or the Investigator is informed that an inspection is to be performed, the other 
party must be informed immediately. Audits/inspection may take place after the end of the study.


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19.
REFERENCES
1
Bowman JM. RhD hemolytic disease of the newborn. N Engl J Med 1998;339:1775-77.
2
Urbaniak SJ, Greiss MA. RhD haemolytic disease of the foetus and the newborn. Blood Rev 
2000;14:44-61.
3
Bowman J. Thirty-five years of Rh prophylaxis. Transfusion 2003;43:1661-66.
4
Avent ND. The rhesus blood group system: insights from recent advances in molecular 
biology. Transfus Med Rev 1999;13:245-266.
5
Debbia M, Brossard Y and Lambin P. Measurement of the affinity of anti-D in the serum of 
immunized mothers and in immunoglobulin preparations with unlabeled antibodies. 
Transfusion 2005;45:975-83.
6
Beliard R, Waegemans T, Notelet D, Massad L, Dhainaut F, Romeuf C, Guemas E, Haazen 
W, Bourel D, Teillaud JL, Prost JF. A human anti-D monoclonal antibody selected for 
enhanced FcRIII engagement clears RhD+ autologous red cells in human volunteers as 
efficiently as polyclonal anti-D antibodies. Br J Haematol 2008;141:109-19.
7
A phase I, double-blind, randomised, placebo-controlled safety, pharmacokinetic and 
absolute bioavailability study of LFB-R593 recombinant monoclonal antiRh(D) antibody 
administered to healthy Rh(D) volunteers. Internal Report (ADNC-0701).
8
A phase II, open-label, randomized, dose-finding study to compare the efficacy (in terms of 
clearance of RhD-positive RBCs and safety of LFB-R593, a monoclonal anti-RhD antibody 
versus Rhophylac, a polyclonal anti-RhD immunoglobulin in healthy RhD negative 
volunteers. Draft Internal Report (ADNC-0726).
9
Bichler J, Schöndorfer G, Pabst G, Andresen I. Pharmacokinetics of anti-D IgG in pregnant 
RhD-negative women. BJOG 2003;110(1):39-45.
10
Mari G. Middle cerebral artery peak systolic velocity for the diagnosis of fetal anemia. 
Ultrasound Obstet Gynecol 2005;25:335-40.
11
Thorpe SJ, Sands D, Rautmann G and Schaffner G. International collaborative study to 
evaluate methods for quantifications of anti-D in immunoglobulin preparations. Vox Sang 
2002;83(1):412-50.
12
Liu W, Avent ND, Jones JW, Scott ML, Voak D. Molecular configuration of RhD epitopes 
as defined by site-directed mutagenesis and expression of mutant Rh constructs in K562 
erythroleukemia cells. Blood 1999;94:3986-96.
13
Kulkarni SS, Vasantha K, Gupte SC, Mohanty D, Ghosh K. Potential of commercial anti-D 
reagents in the identification of partial D variants in Indian population. Indian J Med Res 
2007;125(5):641-4.


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14
Thorpe SJ, Fox B, Turner C, Scott M. Competitive enzyme-linked immunoassay of 
monoclonal immunoglobulin G anti-D preparations. Transfus Med 2003;13(3):153-9.
15
Wing MG, Moreau T, Greenwood J, Smith RM, Hale G, Isaacs J, Waldmann H, Lachmann 
PJ, Compston A. Mechanism of first-dose cytokine-release syndrome by CAMPATH 1-H: 
involvement of CD16 (FcgammaRIII) and CD11a/CD18 (LFA-1) on NK cells. J Clin Invest 
1996;98:2819-26.
16
Dillman RO. Infusion reactions associated with the therapeutic use of monoclonal 
antibodies in the treatment of malignancy. Cancer Metastasis Rev 1999;18:465-71.
17
Sameer Wagle, MBBS, MD; Chief Editor: Ted Rosenkrantz, MD. Hemolytic Disease of 
Newborn Treatment & Management. http://emedicine.medscape.com/article/974349-
treatment.
18
Maayan-Metzger A, Schwartz T, Sulkes J, Merlob P. Maternal anti-D prophylaxis during 
pregnancy does not cause neonatal haemolysis. Arch Dis Child Fetal Neonatal Ed. 2001; 
Jan;84(1):F60-2.
19
Bennardello F, Coluzzi S, Curciarello G, Todros T, Villa S; as Italien Society of Transfusion 
Medicine and Immumnohaematology (SIMTI) and Italian Society of Gynaecology and 
Obstetrics (SIGO) working group. Recommendations for the prevention and treatment of 
haemolytic disease of the foetus and newborn. Blood Transfus 2015;13(1):109-34.
20
Stengel JZ, Arnold HL. Is infliximab safe to use while breastfeeding? World J Gastroenterol 
2008;14(19):3085-7.
21
Yver A, Homery MC, Fuseau E, Guemas E, Dhainaut F, Quagliaroli D, Beliard R, Prost JF 
Pharmacokinetics and safety of roledumab, a novel human recombinant monoclonal anti-
RhD antibody with an optimized Fc for improved engagement of FCγRIII, in healthy 
volunteers. Vox Sang 2012;103(3):213-22.


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20.
APPENDICES
Not applicable.

SAP_003.pdf: 
 
COVER PAGE 
 
 
TITLE: Pharmacokinetics and Safety of Roledumab, a fully human recombinant monoclonal 
anti-RhD antibody, in RhD-negative pregnant women carrying an RhD-positive foetus: a 
phase IIb, multicenter, open-label study. 
 
NCT Number: NCT02287896 
 
 
DOCUMENT: Statistical analysis plan (v2.0)  
 
 
VERSION & DATE OF DOCUMENT: Version 2.0; March 15, 2018 


Statistical Analysis Plan – version 2.0 dated 15 March 2018 
Page 1 / 63 
(PK, Efficacy and Safety) 
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STATISTICAL ANALYSIS PLAN 
 
Pharmacokinetics and Safety of Roledumab, a fully human recombinant monoclonal 
anti-RhD antibody, in RhD-negative pregnant women carrying an RhD-positive foetus: a 
phase IIb, multicentre, open-label study 
 
Statistical Analysis Plan Status: final version 2.0 (PK, Efficacy and Safety) 
Statistical Analysis Plan Date: 15 March 2018 
Study Drug: Roledumab 
Clinical Study Protocol Number: ADNC-1301  
Updated with Amendment 10 and 11 (Protocol v12) 
Covance Study Number: 8368060 
Clinical Phase 2b 
 
 
 
 
Sponsor: 
LFB Biotechnologies 
3, avenue des Tropiques 
BP 40305 
91958 COURTABOEUF Cedex - France 
 
 
Clinical Project Manager: 
  
 
 
Project Statistician: 
 
 
 
 
 
 
 




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2. TABLE OF CONTENTS 
1. 
STATISTICAL ANALYSIS PLAN APPROVAL SIGNATURES ......................................... 2 
2. 
TABLE OF CONTENTS ............................................................................................................. 3 
3. 
ABBREVIATIONS ...................................................................................................................... 5 
4. 
INTRODUCTION ........................................................................................................................ 8 
5. 
STUDY OBJECTIVES ................................................................................................................ 8 
6. 
STUDY DESIGN .......................................................................................................................... 9 
6.1. 
Study plan and assessments ................................................................................................... 9 
6.2. 
Pharmacokinetic Samplings ................................................................................................. 17 
6.2.1. 
Maternal serum sampling ............................................................................................. 17 
6.2.2. 
Milk sampling .............................................................................................................. 18 
6.2.3. 
Cord blood sampling .................................................................................................... 18 
7. 
TREATMENTS .......................................................................................................................... 18 
8. 
SAMPLE SIZE JUSTIFICATION ........................................................................................... 18 
9. 
DEFINITION OF ANALYSIS POPULATIONS .................................................................... 19 
10. 
STUDY ENDPOINTS ............................................................................................................ 20 
10.1. Primary endpoints ................................................................................................................ 20 
10.2. Secondary endpoints ............................................................................................................ 20 
11. 
STATISTICAL METHODOLOGY ..................................................................................... 21 
11.1. General ................................................................................................................................. 21 
11.1.1. 
Definition of Baseline and Change from Baseline ....................................................... 21 
11.1.2. 
Repeat and Unscheduled Readings .............................................................................. 22 
11.2. Study subjects ...................................................................................................................... 22 
11.2.1. 
Subject disposition ....................................................................................................... 22 
11.2.2. 
Protocol deviations ....................................................................................................... 23 
11.3. Demographic and baseline characteristics ........................................................................... 23 
11.3.1. 
Demographics .............................................................................................................. 23 
11.3.2. 
Medical history and surgical history ............................................................................ 24 
11.3.3. 
Prior and concomitant therapies ................................................................................... 25 
11.4. Measurement of treatment compliance ................................................................................ 25 
11.5. Pharmacokinetic Assessment ............................................................................................... 26 
11.5.1. 
Non-Compartmental Pharmacokinetic Analysis (NCA) .............................................. 26 
11.5.2. 
Presentation of Pharmacokinetic Data ......................................................................... 29 
11.6. Efficacy Assessment ............................................................................................................ 29 
11.7. Safety Assessment ................................................................................................................ 30 
11.7.1. 
Extent of exposure........................................................................................................ 30 
11.7.2. 
Adverse events ............................................................................................................. 31 
11.7.3. 
Clinical laboratory evaluation ...................................................................................... 32 


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11.7.4. 
Vital Signs .................................................................................................................... 34 
11.7.5. 
Delivery ........................................................................................................................ 34 
11.7.6. 
Characteristics and general status of the newborn ....................................................... 34 
11.7.7. 
Other observations related to safety ............................................................................. 34 
12. 
STATISTICAL/ANALYTICAL ISSUES ............................................................................ 36 
12.1. Handling of Dropouts and Missing Data ............................................................................. 36 
12.1.1. 
Dropouts ....................................................................................................................... 36 
12.1.2. 
Missing data ................................................................................................................. 36 
12.1.3. 
Missing or incomplete dates......................................................................................... 36 
12.1.4. 
Randomization ............................................................................................................. 36 
12.1.5. 
Data safety monitoring board ....................................................................................... 36 
12.1.6. 
Blind Review ................................................................................................................ 36 
12.1.7. 
Maintaining the blind ................................................................................................... 36 
12.1.8. 
Pooling of Centers ........................................................................................................ 36 
12.1.9. 
Multiplicity issues ........................................................................................................ 37 
12.1.10. 
Subgroup analysis .................................................................................................... 37 
12.1.11. 
Other issues .............................................................................................................. 37 
13. 
INTERIM ANALYSES ......................................................................................................... 37 
14. 
SPECIFIC ANALYSIS RULES............................................................................................ 37 
15. 
VISIT AND TIME WINDOWS ............................................................................................ 38 
16. 
DEFINITION OF DERIVED VARIABLES ....................................................................... 39 
17. 
LABORATORY STANDARDIZATION (FOR LOCAL LABS) ...................................... 40 
18. 
CHANGES FROM THE PROTOCOL SPECIFIED STATISTICAL ANALYSES ....... 40 
18.1. Definition of analysis populations (Section 9) ..................................................................... 40 
18.2. Primary endpoints (Section 10.1) ......................................................................................... 40 
18.3. Other observations related to safety (Section 11.1) ............................................................. 40 
18.4. Other observations related to safety (Section 11.7.5) .......................................................... 40 
19. 
DATA PRESENTATION ...................................................................................................... 41 
20. 
REFERENCES ....................................................................................................................... 42 
21. 
APPENDICES ........................................................................................................................ 43 
21.1. Appendix 1 - Tables, figures and listings............................................................................. 43 
21.1.1. 
Table of contents - Section 14 of CSR ......................................................................... 43 
21.1.2. 
Table of contents - Section 16 of CSR ......................................................................... 46 
21.1.3. 
Table shells................................................................................................................... 49 
21.2. Appendix 2 - Population analysis plan................................................................................. 53 
21.3. Appendix 3 - Tables, Figures, and Listings of PK report (NCA and POPPK) .................... 63 
 


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3. ABBREVIATIONS 
Abbreviations pertain to the SAP only (not the TFLs). 
Abbreviation or 
specialist term 
Definition or explanation 
ADA 
Anti-Drug Antibody 
ADaM 
Analysis data model 
AE 
Adverse event 
AIC 
Akaike’s information criterion 
ALT 
Alanine aminotransferase 
APGAR 
Appearance, Pulse, Grimace, Activity, and Respiration 
aPTT 
Activated partial thromboplastin time 
AST 
Aspartate aminotransferase 
ATC 
Anatomical therapeutic chemical 
AUC 
Area under the plasma drug concentration-time curve 
AUC0-t 
Area under the concentration-time curve from time 0 to time Tlast 
AUC%extrap 
Percentage of AUC that is due to extrapolation from the time of the last 
quantifiable concentration to infinity 
AUC0-∞ 
Area under the concentration-time curve from time zero to infinity 
BLQ 
Below the limit of quantification 
BMI 
Body mass index 
CDISC 
Clinical data interchange standards consortium 
CL 
Total serum clearance 
CL/F 
Apparent total serum clearance 
Cmax 
maximum observed plasma concentration 
CPAP 
Clinical pharmacology analysis plan 
CRCL 
Creatinine clearance 
CRF 
Case report form 
CRP 
C-Reactive protein 
CSR 
Clinical study report 
CV 
Coefficient of variation  
CWRES 
Conditional weighted residuals 
DAT 
Direct antiglobulin test 
DBL 
Database lock 
DSMB 
Data safety monitoring board 
DV 
Dependent variable 
EC 
Early clinical 
ENT 
Ear–nose-throat 
EOS 
End of study 
ETA or  
Random inter-individual effect that is normally distributed with mean 0 and 
variance-covariance matrix  
F 
Bioavailability 
FAS 
Full analysis set 
FMH 
Fetomaternal hemorrhage 
FO 
First-Order estimation method 


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Abbreviation or 
specialist term 
Definition or explanation 
FOCE 
First-order conditional estimation method 
FOCEI 
First-order conditional estimation with interaction method 
GA 
Gestational age 
HBV 
Hepatitis B virus 
HCV 
Hepatitis C virus 
HIV 
Human immunodeficiency virus 
IAT 
Indirect antiglobulin test 
ICH 
International conference on harmonisation 
IIV 
Inter-individual variability 
IM 
Intramuscular 
IMP 
Investigational medicinal product 
IPRED 
ITT 
Individual predicted concentration 
Intent-To-Treat 
IV 
Intravenous 
Ka 
Absorption rate constant 
KB 
Kleihauer-Betke 
z 
Apparent terminal elimination constant 
LLOQ 
Lower limit of quantification 
LLR 
Log likelihood ratio 
LOCF 
Last observation carried forward 
LOESS 
Locally weighed smoothed line 
LRT 
Likelihood ratio test 
MCA-PSV 
Middle cerebral arterial-peak systolic velocity 
MCH 
Mean Corpuscular Hemoglobin 
MCHC 
Mean Corpuscular Hemoglobin Concentration 
MCV 
Mean Corpuscular Volume 
MedDRA 
Medical dictionary for regulatory activities 
MOF 
Minimum value of the objective function 
MoM 
Multiple of medians 
NCA 
Non-compartmental pharmacokinetic analysis 
NM-TRAN 
NONMEM translator 
NONMEM 
Non-linear mixed-effects modeling software 
PAP 
Population analysis plan 
PD 
Protocol deviation 
PK 
Pharmacokinetic or pharmacokinetics 
PKS 
Pharmacokinetics set 
PopPK 
Population pharmacokinetic 
PRED 
Population predicted concentration 
PT 
Preferred term 
Q 
Inter-compartmental clearance 
Q1 
First quartile 
Q3 
Third quartile 
QQ 
Quantile-quantile or QQ-plot 


