Clinical trial NCT04007107
Conditions: Healthy Volunteers Diabetes Mellitus, Type 2 Healthy Volunteers Overweight Healthy Volunteers Obesity
Sponsor: Novo Nordisk A/S
Start date: 2019-06-27; Completion date: 2019-09-04
Enrollment: 103 participants; Allocation: RANDOMIZED; Intervention model: CROSSOVER

Brief summary:
This study in healthy men and women compares the injection site experience of the DV3396 pen to that of the PDS290 pens when both pens are used to deliver 0.25 mg semaglutide subcutaneously (sc, under the skin). Participants will receive 2 single doses of semaglutide 0.25 mg on 1 day. The 2 injections will be given at least 30 minutes apart, one in each side of the stomach. Participants will be in the clinic research center for 1 day. A follow-up phone call will take place between 4 and 5 weeks after the injections were given.

Detailed description:

Arms: arm 1: The 2 treatments will be administered at least 30 minutes apart, one in each side of the stomach arm 2: The 2 treatments will be administered at least 30 minutes apart, one in each side of the stomach
Interventions: intervention 1: Semaglutide (administered by DV3396 pen) intervention 2: Semaglutide (administered by PDS290 pen)
Intervention descriptions: intervention 1: Subjects will receive 1 single dose of semaglutide 0.25 mg on 1 day intervention 2: Subjects will receive 1 single dose of semaglutide 0.25 mg on 1 day

Protocol documents:
Prot_000.pdf: 
Cover Page for Protocol 
Sponsor name: 
Novo Nordisk A/S 
NCT number 
NCT04007107 
Sponsor trial ID: 
INS-4582 
Official title of study: 
A TRIAL TO COMPARE THE INJECTION SITE PAIN 
EXPERIENCE OF 0.25 MGSEMAGLUTIDE SC 
ADMINISTERED BY 2 DIFFERENT PRODUCTS 
Document date: 
17 March 2020 


16.1.1 Protocol
/LVWRIFRQWHQWV
CLINICAL STUDY PROTOCOL, INS-4582/
, VERSION 2.0, 21 JUN 2019 ...................................
16.1.1.1 Filenote_03-Jun-2019 ...........................................................................................................................................
2
59
CONFIDENTIAL
Date:
1 0DU 2020 
Novo Nordisk
Version: 
1.0 
Status:
Final
Semaglutide 
Trial ID: INS-4582
Clinical Trial Report
Appendix 16..
Page:
1 of 
CONFIDENTIAL
 
 
 
 
Redacted protocol 
Includes redaction of personal identifiable information only. 
 
 
 
. 
 
 


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CLINICAL STUDY PROTOCOL
A TRIAL TO COMPARE THE INJECTION SITE PAIN EXPERIENCE OF 0.25 MG 
SEMAGLUTIDE SC ADMINISTERED BY 2 DIFFERENT PRODUCTS
CONFIDENTIAL
Sponsor code: INS-4582
 code:
EudraCT number: 2019-002284-10
Universal Trial Number: U1111-1233-9590
Comparison of injection site pain experience for semaglutide sc 
Investigational product: 
Semaglutide
Clinical phase: 
Phase 3b 2 study
Indication to be studied: 
Not applicable
Sponsor:
Novo Nordisk A/S
Novo Allé
DK-2880 Bagsvaerd
Denmark
Contract Research
Organization and
Clinical Site:
Principal Investigator:
Phone:
Fax:
E-mail:
Version 2.0, 21 Jun 2019
Version 1.0, 03 Jun 2019
This study will be performed in compliance with the principles of Good Clinical Practice.
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CONFIDENTIAL


