An Exploratory PET-study of Deposition, Disposition and Brain Uptake of [11C]Nicotine After Inhalation of 2 Nicotine Formulations Via the myBluTM E-cigarette in Smokers
Phase: NA
Conditions: Healthy Volunteers
Sponsor: Imperial Brands PLC

 
 
AN EXPLORATORY PET-STUDY OF DEPOSITION, DISPOSITION AND BRAIN 
UPTAKE OF [11C]NICOTINE AFTER INHALATION OF 2 NICOTINE 
FORMULATIONS VIA THE MYBLUTM E-CIGARETTE IN SMOKERS 
 
 
NCT# NCT04780815 
 
 
 
 
Study Protocol - FINAL v2.0; 30JAN2020  
 
 
 
 
 

 
 
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Clinical Study Protocol 
 
 
Investigational Product 
myblu™ e-cigarette system 
 
Sponsor study code 
NER 03/001 
 
Protocol Version and Date 
FINAL v2.0; 30JAN2020 
 
 
AN EXPLORATORY PET-STUDY OF DEPOSITION, DISPOSITION AND  
BRAIN UPTAKE OF [11C]NICOTINE AFTER INHALATION OF 2 NICOTINE 
FORMULATIONS VIA THE MYBLU™ E-CIGARETTE IN SMOKERS 
 
Test product and dose 
myblu™ e-cigarette system  
Formulation 1: 2.5% freebase nicotine formulation 
                        (no flavour) 
Formulation 2: 2.5% Nicotine salt formulation 
                        (no flavour) 
Sponsor signatory 
 
Nerudia Ltd, 
Wellington house 
Physics road, Speke 
Liverpool, L24 9HP 
United Kingdom 
Principal Investigator 
 
CTC Clinical Trial Consultants AB 
Uppsala University Hospital, 
Entrance 85, 2nd level  
SE-751 85 Uppsala, Sweden 
Clinical study conduct and 
management 
CTC Clinical Trial Consultants AB 
Uppsala University Hospital, 
Entrance 85, 2nd level  
SE-751 85 Uppsala, Sweden 
PET Centre, 
Uppsala University Hospital, 
Entrance 70, 1st floor, 
SE-751 85 Uppsala, Sweden 
CTC Clinical Trial Consultants AB 
Dag Hammarskjölds väg 10B 
SE-752 37 Uppsala, Sweden 
 
 
 

 
 
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1 
STUDY SYNOPSIS 
Study title 
An exploratory PET-study of deposition, disposition and brain uptake of [11C]nicotine after 
inhalation of 2 nicotine formulations via the myblu™ e-cigarette in smokers 
Study code 
NER 03/001 
Planned study period 
Q1 2020 to Q2 2020 
Principal Investigator  
 
CTC Clinical Trial Consultants AB, Uppsala, Sweden 
Study design 
This is a single centre, exploratory PET-study of deposition, disposition and brain uptake of 
[11C]nicotine when given to smokers as two different formulations via the myblu™ e-
cigarette system. 
The study is divided in two parts: 
PART A: PET 1 and PET 2 (including 5 subjects/formulation, total 10 subjects) 
PART B: PET 3 (including 2 subjects/formulation + 3 optional subjects, total 4-7 subjects) 
Objectives 
Primary objectives PART A 
• To measure the amount of nicotine deposited in the lungs and the oral cavity during 
40 minutes after administration of 11C-labelled nicotine via the myblu™ e-cigarette 
system using PET  
Secondary objectives PART A 
• To describe the single-dose pharmacokinetics of 11C-labelled nicotine, given via the 
myblu™ e-cigarette system, by measuring the radioactivity in arterial blood in discrete 
samples up to 30 minutes following administration 
• To evaluate safety and tolerability of 11C-labelled nicotine inhaled via the myblu™  
e-cigarette system 
Exploratory objectives PART B 
• To measure the amount of nicotine deposited in the brain during 30 minutes after 
administration of 11C-labelled nicotine via the myblu™ e-cigarette system using PET 
• To describe the single-dose pharmacokinetics of 11C-labelled nicotine, given via the 
myblu™ e-cigarette system, by measuring the radioactivity in arterial blood in discrete 
samples up to 30 minutes following administration 
• To evaluate safety and tolerability of 11C-labelled nicotine inhaled via the myblu™  
e-cigarette system 
 