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Abbreviation or 
specialist term 
Definition or explanation 
RBC 
Red blood cells 
RES 
Residual 
SAE 
Serious adverse event 
SAF 
Safety analysis set 
SAP 
Statistical analysis plan 
SD 
Standard deviation 
SDTM 
Study data tabulation model 
SOC 
System organ class 
SOJ 
Site of injection 
t1/2 
Apparent terminal elimination half-life 
Tlast 
Last time point with a measurable concentration 
TEAE 
Treatment-emergent adverse event 
TFLs 
Tables, figures, and listings 
TNF 
Tumor necrosis factor 
Tmax 
Time of maximum observed concentration 
TVP 
Typical value of a parameter P 
ULOQ 
Upper limit of quantification 
V2 or Vc 
Central compartment volume of distribution 
V3 or Vp 
Peripheral compartment volume of distribution 
Vd 
Volume of distribution 
Vz/F 
Apparent volume of distribution during the terminal phase after non-
intravenous administration  
Vz 
Volume of distribution during the terminal phase 
Vss 
Volume of distribution at steady-state (within the body)  
WT 
Body weight (kg) 
 
 
 


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4. INTRODUCTION 
This SAP has been developed consistent with the clinical study protocol (final version 12.0 dated 27 
October 2017) [1]. 
This SAP describes the planned analysis of the efficacy, safety and PK data from this study. A 
detailed description of the planned TFLs to be presented in the Clinical Study Report (CSR) is 
provided in the accompanying TFL shell document. 
The intent of this document is to provide guidance for the statistical analyses of efficacy, PK and 
safety data. In general, the analyses are based on information from the protocol, unless they have 
been modified by agreement between LFB Biotechnologies and Covance Early Clinical (EC) 
Biometrics. A limited amount of information concerning this study (e.g. objectives, study design) is 
given to help the reader’s interpretation. This SAP must be finalized prior to the database lock 
(DBL) for this study.  
This SAP supersedes the statistical considerations identified in the protocol; where considerations 
are substantially different, they will be so identified. If additional analyses are required to 
supplement the planned analyses described in this SAP, they may be performed and will be 
identified in the CSR. Any substantial deviations from this SAP will be agreed upon between LFB 
Biotechnologies and Covance EC Biometrics and identified in the CSR. 
This SAP is written with consideration of the recommendations outlined in the International 
Conference on Harmonisation (ICH) E9 guideline entitled “Guidance for Industry: Statistical 
Principles for Clinical Trials” and the ICH E3 guideline entitled “Guidance for Industry: Structure 
and Content of Clinical Study Reports [2], [3].” 
5. STUDY OBJECTIVES 
Primary Objective 
The primary objective is to assess the pharmacokinetic profile (PK) of roledumab 300 μg 
administered intramuscular (IM) or intravenous (IV) in RhD-negative pregnant women carrying an 
RhD-positive fetus. 
Secondary Objectives 
The secondary objectives are: 
 To assess the safety of roledumab administered IM and IV in RhD-negative pregnant women and 
in RhD-positive fetus and newborn 
 To assess the efficacy of roledumab 300 μg IM and IV to prevent RhD-alloimmunization in RhD-
negative pregnant women carrying an RhD-positive fetus 
 To assess the immunogenicity of roledumab 
 To measure roledumab concentration in first milk and breast milk 
 To measure roledumab concentration in cord blood after delivery 


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6. STUDY DESIGN 
This is a phase 2b, interventional, multicenter, open-label study. The study is not randomized. When 
the target number of subjects was reached in IM part, the subsequent subjects were selected to 
participate in the IV part (sequential). It was initially planned to enroll 35 subjects to be treated with 
300 μg IM (IM arm) and subsequently 25 subjects to be treated with 300 μg IV (IV arm) (See 
Section 8 for the justification of the sample size). 
6.1. Study plan and assessments 
Figure 1: Study Plan (IM and IV arms) 
 
 
 


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Figure 2: Visits schedule in the IM arm 
 


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Figure 3: Visits schedule in the IV arm 
 


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The subjects will attend at least 13 visits in the IM arm and 14 in the IV arm (9 will be performed at 
the hospital unit and 4 (IM) and 5 (IV) at home possibly). Additional visits may occur. 
Additional visits will occur in case of: 
 Sensitizing events requiring IMP administration 
 Potential RhD-alloimmunization requiring follow-up up to 12 months after the IMP 
administration. 
The study consists of a screening period, an antenatal period, a postnatal period and a follow-up 
period: 
 The screening period starting between 12 and 27 weeks gestation age (GA) with the screening 
visit performed as close as possible to the antenatal IMP administration, i.e. within 2 weeks 
 The antenatal period comprises 6 visits (IM) and 7 visits (IV) and lasts about 12 weeks. 
 The postnatal period comprises 2 visits and lasts 3  1 days. 
The follow-up period comprises at least 4 visits (6 months  2 weeks) and when necessary (i.e. in 
case of a positive indirect Coombs test), 2 additional visits. 
Each subject will participate in this study until 9.5 months if the Indirect Antiglobulin Test (IAT) 
result is negative at 6-month post-delivery. Otherwise, the maximum duration of the participation 
for a subject could go up to 18 months. 
 


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Table 1: Schedule of assessments for the IM arm 
Visit (V) 
Day (D) 
Week (W) 
Month (M) 
Screening 
visit 
(V1) 
12-27 GA 
Antenatal period  
Postnatal period  
Follow-up period 
V2 
Week 28 or 29 GA 
(D1) 
V310 
48h24 
V410 
120h24 
V5 
9d3 
V6 
29d4 
V7 
59d4 
V8 
Postnatal Treatment 
72 hours of 
delivery 
V 9 
Discharge 
48h24 
V1010 
72h24 
after post-
natal 
injection* 
V1110 
9d2 after 
postnatal 
injection* 
V12 
6W2W 
V13 
6M2W 
Additional 
AV1 
9M 
2W 
Additional 
AV2 
12 M 
2W 
Inclusion 
Injection 
Before 
Injection* 
Informed Consent 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Eligibility Criteria 
x 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Demographics/ 
Medical History/ 
Life habits 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Serology: hepatitis 
B and C, HIV 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
RhD status 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
ABO status  
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Non-inv. fetal RhD 
genotyping 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Vital Signs1 
x 
x 
x 30 min 
and 6h 
after 
injection 
x 
x 
x 
x 
x 
x 
x 30 min 
after 
injection 
x 
x 
x 
x 
x 
x 
x 
Physical 
Examination 
x 
x2 
 
 
 
 
 
 
x2 
 
 
 
 
 
x2 
x2 
x2 
Laboratory Tests  
x3 
 
 
 
 
 
 
x4 
x4 
 
 
 
 
x4 
x12 
x4 
x4 
Cytokines5 
 
x 
x 
6h after 
injection 
 
 
 
 
 
x 
x 
6h after 
injection 
 
 
 
 
 
 
 
CRP 
 
x 
 
x 
 
 
 
 
x 
 
x 
 
 
 
 
 
 
Obstetric and fetal 
Doppler ultrasound  
 
x 
 
 
 
x 
x 
x 
 
 
 
 
 
 
 
 
 
IAT in mothers  
x 
x 
 
 
 
 
 
 
x 6 
 
 
 
 
 
x 6 
x 6 
x 6 
Anti-D 
Quantification 
 
x 
 
 
 
x 
x 
 
x 
 
 
 
x 
x 
x 
 
 
IM IMP 
administration 
 
 
x 
 
 
 
 
 
 
x 
 
 
 
 
 
 
 
PK sample 
 
x 
 
x 
x 
x 
x 
x 
x 
 
 
x 
x 
x 
 
 
 
KB test  
 
 
 
 
 
 
 
 
x 
x11 
 
 
 
 
 
 
 


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Visit (V) 
Day (D) 
Week (W) 
Month (M) 
Screening 
visit 
(V1) 
12-27 GA 
Antenatal period  
Postnatal period  
Follow-up period 
V2 
Week 28 or 29 GA 
(D1) 
V310 
48h24 
V410 
120h24 
V5 
9d3 
V6 
29d4 
V7 
59d4 
V8 
Postnatal Treatment 
72 hours of 
delivery 
V 9 
Discharge 
48h24 
V1010 
72h24 
after post-
natal 
injection* 
V1110 
9d2 after 
postnatal 
injection* 
V12 
6W2W 
V13 
6M2W 
Additional 
AV1 
9M 
2W 
Additional 
AV2 
12 M 
2W 
Inclusion 
Injection 
Before 
Injection* 
Breast milk sample 
 
 
 
 
 
 
 
 
 
 
 
x 
x 
 
 
 
 
ADA tests 
x13 
x 
 
 
 
x 
x 
 
x 
 
 
 
x 
x 
x 
x 
x 
Adverse events  
 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Concomitant 
treatment  
x 
x 
 
x 
x 
x 
x 
x 
x 
 
x 
x 
x 
x 
x 
x 
x 
Delivery7 
 
 
 
 
 
 
 
 
x 
 
 
 
 
 
 
 
 
Cord blood 
sample8 
 
 
 
 
 
 
 
 
x 
 
 
 
 
 
 
 
 
Newborn RhD 
status9 
 
 
 
 
 
 
 
 
x 
 
 
 
 
 
 
 
 
Newborn ABO 
status14 
 
 
 
 
 
 
 
 
x 
 
 
 
 
 
 
Newborn general 
status 
 
 
 
 
 
 
 
 
x 
 
x 
 
 
x 
 
 
 
1 
Include blood pressure, heart rate and body temperature 
2 
Partial examination (Weight, General Appearance, Skin, Heart, Lungs, Abdomen) 
3 
Hematology, blood chemistry, aPTT, urinalysis 
4 
Hematology, blood chemistry 
5 
Cytokines: IFN-, IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13 and TNF- 
6 
If IAT positive, IA characterization and Anti-D antibodies quantification  
7 
Collection of APGAR score, AEs, Concomitant Medications 
8 
Cord blood sample: DAT and hematocrit, hemoglobin, reticulocytes, total bilirubin including routine tests (RhD status), detection of roledumab 
9 
Only in case of a confirmed alloimmunization: RhD genotyping in cord blood will be performed to search for potential D-variants 
10 
Visit performed at home  
11 
Kleihauer-Betke test to be performed within 24h after IMP administration only if the one prior to the IMP administration was positive. 
12 
Hematology, blood chemistry, urinalysis 
13 
Evaluation of pre-existing antibody presence 
14 
If performed in routine practice during the delivery hospitalization of the mother 
* Postnatal injection should be performed within 72h after delivery. All subsequent visits are based from this time point. 
GA: gestational age refers to the duration of the pregnancy since the 1st day of the last menstrual period and will be expressed in week.s 
 


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Table 2: Schedule of assessments for the IV arm 
 
Visit (V) 
Hour (H) 
Day (D) 
Week (W) 
Month (M) 
Screening 
visit 
(V1) 
12-27 GA 
Antenatal period  
Postnatal period  
Follow-up period 
V2 
Week 28 or 29 GA (D1) 
V2b10 
24h 
2h 
V310 
48h 
6 h 
V410 
96h-
120h 
 
V5 
9d 
3 
V6 
29d4 
V7 
59d4 
V8 
Postnatal 
Treatment <72 
hours of delivery 
V 9 
Discharge 
48h24 
V1010 
72h24 
after 
post-
natal 
injection* 
V1110 
9d2 
after 
post-
natal 
injection* 
V12 
6W2W 
 
V13 
6M2W 
Additional 
AV1 
9M 
2W 
Additional 
AV2 
12 M 
2W 
Inc. Inj. 
(H0) 
30 
5min 
H1 
5min 
H6 
15min 
Before Injection* 
 
Informed Consent 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Eligibility Criteria 
x 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Demographics/ 
Medical History/ 
Life habits 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Serology: hepatitis 
B and C, HIV 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
RhD status  
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
ABO status 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Non-inv. fetal RhD 
genotyping 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Vital Signs1 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 30 min 
after inj. 
x 
x 
x 
x 
x 
x 
x 
Physical 
Examination 
x 
x2 
 
 
 
 
 
 
 
 
 
 
x2 
 
 
 
 
 
x2 
x2 
x2 
Laboratory Tests  
x3 
 
 
 
 
 
 
 
 
 
 
x4 
x4 
 
 
 
 
x4 
x12 
x4 
x4 
Cytokines5 
 
x 
 
 
 
x 
 
 
 
 
 
 
x 
x 
6h after 
inj. 
 
 
 
 
 
 
 
CRP 
 
x 
 
 
x 
 
x 
x 
 
 
 
 
x 
 
x 
 
 
 
 
 
 
Obstetric and fetal 
Doppler ultrasound  
 
x 
 
 
 
 
 
 
 
x 
x 
x 
 
 
 
 
 
 
 
 
 
IAT in mothers  
x 
x 
 
 
 
 
 
 
 
 
 
 
x 6 
 
 
 
 
 
x 6 
x 6 
x 6 
Anti-D 
Quantification 
 
x 
 
 
 
 
 
 
 
x 
x 
 
x 
 
 
 
x 
x 
x 
 
 
IV IMP 
administration 
 
 
x 
 
 
 
 
 
 
 
 
 
 
x 
 
 
 
 
 
 
 
PK sample 
 
x 
 
 
x 
 
x 
x 
x 
 
x 
x 
x 
 
 
x 
x 
x 
 
 
 
KB test  
 
 
 
 
 
 
 
 
 
 
 
 
x 
x11 
 
 
 
 
 
 
 


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Visit (V) 
Hour (H) 
Day (D) 
Week (W) 
Month (M) 
Screening 
visit 
(V1) 
12-27 GA 
Antenatal period  
Postnatal period  
Follow-up period 
V2 
Week 28 or 29 GA (D1) 
V2b10 
24h 
2h 
V310 
48h 
6 h 
V410 
96h-
120h 
 
V5 
9d 
3 
V6 
29d4 
V7 
59d4 
V8 
Postnatal 
Treatment <72 
hours of delivery 
V 9 
Discharge 
48h24 
V1010 
72h24 
after 
post-
natal 
injection* 
V1110 
9d2 
after 
post-
natal 
injection* 
V12 
6W2W 
 
V13 
6M2W 
Additional 
AV1 
9M 
2W 
Additional 
AV2 
12 M 
2W 
Inc. Inj. 
(H0) 
30 
5min 
H1 
5min 
H6 
15min 
Before Injection* 
 
Breast milk 
sample 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
x 
x 
 
 
 
 
ADA tests 
x13 
x 
 
 
 
 
 
 
 
x 
x 
 
x 
 
 
 
x 
x 
x 
x 
x 
Adverse events  
 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Concomitant 
treatment  
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
 
x 
x 
x 
x 
x 
x 
x 
Delivery7 
 
 
 
 
 
 
 
 
 
 
 
 
x 
 
 
 
 
 
 
 
 
Cord blood 
sample8 
 
 
 
 
 
 
 
 
 
 
 
 
x 
 
 
 
 
 
 
 
 
Newborn RhD 
status9 
 
 
 
 
 
 
 
 
 
 
 
 
x 
 
 
 
 
 
 
 
 
Newborn ABO 
status14 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Newborn general 
status  
 
 
 
 
 
 
 
 
 
 
 
 
x 
 
x 
 
 
x 
 
 
 
1 
Include blood pressure, heart rate and body temperature 
2 
Partial examination (Weight, General Appearance, Skin, Heart, Lungs, Abdomen) 
3 
Hematology, blood chemistry, aPTT, urinalysis 
4 
Hematology, blood chemistry 
5 
Cytokines: IFN-, IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13 and TNF- 
6 
If IAT positive, IA characterization and Anti-D antibodies quantification  
7 
Collection of APGAR score, AEs, Concomitant Medications  
8 
Cord blood sample: DAT and hematocrit, hemoglobin, reticulocytes, total bilirubin including routine tests (RhD status), detection of roledumab 
9 
Only in case of a confirmed alloimmunization: RhD genotyping in cord blood will be performed to search for potential D-variants 
10 
Visit performed at home  
11 
Kleihauer-Betke test to be performed within 24h after IMP administration only if the one prior to the IMP administration was positive 
12 
Hematology, blood chemistry, urinalysis 
13 
Evaluation of pre-existing antibody presence 
14 
If performed in routine practice during the delivery hospitalization of the mother 
* Postnatal injection should be performed within 72h after delivery. All subsequent visits are based from this time point. 
Inc: Inclusion / Inj: Injection / h: Hours / Min: Minutes; GA: gestational age refers to the duration of the pregnancy since the 1st day of the last menstrual period and will be expressed 
in weeks 


 
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Table 3: Study flow chart – additional visits in case of sensitizing event for IM/IV arms 
Visit 
Before injection 
6 hours after IMP 
administration 
24 hours after IMP 
administration 
Sensitizing event 
(description) 
x 
 
 
Vital signs 
x 
x 
x 
Partial physical 
examination 
x 
 
 
IAT 
x* 
 
 
Kleihauer-Betke test 
x** 
 
x*** 
Cytokines 
 
x 
 
CRP 
 
 
x 
Adverse Events 
x 
x 
x 
Concomitant 
treatment 
x 
x 
x 
* 
Perform an anti-D quantification by microtitration or ponderal dosage analysis if IAT is positive, to confirm the anti-D 
concentration 
** 
Before and after (if the first one is positive) anti-D administration 
*** The dose will be adapted to the KB test results 
The PK sampling schedule after the antenatal injection should stay the same in case of a sensitizing 
event treated with roledumab. 
6.2. Pharmacokinetic Samplings 
6.2.1. Maternal serum sampling 
Roledumab in maternal serum will be measured using a flow cytometry PK method by Envigo 
(formerly Huntingdon Life Sciences), located at Huntingdon, UK. The method was validated based 
on FDA (2001) and EMA (2012) bioanalytical guidelines. The validated detection range for 
roledumab in whole serum is from 1 to 15 ng/mL and dilutional integrity was demonstrated, 
allowing pre-dilution of study samples when required. All analytical runs will include a calibration 
curve and three levels of quality control samples with predefined acceptance criteria. 
PK parameters calculation (NCA) or estimates (population PK analysis) will be derived from 
roledumab measurements in serum samples of pregnant and post-delivery RhD-negative pregnant 
women carrying an RhD-positive fetus administered IM or IV with roledumab. All samples of a 
specific subject are to be assayed in the same assay run, except for those who will need to be rerun, 
such as results above the upper limit of quantification (ULOQ).  