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AUTHORIZATION OF CLINICAL STUDY PROTOCOL BY CONTRACT 
RESEARCH ORGANIZATION
The Sponsor and the Contract Research Organization agree to conduct the study as outlined in this 
clinical study protocol. Any modification of the clinical study protocol must be agreed upon by the 
Sponsor and the Contract Research Organization and must be documented in writing.
Name/Position:
Date:
Signature:
Contract Research Organization:
………
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CONTACT INFORMATION
Sponsor
Novo Nordisk A/S
Novo Allé
DK-2880 Bagsvaerd
Denmark
Sponsor's Contact
Trial Operations, Insulin & Devices
Phone:
E-mail:
Sponsor’s Medical Specialist
Medical & Science, Devices & Titration
Phone:
email:
Contract Research Organization and Clinical Site
Phone:
Fax:
Clinical Site Contacts
Phone:
Fax:
E-mail:
Phone:
Fax:
E-mail:
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Clinical Laboratory 
Phone:
Fax:
Medical Screening Centers
Phone:
Fax:
Phone:
Study Monitor
Phone:
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SUMMARY OF CHANGES
The following changes have been introduced in this Version 2.0 of the protocol and are given as a 
combination of double underlined and italic text. Deleted text is given as double strikethrough.
x
In the study title and other sections of the protocol, the word ‘pain’ has been added between 
the words ‘injection site’ and ‘experience’ for clarification.
x
The clinical phase of the study has been changed from Phase 3b to Phase 2.
x
The language on subject numbering has been changed.
The changes can be found in:
x
Title Page
x
Synopsis
x
Section 1.3, Study Rationale
x
Section 2, OBJECTIVES
x
Section 4.1.1, Type of Study
x
Section 4.4.3, Method of Assigning Subjects to Treatment Groups
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SYNOPSIS 
Study Title
A TRIAL TO COMPARE THE INJECTION SITE PAIN EXPERIENCE OF 0.25 MG SEMAGLUTIDE SC
ADMINISTERED BY 2 DIFFERENT PRODUCTS
Short Study Title
Comparison of injection site pain experience for semaglutide sc
Study Codes 
Sponsor code
: INS-4582
 code
: 
EudraCT number 
: 2019-002284-10
Universal Trial Number 
: U1111-1233-9590
Sponsor
Novo Nordisk A/S, Novo Allé, DK-2880 Bagsvaerd, Denmark
Sponsor’s contact 
: 
Contract Research Organization and Clinical Site
Principal Investigator
Clinical Phase
: 2
Objectives
The primary objective is to compare, in healthy subjects, the injection site pain experience of a single dose 
of 0.25 mg semaglutide subcutaneously (sc), given as the DV3396 product, to that of the PDS290 product.
Endpoints
Primary Endpoint
Endpoint Title
Time Frame
Unit
Intensity of injection site pain
1 min after each injection (Day 1)
mm on a 100-mm horizontal 
visual-analogue scale (where 
0 mm= no pain, 100 mm = 
unbearable pain)
Exploratory Endpoints
Endpoint Title
Time Frame
Unit
Categorical 
assessment 
of 
injection site pain
Immediately 
after 
rating 
of 
intensity of injection site pain, ie, 
after 1 min plus the time it takes 
to complete the rating of intensity 
of injection site pain by visual 
analogue scale (Day 1)
6-point scale (none – very mild –
mild – moderate – severe – very 
severe)
Moderate or more injection site 
pain
Dichotomous variable based on 
categorical 
assessment 
of 
Yes = moderate, severe, very 
severe
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Endpoint Title
Time Frame
Unit
intensity of injection site pain 
(Day 1) (item above)
No = none, very mild, mild
Quality of pain
Immediately after categorical 
assessment 
of 
intensity 
of 
injection site pain, ie, after 1 min 
plus the time it takes to complete 
the rating of intensity of injection 
site pain and the categorical 
rating of intensity of injection site 
pain (Day 1)
Pain quality items on modified 
SF-MPQ-2 inventory (select all 
that apply):
Throbbing pain տ
Shooting pain տ
Stabbing pain տ
Sharp pain տ
Cramping pain տ
Gnawing pain տ
Hot-burning pain տ
Aching pain տ
Heavy pain տ
Tender տ
Splitting pain տ
Tiring-exhausting տ
Sickening տ
Fearful տ
Punishing-cruel տ
Electric-shock pain տ
Cold-freezing pain տ
Piercing տ
Duration of pain
From time of injection until 
cessation of pain (Day 1)
min and sec
Comparative pain experience
After pain has ended after the 
last injection (Day 1)
5-point scale (the last injection 
hurt much more than the first 
injection – the last injection hurt 
more than the first injection – 
they 
hurt 
about 
the 
same 
[includes: none of the injections 
hurt] – the last injection hurt less 
than the first injection – the last 
injection hurt much less than the 
first injection) 
The DV3396 product hurt less 
than or about the same as the 
PDS290 product
Dichotomous variable based on 
“Comparative pain experience” 
variable above (Day 1)
Yes = “about the same” (includes 
“none of the products hurt” or 
“the DV3396 product hurt less 
than the PDS290 product” or “the 
DV3396 product hurt much less 
than the PDS290 product”)
No = “The DV3396 product hurt 
much more than the PDS290 
product” 
or 
“The 
DV3396 
product hurt more than the 
PDS290 product”
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Design and Treatments
This is a single-center, cross-over, randomized, double-blind study in healthy men and women comparing 
the injection site pain experience of the DV3396 product to that of the PDS290 product when both products 
are used to deliver 0.25 mg semaglutide sc. Subjects will be randomized in a 2×2 scheme evenly to 
4 sequences of product and injection side as in the table below:
Treatment Short Identifier
Treatment Long Identifier
Ratio Required
First injection on right side, Second injection on left side
A
DV3396 (Right) followed by PDS290 (Left)
1
C
PDS290 (Right) followed by DV3396 (Left)
1
First injection on left side, Second injection on right side
B
DV3396 (Left) followed by PDS290 (Right)
1
D
PDS290 (Left) followed by DV3396 (Right)
1
Subjects will receive 2 single doses of semaglutide 0.25 mg on 1 day. The 2 products will be administered 
at least 30 minutes apart, in the anterior aspect of the abdominal wall on opposite sides of the midline.
An overview of the study design is given below:
Study Schedule
Screening
: Between Day -21 and Day -1
Confinement period 
: One day in the clinic: Day 1
Follow-up
: A follow-up phone call will take place between 4 and 5 weeks after drug 
administration on Day 1
Subjects
A total of 104 subjects are planned to be randomized to achieve at least 100 completed subjects. The aim is 
to have an even distribution of male and female subjects in the study; at least 30% of the same sex should 
be randomized in the study.
Key Inclusion Criteria
x
Male or female subjects, aged 18 to 75 years (both inclusive) at the time of signing informed consent.
x
%0,NJP2
x
Considered to be generally healthy based on the medical history, physical examination, and the results 
of vital signs, electrocardiogram (ECG), and clinical laboratory tests performed during the screening visit, 
as judged by the Investigator.
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Key Exclusion Criteria
x
Woman who is pregnant or breast-feeding or intends to become pregnant within 4 weeks after 
administration of the study drug, or is of childbearing potential and not using highly effective contraceptive 
methods with her fertile male sexual partner.
x
Any disorder that in the Investigator’s opinion might jeopardize subject’s safety, evaluation of results, or 
compliance with the protocol.
x
Glycosylated hemoglobin 6.5% at screening.
x
Use of prescription medicinal products or non-prescription drugs or herbal products, except routine 
vitamins, topical medication, contraceptives, and occasional use of paracetamol (paracetamol not 
allowed within 24 hours prior to drug administration), within 14 days prior to drug administration.
x
Average intake of more than 21 units of alcohol per week for male subjects and more than 14 units per 
week for female subjects: 1 unit of alcohol equals approximately 250 mL of beer, 100 mL of wine, or 
35 mL of spirits.
x
Positive drug and alcohol screen (opiates, methadone, cocaine, amphetamines [including ecstasy], 
cannabinoids, barbiturates, benzodiazepines, tricyclic antidepressants, and alcohol) at screening and 
admission to the clinical research center.
x
Use of tobacco and nicotine products, defined as any of the below:
o
Smoking more than 1 cigarette or the equivalent per day on average
o
Not able or willing to refrain from smoking and the use of nicotine substitute products during the 
in-house period
x
Subject is not able to understand and read English or Dutch, or subject is not able to understand and 
comply with the study requirements.
Study Drug
Material Number
Name
Name of Intervention Product
Semaglutide B 1.34 mg/ml PDS290
PDS290 product
Semaglutide C 0.5 mg/ml DV3396
DV3396 product
Variables
Safety
: Intensity of injection site pain on a visual analogue scale (VAS), categorical 
assessment of injection site pain intensity (6-point scale), quality of pain
modified SF-MPQ-2 inventory, duration of pain assessment, comparative pain 
assessment (5-point scale), adverse events, clinical laboratory (if postdose 
assessments are deemed relevant by the Investigator), physical examination (if 
postdose assessments are deemed relevant by the Investigator), 12-lead ECG 
(if postdose assessments are deemed relevant by the Investigator), and vital 
signs
Statistical Methods
Sample size calculation 
: The sample size calculation considers both the primary endpoint and the 
dichotomous endpoints. Considering precision on the proportion of the 
dichotomous endpoints, a sample size of 100 subjects results in a confidence 
interval (CI) with a width of 10% on each side, eg, for a proportion of 60%, the 
CI is 60% (50%-70%).
A standard deviation of within-pair differences in VAS pain score between 
2 subcutaneous injections was obtained from Novo Nordisk clinical trial 
INS-4011. The within-subject variance (adjusting for the effects of speed, 
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volume and injection region) was estimated to 348, corresponding to a standard 
deviation (SD) of the within-pair differencHRI¥Â PP$SURVSHFWLYH
study of acute pain found that the minimum clinically important difference for 
worsening of pain, ie, the difference on a VAS associated with the selection of 
“a little more pain” rather than “about the same” had a mean value of 10 mm.3  
With 100 subjects, a conservative standard deviation of 30 mm, and 90% power, 
it is possible to detect a difference smaller than 10 mm. Hence a sample size of 
100 subjects is considered sufficient to detect a clinically important difference. 
Allowing for 4% dropouts or missing data, a total of 104 subjects will need to be 
randomized.
Analysis of endpoints 
: The primary endpoint will be analyzed by a fixed analysis of variance model with 
VAS score as the dependent variable, and product, injection side (right side, left 
side), injection number (first injection, second injection), and subject as fixed 
effects. From the model, the difference in VAS score between the 2 products is 
estimated and presented with a 95% CI and a p-value. The interpretation of the 
difference in VAS pain scores will be supported and contextualized by the 
results of the analyses of the exploratory endpoints.
Descriptive statistics only will be applied for other safety parameters and 
secondary endpoints.
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Table 1 
Schedule of Assessments
Visit
Screening
Pretreatment
Treatment
Follow-up Phone Call
Study Day
Days
-21 to -1
Day 1
Day 29 to Day 36 
Predose
Dosing and After
Ambulatory
X
X
X
X
Admission
X
Discharge
X
Informed Consent
X
Medical History
X
Previous Injection Experiencea
X
Demographics
X
Physical Examination
X
Xb
Body Height, Body Weight and BMI Calculation
X
Serology (HBsAg, anti-HCV, anti-HIV-1 and -2)
X
Urine Drug and Alcohol Screen
X
X
Urine Pregnancy Test (Females of Childbearing Potential Only)
X
X
Xc
Clinical Laboratoryd
X
Xd
HbA1c
X
12-lead ECGe
X
Xe
Vital Signsf
X
X
Eligibility Check
X
Xg
Randomization and Stratificationh
X
Study Drug Administrationh
X
Intensity of Injection Site Pain VASi
X
Categorical Assessment of Injection Site Pain Intensityj
X
Quality of Pain Modified SF-MPQ-2 Inventoryk
X
Duration of Painl
X
Comparative Pain Assessmentm
X
Previous and Concomitant Medication
X
X
X
X
AE Monitoring
X
X
X
X
Technical Complaintsn
X
X
Drug Accountability
X
AE=adverse event; BMI=body mass index; ECG=electrocardiogram; HbA1c=glycoslated hemoglobin; HBsAg=hepatitis B surface antigen; HCV=hepatitis C virus; 
HIV=human immunodeficiency virus; sc=subcutaneous; SF-MPQ-2=short-form McGill pain questionnaire; VAS=visual analogue scale
a
Previous injection experience as recalled by the subject to the best of their ability: product used (name, dose, type [vial/syringe, prefilled syringe, autoinjector, 
other], indication, frequency of use, injection site, chronological period used).
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b
Physical examination: this will be a focused examination only done if deemed relevant by the Investigator
c
For the check on pregnancy at the follow-up phone call, female subjects of childbearing potential will be handed out a urine pregnancy test on Day 1. They 
will have to do the pregnancy test at the ealiest in the morning of Day 29 and at the latest in the morning of the follow-up phone call.
d
Clinical laboratory tests (including clinical chemistry and hematology): at screening (non-fasting). 
The same tests may be done after the second study drug administration on Day 1 prior to discharge, if deemed relevant by the Investigator.
e
12-lead ECG: at screening.
12-lead ECG may be done after the second study drug administration on Day 1 prior to discharge, if deemed relevant by the Investigator.
f
Vital signs (supine systolic and diastolic blood pressure, and pulse): at screening and after the second study drug administration on Day 1 prior to discharge.
g
Eligibility check on Day 1 before dosing concerns the urine pregnancy test (female subjects of childbearing potentialonly), urine drug and alcohol screen,
and concomitant medication only.
h
Subjects will be randomized in a 2×2 scheme evenly to 4 sequences of product and injection side as in the table below: 
Treatment Short Identifier
Treatment Long Identifier
Ratio Required
First injection on right side, Second injection on left side
A
DV3396 (Right) followed by PDS290 (Left)
1
C
PDS290 (Right) followed by DV3396 (Left)
1
First injection on left side, Second injection on right side
B
DV3396 (Left) followed by PDS290 (Right)
1
D
PDS290 (Left) followed by DV3396 (Right)
1
Subjects will receive 2 single doses of semaglutide 0.25 mg on 1 day. The 2 products will be administered at least 30 minutes apart, in the anterior aspect 
of the abdominal wall on opposite sides of the midline.
i
One minute after each injection, the VAS to rate the intensity of injection site pain will be administered.
j
The categorical assessment of injection site pain intensity (6-point scale) will be started after the VAS to rate the intensity of injection site pain has been 
completed after each injection.
k
The quality of pain modified SF-MPQ-2 inventory will be started after the categorical assessment of injection site pain intensity (6-point scale) has been 
completed after each injection.
l
Subjects have to indicate after each injection when the pain, if any, is gone.
m
The comparative pain assessment (5-point scale) comparing pain between the 2 injections will be completed after the second injection.
n
All technical complaints that occur, from the time of receipt of the product at the site until the time of the last usage of the product, must be collected and 
reported to Novo Nordisk. 
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TABLE OF CONTENTS
TITLE PAGE .......................................................................................................................1
SPONSOR AUTHORIZATION OF CLINICAL STUDY PROTOCOL.................................. 2 
AUTHORIZATION OF CLINICAL STUDY PROTOCOL BY CONTRACT RESEARCH
ORGANIZATION .......................................................................................................... 3 
CONTACT INFORMATION ................................................................................................ 4 
SUMMARY OF CHANGES................................................................................................. 6 
SYNOPSIS.......................................................................................................................... 7 
TABLE OF CONTENTS.................................................................................................... 14 
TABLE OF TABLES......................................................................................................... 16 
TABLE OF FIGURES ....................................................................................................... 17 
LIST OF ABBREVIATIONS .............................................................................................. 18 
1. INTRODUCTION ........................................................................................................ 19 
1.1 
Background.................................................................................................................19 
1.2 
Risk-Benefit Assessment and Risk Mitigation..........................................................20 
1.2.1 Anticipated Clinical Benefits...............................................................................20 
1.2.2 Risks Related to Semaglutide............................................................................20 
1.2.3 Risks Associated with Participation in the Clinical Study....................................21 
1.2.4 Possible Interactions with Concomitant Medical Treatments..............................21 
1.2.5 Risk Mitigation ...................................................................................................21 
1.2.6 Risk-Benefit Rationale .......................................................................................21 
1.3 
Study Rationale...........................................................................................................22 
2. OBJECTIVES ............................................................................................................. 23 
3. ENDPOINTS............................................................................................................... 23 
4. INVESTIGATIONAL PLAN ........................................................................................ 25 
4.1 
Overall Study Design and Plan ..................................................................................25 
4.1.1 Type of Study ....................................................................................................25 
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4.1.2 Screening Period ...............................................................................................25 
4.1.3 Treatment Period...............................................................................................26 
4.1.4 Follow-up...........................................................................................................26 
4.2 
Discussion of Study Design.......................................................................................26 
4.3 
Selection of Study Population....................................................................................27 
4.3.1 Inclusion Criteria................................................................................................27 
4.3.2 Exclusion Criteria...............................................................................................27 
4.3.3 Removal of Subjects from Assessment..............................................................28 
4.4 
Treatments...................................................................................................................29 
4.4.1 Treatments Administered...................................................................................29 
4.4.2 Identity of Investigational Products ....................................................................29 
4.4.3 Method of Assigning Subjects to Treatment Groups ..........................................30 
4.4.4 Selection of Doses in the Study .........................................................................31 
4.4.5 Timing of Doses in the Study .............................................................................31 
4.4.6 Meals During the Study......................................................................................32 
4.4.7 Blinding..............................................................................................................32 
4.4.8 Concomitant Medication and Other Restrictions During the Study.....................33 
4.4.9 Treatment Compliance.......................................................................................33 
4.5 
Safety Measurements and Variables .........................................................................33 
4.5.1 Safety Measurements Assessed and Schedule of Assessments .......................34 
4.5.1.1 
Safety and Tolerability Measurements ...............................................34 
4.5.1.1.1 
Intensity of Injection Site Pain Visual Analogue Scale .....34 
4.5.1.1.2 
Categorical Assessment of Injection Site Pain Intensity...35 
4.5.1.1.3 
Quality of Pain Modified SF-MPQ-2 Inventory .................35 
4.5.1.1.4 
Duration of Pain...............................................................36 
4.5.1.1.5 
Comparative Pain Assessment (5-point scale) ................36 
4.5.1.1.6 
Adverse Events ...............................................................36 
4.5.1.1.7 
Technical Complaints ......................................................39 
4.5.1.1.8 
Clinical Laboratory...........................................................41 
4.5.1.1.9 
Vital Signs .......................................................................42 
4.5.1.1.10 Electrocardiogram ...........................................................42 
4.5.1.1.11 Physical Examination ......................................................42 
4.5.1.2 
Total of Blood Volume........................................................................42 
4.5.2 Appropriateness of Measurements ....................................................................42 
4.5.3 Safety Variables ................................................................................................42 
4.6 
Statistical Procedures and Determination of Sample Size.......................................43 
4.6.1 Analysis Sets.....................................................................................................43 
4.6.1.1 
Safety Set..........................................................................................43 
4.6.1.2 
Full analysis set (FAS).......................................................................43 
4.6.1.3 
Per-Protocol Set ................................................................................43 
4.6.2 Statistical and Analytical Plan for Safety Evaluation...........................................43 
4.6.2.1 
Evaluation of Safety and Tolerability..................................................43 
4.6.2.1.1 
Intensity of Injection Site Pain Visual Analogue Scale .....43 
4.6.2.1.2 
Categorical Assessment of Injection Site Pain Intensity, 
Quality of Pain Modified SF-MPQ-2 Inventory, Duration of 
Pain, and Comparative Pain Assessment........................44 
4.6.2.1.3 
Adverse Events ...............................................................45 
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4.6.2.1.4 
Technical Complaints ......................................................45 
4.6.2.1.5 
Clinical Laboratory...........................................................45 
4.6.2.1.6 
Vital Signs and Electrocardiograms................................. 46 
4.6.2.1.7 
Physical Examination ......................................................46 
4.6.3 Determination of Sample Size............................................................................46 
4.6.4 Interim Analysis ................................................................................................. 46 
4.7 
Data Quality Assurance..............................................................................................46 
5. ETHICS....................................................................................................................... 48 
5.1 
Independent Ethics Committee..................................................................................48 
5.2 
Ethical Conduct of the Study .....................................................................................48 
5.3 
Subject Information and Consent ..............................................................................49 
5.4 
Privacy.........................................................................................................................49 
6. STUDY ADMINISTRATIVE STRUCTURE ................................................................. 50 
6.1 
Distribution of Activities.............................................................................................50 
6.2 
Documentation............................................................................................................50 
6.2.1 Archiving............................................................................................................50 
6.2.2 Recording of Data in Source Documents and eCRFs ........................................50 
7. CONFIDENTIALITY AND PUBLICATION POLICY ................................................... 52 
7.1 
Publication Policy .......................................................................................................52 
7.2 
Dissemination of Clinical Trial Data...........................................................................52 
8. REFERENCES ........................................................................................................... 53 
9. APPENDICES............................................................................................................. 54 
9.1 
Drug Accountability....................................................................................................54 
9.2 
Adverse Events: Definitions and Procedures for Recording, Evaluation, Follow-up, 
and Reporting to Sponsor ..........................................................................................54 
TABLE OF TABLES
Table 1 
Schedule of Assessments.............................................................................................12 
Table 2 
Primary Endpoint...........................................................................................................23 
Table 3 
Exploratory Endpoints ...................................................................................................23 
Table 4 
Product and Side of Injection on Abdomen per Treatment ............................................25 
Table 5 
Products Supplied by Novo Nordisk A/S...................................................................30 
Table 6 
Visual Appearance and Function of the DV3396 Product and PDS290 Product............30 
Table 7 
Number and Volume of Blood Samples and Total Blood Volume Collected per Subject42 
Table 8 
Primary Endpoint........................................................................................................... 44 
Table 9 
Exploratory Endpoints ...................................................................................................44 
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Table 10 Detectable Difference with a Sample Size of 100 Subjects for Different Combinations of 
Power, and Standard Deviation.....................................................................................46 
TABLE OF FIGURES
Figure 1 Overview of the Study Design....................................................................................25 
Figure 2 Decision Tree for Determining the Event Type and the Respective Forms to 
Complete with Associated Timelines ........................................................................38 
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LIST OF ABBREVIATIONS
AE
adverse event 
ALAT (ALT)
alanine aminotransferase
ASAT (AST)
aspartate aminotransferase
BMI
body mass index
CA
Competent Authority
CHMP
Committee for Medicinal Products for Human Use (formerly: Committee for 
Proprietary Medicinal Products [CPMP])
CI
confidence interval
CRF
case report form
CSP
clinical study protocol
CSR
clinical study report
CTD
Clinical Trial Directive
CV
cardiovascular
DUN
dispensing unit number
ECG
electrocardiogram
eCRF
electronic case report form
EDS
Early Development Services
EMA
European Medicines Agency
FAS
full analysis set
GCP
Good Clinical Practice
GDPR
General Data Protection Regulation
GI
gastrointestinal
GLP
glucagon-like peptide
HbA1c
HBsAg
glycosylated hemoglobin
hepatitis B surface antigen
HCV
hepatitis C virus
HIV
human immunodeficiency virus
IB
investigator’s brochure
ICF
informed consent form
ICH
International Council for Harmonisation (formerly: International Conference on 
Harmonisation)
IEC
Independent Ethics Committee
MCID
minimum clinically important difference
MEB 
Medicine Evaluation Board
PCD
primary completion date
PK
pharmacokinetic(s)
RA
receptor agonist
SAE
serious adverse event
SAP
statistical analysis plan
sc
subcutaneous
SD
standard deviation
SF-MPQ-2
Short-form McGill Pain Questionnaire 2
SOP
standard operating procedure
T2D
type 2 diabetes mellitus
TEAE
treatment-emergent adverse event
TMM
trial materials manual
ULN
upper limit of normal
VAS
visual analogue scale
WMA
World Medical Association
WMO
Wet medisch-wetenschappelijk onderzoek met mensen (medical research 
involving human subjects act)
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1.
INTRODUCTION
1.1
Background
Semaglutide is a potent glucagon-like peptide-1 (GLP-1) receptor agonist (RA) 
developed by Novo Nordisk for weight management (Project NN9536). Semaglutide has 
a half-life of approximately 160 hours, making it suitable for once-weekly dosing. 
Semaglutide is currently approved in the EU and other regions for 0.5 and 1 mg weekly 
subcutaneous (sc) administration for treatment of type 2 diabetes (T2D) (Ozempic). 
Weekly administration of semaglutide sc 2.4 mg is currently being evaluated in Phase 
3a trials for weight management.
A 52-week Phase 2 dose-finding trial within weight management (NN9536-4153) has 
been completed. Relative to baseline, the estimated mean weight loss at week 52 was 
13.8% at the highest dose tested (0.4 mg once-daily as an adjunct to diet and exercise) 
compared to the mean weight loss of 2.3% achieved by diet, exercise and placebo alone.
Clinical and nonclinical data indicate that the body weight-reducing effect of semaglutide 
is mainly mediated by a reduced energy intake. No unexpected safety findings were 
identified and the tolerability and safety profile were overall consistent with previous 
findings in the T2D development programme and the GLP-1 RA class in general.
The clinical development program for semaglutide sc was conducted with a cartridge-
based prefilled PDS290 pen-injector.  
For the weight management indication, Novo Nordisk is currently developing the DV3396 
pen-injector, a single-dose pen-injector for sc injection of semaglutide, a new single-dose 
device with an integrated prefilled syringe. 
The DV3396 pen-injector for the 0.25 mg dose (first escalation dose) houses a 1.0-mL 
syringe pre-filled with a 0.5-mL semaglutide solution. A semaglutide concentration of 0.5 
mg/mL is used to obtain a dose of 0.25 mg. The device comes with a 29G regular-walled 
needle and is shield-activated, ie, the injection is activated when pushing the pen against 
the injection site. 
To allow the DV3396 pen-injector to provide the semaglutide dose in a volume of 0.5 mL, 
a new semaglutide formulation has been required compared to the semaglutide 
formulation used for the PDS290 pen-injector. In addition to the change of semaglutide 
concentration, the preservative (phenol) is removed from the new formulation, as it is to 
be used for single injections, and the concentration of propylene glycol has been slightly 
increased to compensate for the absence of phenol and ensure isotonicity.
Detailed information about semaglutide sc, the PDS290 semaglutide pen-injector, and 
the DV3396 pen-injector are available in the investigator’s brochure (IB).1  
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1.2
Risk-Benefit Assessment and Risk Mitigation
1.2.1
Anticipated Clinical Benefits
In the present study, healthy overweight/obese subjects are enrolled and they will 
therefore not benefit from the treatment with 2 single doses of semaglutide.
1.2.2
Risks Related to Semaglutide
The sections below describe identified and potential risks associated with semaglutide 
treatment. For classification and further details of the risks, please refer to the IB.1 The 
identified/potential risks are based on findings in nonclinical studies and clinical trials 
with semaglutide as well as with other GLP-1 RAs. For each of these risks, mitigating 
actions have been implemented to minimize the risks for subjects enrolled in this study.   
Gastrointestinal adverse events
Consistent with findings with other GLP-1 RAs, the most frequently reported adverse 
events (AE) in clinical trials with semaglutide were gastrointestinal (GI) AEs. A low dose
of semaglutide will be implemented in the study to mitigate the risk of GI AEs.
Cholelithiasis
Events of cholelithiasis were the most frequently reported gallbladder events in the
Phase 2 weight management trial (NN9536-4153) and were in a few instances 
co-reported with the event adjudication committee confirmed acute pancreatitis. As a 
precaution, if cholelithiasis is suspected, appropriate clinical follow-up is to be initiated 
at the Investigator’s discretion. 
Acute pancreatitis 
Acute pancreatitis has been observed with the use of GLP-1 RA drug class. As a 
precaution, subjects with a history of chronic pancreatitis or recent acute pancreatitis will 
not be enrolled in the study. 
Medullary thyroid cancer (MTC) 
Expected proliferative thyroid C-cell changes were seen in the mouse and rat 
carcinogenicity studies after daily exposure to semaglutide for 2 years. No hyperplasia 
was observed in monkeys after 52 weeks exposure up to 13-fold above the clinical 
plasma exposure at 2.4 mg/week. In clinical trials with semaglutide, there have been no 
clinically relevant changes in calcitonin levels. The C-cell changes in rodents are 
mediated by the GLP-1 receptor, which is not expressed in the normal human thyroid. 
Accordingly, the risk of GLP-1 receptor-mediated C-cell changes in humans is 
considered to be low. However, as a precaution, exclusion criteria related to medical 
history of multiple endocrine neoplasia type 2 are implemented in the study.
Pancreatic cancer 
There is currently no support from nonclinical studies, clinical trials or postmarketing data 
that GLP-1 RA-based therapies increase the risk of pancreatic cancer, but pancreatic 
cancer has been classified as a potential class risk of GLP-1 RAs by the European 
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Medicines Agency (EMA). As a precaution, subjects with a history of malignant 
neoplasms within the past 5 years prior to screening will be excluded from the study.
Allergic reactions
As is the case with all protein-based pharmaceuticals, subjects treated with semaglutide 
are at risk of developing immunogenic and allergic reactions. As a precaution, subjects 
with known or suspected hypersensitivity to semaglutide or related products will not be 
enrolled in this study.
Pregnancy and fertility (based on nonclinical data) 
Studies in animals have shown reproductive toxicity. There are limited data from the use 
of semaglutide in pregnant women. Therefore, semaglutide should not be used during 
pregnancy. Exclusion and discontinuation criteria related to pregnancy are implemented 
in the study. 
1.2.3
Risks Associated with Participation in the Clinical Study 
In the current study, risks related with venous blood sampling are minimal as no PK 
samples will be collected; only a few blood samples will be taken for clinical laboratory 
evaluations.
1.2.4
Possible Interactions with Concomitant Medical Treatments
Semaglutide delays gastric emptying and has the potential to impact the rate of 
absorption of concomitantly administered oral medicinal products. The potential effect of
semaglutide on the absorption of co-administered oral medications was studied in trials 
at semaglutide 1 mg/week steady-state exposure. No clinically relevant drug-drug 
interactions with semaglutide were observed based on the evaluated medications. 
1.2.5
Risk Mitigation
All subjects in this study will have a thorough medical check before entering the study. 
During the study, they will be followed closely and carefully by qualified medical staff. 
This will mitigate the known and potential risks. 
1.2.6
Risk-Benefit Rationale
The healthy subjects enrolled in the study will not have immediate health benefits from 
participating in this study. Data from this study might, however, benefit future treatment 
of subjects with T2D and overweight/obesity through a more convenient drug 
administration with the DV3396 product than with the PDS290 semaglutide product. The 
safety profile of semaglutide is well documented; AEs were mostly predictable based on 
the known effects of GLP-1 RAs. Thus, based on the large nonclinical and clinical 
development program with semaglutide 1.34 mg/mL and on the usability evaluation of 
the DV3396 product, it is concluded that there are no safety issues that would prohibit 
administration of semaglutide in accordance with the planned clinical study.
More detailed information about the known and expected benefits and risks and 
reasonably expected AEs of semaglutide sc, the PDS290 semaglutide pen-injector, and 
the DV3396 pen-injector are available in the IB.1
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1.3
Study Rationale
Novo Nordisk is currently developing the DV3396 pen-injector for the sc administration 
of semaglutide, the drug that is currently marketed for T2D in the PDS290 pen-injector 
in a different formulation. To evaluate the injection site pain experience after a single 
dose of semaglutide sc 0.25 mg, the present explorative study compares injection pain 
between the DV3396 product and the PDS290 pen-injector used with the Ypsomed 
Clickfine AutoProtect, 8 mm x 29G needle: The Ypsomed Clickfine® AutoProtect, 8 mm 
x 29G needle is chosen for its similarity to the built-in needle of the DV3396 product and 
is larger than the needle with which the PDS290 pen-injector is commercialized. 
Together, the PDS290 pen-injector and the Ypsomed Clickfine® AutoProtect, 8 mm x 
29G needle will be referred to as the PDS290 product.
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2.
OBJECTIVES 
The primary objective is to compare, in healthy subjects, the injection site pain
experience of a single dose of 0.25 mg semaglutide sc, given as the DV3396 product to 
that of the PDS290 product.
3.
ENDPOINTS
Table 2 
Primary Endpoint
Endpoint Title
Time Frame
Unit
Intensity of injection site pain
1 min after each injection
(Day 1)
mm on a 100-mm horizontal 
visual
analogue 
scale 
(where 0 mm = no pain, 
100 mm = unbearable pain)
Table 3 
Exploratory Endpoints
Endpoint Title
Time Frame
Unit
Categorical 
assessment 
of 
injection site pain intensity
Immediately after rating of 
intensity of injection site pain, 
ie, after 1 min plus the time it 
takes to complete the rating of 
intensity of injection site pain by 
visual analogue scale (Day 1)
6-point scale (none – very mild 
– mild – moderate – severe – 
very severe)
Moderate or more injection site 
pain
Dichotomous variable based 
on categorical assessment of 
intensity of injection site pain 
(Day 1) (item above)
Yes = moderate, severe, very 
severe
No = none, very mild, mild
Quality of pain
Immediately after categorical 
assessment of intensity of 
injection site pain, ie, after 1 
min plus the time it takes to 
complete the rating of intensity 
of injection site pain and the 
categorical rating of intensity of 
injection site pain (Day 1)
Pain quality items on modified 
SF-MPQ-2 inventory (select all 
that apply):
Throbbing pain տ
Shooting pain տ
Stabbing pain տ
Sharp pain տ
Cramping pain տ
Gnawing pain տ
Hot-burning pain տ
Aching pain տ
Heavy pain տ
Tender տ
Splitting pain տ
Tiring-exhausting տ
Sickening տ
Fearful տ
Punishing-cruel տ
Electric-shock pain տ
Cold-freezing pain տ
Piercing տ
Duration of pain
From time of injection until 
cessation of pain (Day 1)
min and sec
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Endpoint Title
Time Frame
Unit
Comparative pain experience
After pain has ended after the 
last injection (Day 1)
5-point scale (the last injection 
hurt much more than the first 
injection – the last injection hurt 
more than the first injection – 
they hurt about the same 
[includes: none of the injections 
hurt] – the last injection hurt 
less than the first injection – the
last injection hurt much less 
than the first injection) 
The DV3396 product hurt less 
than or about the same as the 
PDS290 product
Dichotomous variable based 
on 
“Comparative 
pain 
experience” 
variable 
above
(Day 1)
Yes 
= 
“about 
the 
same”
(includes “none of the products 
hurt” or “the DV3396 product 
hurt less than the PDS290 
product” 
or 
“the 
DV3396 
product hurt much less than the 
PDS290 product”)
No = “The DV3396 product hurt 
much more than the PDS290 
product” 
or 
“The 
DV3396 
product hurt more than the 
PDS290 product”
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4.
INVESTIGATIONAL PLAN
4.1
Overall Study Design and Plan
4.1.1
Type of Study
This is a single-center, cross-over, randomized, double-blind study in healthy men and 
women comparing the injection site pain experience of the DV3396 product to that of the 
PDS290 product when both products are used to deliver 0.25 mg semaglutide sc. 
Subjects will be randomized in a 2×2 scheme evenly to 4 sequences of product and 
injection side as in Table 4.
Table 4 
Product and Side of Injection on Abdomen per Treatment
Treatment 
Short Identifier
Treatment Long Identifier
Ratio Required
First injection on right side, Second injection on left side
A
DV3396 (Right) followed by PDS290 (Left)
1
C
PDS290 (Right) followed by DV3396 (Left)
1
First injection on left side, Second injection on right side
B
DV3396 (Left) followed by PDS290 (Right)
1
D
PDS290 (Left) followed by DV3396 (Right)
1
Subjects will receive 2 single doses of semaglutide 0.25 mg on 1 day. The 2 products 
will be administered at least 30 minutes apart, in the anterior aspect of the abdominal 
wall on opposite sides of the midline.
The end of the study is defined as last subject last visit (last follow-up phone contact). 
An overview of the study design is given in Figure 1. 
Figure 1 Overview of the Study Design
4.1.2
Screening Period
Subjects will report to the medical screening facility for the eligibility screening (see 
Section 4.3 for inclusion and exclusion criteria) within 3 weeks prior to drug 
administration. 
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Subjects will sign the study-specific informed consent form (ICF) prior to any 
study-specific screening procedures being performed. The written informed consent will 
be obtained for all subjects, regardless of their eligibility for the study; the signed ICFs 
will be retained and archived at 
, and a copy will be provided 
to the subject.
Eligibility screening will consist of the assessments as presented in the schedule of 
assessments (Table 1).
4.1.3
Treatment Period
Subjects will be in the clinic research center for 1 day, on Day 1. On this day, the subjects 
will be admitted, the drug will be administered twice, and the subjects will be discharged 
after all assessments have been performed.
If the subject admits to bodily pain (eg, headache) at admission (Day 1), then Day 1 
should be rescheduled within the visit window as the existing pain could impact the 
injection pain assessment.
Assessments during the treatment period will be performed as presented in the schedule
of assessments (Table 1). 
4.1.4
Follow-up
A follow-up phone call will take place between 4 and 5 weeks after drug administration 
on Day 1.
4.2
Discussion of Study Design
A total of 104 subjects are planned to be randomized to achieve at least 100 completed 
subjects. The aim is to have an even distribution of males and females in the study; at 
least 30% of the same sex should be randomized in the study.
Subjects with a body mass index (BMI)  kg/m2 but generally healthy will be included 
to minimize inter-subject variability due to frequent comorbidities in the target users of 
semaglutide (T2D and general population with overweight/obesity). It is assumed that 
the ability to detect differences between the 2 formulations does not differ between 
healthy subjects and target users. Only subjects with a BMI greater than or equal to 
25 kg/m2 will be included to reduce the risk of intramuscular injection. Both male and 
female subjects will be included to ensure generalizability to the intended users of both 
genders. Only individuals of age 18 years or older are eligible because semaglutide sc 
is only approved for use in adults with T2D. The upper age limit of 75 years is justified 
by the cognitive demands of the subjects when completing the inventories used in this 
study. 
The length of the cross-over interval is set to be as short as possible to minimize 
systemic effects of semaglutide, especially nausea, between the first and the last 
injection, while being long enough to minimize residual pain and emotional carry-over 
from the first injection. 
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4.3
Selection of Study Population
4.3.1
Inclusion Criteria
Note: Restrictions that apply to the period after admission are described in Section 4.4.8,
except when they concern a statement of willingness.
The following inclusion criteria must be met for a subject to be eligible for inclusion in the 
study:
1.
Informed consent obtained before any study-related activities. Study-related 
activities are any procedures that are carried out as part of the study, including 
activities to determine suitability for the study.
2.
Male or female subjects, aged 18 to 75 years (both inclusive) at the time of signing 
informed consent.
3.
%0,NJP2
4.
Considered to be generally healthy based on the medical history, physical 
examination, and the results of vital signs, electrocardiogram, and clinical 
laboratory tests performed during the screening visit, as judged by the Investigator.  
4.3.2
Exclusion Criteria
Note: Restrictions that apply to the period after admission are described in Section 4.4.8,
except when they concern a statement of willingness.
A subject who meets any of the following exclusion criteria will not be eligible for inclusion 
in the study:
1.
Known or suspected hypersensitivity to the study product or related products. 
2.
Previous participation in this study. Participation is defined as having received 
investigational product.
3.
Woman who is pregnant or breast-feeding or intends to become pregnant within 4
weeks after administration of the study drug, or is of childbearing potential and not 
using highly effective contraceptive methods with her fertile male sexual partner 
(see Section 4.4.8 for details on highly effective contraception).
4.
Participation in a drug study within 60 days prior to drug administration in the 
current study. Participation in more than 4 other drug studies in the 12 months prior 
to drug administration in the current study.
5.
Any disorder that in the Investigator’s opinion might jeopardize subject’s safety, 
evaluation of results, or compliance with the protocol.
6.
Glycosylated hemoglobin (HbA1c) 6.5% at screening.
7.
Supine blood pressure at screening (after resting for at least 5 minutes) outside the 
range of 90 to 160 mmHg for systolic or 45 to 89 mmHg for diastolic. 
8.
Supine pulse rate (as part of vital signs) (after resting for at least 5 minutes) outside 
the range of 40 to 100 beats per minute.
9.
Use of prescription medicinal products or non-prescription drugs or herbal 
products, except routine vitamins, topical medication, contraceptives, and 
occasional use of paracetamol (paracetamol not allowed within 24 hours prior to 
drug administration), within 14 days prior to drug administration.
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10.
Diagnostic test results positive for human immunodeficiency virus (HIV)-1 or HIV-2
infection.
11.
Diagnostic test results positive for hepatitis B or hepatitis C infection. 
12.
Mental incapacity, language barriers, or unwillingness to comply with the
requirements of the protocol, which may preclude adequate understanding or 
cooperation during the study as judged by the Investigator.
13.
Average intake of more than 21 units of alcohol per week for male subjects and 
more than 14 units per week for female subjects: 1 unit of alcohol equals 
approximately 250 mL of beer, 100 mL of wine, or 35 mL of spirits.
14.
Positive drug and alcohol screen (opiates, methadone, cocaine, amphetamines 
[including ecstasy], cannabinoids, barbiturates, benzodiazepines, tricyclic 
antidepressants, and alcohol) at screening and admission to the clinical research 
center.
15.
Use of tobacco and nicotine products, defined as any of the below:
x
Smoking more than 1 cigarette or the equivalent per day on average
x
Not able or willing to refrain from smoking and the use of nicotine substitute 
products during the in-house period
16.
Blood donation, plasma donation or blood draw (as declared by the subject or 
reported in the medical records): 
x
In excess of 400 mL within the past 90 days prior to the day of screening
x
In excess of 50 mL within the past 30 days prior to the day of screening
17.
Personal or first-degree relative(s) history of multiple endocrine neoplasia type 2 
or medullary thyroid carcinoma (as declared by the subject or reported in the 
medical records)  
18.
Subjects with a history of malignant neoplasms within the past 5 years prior to 
screening. 
19.
Presence or history of pancreatitis (acute or chronic; as declared by the subject or 
reported in the medical records).
20.
Subject is not able to understand and read English or Dutch, or subject is not able 
to understand and comply with the study requirements.
21.
Subject depends on the Sponsor, the Investigator, or the study center, or subject 
is the Investigator or any sub-investigator, research assistant, pharmacist, study 
coordinator, other staff, or relative thereof directly involved in the conduct of the 
study. 
22.
Vulnerable subject (eg, person kept in detention) who may have an increased 
likelihood of being wronged or of incurring additional harm.
Please note that subjects should refrain from consumption of any foods containing poppy 
seeds and alcohol within 48 hours (2 days) prior to screening and admission to the 
clinical research center to avoid false-positive drug screen results. In addition, they 
should refrain from strenuous exercise within 96 hours (4 days) prior to screening and 
admission as this could result in abnormal clinical laboratory values.
4.3.3
Removal of Subjects from Assessment
Participation in the study is strictly voluntary. A subject has the right to withdraw from the 
study at any time for any reason.
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The subject may be discontinued at any time during the study at the discretion of the 
Investigator for safety, behavioral, compliance, or administrative reasons.
If a subject is withdrawn from the study, the Sponsor will be informed immediately. If 
there is a medical reason for withdrawal, the subject will remain under the supervision 
of the Investigator until satisfactory health has returned.
Subjects who drop out or withdraw for any reason without completing all screening 
evaluations successfully will be considered screening failures. 
A subject who is withdrawn or voluntarily withdraws from the study for any reason, 
whether related to the study product or not, after having received a subject number, will 
be considered an early-termination subject. Early-termination subjects will not be 
replaced. 
 will make every effort to ensure that early-termination subjects who have received 
study drug complete the safety follow-up assessments.
4.4
Treatments
4.4.1
Treatments Administered
Subjects will be randomized in a 2×2 scheme evenly to 4 sequences of product and 
injection side as in Table 4 in Section 4.1.1. Subjects will receive 2 single doses of 
semaglutide 0.25 mg on 1 day. The 2 products will be administered at least 30 minutes 
apart, in the anterior aspect of the abdominal wall on opposite sides of the midline.
Unblinded members of the 
 staff not otherwise involved in the study procedures will 
be responsible for study drug administration. In each subject, the 2 semaglutide 
injections should be performed by the same staff member.
The 
 staff member will be instructed to select an injection site on the assigned side 
of the midline, that, according to their judgment based on visual inspection and palpation, 
has the maximal thickness of subcutaneous fat. The injection with the DV3396 product 
will be performed as described in the DV3396 pen-injector directions for use, ie, without 
making a lifted skin fold. Injection with the PDS290 product will be performed as 
described in the PDS290 pen-injector directions for use and the Ypsomed Clickfine 
AutoProtect, 8 mm x 29G needle instructions for use. However, to standardize the 
injections, using a lifted skin fold with the PDS290 product will not be an option.  
The unblinded 
 staff will instruct the subject to avoid discussing the administration 
of the study drug with other study staff including the Investigator.  
4.4.2
Identity of Investigational Products 
Novo Nordisk A/S Denmark will supply the following products:
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Table 5 
Products Supplied by Novo Nordisk A/S
Material Number
Name
Name of Intervention Product
Semaglutide B 1.34 mg/ml PDS290
PDS290 product
Semaglutide C 0.5 mg/ml DV3396
DV3396 product
In this study only, the PDS290 product will be used with an Ypsomed Clickfine 
AutoProtect, 8 mm x 29G needle. This needle will be sourced directly by 
For details concerning drug storage and drug accountability see Appendix 9.1. 
The characteristics of the 2 products are shown in Table 6. 
Table 6 
Visual Appearance and Function of the DV3396 Product and PDS290 
Product
* Needle used with the PDS290 product is specific to study INS-4582
All dose administrations will be performed by trained, unblinded staff at the study site. 
Directions for use of all products provided by Novo Nordisk are provided in the Trial 
Materials Manual (TMM) also provided by Novo Nordisk.
4.4.3
Method of Assigning Subjects to Treatment Groups
After obtaining informed consent, subjects will be screened according to the inclusion 
and exclusion criteria. Subjects who have met all eligibility criteria will receive a subject 
number upon inclusion in the study. They receive the subject number just prior to dosing. 
The subject number will ensure identification throughout the study.
The subject number will be the same as the randomization number given on the 
randomization list produced by Novo Nordisk. The randomization list will be transferred 
to the 
 Pharmacy and kept in a restricted area to which only the 
 Pharmacy has 
access. 
Subjects will be randomized in a 2×2 scheme evenly to 4 sequences of product and 
injection side as described in Section 4.1.1. Subjects will receive a randomization 
Appearance (colours and 
labelling not final)
Uses
Length (with cap)
Diameter
Housing colour
Dose volume (0.25 mg)
Needle
Activation mechanism
Needle shield
Dose button
Must be attached before injection: Ypsomed 
Clickfine® AutoProtect, 
8 mm x 29G*
Integrated and hidden, 6 mm, 29 G
0.5 mL
0.19 mL
123 mm
138 mm
17 mm
19 mm
White
Light blue
DV3396 product
PDS290 product
Single
Multiple
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number based on the randomization list produced by Novo Nordisk. The randomization 
number encodes the assignment to one of the 4 sequences. The randomization list will 
be transferred to the 
 Pharmacy and kept in a restricted area to which only the 
 