 
 
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Endpoints 
Primary endpoints PART A 
• Dynamic PET Digital Imaging and Communications in Medicine (DICOM) 
images over the lungs and oral cavity 0-40 minutes after administration 
• Radioactivity vs. time profiles for the lungs and oral cavity 0-40 minutes after 
administration 
Secondary endpoints PART A 
• Radioactivity vs. time profiles for arterial blood at 2, 4, 6, 8, 10, 15, 20 and 30 minutes 
after administration of [11C]nicotine inhalation 
• Frequency, intensity and seriousness of adverse events (AEs) 
• Clinically significant changes in vital signs 
Exploratory endpoints (PART B) 
• Dynamic PET DICOM images over the brain 0-30 minutes after administration 
• Radioactivity vs. time profiles for arterial blood at 2, 4, 6, 8, 10, 15, 20 and 30 minutes 
after administration of [11C]nicotine inhalation 
• Frequency, intensity and seriousness of AEs 
• Clinically significant changes in vital signs 
Number of subjects planned 
Approximately 25 subjects will be screened to achieve a total of 14-17 included and 
evaluated subjects.  
Diagnosis and main eligibility criteria 
Male and female (of non-childbearing potential) smokers, aged 50-65 years, with a body 
mass index (BMI) of ≥ 18.0 and ≤ 30.0 kg/m2 will be considered for participation in the 
study. 
Subjects with a history of any clinically significant disease or disorder which, in the opinion 
of the Investigator, may either put the subject at risk because of participation in the study, or 
influence the results or the subject’s ability to participate in the study will be excluded from 
participation. Subjects, who previously have been exposed to radiation in studies, their 
medical history or their occupation, and who are under regular medical treatment for 
respiratory tract disorders or who use anticoagulantia will also be excluded from 
participation. 
For PART B, subjects with conditions contraindicating magnetic resonance imaging (MRI) 
will be excluded.  

 
 
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Methodology – PART A 
In PART A of the study, two different formulations of [11C]nicotine will each be given to 5 
subjects. The subjects will have two PET-assessments each (PET 1 and PET 2) after 
inhalation of [11C]nicotine via the myblu™ during the same treatment day. PART A will be 
completed for 5 subjects on each formulation before initiation of PART B. 
The IP will be given as inhalation (“puffs”) via the myblu™ e-cigarette system to obtain a 
tracer dose of up to 25 megabecquerel (MBq)/PET-session. 
Subjects will come for 2 visits to the clinic and have 1 End-of Study follow-up telephone 
call. 
Screening (Visit 1) will take place from Day -28 to Day -1 and will include an eligibility 
check, review of health status and inhalation training. 
At Visit 2, subjects will arrive to the clinic in the morning of Day 1 and will remain at the 
clinic until the afternoon. Day 1 will contain pre-PET safety assessment, preparation for the 
PET assessments (including insertion of an arterial catheter), two PET assessment sessions 
with at least 2 hours between the sessions and safety follow-up (including removal of arterial 
catheter) after the second PET assessment. 
The subjects will have breakfast (optional) at the research clinic before the first PET-session. 
Lunch will be served at the research clinic between the two PET-sessions. 
The subjects will be carefully monitored by clinical staff during and after the PET-sessions. 
There is immediate access to equipment and qualified staff in case of emergency.  
A final End-of-Study phone call (Visit 3) will take place on Day 7 (±2) or after early 
withdrawal to follow up on safety (AEs and concomitant medications). 
Methodology – PART B 
In PART B of the study, two different formulations of [11C]nicotine will each be given to 
2 subjects. The subjects will have one PET-assessments each (PET 3) after two inhalations of 
[11C]nicotine via the myblu™. 
PART B will be completed for 2 subjects on each formulation before a decision will be taken 
if 3 additional subjects should be added to PART B. The decision is based on whether the 
results from the first scans show significant, quantifiable uptake of radioactivity in the brain. 
If 3 additional subjects are included, they will be treated with the formulation that showed 
the best uptake profile. 
The IP will be given as inhalation (“puffs”) via the myblu™ e-cigarette system to obtain a 
tracer dose of up to 50 MBq. 
Subjects will come for 2 visits to the clinic, 1 MRI assessment visit and have 1 End-of Study 
follow-up telephone call (Visit 3).  
Screening (Visit 1) will take place from Day -28 to Day -1 and will include an eligibility 
check, review of health status and inhalation training.  
At Visit 2, subjects will arrive to the clinic in the morning of Day 1 and will remain at the 
clinic until the afternoon. Day 1 will contain pre-PET safety assessments, preparation for the 
PET assessments (including insertion of an arterial catheter), one PET assessment session 
and safety follow-up (including removal of arterial catheter) after the PET assessment. 