 
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Samples of subjects enrolled in the ADNC-1301 clinical trial will be drawn within the 24 hours 
preceding injection of roledumab (pre-dose) and at the theoretical timepoints post-first and second 
administration described in Figure 2 and Figure 3 depending on the administration route. 
It is of importance to note that this method of describing the scheduled sampling times is for 
information only. Calculation of the PK parameters will require the use of the actual elapsed 
time from the time of injection to the actual time of sampling. 
6.2.2. Milk sampling 
The validated lower limit of quantification (LLOQ) of roledumab in maternal serum is 1.5 ng/mL. 
Based on current product knowledge, it is expected that only very few of the clinical samples will 
have reported concentrations above the LLOQ. 
If available the ratio between concentration of roledumab in breast milk and maternal serum 
concentration will be calculated. 
6.2.3. Cord blood sampling 
Cord blood at delivery (V8) will be collected for roledumab fetal/neonate concentration. The ratio 
between cord blood concentration of roledumab and maternal serum concentration will be 
calculated. 
7. TREATMENTS 
The following is a list of the study treatment abbreviations and ordering that will be used in the 
TFLs. 
Study Treatment Name 
Abbreviation 
Treatment Order on TFLs 
300 µg IM roledumab 
IM 
1 
300 µg IV roledumab 
IV 
2 
8. SAMPLE SIZE JUSTIFICATION 
No statistical hypothesis will be tested. Therefore no formal sample size calculation is done. The 
expected number of subjects to be recruited was initially as follows: 
 In the IM arm, thirty-five (35) subjects to be screened in order to get 30 evaluable subjects 
 In the IV arm, twenty-five (25) subjects in order to get 20 evaluable subjects.  
 
 


 
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9. DEFINITION OF ANALYSIS POPULATIONS 
The intent-to-treat (ITT) population: 
The intent-to-treat (ITT) population includes all subjects who were enrolled in the study (i.e. who 
signed the ICF and who were not screening failures) and/or who did not complete the EOS form 
before the inclusion visit (V2) (See Section 18.1). 
The full analysis set (FAS): 
The full analysis set (FAS) is a subset of ITT population and will be used for the efficacy analysis. It 
consists of all ITT subjects who have a negative IAT test at inclusion and who delivered an RhD-
positive newborn. 
The safety set (SAF): 
The Safety Set (SAF) will be used for all safety analyses. It consists of all subjects who received at 
least one administration of IMP. 
Pharmacokinetics Sets (PKS): 
PKS1: Analysis population 1 for population PK modeling 
The PKS1 consists of all ITT subjects enrolled in study ADNC-1301 (IM or IV arm), treated at least 
once with roledumab, and providing at least one roledumab available concentration after 
administration (including antenatal and postnatal measurements after 1st or 2nd administration). 
PKS2: Analysis population 2 for NCA IM arm 
The PKS2 consists of all ITT subjects enrolled in the IM arm of study ADNC-1301, who have at 
least one valid PK assessment after the first IMP administration, where valid PK assessment is 
defined as providing at least one evaluable PK parameter. 
PKS3: Analysis population 3 for NCA IV arm 
The PKS3 consists of all ITT subjects enrolled in the IV arm of study ADNC-1301, who have at 
least one valid PK assessment after the first IMP administration, where valid PK assessment is 
defined as providing at least one evaluable PK parameter. 
For the PK analysis, if any of the following elements is missing, all of that subject’s data will be 
excluded from all analyses: 
a. Exact date/time of IMP administration 
b. Dose amount 
Missing date and time of sampling for a subject will lead to a deviation for the specific PK 
timepoint. 
 Additionally, subjects who have complete dosing data but no concentrations associated to 
samplings will be excluded from all PK analysis 
 If either a sampling time or its concentration is missing, then that sample data point will be 
excluded 
 If preanalytical conditions are not met, sample results will be discarded from analysis.  


 
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All protocol deviations that occur during the study will be considered prior to DBL for their 
severity/impact and will be taken into consideration when subjects are assigned to analysis 
populations. Details of subject assignment to the analysis populations will be listed. 
The complete specific criteria defining protocol deviations and their classification (major/minor) 
will be agreed by the study team during the Data Review or during Protocol Deviation discussion 
meeting prior to DBL. The protocol deviation specifications are described in a separate document 
(ADNC-1301 Protocol Deviation Specifications). 
10. 
STUDY ENDPOINTS 
10.1. Primary endpoints 
Serum concentrations of roledumab will be measured at the defined time points as described in the 
schedule of assessments (See Table 1 and Table 2). The PK profile of roledumab will be described 
by: 
1. A primary compartmental population PK model will be performed with 6  primary parameters: 
Central compartment clearance (CL), Central compartment volume of distribution (V2), Inter-
compartmental clearance (Q), Peripheral compartment volume of distribution (V3),  IM route 
bioavailability (F) and IM route Absorption rate constant (Ka); and derived parameter: half-life 
(t1/2) (See Section 18.2 for changes from protocol). 
2. A secondary PK analysis (NCA) will provide the following parameters: Vd, CL, Cmax, Tmax, 
AUCt, AUC0-∞, terminal half-life and elimination rate constant. Performing PK assessment after 
IV administration will further allow estimation of the bioavailability of roledumab and better 
accuracy of the parameters estimation.  
All samples (ante- and postnatal samples) will be used for population PK modeling whereas only 
samples post-first administration (antenatal samples) will be analyzed by NCA for each study arm. 
10.2. Secondary endpoints 
Safety of the mother and the fetus/newborn will be assessed throughout the study by the incidence, 
nature, severity, seriousness, relationship to the IMP of AEs and by changes in physical examination 
findings, and vital sign measurements, clinical laboratory tests and Doppler ultrasound. Safety of 
newborns/infants up to 6 months age will be assessed by recording and reporting any AE and related 
concomitant medications.  
The efficacy of roledumab 300 µg IM/IV to prevent RhD-alloimmunization in RhD-negative 
pregnant women carrying a RhD-positive fetus will be assessed by screening the blood sample 
collected at 6 months (and possibly at 9 and 12 months) after birth of the RhD-positive fetus using 
Indirect Antiglobulin Test (IAT). In case of a positive IAT, the anti-RhD antibodies will be 
quantified centrally by microtitration or ponderal dosage. 
If the IAT is negative at 6 months post-delivery, the subject will be considered as negative at 9 and 
12 months. If the IAT is positive (i.e. alloimmunization confirmed by anti-D quantification) at 6 
months, the subject will be followed until 9 and eventually 12 months in order to distinguish 
residual anti-RhD antibodies from anti-RhD alloantibodies. If the IAT is positive (i.e. confirmed by 


 
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anti-D quantification) at 6 months but negative at 9 months post-delivery, the subject will be 
considered as negative at 12 months.  
The immunogenicity of roledumab will be assessed during the course of the study using the results 
of ADA tests. 
Roledumab breast milk/first milk concentration, maternal serum concentration and cord blood 
concentration after delivery will be also measured. 
11. 
STATISTICAL METHODOLOGY 
11.1. General 
For continuous data, summary statistics will include the arithmetic mean, arithmetic standard 
deviation, median, minimum, maximum, Q1, Q3, number of observations and number of missing 
observations. For log-normal data (e.g. the PK parameters: areas under the concentration-time curve 
[AUCs] and maximum observed concentration [Cmax]), the geometric mean and geometric 
coefficient of variation (CV%) will also be presented. 
Categorical data (binary, nominal and ordinal) will be summarized by number of observations, 
number of missing observations, frequency count and percentages. Percentages will be calculated on 
the observed cases. 
In general, missing values will not be imputed, otherwise it will be specified. Only descriptive 
statistics will be given, no confidence interval will be provided. 
Individual data listings in Section 16.2 of the CSR will be provided for all subjects, including screen 
failures.  
Summary statistics and statistical analyses will be performed for subjects included in the relevant 
analysis populations. 
Data analysis will be performed using SAS v9.4 or latest version in a secure and validated 
environment. All report outputs will be provided to the Sponsor in Microsoft Word version 10. 
Analysis Data Model (ADaM) datasets will be prepared using Clinical Data Interchange Standards 
Consortium (CDISC) ADaM Version 2.1, and CDISC ADaM Implementation Guide Version 1.1. 
Pinnacle 21 Community Validator Version 2.1.2 will be used to ensure compliance with CDISC 
standards. 
Study Data Tabulation Model (SDTM)) datasets will be prepared using Clinical Data Interchange 
Standards Consortium (CDISC) SDTM Version 1.4, and CDISC SDTM Implementation Guide 
Version 3.2 utilized. 
11.1.1. Definition of Baseline and Change from Baseline 
Baseline for each parameter is defined as the last value measured prior to the first IMP injection, 
including repeat (vital signs) and unscheduled (clinical laboratory parameters) readings (see Section 
11.1.2 for definitions of repeat and unscheduled readings). 


 
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Mean change from baseline is the mean of all individual subjects’ change from baseline values. 
Each individual change from baseline will be calculated by subtracting the individual subject’s 
baseline value from the value at the timepoint. The individual subject’s change from baseline values 
will be used to calculate the mean change from baseline. 
11.1.2. Repeat and Unscheduled Readings 
Repeat readings occur when the original vital signs result requires confirmation. Repeat readings are 
labelled as ‘Repeat’ in the listings and replace the original readings in all summaries and changes 
from baseline presentations and calculations. Prior to dosing, all readings taken in addition to the 
original reading are defined as pre-dose repeats. Post-dose repeat readings are defined as readings 
collected within 15 minutes of the actual time of the original reading. 
With the exception of pre-dose results described above, unscheduled readings for vital signs are 
defined as readings collected >15 minutes from the actual time of the original reading. All results 
not taken at a scheduled timepoint for other data types (e.g. clinical laboratory parameters) are 
unscheduled. Unscheduled readings are labelled as ‘Unscheduled’ in the listings. Because 
unscheduled readings are not associated with any scheduled timepoint, they are excluded from all 
summaries (with the exception that they may be used as baseline, as stated in Section 11.1.1). 
11.2. Study subjects 
11.2.1. Subject disposition 
The subject disposition will be summarized as follows: 
 Number of screened subjects 
 Number and percentage of screen failures 
 Number of subjects enrolled into the study (who signed inform consent and not screening 
failure – see Section 18.1) 
 Number and percentage of subjects who received study treatment 
 Number and percentage of mothers treated with RhD-negative fetus 
 Number and percentage of subjects who completed the study 
 Number and percentage of subjects early withdrawn from the study and reasons for study 
discontinuation 
The analysis populations will be summarized with the number and the percentage of subjects 
included in each analysis set with respect to the number of enrolled subjects: 
 Number and percentage of subjects in ITT population 
 Number and percentage of subjects in FAS 
 Number and percentage of subjects in SAF 
 Number and percentage of subjects in PKS1 
 Number and percentage of subjects in PKS2 


 
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 Number and percentage of subjects in PKS3 
For each analysis population, the number and the percentage of subjects who completed the study 
will be displayed. 
A listing of subject disposition will be presented by subject. This listing will present for each subject 
the enrollment status (yes/no), the date of signature of the ICF, the date of screening visit, the date of 
inclusion visit, the date of last visit, date of completion (i.e. date of completion of the EOS form) the 
individual study duration (in days) and the final status of the subject (completed study or not).  
A by-subject table of screen failures will be provided with the reason for excluding subjects during 
the screening phase. Another by-subject listing will provide the inclusion and exclusion criteria by 
subject for all screened subjects. 
A listing of withdrawals will also be displayed with the reasons for discontinuation. 
The disposition of mothers who delivered RhD-negative neonates will also be presented if any.  
A by-subject listing will display the visits done by each subject during the study with the date 
associated to each visit. It will also be indicated if the visit was done at hospital or at home as well 
as the related day (from the first IMP administration). 
In addition, a by-subject listing for each analysis population (ITT, FAS, SAF, PKS1, PKS2 and 
PKS3) will be provided with the reason for exclusion from the considered analysis population. 
11.2.2. Protocol deviations 
All protocol deviations (PD) will be listed in a by-subject listing and will also be summarized in a 
table displayingthe number and percentage of subjects with at least one PD for each protocol 
deviation category and classification (minor/major) on ITT population. 
11.3. Demographic and baseline characteristics  
11.3.1. Demographics 
Summary statistics will be provided for demographic and other baseline characteristics by route of 
administration and overall for the SAF. 
Demographic and other baseline characteristics will be qualitatively or quantitatively described 
(depending on the nature of data). No comparison will be performed between the groups of subjects 
based on the route of administration. 
 
The following demographic characteristics at baseline will be described in a summary table: age 
at enrolment (years), gender, race, weight before pregnancy (kg), weight at screening (kg) and 
BMI before pregnancy (kg/m²). These demographic characteristics will also be described for the 
FAS and PKS populations in addition to the SAF. 
A by-subject listing will display some demographic data (age and race) as well as the date of 
screening, the date of signature of the ICF and the date of birth. 
 