Pharmacy has access.
Subjects who drop out or withdraw for any reason without completing all screening 
evaluations successfully will be considered screening failures. Such subjects, and also 
subjects who are eligible for inclusion in the study but do not receive the study drug, will 
not receive a subject number, and only applicable data will be entered in the electronic 
case report forms (eCRFs). 
4.4.4
Selection of Doses in the Study
The lowest available dose for DV3396 product is 0.25 mg. Hence, to compare the 
DV3396 and PDS290 products, a dose of 0.25 mg has been chosen for both products. 
The subject will receive 2 injections at least 30 minutes apart giving a total dose of 
0.5 mg, which is the highest single dose of semaglutide tolerated without dose escalation 
due to risk of GI AEs.
4.4.5
Timing of Doses in the Study
The first dose will be administered as follows. A blinded 
 staff member who is 
present in the study room will start a timer when the unblinded staff member starts the 
injection. The timer will be placed such that it is not visible to the subject. The subject 
will be instructed to indicate when all pain is gone. If no pain is experienced, “00 min, 00 
s (sec)” will be recorded.
One minute after the injection, the blinded staff member will administer the visual 
analogue scale (VAS) to rate the intensity of injection site pain (not the momentary pain 
at the time of VAS completion) (see Section 4.5.1.1.1).
When the subject has completed the intensity of injection site pain VAS after the first 
injection of the study drug, the subject will complete a categorical assessment of injection 
site pain intensity (6-point scale) (see Section 4.5.1.1.2) and then a quality of pain 
modified SF-MPQ-2 inventory (see Section 4.5.1.1.3). 
Because the completion of the inventories may distract the subject, they will be reminded 
at regular intervals to indicate when the pain, if any, is gone, if they have not already 
done so.
At least 30 minutes after the first injection, after confirmation from the subject that the 
pain is entirely gone (to not contaminate the rating of the last injection), the timer will be 
reset, and the last injection will be given. If the pain is not gone at 30 minutes, another 
attempt will be made at 60 minutes. If pain is still present at that time, no further attempts 
will be made.
Following the last injection, the same measurements will be performed as after the first 
injection, as described above. Subjects will not be allowed to consult their own prior 
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ratings. After confirmation from the subject that the pain is entirely gone, the subjects will 
complete a comparative pain assessment (5-point scale) comparing pain between 
injections (see Section 4.5.1.1.4).
During the 30 minutes after each administration of the study drug the subjects should 
stay in bed or should be seated in a chair, and not walk around.
4.4.6
Meals During the Study
There are no requirements with regard to fed or fasting conditions prior to or after study 
drug administration. Further, no fasting is required before obtaining clinical laboratory 
samples.
With the exception of the restrictions with respect to methylxanthine- and 
alcohol-containing beverages or food, as described in Section 4.4.8, there are no special 
requirements related to food and beverage intake. Meals and snacks (such as 
decaffeinated coffee, herbal tea, fruit, biscuits) will be provided according to 
 