 
 
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The subjects will have breakfast (optional) at the research clinic before the PET-session. 
Lunch (optional) will be served at the research clinic after the PET session. 
The subjects will be carefully monitored by clinical staff during and after the PET-session. 
There is immediate access to equipment and qualified staff in case of emergency. 
A final End-of-Study phone call (Visit 3) will take place on Day 7 (±2) or after early 
withdrawal to follow up on safety (AEs and concomitant medications). 
During the study period, the subjects will also have an MRI-scan. The MRI can be performed 
at any suitable timepoint during the study after the screening visit until the End-of-Study-
visit. 
Investigational Product (IP), dosage and mode of administration 
The IPs will be supplied as the myblu™ e-cigarette system with e-liquid pods: 
The myblu™ e-cigarette will be supplied together with a USB charger. 
E-liquid pods will be supplied as: 
- 
Pods for training purposes, containing 2.5% freebase nicotine formulation (no flavour) 
corresponding to Formulation 1. 
- 
Pods for training purposes, containing 2.5% nicotine salt formulation (no flavour) 
corresponding to Formulation 2.  
- 
Empty pods (for filling at the PET Centre with radiolabelled Formulation 1 and 
Formulation 2) 
Bulk liquid Formulation 1 and Bulk liquid Formulation 2, will be supplied, for radiolabelling 
and filling of empty e-liquid pods at the PET Centre. 
Duration of treatment 
1 day 
Duration of each subject’s involvement in the study 
Each subject is expected to participate in the study for approximately 7 days from the day of 
PET-session (Day 1, Visit 2) to the End-of Study Visit. The Screening Visit will be 
performed within 28 days prior to PET-scanning (Day -28 to Day -1). 
PET assessment 
PET DICOM images over the lungs and oral cavity (PART A) and brain (PART B). 
Radioactivity vs. time profiles for arterial blood after administration of [11C]nicotine 
Safety assessments  
AEs 
Vital signs 
Statistical methods 
No formal sample size calculation has been performed for this study. The proposed sample 
size is considered sufficient to provide adequate information for the study objectives. 
Continuous data will be presented in terms of evaluable and missing observations, arithmetic 
mean, standard deviation (SD), median, minimum and maximum value. Categorical data will 
be presented as counts and percentages. When applicable, summary data will be presented by 

 
 
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treatment, and by assessment time. Individual subject data will be listed by subject number, 
treatment, and, where applicable, by assessment time. All descriptive summaries and 
statistical analyses will be performed using SAS Version 9.4 or later (SAS Institute, Inc., 
Cary, NC). Baseline will be defined as the visit with last data collection point prior to the 
first administration of IP. No imputation of missing data will be performed. 
The PET Centre will be responsible for evaluation of PET data, and for presenting time 
activity data on [11C]nicotine deposition during the dynamic scans over the two body 
positions in PET-scan 1 and 2. For the evaluation of [11C]nicotine kinetics in the lung, 
primary bronchi, oral cavity and airways, volumes of interest (VOIs) will be delineated, and 
time activity data generated utilising PET-image analysis software  
For PET-scan 3 acquired over the brain, the amount of nicotine distributed to the brain will 
be estimated. A whole brain VOI will be delineated on the T1-MR images overlaid on the 
brain PET-image. This part of the study is explorative and dependent on the time course of 
the [11C]nicotine uptake to the blood and the time activity uptake for brain nicotine may be 
difficult to measure with a sufficient signal-to-noise ratio. 
Study reporting 
A clinical study report (CSR) will be prepared after completion of the study. 
 