Other baseline characteristics: 
 Laboratory data at baseline: 


 
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 Serology 
o HBV test (HBs Ag, Anti-HBc Ab, Anti-HBs Ab) 
o HCV test (Anti-HCV Ab) 
o HIV test (Anti-HIV 1+2) 
 Immunogenicity assessment – ADA test 
 Indirect Antiglobulin Test (IAT) 
 Life habits: Substance use (Tobacco Consumption and Alcohol Consumption)  
 Current pregnancy information 
 ABO status for the mother and RhD status for the mother and the fetus 
Laboratory data for serology (positive/negative/not done for each test) at baseline will be displayed 
in summary tables as well as in a by-subject listing. 
The number of subjects with ADA tested at baseline and the associated result (positive/negative) 
will be qualitatively summarized. 
The results of IAT at baseline will be qualitatively summarized (positive/negative). 
Life habits will be summarized as frequency count and percentages. These data will also be listed in 
former and current smokers and/or in subjects with alcohol consumption. In addition, life habits will 
be listed for all subjects in another listing.  
Current pregnancy information will be provided in a by-subject listing (first trimester (11-14 weeks) 
ultrasound performed (yes/no), date of the first trimester ultrasound, gestational age at the inclusion 
visit, type of pregnancy (single/multiple), maternal or fetal abnormality detected (yes/no), 
complication during the current pregnancy (yes/no), anti-RhD+ injection during the current 
pregnancy (yes/no)). 
The ABO status and the RhD status of the mother as well as the RhD status for the fetus will be 
presented in a listing by subject idenfier of the mother. 
11.3.2. Medical history and surgical history 
Significant medical and surgical history 
The MedDRA (V16.0) SOC in decreasing order (based on the overall column) and PT within SOC, 
for medical and surgical history will be summarized by frequency and percentages on the SAF by 
route of administration and overall. 
Medical and surgical history data will also be listed by subject. This listing will contain the actual 
route of IMP administration, the verbatim text, SOC and PT associated to each individual surgical 
and medical history, the start date and the end date (or the on-going status) as well as the intake of a 
concomitant treatment if any (yes/no).   
Obstetrical history 
The obstetrical history will be summarized using appropriate statistics (depending on the nature of 
data) for the SAF by route of administration and overall: 
 Previous pregnancy(ies) (Primigravidae subject: yes/no; if no, number of previous 
pregnancies) and delivery(ies) (number of deliveries >24 weeks of pregnancy; number of 
preterm deliveries (<37 weeks)) 


 
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 Number of induced abortions  
 Number of Spontaneous Miscarriages 
 Number of live births 
 Anti-RhD Injection during previous pregnancy(ies) (yes/no) 
 Any complication during previous pregnancy(ies) (yes/no) 
 Any complication post-partum (yes/no). 
These data will also be presented in a by-subject listing. In addition, a by-subject listing will provide 
details on miscarriages and abortions (date, nature and reason of the event, gestation duration, type 
of delivery, anti-D injection (yes/no) and doses received if applicable). Another by-subject listing 
will provide information on complications during previous pregnancies (date of the event, 
complication post-event (yes/no) and details if any). 
In addition, a by-subject listing will display information on newborn after previous pregnancies 
(date of delivery, gender, height, weight, gestation duration, mode of delivery, anesthesia (yes/no), 
instrumental extraction (yes/no) and details if any). Another by-subject listing will provide 
information on dose received for anti-D injection (anit-D injection (yes/no), dose, unit, antenatal 
dose, postnatal dose) and another listing will provide information on complications during previous 
pregnancies if any (description of the complications, start and end date). 
11.3.3. Prior and concomitant therapies 
Prior/Concomitant treatments will be summarized on the SAF by route of administration and 
overall. 
Prior/Concomitant medications will be coded using WHO Drug Dictionary version 13.0. The 
number and percentage of subjects receiving at least one medication will be presented by generic 
name, ATC code (level 2 and 4).  
A frequency table with number and percentage of subjects (a table for prior medications and another 
table for concomitants medications) with a least one medication will be provided. The results will be 
displayed in the alphabetic order (for the two considered levels). 
Prior medication in this study refers to any medication received (within 1 month before ICF 
signature, 6 months before for blood transfusion (or blood derived products)) and stopped before 
signature of the informed consent. 
Concomitant medication refers to any medication that the subject receives at any time from the 
signature of the informed consent to the last study visit. This includes the screening/baseline period, 
treatment period and follow-up period. 
All prior and concomitant medications will be listed (based on coded drug names) for each subject 
with the associated ATC codes and will be identified as prior or as concomitant medications. 
11.4. Measurement of treatment compliance 
Summary statistics will be provided for treatment compliance for the SAF, by route of 
administration and overall. 
Compliance to the administration schedule will be measured as follows: 


 
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 Number and percentage of subjects with IMP administration within the planned window of 28 or 
29 weeks of gestation for the first administration (yes/no). In case of IMP administration outside 
the window, the number of days the administration is given outside the planned window of 28 or 
29 weeks of gestation (first administration) will be quantitatively summarized 
 Number and percentage of subjects with IMP administration within the planned window of 
delivery <72h (Yes/No). The number of hours the IMP administration given outside the planned 
window of delivery will be quantitatively summarized 
 Number and percentage of subjects with IMP administration within the planned window of 
sensitizing event if any (yes/no). The number of hours the IMP administration given outside the 
planned window will be quantitatively summarized. 
 Overall compliance will assess the number and percentage of subjects with all IMP 
administrations performed within the planned windows for the cases mentioned above. 
All compliance data will also be displayed in a by-subject listing. 
11.5. Pharmacokinetic Assessment 
Pharmacokinetic assessment will be based on two PK analyses: 
 Primary PK analysis will consist in a population PK analysis (PopPK analysis) that will be 
conducted on the analysis population PKS1 (See Section 9)  
 Secondary PK analysis will be a non-compartmental PK analysis (NCA) that will be 
performed on the analysis populations PKS2 and PKS3 (See Section 9) 
The PopPK analysis is detailed in Section 21.2. The non-compartmental PK analysis is presented 
below in Section 11.5.1.  
11.5.1. Non-Compartmental Pharmacokinetic Analysis (NCA) 
The following NCA PK parameters will be determined where possible from the serum roledumab 
concentrations following the first administrations using non-compartmental methods performed 
using Phoenix WinNonlin (Pharsight Corporation, Version 6.4 or higher): 
 
 


 
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IM Arm 
Parameter 
Definition 
AUC0-t 
Area under the concentration-time curve from time 0 to time Tlast, where Tlast is 
the last time point with a measurable concentration, calculated using the linear 
trapezoidal rule for increasing concentrations and the logarithmic rule for 
decreasing concentrations 
AUC0-∞ 
Area under the concentration-time curve extrapolated to infinity 
%AUCextrap 
Percentage of AUC that is due to extrapolation from the last measurable 
concentration to infinity 
Cmax 
Maximum observed serum concentration 
Tmax 
Time of maximum observed serum concentration 
Tlast 
Time of last quantifiable serum concentration 
t1/2 
Apparent terminal elimination half-life 
z 
Apparent terminal elimination constant 
CL/F 
Apparent total serum clearance  
Vz/F 
Apparent volume of distribution during the terminal elimination phase after non-
intravenous administration 
 
IV Arm 
Parameter 
Definition 
AUC0-t 
Area under the concentration-time curve from time 0 to time Tlast, where Tlast is 
the last time point with a measurable concentration, calculated using the linear 
trapezoidal rule for increasing concentrations and the logarithmic rule for 
decreasing concentrations 
AUC0-∞ 
Area under the concentration-time curve extrapolated to infinity 
%AUCextrap 
Percentage of AUC that is due to extrapolation from the last measurable 
concentration to infinity 
Cmax 
Maximum observed serum concentration 
Tmax 
Time of maximum observed serum concentration 
z 
Apparent terminal elimination constant 
t1/2 
Apparent terminal elimination half-life 
CL 
Total serum clearance 
Vz 
Volume of distribution during the terminal elimination phase 
Vss 
Volume of distribution at steady state (within the body) 
 
 
 


 
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Additional PK parameters may be determined where appropriate. 
PK analysis will be carried out using actual post-dose sampling times. Cmax and Tmax will be 
obtained directly from the serum concentration-time profiles following the first dose administration. 
11.5.1. 1. Criteria for handling concentrations below the limit of quantification in NCA analysis 
Concentration values that are below the limit of quantification (BLQ) will be set to zero, with 
defined exceptions as follows: 
 Any embedded BLQ value (between 2 quantifiable concentrations) and BLQ values following the 
last quantifiable concentration in a profile will be set to missing for the purposes of PK analysis 
 If there are late positive concentration values following 2 BLQ concentration values in the 
apparent terminal phase, these values will be evaluated to determine if these are plausible based 
on the PK characteristics of the drug. If these values are considered to be anomalous, they will be 
set to missing 
 If an entire concentration-time profile is BLQ, the profile will be excluded from the PK analysis 
 If a pre-dose concentration is missing, these values will be set to zero 
 
11.5.1. 2. Criteria for the Calculation of an Apparent Terminal Elimination Half-Life 
 At least three data points will be included in the regression analysis and should not include Cmax 
 When assessing terminal elimination phases, the R2 adjusted value will be used as a measure of 
the goodness-of-fit of the data points to the determined line 
 An elimination half-life will only be calculated if the R2 adjusted value of the regression line is 
greater than or equal to 0.7 
 Time period used for the estimation of apparent terminal elimination half-lives, where possible, 
will be over at least two half-lives 
 Where an elimination half-life is estimated over a time period of less than two half-lives, it will 
be flagged in the data listings at the discretion of the pharmacokineticist, and the robustness of 
the value should be discussed in the study report 
11.5.1. 3. Calculation of AUC 
The minimum requirement for the calculation of AUC will be the inclusion of at least three 
consecutive serum concentrations above the LLOQ, with at least one of these concentrations 
following Cmax. 
 AUC0- values where the percentage extrapolation is less than 20% will be reported.  
AUC0- values where the percentage extrapolation is between 20 to 30% will be flagged and 
included in the descriptive statistics, whilst AUC0- values where the percentage extrapolation is 
greater than 30% will be reported but excluded from descriptive statistics.  
11.5.1. 4. Anomalous Values 
If an outlier value is considered to be anomalous due to being inconsistent with the expected PK 
profile, it may be appropriate to exclude this point from the PK analysis. However, the exclusion of 
data must have strong justification and will be documented in the raw data and study report. 


 
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Embedded BLQ values may be considered anomalous depending on the route of administration and 
the characteristics of the drug. 
11.5.2. Presentation of Pharmacokinetic Data 
11.5.2. 1. Presentation of Pharmacokinetic Plasma Drug Concentration Data 
The following rules will be applied if there are values that are BLQ or if there are missing values 
(e.g. no result [NR]) in a plasma concentration data series to be summarized. 
 For the calculation of summary statistics, BLQ values will be set to zero 
 If an embedded BLQ value is considered anomalous within the concentration-time profile, this 
value will be excluded from the summary statistics 
 Where there is NR, these will be set to missing 
 If there are less than three values in the data series, only the min, max and N will be presented. 
The other summary statistics will be denoted as not calculated (NC). BLQ is considered as a 
value. 
 If all the values are BLQ, then the arithmetic mean, arithmetic SD, median, min and max will be 
presented as zero, and the geometric mean and geometric CV% will be denoted as NC 
 If the value of the arithmetic mean or median is below the LLOQ, these values will be presented 
as zero and the geometric mean and geometric CV% will be denoted as NC. 
11.5.2. 2. Presentation of NCA Parameters 
 For the calculation of summary statistics of PK parameters, all NR and NC values in a data series 
will be set to missing 
 The AUC values will be set to NC if they have been calculated using fewer than three 
concentrations, and/or three concentrations if the last is Cmax 
 Derived parameters will be summarized by the appropriate statistics (N, geometric mean, 90% 
confidence intervals, median, min and max, with the exception of Tmax to be summarized by 
median, min and max only) 
11.5.2. 3. Pharmacokinetic Report  
A PK report will be written detailing all methodology followed during the Population PK (PopPK) 
process and will contain the results of both PopPK analysis (Section 21.2) and non-compartmental 
PK analysis (Section 21.3). 
Primary and secondary PK parameters of roledumab in pregnant women will be given in tabular 
format. Results and interpretation of results will be discussed. 
11.6. Efficacy Assessment 
Efficacy analyses will be only descriptive, conducted on the FAS. The results will be displayed by 
route of administration and overall. 
The number and the percentage of subjects with a positive and a negative anti-RhD IgG result (IAT) 
will be displayed by timepoints (V2/V8/V13 and possibly AV1 at 9 months post-delivery/AV2 at 12 
months post-delivery and in case of sensitizing event). In addition, a by-subject listing wil display 
the sampling date and time as well as the corresponding study day.  


 
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In case of positive IAT, the results of the anti-RhD antibodies will be quantitatively analyzed. The 
number and percentage of subjects with passive anti-RhD will also be presented as well as the 
alloimmunization status of subjects. A by-subject listing will also be provided with the following 
information: visit, microtitration/ponderal dosage performed (yes/no), related day (from the first 
IMP administration), study day, sampling date and time, place where sampling was done, 
quantitative anit-RhD IgG result with the original unit, original normal ranges and investigator’s 
assessment.   
IAT results will be listed by visit. The anti-RhD quantification results will also be listed by visit.  
11.7. Safety Assessment 
Safety analyses will be conducted on the SAF. The results will be displayed by route of 
administration and overall. 
Safety on the fetus/newborn will be presented by subgroups of their RhD status (positive / negative) 
as it may happen that child is observed as being RhD-negative at delivery. If all newborns are RhD 
status positive, the safety analyses will be presented on the whole SAF. 
11.7.1. Extent of exposure 
Subjects are planned to receive two injections of roledumab, one between at the 27th and the 29th 
week of gestation (depending on the version of the protocol – See Section 15) and one within 72h 
after delivery. If additional roledumab injection is administered (i.e. in case of sensitizing event), a 
description of the sensitizing event and doses received of roledumab or Rhophylac® will be 
provided. 
The extent of exposure to roledumab will be summarized for:  
 Total number of injections 
 Actual cumulative roledumab dose (µg) 
 Actual roledumab dose per injection (µg) 
Total number of injections (including planned injections and additional injections in case of 
sensitizing event), actual cumulative dose and duration of roledumab exposure (in months) will be 
summarized using descriptive statistics. The duration of exposure will be calculated as shown in 
Section 16. 
Actual IMP dose (per injection), administration duration in seconds, and administration site will be 
summarized using descriptive statistics by visit (V2, V8 and additional visit if applicable). The 
actual IMP dose will be calculated as shown in Section 16. 
The number of subjects who received the antenatal dose, received the postnatal dose and who 
received extra doses of roledumab or Rhophylac® will be displayed and the doses (in mL) received 
will be quantitatively summarized.  
A by-subject listing will display all exposure data by subject ID and visit: Related day (from first 
IMP administration), product injected, batch number, start date and time of injection, administration 
duration (in seconds), volume injected (mL), actual IMP Dose (μg), administration route, 
administration site, description of the sensitizing event if applicable.    