standard operating procedures (SOPs).
4.4.7
Blinding
This is a double-blind study.
Investigators will remain blinded to each subject's assigned product throughout the 
course of the study. Also, all 
 staff not participating in unblinded tasks will be blinded 
to treatment. To maintain this blind, qualified, unblinded members of the 
 staff not 
otherwise involved in the study procedures will be responsible for study drug 
administration according to the randomization list. Technical complaints will also be 
collected by unblinded site staff. The unblinded 
 staff will not be involved in any 
other assessments or safety reporting.
Subjects will be blinded to treatment as well. To maintain blinding of subjects, a visual 
blind will be in place during dose administration. To maintain blinding of 
 staff 
present in the room during administration (for timekeeping, etc.), an acoustic blind will 
be in place (eg, headphones or white noise).
The randomization list will be provided to the pharmacist at 
 for dispensing purposes 
and kept in the pharmacy, accessible to the pharmacist, the pharmacy assistant, and the 
unblinded staff only.
Individual code break envelopes will be provided for all subjects by Novo Nordisk. Each 
sealed envelope containing the sequence will be kept in a medication storage room, 
which is locked with restricted access. To manage the subject’s condition in case of a
medical emergency, the Investigator is allowed to break the code to know which 
semaglutide formulation was administered (using which product). The date when and 
reason why the blind was broken must be recorded in the source documentation. The 
Sponsor (Global Safety department) and unblinded monitor will be informed in case of 
unblinding. 
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4.4.8
Concomitant Medication and Other Restrictions During the Study
Note: Restrictions that apply to the period before admission are described in 
Section 4.3.1 and Section 4.3.2. 
The use of all prescription medicinal products or non-prescription drugs or herbal 
products is not allowed from admission to the clinical research center until discharge. An 
exception is made for routine vitamins, topical medication, and contraceptives, which are 
allowed throughout the study. During the stay in the clinical research center, the 
Investigator may permit a limited amount of paracetamol for the treatment of headache 
or any other pain, but this is only allowed after all pain assessments have been 
completed. Other medication to treat AEs during the stay in the clinical research center 
may only be prescribed if deemed necessary by the Investigator. If medication is used
during the stay in the clinical research center, the name of the drug, the dose, and 
dosage regimen will be recorded in the eCRF. Any medication taken between discharge 
and the follow-up phone call will be documented in a similar fashion.
The use of alcohol, methylxanthine-containing beverages or food (coffee, tea, cola,
chocolate, energy drinks), grapefruit (juice), and tobacco products is not allowed during 
the stay in the clinical research center. 
Strenuous exercise is not allowed during the stay in the clinical research center.
No contraception measures are required for male subjects as the risk of 
teratogenicity/fetotoxicity caused by transfer of semaglutide in seminal fluid is unlikely.
Female subjects of childbearing potential who have a fertile male sexual partner must 
agree to use highly effective contraception from screening up to 4 weeks after drug 
administration. Highly effective contraception is defined as: 
x 
Use of combined (estrogen- and progestogen-containing) hormonal contraception 
associated with inhibition of ovulation (oral, intravaginal, or transdermal [patch]), or
x
Use of progestogen-only hormonal contraception associated with inhibition of 
ovulation (oral or injectable).
The above requirements on contraception do not apply if:
x
The male partner of the female subject is vasectomized and has a confirmed 
post-vasectomy semen analysis.  
x
The female subject practices sexual abstinence during the period mentioned above. 
4.4.9
Treatment Compliance
Each of the single doses will be administered by trained unblinded staff at the clinical 
research center. The exact times of study drug administration will be recorded in the 
eCRF. 
4.5
Safety Measurements and Variables
Only safety parameters will be assessed in this study; the study does not comprise 
efficacy, pharmacokinetic, or pharmacodynamic measurements.
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4.5.1
Safety Measurements Assessed and Schedule of Assessments
A schedule of assessments is presented in Table 1. 
4.5.1.1
Safety and Tolerability Measurements
Safety and tolerability assessments will consist of intensity of injection site pain VAS, 
categorical assessment of injection site pain intensity (6-point scale), quality of pain 
modified SF-MPQ-2 inventory, duration of pain
assessment, comparative pain 
assessment (5-point scale), AEs, clinical laboratory, vital signs, and 12-lead 
electrocardiogram (ECG). In addition, some assessments will only be performed at 
screening or predose for the assessment of eligibility and are also described in this 
section.
All assessments will be performed in accordance with the schedule of assessments
(Table 1). 
4.5.1.1.1
Intensity of Injection Site Pain Visual Analogue Scale
The intensity of injection site pain VAS will be filled in by the subjects starting 1 minute 
after each injection of the study drug to report the pain intensity experienced in the first 
minute after the injection. 
Subjects will be instructed to make a vertical line on the VAS (100 mm); the endpoints 
are “no pain” and “unbearable pain.” The distance (mm) between the endpoint “no pain” 
and the vertical line on the VAS will be recorded in the eCRF.
First injection
English version:
Indicate the intensity of pain you felt during the injection. For this, make a vertical line 
somewhere through the line below, where the endpoints are “no pain” and “unbearable 
pain”.
Last injection
English version:
Indicate the intensity of pain you felt during the last injection only. For this, make a 
vertical line somewhere through the line below, where the endpoints are “no pain” and 
“unbearable pain”.
No pain
Unbearable pain
100 mm
No pain
Unbearable pain
100 mm
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4.5.1.1.2
Categorical Assessment of Injection Site Pain Intensity
The categorical assessment of injection site pain intensity is a 6-point scale in which the 
pain intensity is rated by the subjects as follows: 
Please tick one box describing the pain intensity during the injection:
none
Ƒ
very mild
Ƒ
mild 
Ƒ
moderate 
Ƒ
severe 
Ƒ
very severe
Ƒ
Please tick one box describing the pain intensity during the last injection only:
none
Ƒ
very mild
Ƒ
mild 
Ƒ
moderate 
Ƒ
severe 
Ƒ
very severe
Ƒ
4.5.1.1.3
Quality of Pain Modified SF-MPQ-2 Inventory
The quality of pain will be assessed using the modified short-form McGill pain 
questionnaire (modified SF-MPQ-2 inventory2) to be filled in by the subjects as follows: 
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This questionnaire provides you with a list of words that describe some of the different 
qualities of pain and related symptoms. Please put an X in all boxes that apply:
Throbbing pain
Ƒ
Shooting pain
Ƒ
Stabbing pain
Ƒ
Sharp pain
Ƒ
Cramping pain
Ƒ
Gnawing pain 
Ƒ
Hot-burning pain
Ƒ
Aching pain
Ƒ
Heavy pain
Ƒ
Tender
Ƒ
Splitting pain
Ƒ
Tiring-exhausting 
Ƒ
Sickening
Ƒ
Fearful
Ƒ
Punishing-cruel
Ƒ
Electric-shock pain
Ƒ
Cold-freezing pain
Ƒ
Piercing
Ƒ
4.5.1.1.4
Duration of Pain
The duration of pain as indicated by the subject will be recorded from time of injection 
until cessation of pain in minutes and seconds. 
4.5.1.1.5
Comparative Pain Assessment (5-point scale)
The comparative pain assessment is a 5-point scale in which the subjects will compared 
the pain between the 2 injections.
The subjects will be asked to answer the following question:
Please choose the statement below that best describes your experience:
x
The last injection hurt much more than the first injection.  
x
The last injection hurt more than the first injection.  
x
They hurt about the same (includes: none of the injections hurt). 
x
The last injection hurt less than the first injection. 
x
The last injection hurt much less than the first injection.
4.5.1.1.6
Adverse Events
The definitions of AEs and serious AEs (SAEs) can be found in Appendix 9.2.
The Investigator is responsible for detecting, documenting, recording, and following up 
on events that meet the definition of an AE or SAE.  
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Time period and frequency for collecting AE and SAE information
All AEs will be collected from the first study-related activity after obtaining informed 
consent and until the follow-up visit at the time points specified in the schedule of 
assessments (Table 1).  
All SAEs will be recorded and reported to Novo Nordisk or designee within 24 hours, as 
indicated in Appendix 9.2. The investigator must submit any updated SAE data to Novo 
Nordisk within 24 hours of it being available.
Investigators are not obligated to actively seek AEs or SAEs in former study subjects. 
However, if the Investigator learns of any SAE, including a death, at any time after a 
subject has been discontinued from/completed the study, and the Investigator considers 
the event to be possibly/probably related to the investigational study product or study 
participation, the investigator must promptly notify Novo Nordisk.
The method of recording, evaluating, and assessing causality of AEs and SAEs and the 
procedures for completing and transmitting SAE reports are provided in Appendix 9.2. 
Timelines for reporting of AEs are listed in Figure 2. 
Medication error AEs require additional data collection via a specific event form. For 
definition of medication errors, see Appendix 9.2.
In order for Novo Nordisk to perform a complete investigation of reported SAEs, Novo 
Nordisk might ask the Investigator to complete a technical complaint form. For reporting 
of technical complaints, see Section 4.5.1.1.7.
The reporting timelines are specified in Figure 2. 
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Figure 2 
Decision Tree for Determining the Event Type and the Respective 
Forms to Complete with Associated Timelines
Method of detecting AEs and SAEs
Care should be taken not to introduce bias when detecting AEs and/or SAEs. 
Open-ended and non-leading verbal questioning of the subject is the preferred method 
to inquire about events.
Follow-up on AEs and SAEs
After the initial AE/SAE report, the Investigator is required to proactively follow each 
subject at subsequent visits/contacts. All SAEs will be followed until resolution, 
stabilization, or if the event is otherwise explained (eg, chronic condition), or the subject 
is lost to follow-up. Further information on follow-up procedures is given in Appendix 9.2. 
Regulatory reporting requirements for SAEs
Prompt notification by the Investigator to Novo Nordisk of an SAE (according to Figure 2
and Appendix 9.2) is essential so that legal obligations and ethical responsibilities
towards the safety of subjects and the safety of a study product under clinical 
investigation are met. 
Novo Nordisk has a legal responsibility to notify both the local regulatory authority and 
other regulatory agencies about the safety of a study product under clinical investigation. 
Novo Nordisk will comply with country-specific regulatory requirements relating to safety 
reporting to the regulatory authority, the Independent Ethics Committee (IEC), and 
investigators. 
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Safety reports must be prepared for suspected unexpected serious adverse reactions 
according to local regulatory requirements and Novo Nordisk policy and forwarded to 
regulatory authorities, IEC, and investigators as locally required. 
An Investigator who receives an investigator safety report describing a SAE or other 
specific safety information (eg, summary or listing of SAEs), from Novo Nordisk will 
review and then file it along with the IB and will notify the IEC, if appropriate, according 
to local requirements.
Pregnancies and associated adverse events
Details of pregnancies in female subjects will be collected after the first study-related 
activity after obtaining informed consent and until pregnancy outcome.
If a pregnancy is reported in a female subject, the investigator should inform Novo 
Nordisk within 14 calendar days of learning of the pregnancy and should follow the 
procedures outlined below: 
x
The Investigator will collect pregnancy information on any female subject, who 
becomes pregnant while participating in this study.
x
Information will be recorded on the appropriate form and submitted to Novo Nordisk 
within 14 calendar days of learning of a subject's pregnancy.
x
Subject will be followed to determine the outcome of the pregnancy. The Investigator 
will collect follow-up information on the subject and neonate, which will be forwarded 
to Novo Nordisk. Generally, follow-up will not be required for longer than 1 month 
beyond the delivery date.
x
Any termination of pregnancy will be reported, regardless of fetal status (presence 
or absence of anomalies) or indication for procedure.
x
Although pregnancy itself is not considered to be an AE or SAE, any pregnancy 
complication or elective termination of a pregnancy will be reported as an AE or SAE. 
x
A spontaneous abortion is always considered to be an SAE and will be reported as 
such.
x
Any SAE occurring as a result of a post-study pregnancy that is considered 
possibly/probably related to the study product by the Investigator will be reported to 
Novo Nordisk as described in Appendix 9.2. Although the Investigator is not 
obligated to actively seek this information in former subjects, he or she may learn of 
an SAE through spontaneous reporting. 
Any female subject who becomes pregnant while participating in the study will 
discontinue study product.
Pregnancy outcome should be documented in the subject's medical record. An abnormal 
pregnancy outcome (eg, spontaneous abortion, fetal death, stillbirth, congenital 
anomaly, and ectopic pregnancy) is considered an SAE.
4.5.1.1.7
Technical Complaints
The unblinded 
 staff is responsible for the detection and documentation of technical 
complaints that occur when using the pen-injectors used in this study. 
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Definition of technical complaint:
x
A technical complaint is any written, electronic or oral communication that alleges 
medical product defects and device product defects. The technical complaint may 
be associated with an AE, but does not concern the AE itself.
Examples of technical complaints are as follows:
x
Problems with the physical or chemical appearance of study products (eg, 
discoloration, particles or contamination).
x
Problems with packaging material including labelling.
x
Problems related to pen-injectors (eg, to the injection mechanism, dose setting 
mechanism, push button, or interface between the pen-injector and the needle).
The Investigator must assess whether a technical complaint is related to an AE. 
Time period for detecting technical complaints
All technical complaints that occur from the time of receipt of the product at 
 until 
the time of the last usage of the product must be collected for products predefined on 
the technical complaint form.
Reporting of technical complaints to Novo Nordisk
Technical complaints must be reported on a separate technical complaint form:
x
One technical complaint form must be completed for each affected dispensing unit 
number (DUN). 
x
If a DUN is not available, a technical complaint form for each batch, code, or lot 
number must be completed. 
Timelines for reporting of technical complaints to Novo Nordisk
The Investigator must complete as much information on the form as possible and hand 
it over to the unblinded 
 staff. The unblinded 
 staff must fill in the unblinded 
information on the form (such as the DUN and batch, code, or lot number) and forward 
the technical complaint form to the Customer Complaint Center, Novo Nordisk, within 
the timelines specified below:
x
Technical complaints related to SAEs, within 24 hours
x
All other technical complaints, within 5 calendar days
The technical complaint form must be kept inaccessible for blinded site staff.
Follow-up of technical complaints
The Investigator is responsible for ensuring that new or updated information will be 
recorded on the originally completed form. 
Collection, storage, and shipment of technical complaint samples
The unblinded 
 staff must collect the technical complaint sample and all associated 
parts that were packed in same DUN. The unblinded 
 staff collects the sample and 
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notify the unblinded monitor within 5 calendar days of obtaining the sample at 
 The 
sample and all associated parts must be sent as soon as possible to Customer 
Complaint Center, Novo Nordisk, together with a copy of the completed technical 
complaint form.
The technical complaint sample should contain the batch, code, or lot number and, if 
available, the DUN. If the technical complaint sample is unobtainable, the reason must 
be stated on the technical compliant form.
If several samples are shipped in one shipment, the sample and the corresponding 
technical complaint form should be kept together.
Storage for the technical complaint sample must be done in accordance with the 
conditions prescribed for the product.
Reporting of technical complaints for Novo Nordisk products not included in technical 
complaint form
Technical complaints on Novo Nordisk products not included in the technical complaint 
form should be reported to local Novo Nordisk affiliate with a reference to trial ID.
4.5.1.1.8
Clinical Laboratory 
Blood and urine samples for clinical laboratory assessments will be collected according 
to 
 SOPs. No fasting is required prior to collecting these samples. 
The following parameters will be measured:
x
Clinical chemistry (serum quantitatively):
total bilirubin, alkaline phosphatase, aspartate aminotransferase (ASAT), alanine 
aminotransferase (ALAT), creatinine, albumin, sodium, potassium, and calcium.  
x
Hematology (blood quantitatively):
leukocytes, hemoglobin, and thrombocytes.
HbA1c at screening only.
x
Serology:
hepatitis B surface antigen (HBsAg), anti–hepatitis C virus (HCV), and anti–human 
immunodeficiency virus (HIV)-1 and -2. 
x 
Drug and alcohol screen:
opiates, methadone, cocaine, amphetamines (including ecstasy), cannabinoids, 
barbiturates, benzodiazepines, tricyclic antidepressants, and alcohol in urine. 
x
Hormones (serum quantitatively):
thyroid-stimulating hormone.
x
Pregnancy test (female subjects of childbearing potential only):
ȕ-human chorionic gonadotropin in urine.
In case of unexplained or unexpected clinical laboratory test values, the tests will be 
repeated as soon as possible and followed up until the results have returned to the 
normal range and/or an adequate explanation for the abnormality is found. The clinical 
laboratory will clearly mark all laboratory test values that are outside the normal range, 
and the Investigator will indicate which of these deviations are clinically significant. 
Clinically significant laboratory result deviations after study drug administration will be 
recorded as AEs, and the relationship to the treatment will be indicated (see also 
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Appendix 9.2). Clinically significant laboratory results at screening will not be reported 
as AE as this is considered part of medical history.
4.5.1.1.9
Vital Signs
Systolic and diastolic blood pressure and pulse will be recorded after the subject has 
been resting for at least 5 minutes in the supine position. These assessments will be 
made using an automated device. 
4.5.1.1.10 Electrocardiogram
A standard 12-lead ECG will be recorded after the subject has been resting for at least 
5 minutes in the supine position. The ECG will be recorded using an ECG machine 
equipped with computer-based interval measurements. The following ECG parameters 
will be recorded: heart rate, PR-interval, QRS-duration, QT-interval, QTc-interval 
(Fridericia's), and the interpretation of the ECG profile by the Investigator. 
4.5.1.1.11 Physical Examination
Physical examination will be performed according to 
 SOPs. In addition, body weight 
and height will be measured according to 
 SOPs.
4.5.1.2
Total of Blood Volume
Table 7 presents the number and volume of blood samples and the total volume of blood 
that will be collected throughout the study per subject.
If deemed necessary by the Investigator or the Sponsor, the number and/or volume of 
blood samples per assessment may be increased as long as the total volume of blood 
drawn for a subject does not surpass 500 mL. 
Table 7 
Number and Volume of Blood Samples and Total Blood Volume 
Collected per Subject
Assessment
Maximum # 
Samples
Volume of Blood per 
Sample (mL)
Total Volume of 
Blood (mL)
Clinical Chemistry
2 
3.5
7 
Hematology 
2 
3 
6 
Serology
1 
5 
5 
Total Volume of Blood Drawn
18
4.5.2
Appropriateness of Measurements
The assessments that will be made in this study are standard and generally recognized
as reliable, accurate, and relevant. 
4.5.3
Safety Variables
The safety variables to be measured include but are not limited to the variables as given 
below. A complete list of safety variables will be provided in the statistical analysis plan
(SAP). 
x
Intensity of injection site pain VAS
x
Categorical assessment of injection site pain intensity (6-point scale)
x
Quality of pain modified SF-MPQ-2 inventory
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x
Duration of pain
x
Comparative pain assessment (5-point scale)
x
AEs
x
Clinical laboratory
(if postdose assessments are deemed relevant by the 
Investigator)
x
Vital signs
x
ECG (if postdose assessments are deemed relevant by the Investigator)
x
Physical examination (if postdose assessments are deemed relevant by the 
Investigator)
4.6
Statistical Procedures and Determination of Sample Size
4.6.1
Analysis Sets
4.6.1.1
Safety Set
All subjects who have received at least 1 injection of semaglutide (includes any skin 
contact with study product whether the injection was completed or not).
4.6.1.2
Full analysis set (FAS)
All randomized subjects who have received at least 1 injection of semaglutide (includes 
any skin contact with trial product whether the injection was completed or not). In 
exceptional cases, subjects from the FAS may be excluded. In such cases the exclusion 
will be justified and documented.
4.6.1.3
Per-Protocol Set
All subjects who have received both injections of semaglutide and have completed both 
intensity of injection site pain assessments. 
4.6.2
Statistical and Analytical Plan for Safety Evaluation
An SAP will be generated by the Biostatistics Department of 
 the SAP will be 
finalized prior to database lock. Full details of the analysis to be performed will be 
included in the SAP.
Any deviation from the SAP will be reported in the section “Changes in Planned Analysis” 
in the clinical study report (CSR). 
4.6.2.1
Evaluation of Safety and Tolerability
Safety and tolerability will be assessed through intensity of injection site pain VAS, 
quality of pain modified SF-MPQ-2 inventory, duration of pain assessment, comparative 
pain assessment, AEs, clinical laboratory, vital signs, ECGs, and physical examination 
findings (if results are available after dosing), and any other parameter that is relevant 
for safety assessment.
4.6.2.1.1
Intensity of Injection Site Pain Visual Analogue Scale
The primary endpoint is given in Table 8.
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Table 8 
Primary Endpoint
Endpoint Title
Time Frame
Unit
Intensity of injection site 
pain
1 min after each injection
(Day 1)
mm on a 100 mm horizontal 
visual
analogue 
scale 
(where 0 mm = no pain, 
100 mm = unbearable pain)
The primary endpoint will be analyzed by a fixed analysis of variance model with VAS 
score as the dependent variable, and product, injection side (right side, left side), 
injection number (first injection, second injection), and subject as fixed effects.. From the 
model, the difference in VAS score between the 2 products is estimated and presented 
with a 95% CI and a p-value. The interpretation of the difference in VAS pain scores will 
be supported and contextualized by the results of the analyses of the exploratory 
endpoints.
4.6.2.1.2
Categorical Assessment of Injection Site Pain Intensity, Quality of Pain Modified 
SF-MPQ-2 Inventory, Duration of Pain, and Comparative Pain Assessment
The exploratory endpoints are given in Table 9.
Table 9 
Exploratory Endpoints
Endpoint Title
Time Frame
Unit
Categorical 
assessment 
of 
injection site pain intensity
Immediately after rating of
intensity of injection site pain, 
ie, after 1 min plus the time it 
takes to complete the rating of 
intensity of injection site pain by 
visual analogue scale (Day 1)
6-point scale (none – very mild 
– mild – moderate – severe – 
very severe)
Moderate or more injection site 
pain
Dichotomous variable based 
on categorical assessment of 
intensity of injection site pain 
(Day 1) (item above)
Yes = moderate, severe, very 
severe
No = none, very mild, mild
Quality of pain
Immediately after categorical 
assessment of intensity of 
injection site pain, ie, after 1 
min plus the time it takes to 
complete the rating of intensity 
of injection site pain and the 
categorical rating of intensity of 
injection site pain (Day 1)
Pain quality items on modified 
SF-MPQ-2 inventory (select all 
that apply):
Throbbing pain տ
Shooting pain տ
Stabbing pain տ
Sharp pain տ
Cramping pain տ
Gnawing pain տ
Hot-burning pain տ
Aching pain տ
Heavy pain տ
Tender տ
Splitting pain տ
Tiring-exhausting տ
Sickening տ
Fearful տ
Punishing-cruel տ
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Endpoint Title
Time Frame
Unit
Electric-shock pain տ
Cold-freezing pain տ
Piercing տ
Duration of pain
From time of injection until 
cessation of pain (Day 1)
min and sec
Comparative pain experience
After pain has ended after the 
last injection (Day 1)
5-point scale (the last injection 
hurt much more than the first 
injection – the last injection hurt 
more than the first injection – 
they hurt about the same 
[includes: none of the injections 
hurt] – the last injection hurt 
less than the first injection – the 
last injection hurt much less 
than the first injection) 
The DV3396 product hurt less 
than or about the same as the 
PDS290 product
Dichotomous variable based 
on 
“Comparative 
pain 
experience” 
variable 
above
(Day 1)
Yes 
= 
“about 
the 
same 
(includes none of the products 
hurt” or “the DV3396 product 
hurt less than the PDS290 
product” 
or 
“the 
DV3396 
product hurt much less than the 
PDS290 product”)
No= “The DV3396 product hurt 
much more than the PDS290
product” 
or 
“The 
DV3396 
product hurt more than the 
PDS290 product”
The results will be listed, and they will be presented descriptively, where applicable.
4.6.2.1.3
Adverse Events
A listing of all individual AEs will be provided. Summary tables of treatment-emergent 
adverse events (TEAEs) will be presented by system organ class based on the Medical 
Dictionary for Regulatory Activities terminology list (preferred terms): one containing the 
number of TEAEs (frequency of occurrence, number of subjects experiencing the event) 
by treatment and one containing the number of drug-related TEAEs (frequency of 
occurrence, number of subjects experiencing the event) per treatment. Additional tables 
of total counts by treatment and relationship and by treatment and severity will be given.
4.6.2.1.4
Technical Complaints
A listing of all technical complaints will be provided. 
4.6.2.1.5
Clinical Laboratory
Clinical laboratory data will be listed accompanied by an indication if the parameter is 
outside the reference range. A summary of all data outside the reference range of the 
clinical laboratory will be provided. Clinical laboratory data will be presented 
descriptively, where applicable.
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4.6.2.1.6
Vital Signs and Electrocardiograms
Vital signs and ECG parameters will be listed, and they will be presented descriptively, 
where applicable. 
4.6.2.1.7
Physical Examination
Changes from baseline for physical examination (if available after dosing) will be 
described and listed.
4.6.3
Determination of Sample Size
The sample size calculation considers both the primary endpoint and the dichotomous 
endpoints. Considering precision on the proportion of the dichotomous endpoints, a 
sample size of 100 subjects results in a confidence interval (CI) with a width of 10% on 
each side, eg, for a proportion of 60%, the CI is 60% (50%-70%).
A standard deviation of within-pair differences in VAS pain score between 
2 subcutaneous injections was obtained from Novo Nordisk clinical trial INS-4011. The 
within-subject variance (adjusting for the effects of speed, volume and injection region) 
was estimated to 348, corresponding to a standard deviation (SD) of the within-pair 
GLIIHUHQFH RI ¥Â    PP $ prospective study of acute pain found that the 
minimum clinically important difference (MCID) for worsening of pain, ie, the difference 
on a VAS associated with the selection of “a little more pain” rather than “about the same” 
had a mean value of 10 mm.3  
With 100 subjects, a conservative standard deviation of 30 mm, and 90% power, it is 
possible to detect a difference smaller than 10 mm (Table 10). Hence a sample size of 
100 subjects is considered sufficient to detect a clinically important difference. Allowing 
for 4% dropouts or missing data, a total of 104 subjects will need to be randomized.
Table 10
Detectable Difference with a Sample Size of 100 Subjects for Different 
Combinations of Power, and Standard Deviation
Number of 
Subjects
Power
Standard
Deviation
Difference
100
0.8
25 mm
7.1 mm
100
0.8
30 mm
8.5 mm
100
0.9
25 mm
8.2 mm
100
0.9
30 mm
9.8 mm
4.6.4
Interim Analysis
After the last subject has completed Day 1, all data collected will be released unblinded 
for analysis notwithstanding that the subjects have not completed the safety follow-up
phone call.
4.7
Data Quality Assurance
The study may be audited by the Quality Assurance Department at 
 to assess 
adherence to the clinical study protocol (CSP) and Quality System. During the conduct 
of the study, process-related audits may be performed. An audit certificate will be 
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provided in the appendices of the final CSR outlining any audits and other related 
activities performed. 
The clinical research site will be monitored by the study blinded monitor and unblinded 
monitor to ensure correct performance of the study procedures and assure that the study 
is conducted according to the relevant regulatory requirements. The eCRF entries will 
be verified with the source documentation, if applicable (in some cases there are no 
source pages, therefore verification is not necessary). 
Regulatory authorities, the IEC, and/or the Sponsor’s clinical quality assurance group 
may request access to all source documents, eCRFs, and other study documentation 
for on-site audit or inspection. Direct access to these documents must be guaranteed by 
the Investigator, who must provide support at all times for these activities.
Quality control principles will be applied throughout the performance of this study. 
Review procedures will be followed at 
 for all documents that are generated in 
relation to the study.
An explanation will be given for all missing, unused, and spurious data in the relevant 
sections of the CSR.
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5.
ETHICS
5.1
Independent Ethics Committee
The CSP and the ICFs will be submitted for review and approval by the IEC of the 
foundation “Beoordeling Ethiek Biomedisch Onderzoek” (English translation: “Evaluation 
of Ethics in Biomedical Research”) (
 The 
Netherlands) prior to the eligibility screening. The composition of the IEC is in 
accordance with the recommendations of the World Health Organization, the 
International Council for Harmonisation (ICH) E6 Guideline for Good Clinical Practice 
(GCP)4, and the EU Clinical Trial Directive (CTD) (Directive 2001/20/EC)5 (see below).
will keep the IEC informed about the progress of the study. All changes in research 
activities and all unanticipated problems involving risks to human subjects will be 
immediately reported to the IEC. In accordance with Section 10, Subsection 1, of the 
Dutch law on Medical Research in Human Subjects (WMO, revised December 2015)6, 
will inform the subjects and the IEC if anything occurs on the basis of which it 
appears that the disadvantages of participation may be significantly greater than was 
foreseen in the research proposal, or if further recruitment of subjects in the study has 
been put on hold for that reason, whichever occurs first. The study may be suspended 
pending further review by the IEC, except insofar as suspension would jeopardize the 
subjects’ health. 
will take care that all subjects are kept informed. 
No changes will be made in the study without IEC approval, except when required to 
eliminate apparent immediate hazards to human subjects.
Notification of the end of the study will be sent by 
 to the Competent Authority (CA)
in The Netherlands and to the IEC within 90 days after completion of follow-up for the 
last subject. In case a study is temporarily halted, 
 will notify the IEC immediately, 
including the reason for this. In case a study is ended prematurely, 
 will notify the 
IEC and the CA in The Netherlands within 15 days, including the reasons for the 
premature termination. A summary of the results of the study will be sent by 
 to the 
CA and the IEC within 1 year after the end of the study.
5.2
Ethical Conduct of the Study
This study will be conducted in accordance with the ethical principles that have their 
origin in the World Medical Association (WMA) Declaration of Helsinki, adopted by the 
18th
WMA General Assembly,
Helsinki, Finland, June 1964, and subsequent 
amendments.7
This study is also designed to comply with ICH E6 Guideline for GCP (EMA/Committee 
for Medicinal Products for Human Use [CHMP]/ICH/135/1995)4 and the EU CTD 
Directive 2001/20/EC5, as incorporated into Dutch Law.6
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Guidelines adopted by the ICH and other relevant international guidelines, 
recommendations, and requirements will be taken into account as comprehensively as 
possible, as long as they do not violate local law.
Whenever the term “Investigator” is noted in the CSP text, it may refer to either the
Investigator at the site or an appropriately qualified, trained, and delegated individual of 
the investigational site.
5.3
Subject Information and Consent
All subjects will be informed verbally and in writing regarding the objectives, procedures, 
and risks of study participation. The subjects will have to sign the Dutch or English 
version of the ICF before any study-related procedures are started. The ICF contains 
information about the objectives of the study, the procedures followed during the study,
and the risks and restrictions of the study, with special reference to possible side effects 
of the medication and potential interactions. In addition, insurance coverage provided 
during the study is explained. The elements addressed in the ICF are according to the 
ICH E6 Guideline for GCP (EMA/CHMP/ICH/135/1995).4
5.4
Privacy
All personal details will be treated as confidential by the Investigator and staff at 
and the handling of personal data will be in compliance with the EU General Data 
Protection Regulation (GDPR).8
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6.
STUDY ADMINISTRATIVE STRUCTURE
6.1
Distribution of Activities
Supply of Study Drug
Novo Nordisk A/S Denmark will supply the DV3396 products and the PDS290 products
for use in the study.
The PDS290 product will be used with an Ypsomed Clickfine AutoProtect, 8 mm x 29G 
needle. This needle will be sourced directly by 
.
Laboratory Assessments 
The analysis of clinical laboratory samples will be performed at the 
Clinical 
Laboratory. 
eCRF Design
The eCRF design will be performed with the computer program Oracle Clinical 
 