 

 
 
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2 
TABLE OF CONTENTS 
1 
STUDY SYNOPSIS ............................................................................................... 2 
2 
TABLE OF CONTENTS ...................................................................................... 7 
3 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS .................... 11 
4 
IMPORTANT MEDICAL PROCEDURES TO BE FOLLOWED BY THE 
INVESTIGATOR ............................................................................................................... 13 
4.1 
Medical emergencies contacts ............................................................................. 13 
5 
INVESTIGATOR AND STUDY ADMINISTRATIVE STRUCTURE ............ 13 
6 
INTRODUCTION ............................................................................................... 15 
6.1 
Background ......................................................................................................... 15 
 
myblu™ e-cigarette system ............................................................................ 15 
 
Radiochemistry .............................................................................................. 15 
6.2 
Study rationale .................................................................................................... 16 
6.3 
Risk/benefit assessment ....................................................................................... 16 
7 
STUDY OBJECTIVES AND ENDPOINTS ...................................................... 19 
7.1 
Primary objectives (PART A only)..................................................................... 19 
 
Primary endpoint (PART A only) .................................................................. 19 
7.2 
Secondary objectives (PART A only) ................................................................. 19 
 
Secondary endpoints (PART A only) ............................................................. 19 
7.3 
Exploratory objectives (PART B only) ............................................................... 19 
 
Exploratory endpoints (PART B only) ........................................................... 20 
8 
STUDY DESIGN ................................................................................................. 21 
8.1 
Overall study design and schedule of events ...................................................... 21 
 
PART A: PET 1 and PET 2 ............................................................................ 21 
 
PART B: PET 3 ............................................................................................. 24 
9 
STUDY POPULATION ...................................................................................... 26 
9.1 
Recruitment ......................................................................................................... 26 
9.2 
Screening and enrolment log .............................................................................. 26 
9.3 
Number of subjects ............................................................................................. 26 
9.4 
Inclusion criteria ................................................................................................. 26 
9.5 
Exclusion criteria ................................................................................................ 27 
9.6 
Restrictions during the study .............................................................................. 28 
 
General restrictions ........................................................................................ 28 
 
Prior and concomitant therapy ....................................................................... 29 
9.7 
Screen failures ..................................................................................................... 29 
9.8 
Subject withdrawal ............................................................................................. 29 
 
General withdrawal criteria ............................................................................ 29 
 
Procedures for discontinuation of a subject from the study ............................. 30 
 
Subject replacement ....................................................................................... 30 
9.9 
Randomisation .................................................................................................... 30 
9.10 
Blinding ............................................................................................................... 30 
10 
TREATMENTS .................................................................................................. 31 

 
 
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10.1 
Identity of investigational products .................................................................... 31 
10.2 
Manufacturing, packaging and labelling ........................................................... 31 
10.3 
Conditions for storage ......................................................................................... 31 
10.4 
Preparation and accountability .......................................................................... 31 
10.5 
Treatment administration................................................................................... 31 
10.6 
Continuation of treatment with Investigational Product .................................. 32 
10.7 
Treatment compliance ........................................................................................ 32 
10.8 
Return and destruction of investigational products .......................................... 32 
11 
STUDY ASSESSMENTS .................................................................................... 32 
11.1 
Recording of data ................................................................................................ 32 
11.2 
Demographics and other baseline characteristics .............................................. 32 
 
Informed consent ........................................................................................... 32 
 
Eligibility criteria ........................................................................................... 32 
 
Demographic information .............................................................................. 32 
 
Weight and height .......................................................................................... 33 
 
Medical/surgical history ................................................................................. 33 
 
Prior and concomitant medication .................................................................. 33 
 
HIV and Hepatitis B/C ................................................................................... 33 
 
Urine drug screen ........................................................................................... 33 
 
Alcohol breath test ......................................................................................... 33 
 
Baseline symptoms ........................................................................................ 33 
 