 
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The Rhophylac doses received by the subject in case of sensitizing events will be listed by subject 
if any. 
11.7.2. Adverse events 
All adverse events (AEs) of the mother and/or the fetus/newborn/infants will be coded using the 
latest version of Medical Dictionary for Regulatory Activities (MedDRA) available at the beginning 
of the study at LFB Biotechnologies, i.e. Version 16.0.  
All TEAEs (treatment-emergent adverse events), i.e. adverse events occurring between the first IMP 
administration and the end-of-study (EOS) visit will be described overall and by route of 
administration. In case the EOS visit did not take place as per protocol for any reason and in case of 
early withdrawal for any reason, the date of the EOS form in the e-CRF will be used to identify the 
end of treatment period. 
A summary table will display the number of AEs as well as frequency counts and percentages of 
subjects with at least one AE. AEs will be divided into pre-treatment AEs, TEAEs, serious AEs, 
serious TEAEs and TEAEs leading to withdrawals. TEAEs will be split into severity categories 
(mild, moderate, severe), causal relationship with IMP (yes/no) and outcome (resolved without 
sequelae, resolved with sequelae, recovering, not recovered, fatal, unknown). A summary table will 
be provided for the mother, another one for both mother and fetus and another one for the newborn. 
A summary table of all drug-related TEAEs will also be presented for the mother, another one for 
both mother and fetus and another one for the newborn. The number of TEAEs as well as the 
number and percentages of subjects with at least one drug-related TEAE will be displayed. The 
events will be divided into the following categories: seriousness, severity, outcome, action taken 
with IMP (none, dose reduced, dose increased, discontinuation of IMP, temporary interruption of 
IMP, not applicable), other action taken (none, procedure, withdraw from clinical trial due to AE, 
other), and corrective medication (yes/no). 
Tables for the mother, for both mother and fetus, and for the newborn will list each TEAE, the 
number and percentage of subjects with at least one event by SOC and PT sorted by decreasing 
frequency of SOC and PT within SOC. The events will be classified in terms of severity and 
relationship to IMP (Related/Not related) as shown in Section 21.1.  
Duration of TEAE and Time to onset of TEAE will be calculated as detailed in Section 16 and 
summarized using quantitative statistics. A table will be provided for the mother, another one for 
both mother and fetus, and another one for the newborn. 
All AEs, including pre-treatment AEs (i.e. AEs occurring during the screening period and before the 
first injection of IMP), will be displayed in by-subject listings. For each AE, the following 
information will be given: who is concerned (mother/fetus/newborn), verbatim text, SOC, PT, start 
date and time, stop date and time or ongoing, duration in days, seriousness and seriousness criteria, 
severity, outcome, action taken, corrective medication, relationship with IMP. Among the AEs, the 
TEAEs will be clearly identified. 
Serious AEs will also be listed. For each serious AE, the following information will be given: who is 
concerned (mother/fetus/newborn), PT, start date and time, stop date and time or ongoing, duration 
in days, seriousness (yes/no) and seriousness criteria, severity, outcome, action taken, corrective 
medication, relationship with IMP. The TEAEs will be clearly identified. AEs leading to 
withdrawals and deaths will also be displayed in by-subject listings containing the same information 
as the listing for serious AEs. A listing will also be specifically dedicated to displaying AEs 


 
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occurring in breastfed newborn with the same kind of information for each AE. In addition, the type 
of diet will be indicated (breast-milk/breast-milk and bottle-feeding). 
11.7.3. Clinical laboratory evaluation 
All laboratory data will be described using standard units.  
 Hematology: Red blood cells (RBC), Hematocrit, Leucocytes, Neutrophils, Basophils, 
Eosinophils, Lymphocytes, Monocytes, Platelets, Hemoglobin, Mean corpuscular hemoglobin 
(MCH), Mean corpuscular hemoglobin concentration (MCHC), Mean corpuscular volume 
(MCV), Activated Partial Thromboplastin Time (aPTT) (baseline only), Prothrombin Intl. 
Normalized Ratio (baseline only) 
 Biochemistry: Creatinine, Blood Urea Nitrogen, Glucose, Total Bilirubin, ALT, AST, Alkaline 
Phosphatase, GT, Elecrophoresis (Total proteins, Albumin, Alpha 1, Alpha 2, Beta 1, Beta 2, 
Gamma, Creatinine Clearance, Protein Peak) 
 Urinalysis: Dipstick (Glucose, Nitrite, Occult Blood, Protein) and microscopy in case of positive 
dipstick (Erythrocytes, Leukocytes, Epithelial Cells, bacteria, casts, crystals)  
 Cytokines: IFN-, IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-13 and TNF- 
 C-Reactive Protein (CRP) 
 KB test 
 Laboratory parameters in cord blood: DAT, hematocrit, hemoglobin, reticulocytes and total 
bilirubin performed in cord blood in order to detect fetal anemia 
Normal ranges for SI units will be presented by subject in a listing for the parameters of hematology, 
biochemistry and urnialysis. 
Abnormal laboratory values will be displayed in by-subject tables for hematology and biochemistry, 
including the subject ID, the actual route of administration, the age, the visit, the related day (from 
the first IMP administration), the sampling date and time, the place where sampling was done, the 
results in original unit and in SI unit, the original normal ranges, the measured value for each 
parameter at each visit and the absolute change from baseline in SI unit to each post-baseline time 
point for continuous parameters. In addition, each abnormal value will be qualified as abnormal 
without clinical significance or abnormal with clinical significance. For the biochemistry, the 
parameters included in the table will be the following ones: creatinine, blood urea nitrogen, glucose, 
total bilirubin, ALT, AST, alkaline phosphatase, GT and electrophoresis (only an overall 
assessement of the electrophoresis).   
Measured values and changes from baseline (for continuous parameters) will be quantitatively 
described for each parameter (except for protein peak, normal/abnormal) for each visit (including 
additional visits) in summary tables for hematology, biochemistry and for CRP.  
The investigator’s clinical assessment (normal, abnormal without clinical significance, abnormal 
with clinical significance, not reliable, not done) by visit will be displayed in a separate table. For 
the biochemistry, the parameters included in the table will be the following ones: creatinine, blood 
urea nitrogen, glucose, total bilirubin, ALT, AST, alkaline phosphatase, GT and electrophoresis 
(only an overall assessement of the electrophoresis).  
Shift tables for hematology and biochemistry will display for each parameter the number and the 
percentage of subjects with low, normal or high level over time with respect to baseline. 


 
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All laboratory data for hematology, biochemistry and CRP will be presented in by-subject listings 
containing the following information: subject ID, actual route of administration, age, visit, related 
day (from the first IMP administration), sampling date and time, the place where sampling was 
done, results in original unit and in SI unit, the orignal normal ranges, the measured value for each 
parameter at each visit and the absolute change from baseline in SI unit to each post-baseline time 
point for continuous parameters and the clinical assessment.  
The parameters associated to cytokines will be displayed in a summary table by visit. For each 
parameter and each visit, the number and the percentage of subjects BLQ will be presented. In 
addition, boxplots for each parameter of cytokines by visit will be displayed by route of 
administration. In addition, a by-subject and by-time point listing will describe each parameter based 
on central laboratory with the following information: theoretical time, date and time of sampling, 
original result and unit, specimen type and method of analysis. 
The results of KB test (positive/negative) will be presented in a summary table: at delivery before 
the IMP administration for all subjects and at delivery within 24 hours after the IMP administration 
in case the result obtained prior to the IMP administration was positive; after a sensitizing event, 
before the IMP administration and within 24h after the IMP administration only if the result of the 
first test was positive. 
A by-subject listing will also be provided with the following variables: the route of administration, 
the age, the visit, the related day (from the first IMP administration), the sampling date and time, the 
KB test result as well as the FMH volume. 
Results of urinalysis tests using dipstick will be displayed by visit (V1 and V13) in a summary table. 
The investigator’s clinical assessment (normal, abnormal without clinical significance, abnormal 
with clinical significance, not reliable, not done) for each parameter and by visit will also be 
displayed in another table. These tables will also be presented for urinalysis by microscopy 
examination in case of postivie dipstick. A dipstick test was considered as positive when at least one 
of the four tested parameters was positive.  
A by-subject listing for urinalysis (dipstick) will also be provided by subject ID, route of 
administration and visit, with the following information: age of the subject, study day, related day 
(from the first IMP administration), sampling date, result for each parameter and clinical assessment. 
The same listing will be provided for microscopy test in case of positive dispstick.  
Laboratory parameters in cord blood 
The investigator’s clinical assessment for each parameter in cord blood (normal, abnormal without 
clinical significance, abnormal with clinical significance) will be displayed in a summary table by 
parameter.  
In addition, a listing by route of administration and by subject will display the presence or not of an 
abnormality (yes/no) and the investigator’s assessment (normal, abnormal without clinical 
significance, abnormal with clinical significance) for hemoglobin, hematocrit, reticulocytes, total 
bilirubin in cord blood as well as the associated measured value and lower and upper ranges. The 
results for DAT (anti-RhD IgG results, positive/negative/not done) will be displayed in a separate 
listing. 


 
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Another listing by-subject listing will detail for each parameter of cord blood the date and time of 
delivery, the date and time of sampling, where sampling was done, the name of parameters, the 
associated measured values in original and standard unit for hemoglobin, hematocrit, reticulocytes, 
total bilirubin well as the investigator’s assessment. The results for DAT will be presented in 
separate listing with the anti-RhD IgG results (positive/negative) and the investigator’s assessment. 
11.7.4. Vital Signs 
Vital signs parameters include: systolic and diastolic blood pressure, pulse rate, body temperature 
and weight. 
These parmaters in addition to height and BMI will be described by time point in by-subject listings. 
Measured values and changes from baseline for each parameter will be quantitatively described at 
each visit (including additional visits) in a summary table. In order to determine the changes from 
baseline, two baseline values will be considered: baseline 1 will refer to the last value prior to the 
first IMP administration (antenatal period) and baseline 2 will refer to the last value prior to the IMP 
administration after delivery (postnatal period). Within the antenatal (respectively postnatal) period, 
the change from baseline will be evaluated with respect to baseline 1 (respectively baseline 2).   
Vital signs will also be analyzed and presented by time point graphically in boxplots, except for 
weight. 
11.7.5. Delivery 
Information pertaining to delivery and any associated complication will be described in a by-subject 
listing containing the following variables: subject ID, actual route of IMP administration, date and 
time of delivery, gestational age at birth (in weeks), duration of labor (in hours), anasthesia used 
(yes/no), mode of delivery, instrumental delivery (yes/no), any complication during delivery 
(yes/no), comment on delivery if any.  
11.7.6. Characteristics and general status of the newborn 
Information related to the newborn at birth will be described un a by-subject listing containing the 
following variables: subject id, date and time of birth, gestational age at birth (in weeks), gender of 
the newborn, Apgar score measured once at 1 minute after birth, and again at 5 minutes after birth.  
General status of newborn will also be described in a by-subject listing with the following 
information: subject id, actual route of IMP administration, visit (V8, V9, V12), weight (kg) and 
height (cm) of the newborn, type of diet, first/breast milk sample (yes/no), any clinical significant 
abnormality in the newborn (yes/no), new concomitant treatment given to the newborn (yes/no), 
comment if any.  
In addition, the ABO status and the RhD status of the newborn will also be described in a by-subject 
listing. 
11.7.7. Other observations related to safety 
11.7.7. 1. Anti-roledumab antibodies (immunogenicity) 
Immunogenicity of roledumab (ADA) will be summarized using descriptive statistics by visit 
(screening, V2, V5, V6, V8, V11, V12, V13 and additional visits if applicable). A listing will 


 
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display results of ADA test at each visit for subjects with at least one positive ADA test during the 
course of the study. 
In addition, a by-subject listing will show the ADA test resuls for all subjects by visit.  
In case of retest, the last observed value will be used for the analysis. 
11.7.7. 2. Obstetric and fetal Doppler ultrasound 
Obstetric and fetal Doppler ultrasound results will be displayed in by-subject listings including the 
following variables: actual route of administration, visit (V2, V5, V6, V7 and additional visits if 
any), date of ultrasound examination, any abnormality detected (yes/no), normal fetal cardiac 
activity (yes/no), fetal hydrops detected (yes/no), fetal hydrops assessment, sign of polyhydro-
amnios (yes/no), diameter of the largest pocket (cm), MCA-PSV (cm/s), Multiple of Medians 
(MoM), calculated MoM, MoM >1.5 (yes/no), Clinical assessment (by the investigator on MoM).  
11.7.7. 3. Physical examination 
A complete physical examination of the mother will be performed at the screening visit and includes 
the following assessments: height, weight, general appearance, skin, head, eye, Ear-Nose-Throat 
(ENT), lymph nodes, heart, lungs, abdomen, extremities/joints, neurological and mental status. A 
partial physical examination is performed at V2, V8, end of study visit and additional visits and 
includes weight, general appearance, skin, heart, lungs and abdomen.  
A listing by patient and by visit will display when a physical examination was performed and if a 
clinically significant abnormalit was detected since the signature of ICF. Clinically significant 
physical examination abnormalities observed at screening will be reported as medical history and 
those at other visits will be reported as adverse events. 
11.7.7. 4. Roledumab concentration in milk and cord blood 
The concentrations of roledumab in first milk/breast milk and in maternal serum after postnatal 
administration as well as the ratio between these concentrations in milk and in blood will be 
presented using quantitative statistics at each visit (V10, V11) and a by-subject listing. This listing 
will display the following variables: actual route of administration, date and time of sampling, 
roledumab concententration in the milk (ng/mL), method of analysis, maternal concententration at 
V8 (ng/mL), value of the ratio between milk and maternal serum at V8. 
The concentrations of roledumab in the cord blood and in the maternal serum as well as the ratio 
between these two concentrations will be presented using quantitative statistics and in a by-subject 
listing. This listing will display the following variables: actual route of administration, date and time 
of sampling, roledumab concententration in the cord blood (ng/mL), method of analysis, maternal 
concententration at V8 (ng/mL), value of the ratio between cord blood and maternal serum at V8. 


 
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12. 
STATISTICAL/ANALYTICAL ISSUES  
12.1. Handling of Dropouts and Missing Data 
12.1.1. Dropouts 
No general/specific rules will be formed to handle missing data except for PK purposes. All missing 
data will be treated as missing. Every effort will be made to collect data on all study parameters 
even in case of dropout (up to EOS visit). 
12.1.2. Missing data 
In general, missing data will not be imputed. Exceptions will be made for data reported as "" or "" 
x; for quantitative statistics, these indeterminate values will be replaced by the x value itself. 
If either a sampling time or its concentration is missing, then that sample data point will be excluded 
from PK sets as described in the CPAP. 
12.1.3. Missing or incomplete dates 
In general, missing data for dates will not be imputed except for concomitant medications or AEs for 
which they will be imputed to the earliest possible start date or the latest possible end date. If the 
first possible earliest start date (e.g. if only the year is available) is before the first infusion date then 
to be on a conservative side the start date of the AE will be imputed as the first infusion date. 
12.1.4. Randomization 
Not applicable. 
12.1.5. Data safety monitoring board 
An external independent data safety monitoring board (DSMB) will monitor safety outcomes and 
study conduct and will provide the Sponsor with recommendations regarding continuing or stopping 
the study for all subjects or subgroups of subjects all along the study. The reporting made for the 
DSMB is not discussed in the current SAP. 
12.1.6. Blind Review  
Not applicable as it is an open-label study. 
12.1.7. Maintaining the blind 
Not applicable as it is an open-label study. 
12.1.8. Pooling of Centers 
For the analysis, centers will be pooled by route of administration. The center will not be taken into 
account in the statistical description. 


 
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12.1.9. Multiplicity issues  
No multiple testing will be performed in this study.  
12.1.10. 
Subgroup analysis 
No subgroup analysis is planned for the efficacy and safety. However all available data will be 
presented as follows: 
 IM arm 
 IV arm 
 Total (if appropriate) 
12.1.11. 
Other issues 
12.1.11. 1. 
Significance testing and estimation 
For the efficacy and safety, no formal statistical hypothesis testing will be performed. 
12.1.11. 2. 
Experimental units 
For this study the experimental unit is the mother and/or the newborn depending on examination. 
13. 
INTERIM ANALYSES 
No interim analysis will be performed. However and as the primary PK endpoint will be available 
before the last subject withdraws from study- the final analysis of the primary PK endpoint will be 
performed before the final DBL. 
Consequently, all PK-related data will be frozen and this freezing will be documented. 
14. 
SPECIFIC ANALYSIS RULES  
For descriptive statistics, the following number of decimal places (dps) will generally be applied: 
arithmetic mean, SD, Q1, median and Q3 to 1 more dps than the raw data; minimum and maximum 
to the same number of dps as the raw data. Percentages should be presented to 1 dp. 
The information and explanatory notes to be provided in the “footer” or bottom of each table will 
include the following information: date and time of output generation, SAS program name, any 
other output specific details that require further elaboration. All abbreviations used in the table will 
be defined. 
After DBL if there is no change in the SDTM datasets used for the PK analysis (NCA and PopPK), 
the PK analysis done on freeze data will be considered as final. 
 