 by the Database Programming Department 
of 
Data Management
Data management will be performed with the computer programs Oracle Clinical 
 by the Data Management Department of 
.
Statistics
An SAP will be generated by the Biostatistics Department of 
 The safety analysis 
will be conducted by the Biostatistics Department of 
 Statistical analysis will be 
performed with the computer program SAS (
). 
CSR Writing
The CSR, structured in accordance with the guideline “Structure and Content of Clinical 
Study Reports - ICH E3”9, will be written by 
6.2
Documentation
6.2.1
Archiving
All documents concerning the study will be kept on file in the Central Archives of 
 
for at least 15 years after conduct of the study. The Sponsor will receive the completed 
eCRFs (upon request, as PDF file).
6.2.2
Recording of Data in Source Documents and eCRFs
Wherever possible, all data will be entered directly into the eCRFs. Source documents 
will be used in some cases.
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A data management plan will be written by the Data Management Department of 
, 
which will be finalized prior to performing any data validation. An appendix to the data 
management plan (Origin of Source Data List for Data Entry) will identify any data to be 
recorded directly in the eCRF (ie, no prior written or electronic record of data) and which 
data should be considered source data. 
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7.
CONFIDENTIALITY AND PUBLICATION POLICY
By signing this CSP, the Investigator reaffirms to the Sponsor that he will maintain in 
confidence all information furnished to him or resulting from this study. The Investigator
will only divulge such information as may be necessary to the IEC, the members of the 
staff, and the subjects who are involved in this study.
The primary completion date (PCD) is the last assessment of the primary endpoint, and 
is for this study Day 1. If the last subject is withdrawn early, the PCD is considered the 
date when the last subject would have completed Day 1. The PCD determines the 
deadline for results disclosure at clinicaltrials.gov according to the Food and Drug 
Administration Amendments Act. 
7.1
Publication Policy
Novo Nordisk commits to communicate and disclose results of trials regardless of 
outcome. Disclosure includes publication of a manuscript in a peer-reviewed scientific 
journal, abstract submission with a poster or oral presentation at a scientific meeting or 
disclosure by other means. The results of this trial will be subject to public disclosure on 
external web sites according to international and national regulations. Novo Nordisk 
reserves the right to defer the release of data until specified milestones are reached, for 
example when the clinical trial report is available. This includes the right not to release 
the results of interim analyses, because the release of such information may influence 
the results of the entire trial. At the end of the trial, one or more scientific publications 
may be prepared collaboratively by the investigator(s) and Novo Nordisk. Novo Nordisk 
reserves the right to postpone publication and/or communication for up to 30 days to 
protect intellectual property. In all cases the trial results will be reported in an objective, 
accurate, balanced and complete manner, with a discussion of the strengths and 
limitations. In the event of any disagreement on the content of any publication, both the 
investigators’ and Novo Nordisk opinions will be fairly and sufficiently represented in the 
publication.
7.2
Dissemination of Clinical Trial Data
Information of the trial will be disclosed at clinicaltrials.gov and novonordisk-trials.com. 
It will also be disclosed according to other applicable requirements such as those of the 
International Committee of Medical Journal Editors, the Food and Drug Administration 
Amendments Act, European Commission Requirements, and other relevant 
recommendations or regulations. If a subject requests to be included in the trial via the 
Novo Nordisk e-mail contact at these websites, Novo Nordisk may disclose the 
Investigator’s contact details to the subject. As a result of increasing requirements for 
transparency, some countries require public disclosure of Investigator names and their 
affiliations.
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8.
REFERENCES
1.
Novo Nordisk A/S. Investigator's Brochure, semaglutide s.c. for weight 
management and non-alcoholic steatohepatitis, projects NN9536 and NN9931 
(edition 4). Mar 2019.
2.
Dworkin RH, Turk DC, Revicki DA, et al. Development and initial validation of an 
expanded and revised version of the Short-form McGill Pain Questionnaire (SF-
MPQ-2). Pain. 2009;144:35-42.
3.
Todd KH, Funk KG, Funk JP, et al. Clinical significance of reported changes in 
pain severity. Ann Emerg Med.1996;27:485-89.
4.
International Council for Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, 
E6 
(R2): 
Guideline 
for 
Good 
Clinical 
Practice 
(EMA/CHMP/ICH/135/1995), 15 December 2016.
5.
Directive 2001/20/EC of the European Parliament and of the Council of 4 April 
2001 on the approximation of the laws, regulations and administrative provisions 
of the Member States relating to the implementation of good clinical practice in 
the conduct of clinical trials on medicinal products for human use. 
6.
Medical 
Research 
Involving 
Human 
Subjects 
Act 
(WMO, 
Wet 
Medisch-Wetenschappelijk Onderzoek met Mensen), revision December 2015.
7.
WMA Declaration of Helsinki (18th WMA General Assembly 1964), revised at 
64th World Medical Association General Assembly, Fortaleza, Brazil, October 
2013. 
8.
The General Data Protection Regulation (GDPR). Regulation (EU) 2016/679 of 
the European Parliament and the Council of the European Union, 27 April 2016. 
9.
International Council
for
Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, 
E3: 
Structure 
and 
Content 
of 
Clinical 
Study 
Reports 
(CPMP/ICH/137/95), November 1995. 
10.
International Council
for
Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, E2A: Clinical Safety Data Management: Definitions and Standards for 
Expedited Reporting, Note for Guidance on Clinical Safety Data Management, 
June 1995. 
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9.
APPENDICES
9.1
Drug Accountability
x
Study product storage, in-use conditions and in-use time will be available on the label 
and in the TMM. The Investigator must confirm that appropriate temperature 
conditions have been maintained during transit for all study products received and 
any discrepancies are reported and resolved before use of the study products.
x
All study products must be stored in a secure, controlled, and monitored (manual or 
automated) area in accordance with the labelled storage conditions with access 
limited to the pharmacist and pharmacy assistant. 
x
The Investigator must inform Novo Nordisk immediately if any study product has 
been stored outside specified conditions. Additional details regarding handling of 
temperature deviations can be found in the TMM. 
x
Study product that has been stored improperly must not be dispensed to any subject 
before it has been evaluated and approved for further use by Novo Nordisk.
x
The Investigator is responsible for drug accountability and record maintenance (ie, 
receipt, accountability and final disposition records).
x
For study products packed unblinded in a blinded study: Drug accountability will be 
performed by an unblinded person who will ensure that unblinding information such 
as batch numbers are not revealed to personnel that should remain blinded).
x
Drug accountability should be performed at dose level unless this is not possible (eg, 
products in pen-injectors), then describe and explain the actual level of 
accountability.
9.2
Adverse Events: Definitions and Procedures for Recording, Evaluation, 
Follow-up, and Reporting to Sponsor
AE definition
x
An AE is any untoward occurrence in a clinical trial subject that is temporally associated 
with the use of a medicinal product, whether or not considered related to the medicinal 
product.
x
An AE can be any unfavorable and unintended sign, including an abnormal laboratory
finding, symptom, or disease (new or exacerbated) temporally associated with the use of 
a medicinal product. 
Events meeting the AE definition
x
Any abnormal laboratory test results or safety assessments, including those that worsen 
from baseline, considered clinically significant in the medical and scientific judgment of the 
investigator.
x
Exacerbation of a chronic or intermittent pre-existing condition including either an increase 
in frequency and/or intensity of the condition.
x
Signs, symptoms, or the clinical sequelae of a suspected drug-drug interaction.
x
Signs, symptoms, or the clinical sequelae of a suspected overdose of trial product 
regardless of intent.
Events NOT meeting the AE definition 
x
Pre-existing conditions, anticipated day-to-day fluctuations of pre-existing conditions, 
including those identified during screening or other trial procedures performed before 
exposure to trial product. 
Note: Pre-existing conditions should be recorded as medical history/concomitant illness.
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x
Pre-planned procedures, unless the condition for which the procedure was planned has 
worsened from the first trial-related activity after the subject has signed the informed 
consent.
x
Pain: Non-serious pain will be collected on specific pain-rating schemes please refer to 
the schedule of assessments (Table 1) and Section 4.5. 
If the event of pain fulfils the criteria for an SAE then in addition to the above, an AE form 
and a safety information form must also be filled in.
Definition of an SAE
An SAE is an AE that fulfils at least one of the following criteria:
x
Results in death
x
Is life-threatening
The term 'life-threatening' in the definition of 'serious' refers to an event in which the subject 
was at 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.
x
Requires inpatient hospitalisation or prolongation of existing hospitalisation
x
Hospitalization signifies that the subject has been detained at the hospital or 
emergency ward for observation and/or treatment that would not have been 
appropriate in the physician’s office or outpatient setting. Complications that occur 
during hospitalisation are AEs. If a complication prolongs hospitalization or fulfils any 
other serious criteria, the event is serious. When in doubt as to whether 
“hospitalization” occurred or was necessary, the AE should be considered serious.
x
Hospitalization for elective treatment of a pre-existing condition that did not worsen 
from baseline is not considered an AE.
Note:
x
Hospitalizations for administrative, trial-related and social purposes do not constitute 
AEs and should therefore not be reported as AEs or SAEs.  
x
Hospital admissions for surgical procedures, planned before trial inclusion, are not 
considered AEs or SAEs. 
x
Results in persistent disability/incapacity
x
The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions.
x
This definition is not intended to include experience of relatively minor medical 
significance such as uncomplicated headache, nausea, vomiting, diarrhoea, influenza, 
and accidental trauma (eg, sprained ankle), which may interfere with or prevent 
everyday life functions but do not constitute a substantial disruption.
x
Is a congenital anomaly/birth defect
x
Including events leading to fetal distress or fetal death
x
Important medical event:
x
Medical or scientific judgment should be exercised in deciding whether SAE reporting 
is appropriate in other situations. This includes important medical events that may not 
be immediately life-threatening or result in death or hospitalization, but may jeopardize
the subject or may require medical or surgical intervention to prevent one of the other 
outcomes listed in the above definition. These events should usually be considered 
serious and reported as SAEs using the important medical event criterion.
x
The following AEs must always be reported as SAEs using the important medical 
event criterion, if no other seriousness criteria are applicable:
x
suspicion of transmission of infectious agents via the trial product. 
x
risk of liver injury defined as alanine aminotransferase (ALT) or aspartate 
aminotransferase (AST) >3 x the upper limit of normal (ULN) and total bilirubin >2 x 
ULN, where no alternative etiology exists (Hy's law).
x
events that might have led to or might lead directly or indirectly to death or a 
serious deterioration in state of health
These are handled under the SAE reporting system.
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Description of AEs requiring additional data collection (via specific event form) 
Medication error:
A medication error concerning trial products is defined as:
x
Administration of wrong drug or use of wrong device.
Note: Use of wrong DUN is not considered a medication error unless it results in a 
confirmed administration of wrong drug.
x
Wrong route of administration, such as intramuscular instead of sc 
x
Accidental administration of a lower or higher dose than intended. The administered dose 
must deviate from the intended dose to an extent where clinical consequences for the trial 
subject were likely to happen as judged by the investigator, although they did not 
necessarily occur.
AE and SAE recording
x
The investigator will record all relevant AE information in the eCRF and SAE information 
on paper forms. 
x
The investigator will attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information. In such cases, the diagnosis (not the 
individual signs/symptoms) will be documented as the AE/SAE.
x
When an AE/SAE occurs, it is the responsibility of the investigator to review all 
documentation (eg, hospital progress notes, laboratory and diagnostics reports) related to 
the event.
x
For all non-serious AEs the applicable forms should be signed when the event is resolved 
or at the end of the trial at the latest. For sign-off of SAE-related forms, refer to “SAE 
reporting via paper CRF” later in this section.
x
Novo Nordisk products used as concomitant medication: if an AE is considered to have a 
causal relationship with a Novo Nordisk marketed product used as concomitant 
medication in the trial, it is important that the suspected relationship is reported to Novo 
Nordisk, eg, in the alternative etiology section on the safety information form. Novo 
Nordisk may need to report this adverse event to relevant regulatory authorities.
Assessment of severity
The investigator will assess intensity for each event reported during the trial and assign it to 
one of the following categories: 
x
Mild: An event that is easily tolerated by the subject, causing minimal discomfort and not 
interfering with everyday activities.
x
Moderate: An event that causes sufficient discomfort and interferes with normal everyday 
activities.
x
Severe: An event that prevents normal everyday activities. 
Note: Severe is a category used for rating the intensity of an event; and both an AE and 
SAE can be assessed as severe. An event is defined as ‘serious’ when it meets at least 
one of the outcomes described in the definition of an SAE and not when it is rated as 
severe.
Assessment of causality
The investigator is obligated to assess the relationship between trial product and the 
occurrence of each AE/SAE.
Relationship between an AE/SAE and the relevant trial product(s) should be assessed as: 
x
Probable - Good reason and sufficient documentation to assume a causal relationship.
x
Possible - A causal relationship is conceivable and cannot be dismissed.
x
Unlikely - The event is most likely related to etiology other than the trial product.
Alternative etiology, such as underlying disease(s), concomitant medication, and other risk 
factors, as well as the temporal relationship of the event to trial product administration will be 
considered and investigated.
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The investigator should use the investigator’s brochure for the assessment. For each 
AE/SAE, the investigator must document in the medical records that he/she has reviewed the 
AE/SAE and has provided an assessment of causality.
There may be situations in which an SAE has occurred and the investigator has minimal 
information to include in the initial report. However, it is important that the investigator 
always makes an assessment of causality for every event before the initial 
transmission of the SAE data.
The investigator may change his/her opinion of causality in light of follow-up information and 
send a follow-up report with the updated causality assessment.
The causality assessment is one of the criteria used when determining regulatory reporting 
requirements.
Final outcome
The investigator will select the most appropriate outcome:
x
Recovered/resolved: The subject has fully recovered, or by medical or surgical treatment 
the condition has returned to the level observed at the first trial-related activity after the 
subject signed the informed consent.
x
Recovering/resolving: The condition is improving, and the subject is expected to recover 
from the event. This term is only applicable if the subject has completed the trial or has 
died from another AE.
x
Recovered/resolved with sequelae: The subject has recovered from the condition, but 
with lasting effect due to a disease, injury, treatment, or procedure. If a sequelae meets 
an SAE criterion, the AE must be reported as an SAE.
x
Not recovered/not resolved: The condition of the subject has not improved and the 
symptoms are unchanged or the outcome is not known.
x
Fatal: This term is only applicable if the subject died from a condition related to the 
reported AE. Outcomes of other reported AEs in a subject before he/she died should be 
assessed as “recovered/resolved,” “recovering/resolving,” “recovered/resolved with 
sequelae,” or “not recovered/not resolved.” An AE with a fatal outcome must be reported 
as an SAE.
x
Unknown: This term is only applicable if the subject is lost to follow-up.
Follow-up of AE and SAE
The investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as medically indicated or as requested by Novo Nordisk to 
elucidate the nature and/or causality of the AE or SAE as fully as possible (eg, severe 
hypersensitivity reactions). This may include additional laboratory tests (eg, skin prick test) or 
investigations, histopathological examinations, or consultation with other health care 
professionals.
If a subject dies during participation in the trial or during a recognized follow-up period, the 
investigator should provide Novo Nordisk with a copy of autopsy report including 
histopathology.
New or updated information will be recorded in the CRF.
SAE reporting via paper CRF
x
Relevant CRF forms (AE and safety information forms) must be forwarded to Novo 
Nordisk.
x
The Investigator needs to complete the AE and safety information form within the 
designated reporting time frames (as illustrated in Figure 2). 
x
If the eCRF is unavailable for more than 24 hours, then the site will use the paper AE 
form.
Contact details for SAE reporting can be found in the investigator trial master file.
Page 58 of 59