Clinical laboratory profile .............................................................................. 34 
 
Physical examination ..................................................................................... 34 
 
Resting 12-lead ECG ..................................................................................... 34 
11.3 
Assessments related to PET-endpoints ............................................................... 35 
 
Inhalation training .......................................................................................... 35 
 
Insertion/removal of arterial catheter .............................................................. 35 
 
Assessment of blood radioactivity .................................................................. 35 
 
PET/CT (PART A) ........................................................................................ 36 
 
PET/MR (PART B) ....................................................................................... 36 
11.4 
Assessments related to secondary endpoints ...................................................... 37 
 
PET endpoints ............................................................................................... 37 
 
Adverse events............................................................................................... 37 
 
Vital signs ...................................................................................................... 42 
11.5 
Appropriateness of measurements ..................................................................... 42 
12 
PROCEDURES FOR BIOLOGICAL SAMPLES............................................. 42 
12.1 
Sample collection ................................................................................................. 42 
12.2 
Volume of blood .................................................................................................. 42 
12.3 
Handling, storage and destruction of laboratory samples ................................. 43 
12.4 
Chain of custody of biological samples ............................................................... 43 
12.5 
Withdrawal of informed consent for donated biological samples ..................... 43 
13 
QUALITY MANAGEMENT, QUALITY ASSURANCE AND QUALITY 
CONTROL ......................................................................................................................... 43 
13.1 
Quality management: critical process, system and data identification ............. 43 
13.2 
Quality assurance and quality control ............................................................... 43 
14 
ETHICAL AND REGULATORY REQUIREMENTS ..................................... 44 

 
 
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14.1 
Ethical conduct of the study ............................................................................... 44 
14.2 
Ethics review ....................................................................................................... 44 
14.3 
Subject information and consent ........................................................................ 44 
14.4 
Subject data protection ....................................................................................... 45 
14.5 
Changes to the approved clinical study protocol ............................................... 45 
14.6 
Audits and inspections ........................................................................................ 45 
14.7 
Insurance ............................................................................................................. 46 
15 
STUDY MANAGEMENT .................................................................................. 46 
15.1 
Training of study site personnel ......................................................................... 46 
15.2 
Clinical monitoring ............................................................................................. 46 
15.3 
Source data documents ....................................................................................... 47 
15.4 
Study agreements ................................................................................................ 47 
15.5 
Study timetable and end of study ....................................................................... 47 
15.6 
Termination of the study .................................................................................... 48 
15.7 
Reporting and publication .................................................................................. 48 
 
Clinical study report....................................................................................... 48 
 
Confidentiality and ownership of study data................................................... 48 
 
Publication ..................................................................................................... 48 
15.8 
Archiving ............................................................................................................. 48 
16 
DATA MANAGEMENT .................................................................................... 49 
16.1 
The web-based eCRF .......................................................................................... 49 
16.2 
The entering of data into the eCRF .................................................................... 49 
16.3 
The query process ............................................................................................... 49 
16.4 
Audit trail ............................................................................................................ 50 
16.5 
External data ....................................................................................................... 50 
16.6 
Medical coding .................................................................................................... 50 
16.7 
Database lock ...................................................................................................... 50 
17 
STATISTICAL METHODS AND DETERMINATION OF SAMPLE SIZE .. 50 
17.1 
General ................................................................................................................ 50 
17.2 
Determination of sample size .............................................................................. 51 
17.3 
Analysis data sets ................................................................................................ 51 
 
Full analysis set ............................................................................................. 51 
 
Safety analysis set .......................................................................................... 51 
 
Per protocol set .............................................................................................. 51 
17.4 
Description of study population.......................................................................... 51 
 
Demographics and baseline characteristics ..................................................... 51 
 
Medical/surgical history and prior/concomitant medication ............................ 51 
 
Treatment compliance .................................................................................... 52 
 
Physical examination ..................................................................................... 52 
 
12-lead ECG .................................................................................................. 52 
 
Clinical laboratory analyses ........................................................................... 52 
17.5 
Analysis of PET endpoints .................................................................................. 52 
17.6 
Analysis of safety endpoints ................................................................................ 53 
 