 


 
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15. 
VISIT AND TIME WINDOWS 
Visit 
Time windows 
V1 
> or = signature date of ICF and in the range of 12 to 27 weeks of GA (applicable from Protocol V8.0) 
 > or = signature date of ICF and in the range of 12 to 26 weeks of GA (applicable from protocol V5.0 to 
protocol V7.0) 
V2 
> date of V1 and in the range of 28 to 29 weeks of GA  
V2b 
24h  2h after time of IMP injection IV arm only (from protocol V8.0) 
V3 
For IV: 48h  6h after time of IMP injection/ For IM: 48h  24h (applicable from Protocol V8.0) 
2 days ± 1 day after time of IMP injection (applicable from protocol V5.0 to protocol V7.0) 
V4 
For IV: 96h to 120h after time of IMP injection/ For IM: 120h  24h (applicable from Protocol V8.0) 
5 days  1 day after time of IMP injection (applicable from protocol V5.0 to protocol V7.0) 
V5 
9 days  3 days after time of IMP injection 
V6 
29 days  4 days after time of IMP injection  
V7 
59 days  4 days after time of IMP injection  
SE 
> date of V2 and within 72h after start of SE  
SE-FU 
24h after time of IMP injection (no time window) 
V8 
Within 72h after time of newborn birth 
V9 
48h  24h after date of newborn birth (applicable from Protocol V8.0) 
2 days  1 days after date of newborn (applicable from protocol V5.0 to protocol V7.0) 
V10 
72h  24h after time of 1st Postnatal IMP injection (applicable from Protocol V8.0) 
3 days  1 day after time of 1st Postnatal IMP injection (applicable from protocol V5.0 to protocol V7.0) 
V11 
9 days  2 days after day of 1st Postnatal IMP injection  
V12 
6 weeks  2 weeks after day of 1st Postnatal IMP injection 
V13 
6 months + 2 weeks after day of 1st Postnatal IMP injection 
AV1 
9 months  2 weeks after day of 1st Postnatal IMP injection 
AV2 
12 months  2 weeks after day of 1st Postnatal IMP injection 
 
 
 


 
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Timepoint 
Time window 
Measurements at screening V1 
> or = date of ICF 
H0 or Before start of injection 
Before time of IMP administration 
T 30 after IMP administration 
30 min  5 min after end of injection (= time of IMP administration + 1 min) 
T H1 
1 hour  5 min after end of injection 
T H6 
6 hours  15 min after end of injection 
After the end of IMP administration 
> time of IMP administration + 1 min 
IMP injection 
Time window 
V2 
In the range of 28 to 29 weeks of GA (applicable from Protocol v6.0) 
 
In the range of 27 to 29 weeks of GA (applicable before Protocol v6.0) 
SE: 1st injection 
Within 72h after start of SE and > date of V2 
SE-FU 
24h after time of IMP injection (no time window) 
V8 
Within 72h after time of newborn birth 
 
16. 
DEFINITION OF DERIVED VARIABLES 
 Body Mass Index (BMI, kg/m2), will be derived as follows: 
BMI = (Weight / Height2)  
with weight in kg and height in m 
 Actual IMP dose (µg) = volume injected (mL) x 300 (concentration of 300 µg/mL) 
 Duration of IMP exposure (in months) = (Date of last injection - Date of the first injection +1)/30.4375 
 Follow-up after first IMP injection (in months) = (Date end of study assessment - Date of the first day of 
the IMP administration +1)/30.4375 
 Duration in the study (in months) = (Date of end of study assessment - Date of the informed consent 
+1)/30.4375 
 Duration of an AE in days = (AE end datetime - AE start datetime) +1 if duration >24h and /24 if duration 
24h 
 Time to onset of an AE=(AE start datetime after the last administration of IMP - Datetime of the last 
administration of IMP) in days (xx days) if duration >24h and /24 if duration 24h 
 Calculated MoM 
MoMc= Observed MCA-PSV / Expected_MCA-PSV 
Where Expected_MCA-PSV = e2 30921 + 0 0463954*GA 
with GA=gestational age in weeks calculated as follows: 
GA=Integer ((datepart(Visit date)- datepart(Pregnancy date) +14 days)/7 days). 
 The creatinine clearance (CRCL) will be calculated with the Cockcroft-Gault formula [4] 
 For female: 
 
 
 
85
.0
(mg/dL)
 
creatinine
 
72
(kg)
t 
body weigh
age)
-
(140
(mL/min)
 
clearance
 
Creatinine






 
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 Ratio of roledumab in first milk/breast milk concentration to maternal serum concentration= 
Measurement of LFB-R593 in first milk/breast milk (ng/mL) at Vxx** / measurement of LFB-
R593 in maternal serum (concentration at Vxx visit from PK sample*). 
*: sample coming from central measurement (Envigo) 
**: sample coming from central measurement (SGS) 
Vxx: measurement will be performed twice (at V10 and V11) 
 
17. 
LABORATORY STANDARDIZATION (FOR LOCAL LABS) 
No standardization of local labs will be done. 
 
18. 
CHANGES FROM THE PROTOCOL SPECIFIED STATISTICAL ANALYSES 
Not applicable. 
18.1. Definition of analysis populations (Section 9) 
In order to adopt a more standard definition, an enrolled subject will be a subject who signed the 
ICF and who was not screening failure. 
The ITT population is then defined as follows: all subjects who were enrolled in the study (ICF 
signed and not screening failures) and/or who did not complete the EOS form before the inclusion 
visit (V2). 
18.2. Primary endpoints (Section 10.1) 
The population based models in previous Phases I and IIa, and current Phase IIb showed to be a 
two-compartment model instead of one-compartment model. Therefore, there should be two 
additional parameters Q and V3 due to the second compartment in two-compartment model, 
compared with only CL and V2 in one-compartment model. 
In addition, only the derived parameter t1/2 will be calculated. The parameters Cmax, Tmax, AUC0-∞ 
will be reported in secondary PK analysis (NCA) instead of population PK analysis. 
18.3. Other observations related to safety (Section 11.1) 
Except if stated in analyses sections above, standard error (Protocol §12.7.1) [1] will not be reported 
in descriptive tables. 
18.4. Other observations related to safety (Section 11.7.5) 
Physical examination findings: no specific listing will be provided for clinically significant 
abnormalities as they are reported either in medical history or adverse events and so will be analyzed 
in those sections. 


 
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19. 
DATA PRESENTATION 
Some of the TFLs may not have sufficient numbers of subjects or data for presentation. If this 
occurs, the blank TFL shell will be presented with a message printed in the center of the table, such 
as, “NO DATA TO DISPLAY”. 
 
 
 


 
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20. 
REFERENCES 
1 
Clinical Study Protocol - ADNC-1301 (phase 2b) EUDRACT number 2013-000269-35 
Pharmacokinetics and Safety of roledumab, a fully human recombinant monoclonal anti-
RhD antibody in RhD-negative pregnant women carrying an RhD-positive fetus: a phase 
2b, multicenter, open-label study. 
2 
International Conference on Harmonisation of Technical Requirements for Registration 
of Pharmaceuticals for Human Use, ICH Harmonised Tripartite Guideline, Statistical 
Principles for Clinical Trials (E9), 5 February 1998. 
3 
International Conference on Harmonisation of Technical Requirements for Registration 
of Pharmaceuticals for Human Use, ICH Harmonised Tripartite Guideline, Structure and 
Content of Clinical Study Reports (E3), 30 November 1995. 
4 
Cockcroft DW, Gault MH. Prediction of creatinine clearance from serum creatinine. 
Nephron 1976;16(1):31-41. 
 
 
 


 
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21. 
APPENDICES 
21.1. Appendix 1 - Tables, figures and listings 
21.1.1. Table of contents - Section 14 of CSR 
 
TABLES, FIGURES AND GRAPHS REFERRED TO BUT NOT INCLUDED IN THE TEXT 
 
14.1 
DEMOGRAPHIC AND BASELINE DATA 
14.1.1 
Table: Subject Disposition 
14.1.2.1   Table: Summary of protocol deviations 
14.1.2.2 
Table: Subject Populations 
14.1.3 
Listing: Inclusion / Exclusion criteria 
14.1.4 
Listing: Reasons of screening failures 
14.1.5.1 
Table: Demography – (1/3) 
14.1.5.2  
Table: Demography – (2/3) 
14.1.5.3 
Table: Demography – (3/3) 
14.1.5.4.1  Table: Serology for HBV 
14.1.5.4.2  Table: Serology for HCV and HIV 
14.1.5.4.3  Table: Results for ADA test  
14.1.5.4.4  Table: Results for IAT  
14.1.5.5.1  Table: Life habits: summary of tobacco and alcohol consumption 
14.1.5.5.2 Listing: Life habits: details of tobacco consumption and alcohol consumption in 
former and current smokers and/or in subjects with alcohol consumption  
14.1.6 
Table: Medical and surgical history by SOC and PT 
14.1.7 
Table: Summary of obstetrical history 
14.1.8.1 
Table: Summary of prior medications 
14.1.8.2 
Table: Summary of concomitant medications 
 
14.2 
EFFICACY DATA 
14.2.1  
Table: Summary of IAT results 
14.2.2 
 Table: Summary of anti-RhD quantification – FAS – Subset of subjects with a 
positive IAT 
 
 
14.3 
SAFETY DATA 
14.3.1 Displays of Adverse Events 
14.3.1.1 
Table: Summary of adverse events – Mothers 
14.3.1.2 
Table: Summary of adverse events –Mother & Fetus 
14.3.1.3 
Table: Summary of adverse events –Newborn 
14.3.1.4 
Table: Summary of drug-related TEAE – Mothers 


 
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14.3.1.5 
Table: Summary of drug-related TEAE –Mother & Fetus 
14.3.1.6 
Table: Summary of drug-related TEAE –Newborn 
14.3.1.7 
Table: TEAE by severity and relationship by SOC and PT – Mothers - IM 
14.3.1.8 
Table: TEAE by severity and relationship by SOC and PT –Mother & Fetus - IM 
14.3.1.9 
Table: TEAE by severity and relationship by SOC and PT –Newborn - IM 
14.3.1.10 Table: TEAE by severity and relationship by SOC and PT – Mothers - IV 
14.3.1.11 Table: TEAE by severity and relationship by SOC and PT – Mother & Fetus - IV 
14.3.1.12 Table: TEAE by severity and relationship by SOC and PT –Newborn - IV 
14.3.1.13 Table: TEAE by severity and relationship by SOC and PT – Mothers - ALL 
14.3.1.14 Table: TEAE by severity and relationship by SOC and PT – Mother & Fetus - 
ALL 
14.3.1.15 Table: TEAE by severity and relationship by SOC and PT –Newborn – ALL 
14.3.1.16 Table: Summary of duration and time to onset (days) of TEAE – Mothers 
14.3.1.17 Table: Summary of duration and time to onset (days) of TEAE –Mother&Fetus 
14.3.1.18 Table: Summary of duration and time to onset (days) of TEAE –Newborn 
 
14.3.2 Listings of Deaths, Other Serious and Significant Adverse Events 
14.3.2.1 
Listing of Serious TEAE - Mothers - IM 
14.3.2.2 
Listing of Serious TEAE – Mother  & Fetus - IM 
14.3.2.3 
Listing of Serious TEAE - Newborn - IM 
14.3.2.4 
Listing of Serious TEAE - Mothers - IV 
14.3.2.5 
Listing of Serious TEAE – Mother & Fetus - IV 
14.3.2.6 
Listing of Serious TEAE - Newborn - IV 
14.3.2.7 
Listing of Serious TEAE – Mothers - ALL 
14.3.2.8 
Listing of Serious TEAE – Mother & Fetus- ALL 
14.3.2.9 
Listing of Serious TEAE - Newborn - ALL 
 
14.3.3 Narratives of Deaths, Other Serious and Certain Other Significant Adverse Events 
Not applicable. 
 
14.3.4 Abnormal laboratory value listing (each subject) 
14.3.4.1 
Table: Hematology - Subjects with at least one abnormality 
14.3.4.2 
Table: Biochemistry - Subjects with at least one abnormality 
 
14.3.5 Laboratory Measurements 
14.3.5.1 
Table: Hematology - Summary statistics on measured values & change from 
baseline by visit 
14.3.5.2 
Table: Hematology – Investigator assessment of abnormalities by visit  


 
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14.3.5.3 
Table: Hematology – Shift table from baseline by visit 
14.3.5.4 
Table: Biochemistry - Summary statistics on measured values & change from 
baseline by visit 
14.3.5.5 
Table: Biochemistry – Investigator assessment of abnormalities by visit  
14.3.5.6 
Table: Biochemistry – Shift table from baseline by visit  
14.3.5.7 
Table: Summary of cytokines by visit 
14.3.5.8.1 Boxplot of cytokines by visit – IFN-γ - Mother – IM/IV  
14.3.5.8.2 Boxplot of cytokines by visit – IL-10 - Mother – IM/IV  
14.3.5.8.3 Boxplot of cytokines by visit – IL-12 - Mother – IM/IV  
14.3.5.8.4 Boxplot of cytokines by visit – IL-13 - Mother – IM/IV  
14.3.5.8.5 Boxplot of cytokines by visit – IL-1β - Mother – IM/IV  
14.3.5.8.6 Boxplot of cytokines by visit – IL-2- Mother – IM/IV  
14.3.5.8.7 Boxplot of cytokines by visit – IL-4 - Mother - IM/IV  
14.3.5.8.8 Boxplot of cytokines by visit – IL-6 - Mother - IM/IV  
14.3.5.8.9 Boxplot of cytokines by visit – IL-8 - Mother - IM/IV  
14.3.5.8.10 Boxplot of cytokines by visit – TNF-α - Mother – IM/IV  
14.3.5.9 
Table: Cord blood test: Investigator assessment 
14.3.5.10.1 Listing: Subject with at least one abnormality of cord blood test results -
Hemoglobin, hematocrit, reticulocytes, total bilirubin 
14.3.5.10.2 Listing: Subject with at least one abnormality of cord blood test results - Direct 
antiglobulin test (DAT) 
14.3.5.11.1 Table: summary of KB test results at delivery 
14.3.5.11.2 Table: summary of KB test after a sensitizing event 
14.3.5.12 Table: CRP - Summary statistics on measured values & change from baseline by 
visit 
14.3.5.13 Table: Urinalysis (dipstick) – Summary statistics by visit 
14.3.5.14 Table: Urinalysis (dipstick) – Investigator assessment of abnormalities by visit 
14.3.5.15 Table: Urinalysis (microscopy) – Summary statistics by visit – Subset of subjects 
with a positive dipstick 
14.3.5.16 Table: Urinalysis (microscopy) – Investigator assessment of abnormalities by visit 
– Subset of subjects with a positive dipstick 
 
14.3.6 Other Safety Data 
14.3.6.1 
Table: Vital signs - Summary statistics on measured values & change from 
baseline by visit 
14.3.6.2.1 Boxplot of systolic blood pressure by visit 


 
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14.3.6.2.2 Boxplot of diastolic blood pressure by visit 
14.3.6.2.3 Boxplot of pulse rate by visit 
14.3.6.2.4 Boxplot of body temperature by visit 
14.3.6.3 
Listing: Abnormlities detected by obstretric and fetal Doppler ultrasound (1/2) 
14.3.6.4 
Listing: Abnormlities detected by obstretric and fetal Doppler ultrasound (2/2) 
14.3.6.5 
Listing: Description of sensitizing events 
14.3.6.6 
Table: Summary of ADA test results by visit 
14.3.6.7 
Listing: Subjects with at least one positive ADA test result by visit 
14.3.6.8 
Table: Summary of roledumab concentration in cord blood 
14.3.6.9 
Table: Summary of concentration in first/breast milk and maternal serum 
concentration 
14.3.6.10.1 Table: Exposure: summary of total number of injections and time between first 
and last administrations 
14.3.6.10.2 Table: Exposure: summary of doses received 
14.3.6.10.3 Table: Exposure: extent of exposure – actual administered doses by visit 
14.3.6.11 Table: Compliance to the IMP administration schedule 
 
21.1.2. Table of contents - Section 16 of CSR 
 
16.2. SUBJECT DATA LISTINGS 
16.2.1 Discontinued Subjects 
Listing 16.2.1.1: 
Subject Disposition – All Subjects 
Listing 16.2.1.2: 
Subject Disposition – Study Withdrawals 
Listing 16.2.1.3: 
Inclusion and Exclusion Criteria  
Listing 16.2.1.4: 
Study visits 
16.2.2 Protocol deviations 
Listing 16.2.2.1: 
Eligible subjects with Protocol Deviations 
16.2.3 Subjects excluded from the different populations  
Listing 16.2.3.1: 
Subjects Excluded from the ITT 
Listing 16.2.3.2: 
Subjects Excluded from the FAS 
Listing 16.2.3.3: 
Subjects Excluded from the SAF 
Listing 16.2.3.4: 
Subjects Excluded from the PKS1 
Listing 16.2.3.5: 
Subjects Excluded from the PKS2 
Listing 16.2.3.6: 
Subjects Excluded from the PKS3 
 