Memo to File 
Page 1 of 1
Memo to File 
To:
File
From: 
Date:
RE:
Site Affiliation 
The following Investigators are affiliated with
Name 
Signature
Date (dd-Mmm-yyyy) 
Page 59 of 59

SAP_001.pdf: 
Cover Page for Statistical Analysis Plan 
Sponsor name: 
Novo Nordisk A/S 
NCT number 
NCT04007107 
Sponsor trial ID: 
INS-4582 
Official title of study: 
A TRIAL TO COMPARE THE INJECTION SITE PAIN 
EXPERIENCE OF 0.25 MGSEMAGLUTIDE SC 
ADMINISTERED BY 2 DIFFERENT PRODUCTS 
Document date: 
17 March 2020 


16.1.9 Documentation of Statistical Methods 
/LVWRIFRQWHQWV
16.1.9.1 Statistical Analysis Plan_09-Aug-2019 ..................................................................................................................
16.1.9.2 Statistical Methods_-0DU-20 ........................................................................................................................
2
29
CONFIDENTIAL
Date:
1 0DU 2020 
Novo Nordisk
Version: 
1.0 
Status:
Final
Semaglutide 
Trial ID: INS-4582
Clinical Trial Report
Appendix 16..
Page:
1 of 
CONFIDENTIAL
Redacted VWDWLVWLFDODQDO\VLVSODQ 
Includes redaction of personal identifiable information only. 
. 


Page 2 of 34


Statistical Analysis Plan
Protocol: INS-4582
Version Date: 09-Aug-2019
Page 2 of 27
2.0 Table of Contents
1.0 Approvals ................................................................................................................................................1 
2.0 Table of Contents....................................................................................................................................2 
3.0 Introduction .............................................................................................................................................4 
4.0 Changes from Previous Version of Approved SAP ................................................................................4 
5.0 Study Objectives .....................................................................................................................................4 
5.1 Primary................................................................................................................................................4 
5.1.1 Primary Endpoint..........................................................................................................................4 
5.2 Exploratory..........................................................................................................................................4 
5.2.1 Exploratory Endpoint....................................................................................................................4 
5.3 Study Hypothesis ................................................................................................................................6 
6.0 Study Design...........................................................................................................................................6 
6.1 Sample Size Considerations...............................................................................................................6 
6.2 Randomization ....................................................................................................................................7 
7.0 Overview of Planned Analysis ................................................................................................................7 
7.1 Changes from Protocol .......................................................................................................................7 
7.2 Interim Analysis and Key Results .......................................................................................................7 
7.3 Final Analysis ......................................................................................................................................7 
8.0 Data Review............................................................................................................................................7 
8.1 Data Management...............................................................................................................................7 
8.2 Acceptance of Data for Summarization ..............................................................................................7 
9.0 Definitions and General Analysis Methods .............................................................................................8 
9.1 Analysis Data Presentation.................................................................................................................8 
9.1.1 Rounding......................................................................................................................................8 
9.1.2 Imputation ....................................................................................................................................8 
9.1.3 Descriptive Statistics....................................................................................................................8 
9.1.4 Pooling .........................................................................................................................................8 
9.1.5 Unscheduled Measurements .......................................................................................................8 
9.2 Analysis Data Definitions ....................................................................................................................8 
9.2.1 Baseline Definition .......................................................................................................................8 
9.2.2 Treatment/Subject Grouping........................................................................................................9 
9.2.3 Common Variable Derivations .....................................................................................................9 
9.2.4 QC................................................................................................................................................9 
9.2.5 ADaM Datasets and Metadata.....................................................................................................9 
9.3 Software ..............................................................................................................................................9 
9.4 Statistical Methods..............................................................................................................................9 
9.4.1 Statistical Outlier Determination...................................................................................................9 
9.4.2 Predetermined Covariates and Prognostic Factors .....................................................................9 
9.4.3 Hypothesis Testing.......................................................................................................................9 
9.5 TFL Layout ........................................................................................................................................10 
10.0 Analysis Sets.......................................................................................................................................10 
10.1 Safety Set........................................................................................................................................10 
10.2 Full Analysis Set (FAS) ...................................................................................................................10 
10.3 Per-Protocol Set..............................................................................................................................11 
11.0 Subject Disposition..............................................................................................................................11 
12.0 Protocol Deviations and Violations .....................................................................................................11 
13.0 Demographic and Baseline Characteristics........................................................................................11 
13.1 Demographics .................................................................................................................................11 
13.2 Medical History................................................................................................................................11 
13.3 Other Baseline Characteristics .......................................................................................................11 
14.0 Concomitant Medications....................................................................................................................11 
15.0 Treatment Compliance and Exposure ................................................................................................12 
16.0 Safety Analyses ..................................................................................................................................12 
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16.1 Safety Variables..............................................................................................................................12 
16.1.1 Injection Site Pain Experience .................................................................................................12 
16.1.2 Adverse Events ........................................................................................................................15 
16.1.3 Deaths and Serious Adverse Events .......................................................................................16 
16.1.4 Laboratory Data .......................................................................................................................16 
16.1.5 Vital Signs ................................................................................................................................16 
16.1.6 Electrocardiograms ..................................................................................................................16 
16.1.7 Other Observations Related to Safety .....................................................................................16 
17.0 References..........................................................................................................................................17 
Appendix 1: Glossary of Abbreviations .......................................................................................................18 
Appendix 2: Schedule of Assessments.......................................................................................................20 
Appendix 3: List of In-Text Tables ..............................................................................................................22 
Appendix 4: List of End of Text Outputs .....................................................................................................23 
Document History........................................................................................................................................27 
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3.0 Introduction
This Statistical Analysis Plan (SAP) describes the statistical methods that will be used during the analysis 
and reporting of data collected under Novo Nordisk Protocol Version 2.0.
This SAP should be read in conjunction with the study protocol and electronic case report form (eCRF). 
This version of the SAP has been developed using the protocol dated 21-Jun-2019 (including all 
amendments up to this protocol date) and the final eCRF(s) dated 26-Jun-2019.
An approved and signed SAP is a requirement for database lock. An approved SAP is also required for 
unblinding of the study treatments.
This SAP only covers the results that will be processed by the 
 Early Development Services (EDS) 
Biostatistics Department.
 EDS will evaluate the safety and tolerability analysis with focus on the injection site pain experience 
of two different pen-injectors.
This SAP supersedes the statistical considerations identified in the protocol; where considerations are 
substantially different, they will be so identified. Any post-hoc or unplanned analyses, or significant 
changes from the planned analysis in this SAP performed to provide results for inclusion in the clinical 
study report (CSR) but not included in this SAP, will be clearly identified in Section 9.8.2 of the CSR. 
Changes to planned analyses after unblinding do not require an updated SAP but should be included in 
the CSR if significant.
4.0 Changes from Previous Version of Approved SAP
This is the first version of the SAP.
5.0 Study Objectives
5.1 Primary
The primary objective is to compare, in healthy subjects, the injection site pain experience of a single 
dose of 0.25 mg semaglutide subcutaneously (sc), given as the DV3396 product, to that of the PDS290 
product
5.1.1 Primary Endpoint
Endpoint Title
Time Frame
Unit
Intensity of injection site pain
1 min after each injection (Day 
1)
Millimeter (mm) on a 100-mm 
horizontal visual-analogue scale 
[VAS] (where 0 mm= no pain, 
100 mm = unbearable pain)
5.2 Exploratory
Comparison of injection site pain experience of the two products will be further explored by:
x
Categorial assessments by means of a 6-point scale, and a derived dichotomous scale; 
x
Quality of pain;
x
Duration of pain;
x
Comparative pain experience by means of a 5-point scale, and a derived dichotomous scale; 
5.2.1 Exploratory Endpoint
Endpoint Title
Time Frame
Unit
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Endpoint Title
Time Frame
Unit
Categorical 
assessment 
of 
injection site pain intensity
Immediately 
after rating of 
intensity of injection site pain, 
ie, after 1 min plus the time it 
takes to complete the rating of 
intensity of injection site pain 
by visual analogue scale (Day 
1)
6-point scale (none – very mild 
– mild – moderate – severe –
very severe)
Moderate or more injection site 
pain
Dichotomous variable based 
on categorical assessment of 
intensity of injection site pain 
(Day 1) (item above)
Yes = moderate, severe, very 
severe
No = none, very mild, mild
Quality of pain
Immediately after categorical 
assessment 
of 
intensity 
of 
injection site pain, ie, after 1 
min plus the time it takes to 
complete the rating of intensity 
of injection site pain and the 
categorical rating of intensity of 
injection site pain (Day 1)
Pain quality items on modified 
SF-MPQ-2 inventory (select all 
that apply):
Throbbing pain տ
Shooting pain տ
Stabbing pain տ
Sharp pain տ
Cramping pain տ
Gnawing pain տ
Hot-burning pain տ
Aching pain տ
Heavy pain տ
Tender տ
Splitting pain տ
Tiring-exhausting տ
Sickening տ
Fearful տ
Punishing-cruel տ
Electric-shock pain տ
Cold-freezing pain տ
Piercing տ
Duration of pain
From time of injection until 
cessation of pain (Day 1)
min and sec
Comparative pain experience
After pain has ended after the 
last injection (Day 1)
5-point scale (the last injection 
hurt much more than the first 
injection – the last injection hurt 
more than the first injection –
they 
hurt 
about 
the 
same 
[includes: none of the injections 
hurt] – the last injection hurt 
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Endpoint Title
Time Frame
Unit
less than the first injection – the 
last injection hurt much less 
than the first injection)
The DV3396 product hurt less 
than or about the same as the 
PDS290 product
Dichotomous variable based 
on 
“Comparative 
pain 
experience” 
variable 
above 
(Day 1)
Yes 
= 
“about 
the 
same” 
(includes “none of the products 
hurt” or “the DV3396 product 
hurt less than the PDS290 
product” or “the DV3396 product 
hurt much less than the PDS290 
product”)
No = “The DV3396 product hurt 
much more than the PDS290 
product” 
or 
“The 
DV3396 
product hurt more than the 
PDS290 product”
5.3 Study Hypothesis
Not applicable.
6.0 Study Design
This is a single-center, cross-over, randomized, double-blind study in healthy men and women comparing 
the injection site pain experience of the DV3396 product to that of the PDS290 product when both 
products are used to deliver 0.25 mg semaglutide sc. A total of 104 subjects are planned to be 
randomized to achieve at least 100 completed subjects. The aim is to have an even distribution of males 
and females in the study; at least 30% of the same sex should be randomized in the study. Subjects will 
be randomized in a 2×2 scheme evenly to 4 sequences A,B,C or D of product and injection side as in
Table 1.
Table 1 Product and site of injection on abdomen per treatment
Sequence 
Sequence Description
Ratio Required
First injection on right side, Second injection on left side
A
DV3396 (Right) followed by PDS290 (Left)
1
C
PDS290 (Right) followed by DV3396 (Left)
1
First injection on left side, Second injection on right side
B
DV3396 (Left) followed by PDS290 (Right)
1
D
PDS290 (Left) followed by DV3396 (Right)
1
Subjects will receive 2 single doses of semaglutide 0.25 mg on 1 day. The 2 products will be administered 
at least 30 minutes apart, in the anterior aspect of the abdominal wall on opposite sides of the midline. If
pain experience still exists after 60 minutes, the second injection will not be administered. A follow-up
phone call will take place between 4 and 5 weeks after drug administrations on Day 1.
6.1 Sample Size Considerations
The sample size calculation considers both the primary endpoint and the dichotomous endpoints. 
With 100 subjects, a conservative standard deviation of 30 mm on the VAS, and 90% power, it is possible 
to detect a difference smaller than 10 mm. Hence a sample size of 100 subjects is considered sufficient to 
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detect a clinically important difference. Allowing for 4% dropouts or missing data, a total of 104 subjects 
will need to be randomized.
Considering precision on the proportion of the dichotomous endpoints, a sample size of 100 subjects 
results in a confidence interval (CI) with a width of 10% on each side, e.g., for a proportion of 60%, the CI 
is 60% (50%-70%).
6.2 Randomization
Subjects who have met all eligibility criteria will receive a subject number upon inclusion in the study. 
They receive the subject number just prior to dosing. The subject number will ensure identification 
throughout the study. The subject number will be the same as the randomization number given on the 
randomization list produced by Novo Nordisk.
The randomization list will be transferred to the 
 Pharmacy and kept in a restricted area to which only 
the 
 Pharmacy has access.
7.0 Overview of Planned Analysis
7.1 Changes from Protocol
Additional statistical analyses will be performed on the exploratory endpoints. See Sections 16.1.1.2,
16.1.1.4, and 16.1.1.5 for more details on these analyses.
7.2 Interim Analysis and Key Results
A set of topline tables, figures and listings (TFLs) will be provided after Database (DB) lock-1 (without 
follow up data). The study will be unblinded after this DB lock-1. These TFLs are indicated with a * in 
Appendix 4: List of End of Text Outputs .
7.3 Final Analysis
All TFLs will be provided after DB lock-2 (all data included).
8.0 Data Review
8.1 Data Management
Data handling and transfer will take place under the 
 Data Management Plan for the study. Two DB 
locks are planned: 
DB lock-1 : includes all data except Follow up data. These data will be used for Topline TFLs.
DB lock-2 : includes all data. These data will be used for draft and final TFLs.
8.2 Acceptance of Data for Summarization
Programming of analysis datasets and TFLs may be ongoing during the data management of the study. 
However, programming of analysis datasets and TFLs will be completed and quality controlled (QC’d) 
after each DB lock. Only quality assured (QA’d) results released by the Safety Laboratory will be used for 
the programming of analysis datasets and TFLs for the final report. Any data values requiring 
investigation or corrections that are identified while programming the analysis datasets and TFLs will be 
sent to the project Data Manager. If the issue affects the TFLs the Programmer or Statistician who 
identified the issue will follow it to resolution.
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9.0 Definitions and General Analysis Methods
9.1 Analysis Data Presentation
9.1.1 Rounding
In listings, data will be presented with the same precision as the original data.  Derived data will be 
rounded for presentation purposes.
For all summaries, the mean, standard deviation (SD) and median will be presented to one decimal place 
greater than the data, and the minimum (min) and maximum (max) will be presented to the same number 
of decimal places as the data. Frequency percentages will be presented with one decimal.
The above rule can be applied directly to collected data. For derived data rounding will occur prior to 
summarization so a specific number of decimal places will have to be assumed to apply the above 
rounding rules for summary statistics.
P-values will be reported to four decimal places; p-value less than 0.0001 will be reported as p <0.0001.
9.1.2 Imputation
Unless otherwise noted, data will not be imputed.
9.1.3 Descriptive Statistics
Unless otherwise indicated, continuous variables will be summarized with the following descriptive 
statistics: n (number of observations), (arithmetic) mean, SD, min, median and max.
Categorical data will be summarized with frequencies and percentages. Percentages by categories will be 
based on the number of subjects exposed within a treatment.
For categorical data the categories will be presented in the tables exactly as they appear in the CRF / 
Database, unless otherwise indicated. For injection pain experience categories, categories will be 
combined to create dichotomic categories in addition to the original scale.
9.1.4 Pooling
Not applicable.
9.1.5 Unscheduled Measurements
Unscheduled measurements will be included in the listings. With the exception of unscheduled 
measurements used for baseline, unscheduled measurements will be excluded from the descriptive 
statistics and statistical analysis.
9.2 Analysis Data Definitions
9.2.1 Baseline Definition
Unless otherwise stated, baseline for post-dose evaluations is defined as the last observation recorded 
before the first study drug administration. The last observation can be an unscheduled / repeated 
measurement.
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9.2.2 Treatment/Subject Grouping 
Label
Grouping
Study Treatment 
0.25 mg semaglutide sc - DV3396 product
0.25 mg semaglutide sc - PDS290 product 
Treatment
DV3396, PDS290
Dose Level
0.25 mg
9.2.3 Common Variable Derivations
Variable
Data Type
Definition/Calculation
Note
Analysis Study Day 
(Post Dose)
All
Date of Measurement 
minus Dose Date +1
9.2.4 QC
The analysis datasets and the TFLs will be QC’d according to the 
 EDS QC plan.
9.2.4.1 Critical Data
The QC plan requires datasets be classified as critical or non-critical. As the primary objective is to 
compare the injection site pain experience, the datasets considered critical are subject level analysis data 
(ADSL), adverse events analysis data (ADAE), and questionnaire analysis data (ADQS) [containing all 
injection site pain experience data]. As these are related to the primary objectives these datasets will be 
double programmed per the QC Plan.
9.2.5 ADaM Datasets and Metadata
The analysis datasets will be generated in accordance with Clinical Data Interchange Standard 
Consortium (CDISC) Analysis Data Model (ADaM) Version 2.1. 
ADaM compliant datasets will be delivered to the sponsor. A define.xml file version 2 with the 
corresponding metadata will be included. Analysis results metadata are excluded.
9.3 Software
The statistical analysis and reporting will be done using SAS® for WindowsTM Version 9.4 or higher (SAS 
Institute, Inc.).
9.4 Statistical Methods
9.4.1 Statistical Outlier Determination
No statistical outlier analysis is planned.
9.4.2 Predetermined Covariates and Prognostic Factors
There are no predetermined covariates or prognostic factors.
9.4.3 Hypothesis Testing
No formal hypothesis testing will be done.
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9.5 TFL Layout
Report layout will be according to the 
 EDS – ICH E3 compliant – CSR Template. The layout of TFLs 
will be according to the 
 EDS standards.
No table shells will be provided. The TFLs will be provided in Adobe PDF format.
Format:
x
Page size: A4
x
Data in listings will be sorted by subject number and number of injection. Side and treatment 
(product) will be added to the listings.
x
For presentation of  the tables, please refer to the sections in this SAP.
x
Column titles will be in title case letters.
x
All tables and listings will be in landscape format.
x
The following headers and labels will be used in the TFLs:
Treatment:
Injection Number
Injection Site 
o
DV3396 
o
Inj. 1
o
Left
o
PDS290
o
Inj. 2
o
Right
10.0 Analysis Sets
Analyses
All 
Subjects
Safety Set
Full Analysis Set
Per Protocol Set
Disposition
Summaries
3
Safety Assessments
3
Baseline 
Characteristics
3
3
Endpoint Analyses

3
Both the full analysis set and the per protocol set will be used for summaries. All analyses will be based 
on the per protocol analysis set except if it is specifically stated otherwise. Since the safety set and the full 
analysis set are the same, only the safety set will be used for the safety summaries.
The analysis sets are established before DB hardlock-1.
10.1 Safety Set
All subjects who have received at least 1 injection of semaglutide (includes any skin contact with study 
product whether the injection was completed or not).
10.2 Full Analysis Set (FAS)
All randomized subjects who have received at least 1 injection of semaglutide (includes any skin contact 
with trial product whether the injection was completed or not). In exceptional cases, subjects from the 
FAS may be excluded. In such cases the exclusion will be justified and documented before DB hard lock.
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10.3 Per-Protocol Set
All subjects who have received both injections of semaglutide and have completed both intensity of 
injection site pain assessments of the primary endpoint.
11.0 Subject Disposition
The number and percentage of subjects randomized, dosed, and members of each analysis set will be 
presented. The number and percentage of subjects who withdrew from the study prematurely and a 
breakdown of the corresponding reasons for withdrawal will also be presented.
Individual data on randomization, dosing and completion will be listed. 
12.0 Protocol Deviations and Violations
Protocol deviations/violations are collected throughout the study. Protocol deviations for the analysis sets 
will be described in the CSR.
13.0 Demographic and Baseline Characteristics
13.1 Demographics
Individual demographic data will be listed.
Subject demographics at screening will be summarized descriptively for all subjects. All data in this 
summary will be collected at screening and will include the subjects’ age (in years), gender, race, 
ethnicity, weight (in kg), height (in cm), body mass index (BMI) [in kg/m2].
Demographics will be summarized for the safety, FAS, and per protocol sets.
13.2 Medical History
Medical history (including coding according to Medical Dictionary for Regulatory Activities [MedDRA], 
version 22.0), will be listed.
13.3 Other Baseline Characteristics
Previous medications (i.e., any recorded medications that start and end before administration of the study 
drug) will be listed.
Previous injection experience as recalled by the subject to the best of their ability will be listed: product 
name, dose, type [vial/syringe, prefilled syringe, autoinjector, other], indication, frequency of use, injection 
site, chronological period used.
Previous medications and previous injection experience will be coded by using the World Health 
Organisation (WHO) Drug Dictionary Enhanced (DDE). The version described in the Coding Conventions
will be used (version 2019MAR01 B3).
Non-compliance to inclusion or exclusion criteria (if any) will be listed.
14.0 Concomitant Medications
Concomitant medication will be listed including coding (WHO-DDE version 2019MAR01 B3).
Concomitant medications are defined as those taken by the patient at any time between the date of study 
drug administration and study completion/discontinuation. Medication with start date being partially or 
completely missing will be assumed to be concomitant if it cannot be definitively shown that the 
medication did not occur during treatment.
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In addition, non-drug treatments will be listed.
15.0 Treatment Compliance and Exposure
Study drug administration data including location of the injection will be listed by subject.
16.0 Safety Analyses
16.1 Safety Variables
The following safety variables will be summarized:
x
Injection site pain experience:
o
Intensity of injection site pain VAS
o
Categorical assessment of injection site pain intensity (6-point scale; dichotomous scale)
o
Quality of pain modified SF-MPQ-2 inventory;
o
Duration of pain assessment
o
Comparative pain assessment (5-point scale, dichotomous scale);
x
Adverse Events (AEs) 
The following safety variables will be listed:
x
Vital Signs (supine systolic and diastolic blood pressure and pulse rate)
x
Electrocardiograms (ECG) (if post dose assessments are deemed relevant by the investigator)
o
Heart Rate
o
PR Interval
o
QRS-Duration
o
QT Interval
o
QTc (Fridericia’s) Interval
x
Clinical Laboratory Evaluations (if post dose assessments are deemed relevant by the 
investigator)
o
Clinical Chemistry 
o
Hematology
o
Hormones
x
Physical Examination (if post dose assessments are deemed relevant by the investigator)
16.1.1 Injection Site Pain Experience
Summaries will be presented for the full analysis set, and in addition for the per protocol set if the sets are 
different from each other. 
16.1.1.1 Intensity of injection Site Pain – VAS
The pain score will be measured on a visual analogue scale, from ‘no pain’ (0 mm) to ’unbearable pain’ 
(100 mm). VAS score will be listed by subject and injection number, side and treatment will be added to 
the listing.
The following descriptive statistics will be presented: number of subjects, arithmetic mean, SD, coefficient 
of variation (CV), median, Min and Max. The results will be summarized by treatment, by injection 
number, by side, by treatment and injection number, and by treatment and side.
Ping pong plots with individual VAS scores will be presented for each treatment sequence (i.e. by 
treatment and side).
In addition, the primary endpoint will be analyzed for the per protocol population by a fixed analysis of 
variance model with VAS score as the dependent variable, and product, injection side (right side, left 
side), injection number (first injection, second injection), and subject as fixed effects. 
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From the model, the difference in VAS score between the 2 products is estimated and presented with a 
95% CI and a p-value.
Model in SAS:
PROC MIXED data=VAS; /*for per protocol population*/ 
class product inj_number inj_side subject;
/*inj_number: first or second injection*/
/*inj_side: left or right*/
model VAS = product inj_number inj_side subject / ddfm=satterthwaite;
lsmeans product/ diff cl;
RUN;
The interpretation of the difference in VAS pain scores will be supported and contextualized by the results 
of the analyses of the exploratory endpoints.
16.1.1.2 Intensity of Injection Site Pain – Categorial Assessments
Categorical assessment of injection site pain intensity
The categorial pain score will be measured by a 6-point scale in which the pain intensity is rated by the 
subjects as follows: none - very mild – mild - moderate – severe – very severe. 
Individual results will be listed by subject and injection number, side and treatment will be added to the 
listing.
The results will be summarized descriptively, by counting the number (and percentage) of subjects in 
each category. Missing scores will be a separate category. The results will be summarized by treatment.
Moderate or more injection site pain
The 6-point scale will be dichotomized in two categories to evaluate the experience of ‘moderate or more 
injection site pain’. The two categories are:
x
No: none - very mild - mild
x
Yes: moderate – severe – very severe.
The results will be summarized descriptively by counting the number (and percentage) of subjects in each 
category. Missing scores will be a separate category. The results will be summarized by treatment, by 
injection number, by side, by treatment and injection number, and by treatment and side.
In addition, a statistical analysis will be conducted using a McNemar mid-p test (Fagerland et al. 2013) 
with Bonett-Price hybrid score (Fagerland et al. 2014) 95% confidence intervals comparing the two 
products on the moderate or more pain score (No vs Yes).
Model in SAS:
PROC FREQ data=Pain score;
tables DV3396* PDS290 / alpha=0.05;
exact mcnem / midp ;
RUN;
16.1.1.3 Quality of Pain - Modified SF-MPQ-2 Inventory
The quality of pain will be assessed using the modified short-form McGill pain questionnaire (modified 
SF-MPQ-2 inventory).
Individual results will be listed by subject and injection number, side and treatment will be added to the 
listing.
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The results will be summarized descriptively by counting the number (and percentage) of subjects for 
each description of pain quality. Missing scores will be a separate category. The results will be 
summarized by treatment.
16.1.1.4 Duration of Pain Assessment
The duration of pain as indicated by the subject will be recorded from time of injection until cessation of 
pain in minutes and seconds.
Individual results will be listed by subject and injection number, side and treatment will be added to the 
listing.
The results will be summarized with the following descriptive statistics: number of subjects, arithmetic 
mean, SD, CV, median, Min and Max. The results will be summarized by treatment, by injection number, 
by side, by treatment and injection number, and by treatment and side.
In addition, the conditional duration of pain, given there is pain, will be analyzed for subjects in the per 
protocol set who has a non-missing and non-zero duration of pain on both injections, by a fixed analysis 
of variance model with duration of pain as the dependent variable, and product, injection side (right side, 
left side), injection number (first injection, second injection), and subject as fixed effects. 
From the model, the difference in VAS score between the 2 products is estimated and presented with a 
95% CI and a p-value.
Model in SAS:
PROC MIXED data=Duration of pain; /*for per protocol population*/ 
class product inj_number inj_side subject;
/*inj_number: first or second injection*/
/*inj_side: left or right*/
model  duration = product inj_number inj_side subject / ddfm=satterthwaite;
lsmeans product/ diff cl;
RUN;
16.1.1.5 Comparative Pain Assessment 
Comparative pain experience
The comparative pain assessment is a 5-point scale in which the subjects will compare the pain between 
the first and second injection to preserve blinding. After unblinding the scoring will be translated into the 
following 5-point scale:
-
The DV3396 product hurt much more than the PDS290 product.
-
The DV3396 product hurt more than the PDS290 product.
-
The DV3396 and PDS290 products hurt about the same (includes: none of the products hurt).
-
The DV3396 product hurt less than the PDS290 product.
-
The DV3396 product hurt much less than the PDS290 product.
Individual results will be listed by subject.
The results will be summarized descriptively by counting the number (and percentage) of subjects in each 
category. Missing scores will be a separate category.
The DV3396 product hurt less than or about the same as the PDS290 product
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The 5-point scale will be translated in two categories to evaluate if the DV3396 product hurt less than or 
about the same as the PDS290 product. The two categories are:
x
Yes: This includes “none of the products hurt” or “the DV3396 product hurt less than the PDS290 
product” or “the DV3396 product hurt much less than the PDS290 product”.
x
No: This includes “the DV3396 product hurt much more than the PDS290 product” or “the 
DV3396 product hurt more than the PDS290 product”.
The results will be summarized descriptively by counting the number (and percentage) of subjects in each 
category. Missing scores will be a separate category.
In addition, a statistical analysis will be conducted presenting the proportion with Clopper Pearson 95% 
confidence interval of “DV3396 product hurt less than or about the same as the PDS290 product”.
Model in SAS:
PROC FREQ data=comparative_pain;
tables score / binomial(exact) alpha=.05;
RUN;
16.1.2 Adverse Events
A treatment-emergent AE (TEAE) is defined as an event that has onset date on or after the day of the first 
injection and no later than 28 days after the day of the last injection (both injections are scheduled on day 
1), or has onset date before the day of the first injection and increases in severity in the period up to 28 
days after the last injection. If the time is missing for an AE on a dosing day, the AE will be categorized as 
TEAE.
In general, TEAEs will not be assigned to one of the products (PDS290 or DV3396) because time 
between dosing is 30-60 minutes. Local tolerability TEAEs will be categorized by product (PDS290 or 
DV3396), based on the location of the injection site which is indicated for these TEAEs.
AE summaries will include TEAEs only, one table will present all TEAEs (all TEAEs), and one table will 
present TEAEs related to local tolerability only (categorized by product).
A breakdown of the number of events and number and percentage of subjects reporting each AE,
categorized by body system and preferred term coded according to the (MedDRA, version 22.0), will be 
presented for all local tolerability TEAEs, categorized by product (PDS290 or DV3396). Subjects will only 
be counted once within each body system or preferred term per product. A similar table will be provided 
for all TEAEs, subject will only be counted once within each body system or preferred term.
The summary tables will present the counts in descending order by system organ class and preferred 
term (within a system organ class) based on the number of subjects experiencing the event. 
Additional tables for all TEAEs and for local tolerability TEAEs (number of events and number and 
percentage of subjects) by severity (mild/moderate/severe) for all TEAEs will be provided.
All adverse events (including non-treatment-emergent events) recorded on the eCRF will be listed.
A listing of adverse events leading to study discontinuation will be provided.
The following missing data will be imputed as defined (for calculations only / will not be presented):
x
Missing AE start and / or end times for the calculation of onset and duration will be assumed to be 
at 00:01 for a start time and 23:59 for end times
x
Missing AE severity or relationship will be classified as missing severity or missing relationship.
x
Missing AE start times for the determination of treatment emergence will be assumed to occur 
after treatment unless partial date documents the AE as happening prior to treatment
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Statistical Analysis Plan
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Version Date: 09-Aug-2019
Page 16 of 27
x
Missing AE start date will be assumed to be after treatment for the determination of TEAE and on 
treatment for single treatment studies but will not be attributed to treatment in studies with 
multiple treatments
16.1.3 Deaths and Serious Adverse Events
A listing of deaths and other serious adverse events (SAE) will be provided by subject.
16.1.4 Laboratory Data
Clinical laboratory data will be presented using Système International (SI) units (also used in the Study 
data tabulation model [SDTM] Controlled Terminology).
All laboratory data will be listed by subject and scheduled time. Normal ranges (with units corresponding 
to the reported SI units) will be used directly from the clinical laboratory and will be included in the listings. 
Clinical laboratory results outside the reference range will be flagged. In case the out of reference range 
values were considered clinically significant by the investigator, this will also be indicated in the listings as 
well as in the AE. Comments with regard to the laboratory test results will be shown in a separate listing.
A summary listing with all out-of-range values will also be provided. 
16.1.5 Vital Signs
All vital signs measurements will be listed by subject and timepoint. 
16.1.6 Electrocardiograms
All ECG measurements will be listed by subject and timepoint. The physician’s interpretation and the 
specification of any abnormalities will be provided in a separate listing by subject and timepoint.
16.1.7 Other Observations Related to Safety
17.1.6.1 Physical Examination 
Findings at screening and changes from screening (if applicable) for the physical examination will be 
listed.
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Statistical Analysis Plan
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Version Date: 09-Aug-2019
Page 17 of 27
17.0 References
SAS Institute, Inc., SAS® Version 9.4 software, Cary, NC.
Clinical Study Protocol. A TRIAL TO COMPARE THE INJECTION SITE PAIN EXPERIENCE OF 0.25 MG 
SEMAGLUTIDE SC ADMINISTERED BY 2 DIFFERENT PRODUCTS. Version 2.0, Final, 21 June 2019.
Clinical Data Interchange Standards Consortium, Inc. , Analysis Data Model (ADaM), Version 2.1, 2009.
Clinical Data Interchange Standards Consortium, Inc., Analysis Data Model (ADaM) Implementation 
Guide, Version 1.1, 2016
Fagerland MW, Lydersen S, Laake P. The McNemar test for binary matched-pairs data: mid-p and 
asymptotic are better than exact conditional. BMC Medical Research Methodology 2013; 13:91.
Fagerland MW, Lydersen S, Laake P. Recommended tests and confidence intervals for paired binomial 
proportions. Statistics in Medicine 2014; 33:2850–2875.
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Version Date: 09-Aug-2019
Page 18 of 27
Appendix 1: Glossary of Abbreviations
ADaM
Analysis data model
ADAE
Subject level analysis dataset
ADSL
Adverse events analysis dataset
AE
Adverse event
ADQS
Questionnaires analysis dataset
BMI
Body mass index
CDISC
Clinical Data Interchange Standard Consortium
CSR
Clinical study report
CI
Confidence interval
CV
Coefficient of variation
DB
Database
ECG
Electrocardiogram
eCRF
Electronic case report form
EDS
Early Development Services
FAS
Full analysis set
HBA1c
Hemoglobin A1c
HBsAg
Hepatitis B surface antigen
HCV
Hepatitis C virus
HIV
Human immunodeficiency virus
ICH
The International Conference on Harmonization of Technical Requirements 
for Registration of Pharmaceuticals for Human Use
Max
Maximum
MedDRA
Medical Dictionary for Regulatory Activities
Min
Minimum 
mm
millimeter
N
Number of observations
PDF
Portable Document Format
QA’d
Quality assured
QC’d
Quality controlled
SAE
Serious adverse event
SAP
Statistical analysis plan
sc
Subcutaneous(ly)
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Page 19 of 27
SD
Standard deviation
SDTM
Study data tabulation model
SI
Système International
TEAE
Treatment-emergent adverse event
TFLs
Tables, figures and listings
VAS
Visual analogue scale
WHO-DDE
World Health Organization – Drug Dictionary Enhanced
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Statistical Analysis Plan
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Version Date: 09-Aug-2019
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Appendix 2: Schedule of Assessments
Visit
Screening
Pretreatment
Treatment
Follow-up Phone 
Call
Study Day
Days
-21 to -1
Day 1
Day 29 to Day 36
Predose
Dosing and 
After
Ambulatory
X
X
X
X
Admission
X
Discharge
X
Informed Consent
X
Medical History
X
Previous Injection Experiencea
X
Demographics
X
Physical Examination
X
Xb
Body Height, Body Weight and BMI Calculation
X
Serology (HBsAg, anti-HCV, anti-HIV-1 and -2)
X
Urine Drug and Alcohol Screen
X
X
Urine Pregnancy Test (Females of Childbearing Potential 
Only)
X
X
Xc
Clinical Laboratoryd
X
Xd
HbA1c
X
12-lead ECGe
X
Xe
Vital Signsf
X
X
Eligibility Check
X
Xg
Randomization and Stratificationh
X
Study Drug Administrationh
X
Intensity of Injection Site Pain VASi
X
Categorical Assessment of Injection Site Pain Intensityj
X
Quality of Pain Modified SF-MPQ-2 Inventoryk
X
Duration of Painl
X
Comparative Pain Assessmentm
X
Previous and Concomitant Medication
X
X
X
X
AE Monitoring
X
X
X
X
Technical Complaintsn
X
X
Drug Accountability
X
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Version Date: 09-Aug-2019
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AE=adverse event; BMI=body mass index; ECG=electrocardiogram; HbA1c=glycoslated hemoglobin; HBsAg=hepatitis B surface antigen; 
HCV=hepatitis C virus; HIV=human immunodeficiency virus; sc=subcutaneous; SF-MPQ-2=short-form McGill pain questionnaire; VAS=visual 
analogue scale
a
Previous injection experience as recalled by the subject to the best of their ability: product used (name, dose, type [vial/syringe, prefilled 
syringe, autoinjector, other], indication, frequency of use, injection site, chronological period used).
b
Physical examination: this will be a focused examination only done if deemed relevant by the Investigator
c
For the check on pregnancy at the follow-up phone call, female subjects of childbearing potential will be handed out a urine pregnancy test 
on Day 1. They will have to do the pregnancy test at the ealiest in the morning of Day 29 and at the latest in the morning of the follow-up 
phone call.
d
Clinical laboratory tests (including clinical chemistry and hematology): at screening (non-fasting). 
The same tests may be done after the second study drug administration on Day 1 prior to discharge, if deemed relevant by the Investigator.
e
12-lead ECG: at screening.
12-lead ECG may be done after the second study drug administration on Day 1 prior to discharge, if deemed relevant by the Investigator. 
f
Vital signs (supine systolic and diastolic blood pressure, and pulse): at screening and after the second study drug administration on Day 1 
prior to discharge.
g
Eligibility check on Day 1 before dosing concerns the urine pregnancy test (female subjects of childbearing potentialonly), urine drug and 
alcohol screen, and concomitant medication only.
h
Subjects will be randomized in a 2×2 scheme evenly to 4 sequences of product and injection side as in the table below: 
Treatment Short Identifier
Treatment Long Identifier
Ratio Required
First injection on right side, Second injection on left side
A
DV3396 (Right) followed by PDS290 (Left)
1
C
PDS290 (Right) followed by DV3396 (Left)
1
First injection on left side, Second injection on right side
B
DV3396 (Left) followed by PDS290 (Right)
1
D
PDS290 (Left) followed by DV3396 (Right)
1
Subjects will receive 2 single doses of semaglutide 0.25 mg on 1 day. The 2 products will be administered at least 30 minutes apart, in the 
anterior aspect of the abdominal wall on opposite sides of the midline.
i
One minute after each injection, the VAS to rate the intensity of injection site pain will be administered.
j
The categorical assessment of injection site pain intensity (6-point scale) will be started after the VAS to rate the intensity of injection site pain 
has been completed after each injection.
k
The quality of pain modified SF-MPQ-2 inventory will be started after the categorical assessment of injection site pain intensity (6-point scale) 
has been completed after each injection.
l
Subjects have to indicate after each injection when the pain, if any, is gone.
m
The comparative pain assessment (5-point scale) comparing pain between the 2 injections will be completed after the second injection.
n
All technical complaints that occur, from the time of receipt of the product at the site until the time of the last usage of the product, must be 
collected and reported to Novo Nordisk. 
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Version Date: 09-Aug-2019
Page 22 of 27
Appendix 3: List of In-Text Tables
List of In-Text Tables and  Figures:
Output
Title 
Analysis Set
Table
Summary of Subject Disposition
All Subjects
Table 
Summary of Demographics
Safety
Table
Summary of Treatment Emergent Adverse Events by System Organ 
Class and Preferred Term - Local Tolerability
Safety
Table
Summary of Treatment Emergent Adverse Events by Severity for 
Treatment Emergent Adverse Events - Local Tolerability
Safety
Table
Summary of Intensity of Injection Site Pain - VAS
Per Protocol
Figure
Ping-Pong Plots of Individual VAS scores by treatment sequence
Per Protocol
Table
Statistical Analysis on Injection Site Pain - VAS
Per Protocol
Table
Summary of Intensity of Injection Site Pain - Moderate or more 
injection site pain
FAS
Table
Statistical Analysis on Intensity of Injection Site Pain - Moderate or 
more injection site pain
Per Protocol
Table
Summary of Quality of Pain - Modified SF-MPQ-2 Inventory
FAS
Table
Summary of Duration of Pain
FAS
Table
Statistical Analysis on Duration of Pain
Per Protocol
Table
Summary of Comparative Pain - 5-Point Scale
FAS
Table
Summary of Comparative Pain - DV3396 product hurt less than or 
about the same as the PDS290 product
FAS
Table
Statistical Analysis on Comparative Pain - DV3396 product hurt less 
than or about the same as the PDS290 product
Per Protocol
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Statistical Analysis Plan
Protocol: INS-4582
Version Date: 09-Aug-2019
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Appendix 4: List of End of Text Outputs
TFLs indicated with a * will be presented in Topline results.
List of End of Text Tables and Figures:
Output
Title 
Population 
Set
Section 14.1 – Disposition and Demographic Data
Table 14.1.1
Summary of Subject Disposition
All subjects
Table 14.1.2 *
Summary of Demographics
Safety, FAS, 
Per Protocol
Table 14.1.3
Summary of Dosing
Safety
Section 14.2 – Pharmacokinetic Data - Not Applicable
Section 14.3 – Safety Data
Table 14.3.1.1  
Summary of Treatment Emergent Adverse Events by System 
Organ Class and Preferred Term – all TEAEs
Safety
Table 14.3.1.2
Summary of Treatment Emergent Adverse Events by Severity 
for Treatment Emergent Adverse Events – all TEAEs
Safety
Table 14.3.1.3
Summary of Treatment Emergent Adverse Events by System 
Organ Class and Preferred Term - Local Tolerability
Safety
Table 14.3.1.4
Summary of Treatment Emergent Adverse Events by Severity
for Treatment Emergent Adverse Events - Local Tolerability
Safety
Table 14.3.2
Listing of Deaths and Other Serious Adverse Events 
All Subjects
Table 14.3.3
Not part of TFL – Reserved for Narratives in CSR
Table 14.3.4
Listing of Abnormal Laboratory Values
Safety
Table 14.3.5 *
Summary of Intensity of Injection Site Pain - VAS
Per Protocol, 
FAS
Figure 14.3.6 *
Ping-Pong Plots of Individual VAS Scores by Treatment 
sequence
Per Protocol
Table 14.3.7 *
Statistical Analysis on Injection Site Pain - VAS
Per Protocol
Table 14.3.8 *
Summary of Intensity of Injection Site Pain - 6-Point Categorial 
Scale
Per Protocol, 
FAS
Table 14.3.9 *
Summary of Intensity of Injection Site Pain - Moderate or more 
injection site pain
Per Protocol, 
FAS
Table 14.3.10 *
Statistical Analysis on Intensity of Injection Site Pain -
Moderate or more injection site pain
Per Protocol
Table 14.3.11 *
Summary of Quality of Pain - Modified SF-MPQ-2 Inventory
Per Protocol, 
FAS
Table 14.3.12*
Summary of Duration of Pain
Per Protocol, 
FAS
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Table 14.3.13 *
Statistical Analysis on Duration of Pain
Per Protocol
Table 14.3.14 *
Summary of Comparative Pain - 5-Point Scale
Per Protocol, 
FAS
Table 14.3.15 *
Summary of Comparative Pain - DV3396 product hurt less 
than or about the same as the PDS290 product
Per Protocol, 
FAS
Table 14.3.16 *
Statistical Analysis on Comparative Pain - DV3396 product 
hurt less than or about the same as the PDS290 product
Per Protocol
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List of End of Text Listings:
Output
Title 
Section 16.2.1  Disposition
Listing 16.2.1.1
Subject Disposition
Listing 16.2.1.2
Deviation from Eligibility Criteria
Section 16.2.2  Protocol Deviations
Listing 16.2.2.1 
Reference to CSR
Section 16.2.3 Allocation to Analysis Sets
Listing 16.2.3.1 *
Overview Analysis Sets
Section 16.2.4  Demographics Data and Other Baseline Characteristics
Listing 16.2.4.1 *
Subject Demographics
Listing 16.2.4.2
Medical History
Listing 16.2.4.3
Previous Medications
Listing 16.2.4.4
Previous Injection Experience
Section 16.2.5 Compliance and/or Drug Concentration Data
Listing 16.2.5.1
Study Dates
Listing 16.2.5.2 *
Study Drug Administration
Section 16.2.6  Injection Pain Experience Data
Listing 16.2.6.1 *
Intensity of Injection Site Pain – VAS
Listing 16.2.6.2 *
Intensity of Injection Site Pain – Categorial Scale
Listing 16.2.6.3 *
Quality of Pain - Modified SF-MPQ-2 Inventory
Listing 16.2.6.4 *
Duration of Pain 
Listing 16.2.6.5 *
Comparative Pain Assessment
Section 16.2.7 Adverse Events Data
Listing 16.2.7.1
Adverse Events
Listing 16.2.7.2
Adverse Events Leading to Study Drug Discontinuation
Listing 16.2.7.3
Concomitant Medications and Non Drug Therapies 
Section 16.2.8 Laboratory Data
Listing 16.2.8.1
Clinical Laboratory Results – Chemistry
Listing 16.2.8.2
Clinical Laboratory Results – Hematology
Listing 16.2.8.3
Clinical Laboratory Results – Comments
Section 16.2.9 Other Safety Data
Listing 16.2.9.1
Vital Signs
Listing 16.2.9.2
12-Lead Electrocardiogram Results – Individual Parameters
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Listing 16.2.9.3
12-Lead Electrocardiogram Results – Investigator’s Interpretation and 
Specification of ECG Abnormalities
Listing 16.2.9.4
Physical Examination Findings
Other Appendix Outputs:
Output
Title 
Appendix 16.1.7
Randomization
Appendix 16.1.9.2
Statistical Appendices
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Statistical Analysis Plan
Protocol: INS-4582
Version Date: 09-Aug-2019
Page 27 of 27
Document History
Version Date
Modified/Reviewed 
By
Brief Summary of Changes (if created from a 
template, include template code)
Draft for internal review
Draft for NovoNordisk review
Final Draft for NovoNordisk review
Final version
Final version
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Statistical Methods
Version Date: 16Mar20
Page 1 of 6
STATISTICAL METHODS AND ANALYSIS OUTPUT
A TRIAL TO COMPARE THE INJECTION SITE PAIN EXPERIENCE OF 0.25 MG 
SEMAGLUTIDE SC ADMINISTERED BY 2 DIFFERENT PRODUCTS
CONFIDENTIAL
 code: 
Sponsor code: INS-4582
SPONSOR
Novo Nordisk A/S
AUTHOR
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Statistical Methods
Version Date: 16Mar20
Page 2 of 6
TABLE OF CONTENTS
1. DEVIATIONS FROM THE SAP .................................................................................... 3 
2. ANALYSIS POPULATIONS ......................................................................................... 3 
3. STATISTICAL ANALYSIS OUTPUT ............................................................................ 3 
3.1 
Statistical Analysis VAS ............................................................................................................... 3 
3.2 
Statistical Analysis Duration of Pain........................................................................................... 4 
3.3 
Statistical Analysis Comparative Pain ........................................................................................ 5 
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Statistical Methods
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Page 3 of 6
1.
Deviations from the SAP
-Previous injection Experience is listed together with Previous Medications in the Previous Medication Listing 
(16.2.4.3).
-Listing overview of pain Endpoints (16.2.6.6) has been added.
-Statistical Analysis on Intensity of Injection Site Pain - Moderate or more injection site pain: 
Since none of the subjects receiving the PDS290 product reported ‘moderate or more’ injection site pain the 
planned statistical analysis (McNemar mid-p with Bonett-Price hybrid score) was not possible. Table 13.3.10 
has not been created. 
-Plots Intensity of Injection Pain - 6-Point Categorical Scale have been added as figure 14.3.10.
-Table 14.3.12 has been split in 14.3.12.1 Summary of Duration of Pain and 14.3.12.2 Plots Duration of Injection 
Pain. 
2.
Analysis Populations
All 103 subjects were in the safety set and full analysis set.  Subject 185 was excluded from the per protocol set.
 