16.2.4 Demographic data 


 
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16.2.4.1 Demographics 
Listing 16.2.4.1.1: Demographic data at screening 
Listing 16.2.4.1.2: Life habits 
Listing 16.2.4.1.3: Current pregnancy information 
Listing 16.2.4.1.4: ABO /RhD status for the mother and RhD status for the fetus 
16.2.4.2 Obstetrical/pregnancy history and current preganancy 
Listing 16.2.4.2.1: Obstetrical history 
Listing 16.2.4.2.2: Miscarriage – Abortion history (1/2) 
Listing 16.2.4.2.3: Miscarriage – Abortion history (2/2) 
Listing 16.2.4.2.4: Pregnancy history (1/2) 
Listing 16.2.4.2.5: Pregnancy history (2/2) 
Listing 16.2.4.2.6: Pregnancy history - Complications 
16.2.4.3 Medical and Surgical History 
Listing 16.2.4.3: 
Significant Medical and Surgical History 
16.2.4.4 Prior and concomitant medications and non-drug therapies 
Listing 16.2.4.4.1: Prior and concomitant medications (1/2) 
Listing 16.2.4.4.2: Prior and concomitant medications (2/2) 
 
16.2.5 Compliance and/or drug concentration data (if available) 
Listing 16.2.5.1.1: Treatment compliance 
Listing 16.2.5.1.2: IMP injection 
 
16.2.6 Individual efficacy response data 
Listing 16.2.6.1: 
IAT results 
Listing 16.2.6.2: 
Anti-RhD quantification results – Subset of subjects with a positive 
IAT 
 
16.2.7 Adverse event listings (each subject) 
Listing 16.2.7.1.1: Adverse Events (1/2) 
Listing 16.2.7.1.2:  Adverse Events (2/2) 
Listing 16.2.7.2: 
Serious Adverse Events  
Listing 16.2.7.3: 
Adverse Events leading to Withdrawal 
Listing 16.2.7.4: 
Deaths 
Listing 16.2.7.5: 
AEs occurring in breastfed newborns 
 
16.2.8 Listing of individual laboratory measurements by subject 
Listing 16.2.8.1: 
Normal Ranges for SI Units 
Listing 16.2.8.2: 
Hematology (SI Units)  
Listing 16.2.8.3: 
Biochemistry (SI Units) 


 
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Listing 16.2.8.4.1: Urinalysis tests (Dipstick) 
Listing 16.2.8.4.2: Urinalysis tests (Microscopy) - Subset of subjects with positive 
dipstick 
Listing 16.2.8.5: 
Viral serology results at screening 
Listing 16.2.8.6: 
ADA test results by visit 
Listing 16.2.8.7: 
C-Reactive Protein 
 
16.2.9 Listing of other safety data  
Listing 16.2.9.1.1: Vital signs: height, weight and BMI 
Listing 16.2.9.1.2: Vital signs: blood pressure, pulse rate and body temperature 
Listing 16.2.9.2: 
Physical Examination at screening 
Listing 16.2.9.3.1: Delivery 
Listing 16.2.9.3.2: Cord blood tests - hemoglobin, hematocrit, reticulocytes, total 
bilirubin 
Listing 16.2.9.3.3: Cord blood tests - Direct Antiglobulin Test (DAT) 
Listing 16.2.9.3.4: Cord blood tests – rodelumab measurement in cord blood – Central 
assessment 
Listing 16.2.9.3.5: First/breast milk sample 
Listing 16.2.9.3.6: Cytokines – Central laboratory 
Listing 16.2.9.3.7: Kleihauer-Betke test 
 
16.2.10 Characteristics and general status of the newborn 
Listing 16.2.10.1: Newborn demographics 
Listing 16.2.10.2: Newborn general status 
Listing 16.2.10.3: ABO status and RhD status of the newborn 
 
 
 
 


 
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21.1.3. Table shells 
 
Tables will be presented using the following shell as an example: 
 
 
Table x.x <title of the table> 
<Population> 
 
Characteristic 
Statistics 
Roledumab 
300 µg 
IM 
Roledumab 
300 µg 
IV 
Roledumab 
300 µg 
Overall 
Number of subjects 
 
 
 
 
<Numeric variable, unit> 
N 
xx 
xx 
xx 
 
Mean (SD) 
xx.x (x.x) 
xx.x (x.x) 
xx.x (x.x) 
 
Median 
xx.x 
xx.x 
xx.x 
 
Min - Max 
xx - xx 
xx - xx 
xx - xx 
 
Missing 
x 
x 
x 
 
Q1 - Q3 
xx - xx 
xx - xx 
xx - xx 
<Categorical variable> 
 
 
 
 
Category 1 … 
n (%) 
x (xx.x) 
x (xx.x) 
x (xx.x) 
Category 2 
n (%) 
x (xx.x) 
x (xx.x) 
x (xx.x) 
Category 3 
n (%) 
x (xx.x) 
x (xx.x) 
x (xx.x) 
… 
n (%) 
x (xx.x) 
x (xx.x) 
x (xx.x) 
 
 


 
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Table x.x Summary of Adverse Events – Mothers 
<Population> 
 
Characteristic 
Roledumab 300 µg IM 
Roledumab 300 µg IV 
Roledumab 300 µg Overall 
 
E 
N (%) 
E 
N (%) 
E 
N (%) 
Number of subjects 
 
xx 
 
xx 
 
xx 
Any AE 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
Any Pre-Treatment AE 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
Any Serious AEs 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
Any TEAEs 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
Any Serious TEAEs 
xx  
xx (xx.x%) 
xx  
xx (xx.x%) 
xx  
xx (xx.x%) 
Any TEAE 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
Severity 
 
 
 
 
 
 
Mild 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
Moderate 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
Severe 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
Causal relationship with IMP 
 
 
 
 
 
 
Yes 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
No 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
xx 
xx (xx.x%) 
N: Number of subjects, E: Number of events 
 
Study: ADNC-1301, Program: xxx.sas/ddmmmyyyy:hh:mm:ss  
 
Same Table for Both Mother & Fetus and Fetus/Newborn  
 
 


 
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Table x.x Number of subjects with TEAE by intensity and relationship – Mothers 
<Population> 
 
 
Roledumab 300 µg IM (N*=) 
 
SOC/PT 
Mild 
Moderate 
Severe 
Missing 
Total 
 
R 
NR 
Missing 
R 
NR 
Missing 
R 
NR 
Missing 
R 
NR 
Missing 
R 
NR 
Missing 
 
E 
N(%) 
E 
N(%) 
E N(%) 
E 
N(%) 
E 
N(%) 
E N(%) 
E 
N(%) 
E 
N(%) 
E N(%) E N(%) E N(%) E N(%) 
E 
N(%) 
E 
N(%) 
E N(%) 
Any TEAE 
xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx xx(%) xx xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) 
SOC 1 
xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx xx(%) xx xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) 
Preferred 
term 1 
xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx xx(%) xx xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) 
Preferred 
term 2 
xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx xx(%) xx xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
SOC 2 
xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx xx(%) xx xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) 
Preferred 
term 1 
xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx xx(%) xx xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) 
Preferred 
term 2 
xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) xx xx(%) xx xx(%) xx xx(%) xx 
xx(%) 
xx 
xx(%) xx xx(%) 
Each AE is counted only once for each category 
*: used as denominator for the calculation of percentages 
NR: Not Related, R: Related 
SOC: System Organ Class, PT: Preferred Term, MedDRA (V16.0) 
N: Number of subjects, E: Number of events 
 
Study: ADNC-1301, Program: xxx.sas/ddmmmyyyy:hh mm:ss 
Same Table for Both IV and Overall (IM + IV) 


 
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Listings shells 
All individual data collected and derived will be listed in section 16.2 (to be in line with the ICH E3 
CSR guideline). 
Listings will be presented using the following shell as an example: 
 
Listing x.x.x <title> 
<Population> 
Page x/y 
 
Subject 
Identifier 
Group 
Class1 
Var 1 
Var 2 
Var 3 
Var 4 
Var 5 
Var 6 
… 
xxxx 
IM/IV 
Yes/No 
xxxxx 
xxxx 
xxxxx 
xxxx 
xxxx 
xxxx 
… 
 
 
 
 


 
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21.2. Appendix 2 - Population analysis plan 
BACKGROUND OF POPULATION PK ANALYSIS 
The PK data for roledumab are available from previous two clinical trials. The PopPK analyses were 
conducted to evaluate PK and absolute bioavailability of roledumab in healthy RhD-negative 
subjects after IM and IV administration in Phase 1 study (ADNC-0701), and to identify the IV and 
IM effective dose of roledumab required to effectively clear 15 mL of RhD-positive RBCs pre-
injected to healthy RhD-negative subjects in Phase 2 study (ADNC-0726). A two-compartment 
disposition model was used as with bolus input for the IV dose and first-order absorption for the IM 
dose with first-order elimination. 
In the Phase 1 clinical trial ADNC-0701 on healthy RhD-negative subjects, the PK of roledumab 
was nonlinear at doses ranging from 30 to 3000 µg. Exposures (Cmax and AUCs) increased more 
than proportionally with the dose. Clearance (CL) and volume decreased with the dose. At 300 µg, 
the typical values for central volume (Vc) and peripheral volume were and 3.66 L and 3.26 L with 
between-subjects variability of 35% and 13%, respectively; the typical value for CL at was 
0.0113 L/h with between-subjects variability of 28%; the absorption rate constant (Ka) after IM was 
0.00894 h-1 with large between-subjects variability of 78%; the elimination half-life after IV and IM 
was similar and estimated to be 17.9 days. 
In the Phase 2 dose-finding study ADNC-0726, the dataset was enriched using 17 healthy RhD-
negative subjects treated with IV or IM roledumab at doses from 100 to 300 µg from the Phase 1 
study ADNC-0701 to reduce standard errors for all fixed and random effects parameters due to the 
limited time span for blood samples collection in study ADNC-0726. The PK of roledumab was 
nonlinear at doses ranging from 100 to 300 µg. The result showed that clearance and volumes of 
distribution or roledumab increased with subjects’ body weight. The typical weight is 79 kg. The 
typical values for central volume (Vc) and peripheral volume at 300 µg were and 2.94 L and 3.10 L 
respectively, with between subjects variability of 24.9%. The typical value for clearance (CL) at 
300 µg was 0.0113 L/h with between subjects variability 54.6%. The absorption rate constant (Ka) 
after 300 µg IM was 0.00890 h-1 with between subjects variability 80.6%. The elimination half-lives 
after IV and IM were 15.4 and 20.1 days. 
These results confirmed the comparability of the PK parameters observed in the Phases 1 and 2. 
Although a difference in bioavailability between IM and IV exists (76% vs. a presumed 100%, 
respectively at dose of 300 µg), the exposure after 18 days was similar between the two routes of 
administration, indicating that the same dose of 300 µg IM and IV in this Phase 2b clinical study 
will lead to the same longer term protection against alloimmunization. 
This Population Analysis Plan (PAP) contains a detailed description of the methodology and 
procedures that will be used in the planned Population Pharmacokinetic (PopPK) analysis using data 
from ADNC-1301 a phase 2b, multicenter, open-label study to evaluate PK and safety of roledumab, 
a fully human recombinant monoclonal anti-RhD antibody, in Rh-D negative pregnant women 
carrying an RhD-positive fetus. 
The specific activity includes development and evaluation of a population PK model (PopPK). The 
objective of this PopPK modeling and simulation analysis is to obtain a quantitative understanding 
of the PK profile of roledumab 300 µg IV or IM in RhD-negative pregnant women carrying an RhD-


 
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positive fetus in order to confirm the suitability of this dose to ensure adequate anti-D level coverage 
up to delivery. 
This plan may be updated to reflect changes in the planned analysis which may arise due to 
unexpected issues during the study execution provided that the changes occur prior to database lock. 
Since PopPK analysis is fundamentally exploratory in nature, revisions may be data driven and 
based on results obtained during the course of the PopPK analysis; all revisions will be documented 
in the PopPK report. 
 
OBJECTIVES OF POPULATION PK ANALYSIS  
The objectives of PopPK analysis are: 
 To develop and evaluate the PopPK model for roledumab in RhD-negative pregnant women 
carrying an RhD-positive fetus 
 To confirm the suitability of dose for adequate anti-D level coverage up to delivery 
 To quantify the variability in the PK of roledumab 
 To identify significant covariate(s) in the PK of roledumab 
 
SOURCE DATA AND STUDY DESIGN FOR POPULATION PK ANALYSIS 
Plasma concentration data of roledumab from Phase 2b study ADNC-1301 will be used in this 
PopPK analysis. 
Study No. 1 (Phase 2b) 
This is a phase 2b, interventional, multicenter, open-label study of roledumab with IV and IM arms 
in RhD-negative pregnant women carrying an RhD-positive fetus. 
It was planned that the first 35 subjects would be administered a first dose of roledumab 300 µg IM 
during the second visit (V2) at 28 or 29 weeks (gestational age) and a second dose of 300 µg IM 
within 72h of birth during V8. Additional administration according to Kleihauer-Betke chart should 
be applied with 72 hours of procedure in case of sensitizing event in the antenantal period. The 
following 25 subjects were allocated to the IV arm with same dosage regimen. More than one site 
took part in the study in order to include 35 subjects for 30 evaluable subjects in the IM arm and 25 
for 20 evaluable subjects in the IV arm. The subjects should attend 13 visits for IM arm and 14 visits 
for IV arm, and further 1 or 2 follow-up visits if needed to document alloimmunization in both arms. 
A total of 10 blood samples in IM arm and 11 blood samples in IV arm were collected for PopPK 
analysis at the following time points: 
 IM arm: Antenatal period: pre-dose (V2), 21 days (V3), 51 days (V4), 93 days (V5), 294 
days (V6), and 594 days (V7) 
 
Postnatal period: pre-dose (V8, within 72 hours after delivery), 31 days (V10), 92 
days (V11), and 62 weeks (V12) 


 
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 IV arm: Antenatal period: pre-dose (V2), 1 hour5 min (V2), 24 hours2 min (V2b), 486 
hours (V3), 96-120 hours (V4), 294 days (V6), and 594 days (V7) 
 
Postnatal period: pre-dose (V8, within 72 hours after delivery), 3±1 days (V10), 9±2 
days (V11), and 6±2 weeks (V12) 
Roledumab first milk, breast milk, maternal serum concentration, and cord blood concentration for 
assessment of safety of the mother and the fetus/newborn were collected. 
 