3.
Statistical Analysis Output  
3.1
Statistical Analysis VAS  
The Mixed Procedure 
          Number of Observations 
Number of Observations Read             204 
Number of Observations Used             204 
Number of Observations Not Used           0 
Covariance Parameter 
      Estimates 
Cov Parm     Estimate 
Residual       218.98 
        Type 3 Tests of Fixed Effects 
              Num     Den 
Effect         DF      DF    F Value    Pr > F 
TRTA            1      99     218.89    <.0001 
APERIOD         1      99       2.45    0.1205 
EXLAT           1      99       0.66    0.4187 
SUBJID        101      99       1.69    0.0048 
                                            Estimates 
                         Standard 
Label           Estimate    Error    DF    t Value    Pr > |t|     Alpha    Lower    Upper 
DV3396 - PDS290  30.6631   2.0725    99      14.79      <.0001      0.05  26.5507  34.7755 
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Statistical Methods
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 Least Squares Means 
           Actual                                      Standard 
Effect     Treatment    Laterality  Period Estimate       Error      DF    t Value  Pr > |t| 
TRTA       DV3396                           35.0619      1.4654      99      23.93    <.0001 
TRTA       PDS290                            4.3989      1.4654      99       3.00    0.0034 
APERIOD                             1       18.1078      1.4652      99      12.36    <.0001 
APERIOD                             2       21.3529      1.4652      99      14.57    <.0001 
EXLAT                   LEFT                18.8889      1.4654      99      12.89    <.0001 
EXLAT                   RIGHT               20.5719      1.4654      99      14.04    <.0001
Table Statistical Analysis on Injection Site Pain - VAS 
Per Protocol Set 
_________________________________________________________________________________________
                               ______LSmeans_______Difference Test/Reference_
Treatment Comparison           ________  _________               ___95% CI___  _p value__ 
(Test vs. Reference) Parameter Test      Reference Estimate      Lower  Upper     product 
_________________________________________________________________________________________
DV3396 - PDS290      VAS (mm)    35.1       4.40      30.7        26.6   34.8      <.0001 
_________________________________________________________________________________________
Model: ANOVA with a fixed effect for product (DV3396 and PDS290), injection number (1st 
2nd), injection side (left and right), and subject.
3.2
Statistical Analysis Duration of Pain
The Mixed Procedure 
          Number of Observations 
Number of Observations Read              92 
Number of Observations Used              92 
Number of Observations Not Used           0 
Covariance Parameter 
      Estimates 
Cov Parm     Estimate 
Residual        15006 
        Type 3 Tests of Fixed Effects 
              Num     Den 
Effect         DF      DF    F Value    Pr > F 
TRTA            1      43      17.25    0.0002 
APERIOD         1      43       4.01    0.0516 
EXLAT           1      43       1.71    0.1978 
SUBJID         45      43       1.42    0.1242 
                                               Estimates 
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Statistical Methods
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                          Standard 
Label          Estimate   Error   DF  t Value    Pr > |t|   Alpha     Lower  Upper 
DV3396 - PDS290  116.93 28.1562   43     4.15   0.0002    0.05   60.1466 173.71 
                                        Least Squares Means 
           Actual                                  Standard 
Effect     Treatment Laterality Period Estimate    Error  DF  t Value  Pr > |t| 
TRTA       DV3396                        188.65  19.0079  43     9.92    <.0001 
TRTA       PDS290                       71.7203  19.0079  43     3.77    0.0005 
APERIOD                         1        157.69  18.7580  43     8.41    <.0001 
APERIOD                         2        102.68  18.7580  43     5.47    <.0001 
EXLAT                LEFT                148.60  19.0079  43     7.82    <.0001 
EXLAT                RIGHT               111.77  19.0079  43     5.88    <.0001 
Table Statistical Analysis on Duration of Pain 
Per Protocol Set (conditional dataset) 
___________________________________________________________________________________________
                                    ______LSmeans______ _Difference Test/Reference_
Treatment Comparison                   ________  _________            ___95% CI__ _p value_ 
(Test vs. Reference) Parameter        Test    Reference  Estimate  Lower  Upper     product 
___________________________________________________________________________________________
DV3396 - PDS290    Pain Duration (sec) 189        71.7       116.9    60.1  173.7    0.0002 
__________________________________________________________________________________________
Model: ANOVA with a fixed effect for product (DV3396 and PDS290), injection number (1st 
2nd), injection side (left and right), and subject.
Note: The conditional duration of pain, given there is pain, is analyzed for subjects in the 
per protocol set who have a non-missing and non-zero duration of pain on both injections.
3.3
Statistical Analysis Comparative Pain
analysis proportion DV3396 hurt less/same as PDS290 with Clopper Pearson 95% CI 
The FREQ Procedure 
                  Analysis Value Category 1 
                                     Cumulative    Cumulative 
AVALCAT1    Frequency     Percent     Frequency      Percent 
________________________________________________________________
Yes                5        4.90             5         4.90
No                97       95.10           102       100.00
 Binomial Proportion 
    AVALCAT1 = Yes 
Proportion      0.0490 
ASE             0.0214 
Confidence Limits for the Binomial Proportion 
              Proportion = 0.0490 
Type                       95% Confidence Limits 
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Statistical Methods
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Clopper-Pearson (Exact)    0.0161         0.1107 
  Test of H0: Proportion = 0.5 
ASE under H0              0.0495 
Z                        -9.1094 
One-sided Pr <  Z         <.0001 
Two-sided Pr > |Z|        <.0001 
Sample Size = 102 
Table Statistical Analysis on Comparative Pain - DV3396 versus PDS290 
Per Protocol Set 
________________________________________________________________
_______________DV3396 hurts less/same as PDS290 *_______________ 
                                Proportion
 Yes           No               #
  n   (%)     n   (%)             Yes          95% CI
________________________________________________________________
  5 (4.9)    97 (95.1)           0.05       0.02 - 0.11
________________________________________________________________
*: DV3396 product hurt less than or about the same as the PDS290 product.
#: Proportion "Yes" and Clopper-Pearson (Exact) 95% CI.
Page 34 of 34