ANALYSIS POPULATION FOR POPPK ANALYSIS 
PKS1: Analysis population 1 for popPK modeling 
The PKS1 consists of all ITT subjects enrolled in study ADNC-1301 (IM or IV arm), treated at least 
once with roledumab, and providing at least one roledumab available concentration after 
administration (including antenatal and postnatal measurements after 1st or 2nd administration). 
So all enrolled subjects who have received at least one dose of roledumab and have the following 
information entered will be included in PKS1: 
 At least one measurable concentration with adequate information on the dose amount 
administered 
 The date/time of drug administration, and 
 The date/time of sample collection 
 
DATA TRANSFER AND HANDLING 
Dataset Preparation 
Dataset preparation for PopPK analysis will be performed by Covance following the database lock. 
Concentrations, sampling times, dosing times, demographic and other covariate information 
supplied as SAS data transport files will be used to prepare PopPK analysis dataset. A SAS program 
(SAS version 9.3 or higher) will be used to assemble the analysis dataset from source data to be 
compatible with NONMEM translator (NM-TRAN). The analysis dataset will also be formatted for 
PDx-Pop (Version 5.2) and then saved as a comma delimited ASCII text file (*.csv file). 
Diagnostics will be performed to check for any gross errors during the compilation of the datasets. 
Missing PK Data 
If the date and/or time records of dosing are found to be missing at a specified scheduled visit, the 
drug level concentrations related to that time point will be excluded from analysis and an 
appropriate explanatory comment will be provided. 
Drug level concentrations related to missing sample collection date and/or time records will be 
commented upon and excluded from the analysis. Missing drug concentrations will be excluded 
from the analysis. 
Missing Covariate Data 


 
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For missing covariate values, the following methods may be used: 
 Use baseline (last non-missing value prior to study drug administration) covariate value for all 
dosing and observation records for non-time varying covariates. 
 Impute a value for a non-time varying missing covariate using 
 Median value of continuous covariate (i.e. body weight) 
 Mode of categorical covariate (i.e. route of delivery) 
 Stratification by study/treatment group for imputations will be considered 
 Interpolation between time points may be considered for time varying covariates 
BLQ Handling 
No imputation will be performed for the concentrations that are missing or below the assay’s 
quantification limit (BLQ). BLQ data at pre-dose will be set to zero. BLQ values at early time 
points, when appreciable absorption (IM) does not have occurred, will be set to zero. The BLQ data 
at the end of the PK profile will be set to missing. The one BLQ value occurring between two 
adjacent quantifiable values will be excluded from the analysis (treated as missing). When two 
consecutive BLQ values are encountered, all subsequent quantifiable values will be excluded from 
the analysis. 
Outliers 
Prior to any modeling, individual concentration-time profiles will be examined for any potential PK 
outliers (data points discordant from the bulk of the data within the same time interval) by visual 
inspection. Anomalous or extreme PK values (i.e. positive concentrations prior to the first dose, or 
concentrations that is outside of mean 3 SD at each timepoint for rest of subjects via the same 
route) that have a disproportionate effect on parameter estimates will be explained with a comment 
and excluded from the analysis. All data exclusions will be documented in the analysis report. 
Deleted Data 
Drug concentration measurements from the samples that are documented to be processed, stored or 
shipped under inadequate conditions will not be included in the PopPK analysis. 
Data Items Included in Datasets 
The NM-TRAN dataset will include data items such as study number, subject identification, 
actual treatment, actual dose amount, actual dosing day/time, actual sampling day/time, 
plasma concentrations of study drug and baseline demographic, and creatinine clearance. The 
covariates listed in the following table (Table 4) may be tested in the PopPK analysis, depending on 
availability of data. 
Table 4: Potential Covariates to be evaluated in the PopPK Analysis 
Abbreviation 
Covariate 
Code/unit 
AGE 
Age 
years 
GA 
Gestational age 
weeks 
SOJ 
Site of injection 
IV=0, IM=1 


 
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WT 
Body weight 
kg 
CRCL 
Creatinine clearance 
mL/min 
if CRCL >150, CRCL=150 
DLV 
Route of delivery 
Natural=0, C-section=1 
 
Partially missing continuous covariates will be entered using the last observation carried forward 
(LOCF) approach and completely missing continuous covariates will be replaced with the 
population median values. In the case of missing weight, specific median values will be used. 
Categorical covariates (e.g. route of delivery) missing for a subject will be coded as –6. 
Software 
The NONMEM software program (version 7.3 or higher, ICON Development Solutions, MD, USA) 
will be used for model fitting. NONMEM VI may be interfaced with PDxPop Version 3.0 Tools to 
allow for Expediting Population Analysis (ICON Development Solutions, MD, USA). The package 
is installed on a PC platform using Inter FORTRAN Standard Edition –Version 9 under 
Microsoft Windows 7. 
Dataset preparation, exploration and visualization will be performed using SAS and S-PLUS version 
8.2 Professional release 3 for Windows (Insightful, Seattle, WA, USA). The S-PLUS version 7.0 
package, SAS, or R version 3.0.2 or higher may also be used for the statistical analysis and graphical 
display of all NONMEM output. 
 
PHARMACOKINETIC ANALYSES 
The popPK analysis will be performed using the non-linear mixed effects modeling approach. This 
approach estimates the typical (mean) value of parameters as well as associated inter-individual 
variances. 
Estimation methods in NONMEM will be first-order conditional (FOCE) and first-order conditional 
with interaction (FOCEI). If supported by the data, in case of prohibitive computer intensive runs 
with FOCE, the first order (FO) estimation method may be used during model development. In such 
case, critical modeling steps will be reassessed using FOCE (with or without interaction). 
Alternative estimation methods implemented in NONMEM VII may be used, if needed. 
Stability of NONMEM models will be assessed on the basis of: 
 Acceptable goodness-of-fit via basic plots 
 Number of significant digits >=3 for all Thetas 
 Estimates of Theta’s not close to boundary 
 Condition number < 1000 
 Correlation less than 0.95 between any parameters 
 Stable parameter estimates with altering initial values to ensure global minimum being achieved 
Model selection will be based on: 


 
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 The comparison of full vs. reduced models based on the Log-Likelihood Criterion: the difference 
in the minimum value of the objective function (MOF) 
 Decrease in unexplained variability 
 Change towards conditional weighted residuals over time randomly distributed around zero 
 Scientific plausibility of the model 
Exploratory Data Analysis 
Prior to the PopPK analysis, the following pre-modeling analyses will be produced as part of the 
exploratory data analyses: 
 Graphical presentation of the concentration-time data to examine the basic model structure and 
facilitate outlier identification. Pooled plots of study drug concentrations vs. elapsed time (all 
subjects in one panel) for each treatment will be presented in the linear and semi-logarithmic 
scale. 
 Examination of covariate correlations which may help in building a covariate sub model and 
avoid confounds. For quantitative covariates, histograms of density versus residuals, box-whisker 
plots and plots of quartiles of covariate distribution versus quartiles of standard normal 
distribution (Quantile-Quantile plot) may be used in exploratory analysis. Moreover, scatter plots 
will be evaluated for potential correlations between quantitative covariate and box-whisker plots 
will be evaluated for every quantitative covariate against every categorical covariate. 
 Descriptive statistics or noncompartmental analyses 
Depending on the nature of the study data, further exploratory analyses may be conducted. These 
will be fully described in the report. 
 
Population Pharmacokinetic Model Development 
 Model Selection Criteria 
The following criteria will be evaluated to aid model selection: 
 A significant reduction in the minimum values of the objective function (MOF) from the base 
model using the extended least squares method, equivalent to -2 times the maximum log of the 
likelihood of the data (-2LL). The log likelihood ratio (LLR) test will be used to compare 
between base and covariate model. This provides an overall summary of how closely the 
model predictions match the data. The Akaike information Criterion (AIC) may also be used 
to evaluate and rank models based on goodness-of-fit 
 Goodness-of-fit measures including:  
A uniform distribution (i.e. unbiased in the predictions) across the identity line in the diagnostic 
plots of observed vs. predicted concentration values: including both IPRED (individual predicted) 
vs. DV (observed), and PRED vs. DV (observed) 
A more random (less systematic) distribution in the diagnostic plots of both residual (RES) and 
conditional weighted residuals (CWRES) against predicted concentration (PRED) and time after 
dosing to assess unaccounted heterogeneity in the data 
A decrease in the inter-individual variability (IIV) of PK parameters 


 
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A decrease in the residual error 
A decrease in the standard error estimate for model parameters 
 Base Model Development 
 
Structural Pharmacokinetic Model 
Standard structural PK models will be explored, starting with first order absorption and elimination 
rate will be explored and compared to each other. Goodness-of-fit of the structural model will be 
assessed by diagnostic plots: 
 Observed vs. population and individual predictions plots 
 Population and individual conditional weighted residuals (CWRES) vs. time 
 Plots of observations vs. time with population and individual fits 
The model with best fit according to model selection criteria will be chosen as the final structural 
model. 
 
Inter-individual Variability of Model Parameters 
Initially, exponential error models will be utilized for between subject variability in all PK 
parameters and implemented as a diagonal variance matrix for random effects (p). PK parameters 
will be modeled assuming the log-normal distribution as follows: 
Pi = TVP * exp(Pi) 
Where Pi is a parameter of interest for individual i, TVP is the typical value (population 
estimate) of the parameter and Pi is normally distributed with mean zero and variance P2. 
 
Residual Error 
The residual error will start with a combined additive and proportional (constant coefficient of 
variation) error model. 
Cij = 
ij
Cˆ
 (1 + 1ij) + 2ij 
where Cij is the measured concentration j in individual i, 
ij
Cˆ  is the model predicted value 
corresponding to measurement j in individual i, 
1ij
ε  is the proportional component of residual 
error for measurement j in individual i (normally distributed with mean zero and variance 
2
1
σ ), 
2ij
ε
, is the additive component of residual error for measurement j in individual i (normally 
distributed with mean zero and variance 
2
2
σ ). Other residual error models: additive, log-
additive, and proportional error models will also be explored. 
Evaluation of Covariate Effects on model parameter estimates 
The process of covariate screening will be started by visual inspection of posterior Bayes estimates 
of PK parameters’ P plotted versus each of the covariates. For continuous covariates, scatter 
covariate-P plots will be overlaid with a non-parametric locally weighted smoothed line (LOESS) 


 
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to help identify any potential trend. For categorical covariates, box-whisker plots of P for each 
category will be compared side-by-side to identify any potential differences. Covariate-parameter 
relationships indicated by a clear trend will then be further tested. 
 
Covariates 
Reason for investigation 
Parameter 
WT 
CL, and Vc are likely to be 
weight dependent 
CL, Vc 
CRCL 
Creatinine 
clearance 
may 
impact CL and Vc 
CL and Vc 
GA 
Gestational age may impact 
CL and Vc 
CL, Vc and Vp 
AGE 
Age may impact CL and Vc 
CL, Vc and Vp 
 
Continuous covariates may be included in the model using a linear or multiplicative function after 
centering on the median: 


y
θ
x
Covariate
Median
Covariate
θ
TVP








 
or 


y
θ
x
Covariate
Median
Covariate
θ
TVP







 
where TVP is the typical value of a PK parameter, x is a to be estimated covariate 
independent fixed-effect component and y is a covariate dependent fixed-effect component of 
the relationship. 
Categorical covariates will only be tested if at least 5% (or 15 individuals – whichever is smaller) of 
subjects belong to that category. Categorical covariates will be incorporated into the model using 
index coding: 
TVP = x(1 + yIndex) 
where TVP is the typical value of a PK parameter, x is a covariate independent fixed-effect 
component and y is a covariate dependent fixed-effect component of the relationship. Index 
has a value of 1 in the presence of the covariate, otherwise it has a value of 0. 
Assessment of the significance for covariate will use the forward addition and backward elimination 
approach. The MOF based on Likelihood ratio test (LRT) will be a primary evaluation for a 
covariate-parameter relationship to be considered statistically significant based on the alpha level 
specified prior to the analysis. For the addition of a covariate that decreases the MOF by more than 
3.84 (2, p <0.05; df=1) and the removal of a covariate that increases the MOF by more than 10.83 
(2, p <0.001; df=1) will be considered statistically significant. For the purposes of documenting the 
process of covariate screen, the MOF values will be tabulated in accordance with the covariates 
included in the models at each step. 
 
Random Effect Correlations 


 
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Correlations between random effects will be tested after inclusion of significant fixed effects of 
covariates in the model. Pair-wise plots of posterior random effects produced by the POSTHOC step 
of a NONMEM run will be examined visually and correlation coefficients will be calculated. The 
cases with the highest correlations will be tested for significance by including such correlations in 
the population model. The LRT will be applied and the same criteria as for fixed effects will be 
used. 
 Final Model 
Key modeling results including summary of parameters’ estimates from final model, description of 
covariate selection and Goodness of Fit information will be presented to show how well the model 
describe the data. Any limitation on the analysis conducted will be discussed. The assumptions 
during the modeling process will be justified. 
 Model Evaluation 
 A visual predictive check will be performed to evaluate performance of the final PopPK model. 
Simulation using final PK and variability parameters and dosing histories, sampling times and 
covariate information will be obtained for the observed study drug concentration. A 90% 
prediction interval (5th to 95th percentile) will be generated from the simulated values. The 
number of observed concentrations within the prediction intervals will be determined. If this 
number is greater than 80%, the model will be considered suitable for predictive purposes. 
 The standard errors of the final model parameter values will be estimated using the non-
parametric bootstrap approach. For the final model approximately 500-1000 bootstrap datasets 
will be generated. The mean and 95% confidence intervals for the PopPK parameter estimates 
will be calculated and compared with the estimates from the final model. The bootstrap 95% 
confidence interval will be calculated based on the percentile of the empirical distribution of the 
estimated parameters from the bootstrap runs. 
 
Reporting of PopPK analysis 
At the completion of the PopPK analysis, a report will be drafted, reviewed, and revised if 
necessary, and finalized. The final report along with the relevant models, datasets, and other related 
information will be archived. The final report will include a tabular summary of all models explored 
during model development. For the key models a complete set of goodness-of-fit plots will be 
included. 
 
References 
 
 Beal SL, Sheiner LB; NONMEM Project Group C255, University of California at San 
Francisco. NONMEM users guides – Part I-VIII. San Francisco; 1988-1998. 
 Bachman W, et al. PDx Pop Version 3.0 User Manual. 2007. 
 Guidance for Industry PopPK, U.S. Department of Health and Human Services Food and Drug 
Administration, CDER and CBER, February 1999. 


 
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 Guideline on Reporting the Results of PopPK Analyses, European Medicines Agency, CHMP, 
21 June 2007. 
 
 
 
 


 
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21.3. Appendix 3 - Tables, Figures, and Listings of PK report (NCA and POPPK) 
 
Tables 
Table 1: Data Item Abbreviations and Descriptions as Implemented in NONMEM Datasets 
Table 2: Summary of Number of Patients Available in the Database and the Number of Patients Included in the 
Analysis 
Table 3: Summary of Categorical Covariates 
Table 4: Summary of Continuous Covariates 
Table 5: Data Points Removed From the Database, which are considered to be Outliers 
Table 6: Parameter Estimates of Final Base PK Model 
Table 7: Covariate PK Model Development (Forward Addition Steps) 
Table 8: Covariate PK Model Development (Backward Deletion Steps) 
Table 9: Parameter Estimates of Final PK Model 
Figures 
Figure 1: 
Distributions of Quantitative Covariates 
Figure 2: 
Box-Whisker Plots of Continuous Covariates by Category of Categorical Covariates 
Figure 3: 
Scatter-Plot of Continuous Covariates versus Continuous Covariates  
Figure 4: 
plots of quartiles of covariate distribution versus quartiles of standard normal distribution (QQ plot) 
Figure 5: 
Observed Plasma Concentrations versus Time 
Figure 6: 
Population and Individual Predicted vs. Observed Plasma Concentrations (Base Model) 
Figure 7: 
Population Residuals vs. Population Predicted Concentrations and Time (hr) (Base Model) 
Figure 8: 
Weighted Population Residuals vs. Population Predicted Concentrations and Time (hr) (Base Model) 
Figure 9: 
Inter-Subject Random Effect on PK Parameters vs. Covariates 
Figure 10: Population and Individual Predicted vs. Observed Plasma Concentrations (Final Model) 
Figure 11: Population Residuals vs. Population Predicted Concentrations and Time (hr) (Final Model)  
Figure 12: Weighted Population Residuals vs. Population Predicted Concentrations and Time (hr) (Final Model) 
Figure 13: Population Predicted and Observed Plasma Concentrations vs. Time after Dose (hr) for Study (Final Model) 
Figure 14: Histograms of Weighted Residuals in Final Model 
Figure 15: Posterior Bayes Estimates of ETA on CL versus covariates  
Figure 16: Visual predictive check 
Listings 
 Population PK Analysis Dataset 
NONMEM Outputs 
 NONMEM Control file and Output for Base PK Model 
 NONMEM Control file and Output for Final PK Model 
 Individual Panel Plots for Goodness of Fit