Adverse events............................................................................................... 53 
 
Vital signs ...................................................................................................... 53 
18 
REFERENCES.................................................................................................... 54 

 
 
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19 
SIGNATURES .................................................................................................... 55 
19.1 
Principal Investigator statement ........................................................................ 55 
19.2 
Signature page (approval of the clinical study protocol) ................................... 56 
 
List of Tables 
Table 8.1-1 
Schedule of events PART A .......................................................................... 23 
Table 8.1-2 
Schedule of events PART B .......................................................................... 25 
Table 11.2-1 
Clinical laboratory parameters ................................................................... 34 
Table 12.2-1 
Estimated blood volumes ........................................................................... 42 
 
List of Figures 
Figure 6.1-1 
Synthetisation of S-[Methyl-11C]nicotine ....................................................... 15 
Figure 8.1-1 
Overview of the study design ........................................................................ 21 
 
 

 
 
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3 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS 
Abbreviation or term                   Explanation 
ADL 
Activities of daily living 
AE 
Adverse event 
ALT 
Alanine aminotransferase 
AST 
Aspartate aminotransferase 
ATC 
Anatomical therapeutic chemical  
Bq 
Becquerel 
BMI 
Body mass index 
CSP 
Clinical study protocol 
CSR 
Clinical study report 
CT 
Computerised tomography 
CTC 
Clinical Trial Consultants AB 
CTCAE 
Common terminology criteria for adverse events 
DICOM 
Digital imaging and communications in medicine 
DMP 
Data management plan 
ECG 
Electrocardiogram 
eCRF 
Electronic case report form 
EDC 
Electronic data capture 
EEA 
European Economic Area 
FAS 
Full analysis set 
FSH 
Follicle stimulating hormone 
GCP 
Good clinical practice 
GDPR 
General data protection regulation 
Hb 
Haemoglobin 
HIV 
Human immunodeficiency virus 
HPLC 
High-pressure liquid chromatography 
ICF 
Informed consent form 
ICH 
International Council for Harmonisation of 
Technical Requirements for Pharmaceuticals for 
Human Use  
IEC 
Independent ethics committee 
IME 
Important medical event 
IP 
Investigational product 

 
 
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ISF 
Investigator site file 
mAh 
Milliampere hour 
MBq 
Megabecquerel 
MedDRA 
Medical dictionary for regulatory activities 
mGy 
Milligray 
min 
minute 
mL 
Millilitre  
MRI 
Magnetic resonance imaging 
mSv 
millisievert 
PET 
Positron emission tomography 
PII 
Personally Identifiable Information 
PK 
Pharmacokinetics 
PPS 
Per protocol analysis set 
PT 
Preferred term 
PV 
Pharmacovigilance 
QA 
Quality assurance 
QC 
Quality control 
RBC 
Red blood cell 
RBM 
Risk based monitoring 
SAE 
Serious adverse event 
SAP 
Statistical analysis plan 
SD 
Standard deviation 
SDV 
Source data verification 
SOC 
System organ class 
SOP 
Standard operating procedures 
SPE 
Solid phase extraction 
TMF 
Trial master file 
ULDCT 
Ultra low dose computerised tomography 
V 
Volt 
VOI 
Volume of interest 
WBC 
White blood cell 
WHO 
World Health Organisation 

 
 
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Biostatistician 
 
 
 
 
Medical Writer 
 
 
 
 
Medical Monitor 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
  
 
 
 
 
 
 
 provided in Section 19. 

 
 
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6 
INTRODUCTION 
6.1 
Background 
E-cigarettes have become a popular alternative to cigarette smoking and are garnering 
significant attention as potentially reduced exposure products and smoking cessation products. 
E-cigarettes consist of a battery, heating component, and a reservoir (often referred to as a 
pod, cartridge, or tank) containing tobacco-derived nicotine in a solution composed of 
glycerine and/or propylene glycol and flavourings. Upon activation, the heating element heats 
the solution and the consumer inhales the resulting vapour. 
E-liquid formulations aim to provide smokers with an experience that would allow for 
maximal displacement of combustible cigarettes while at the same time not serving as a 
gateway to other more toxic products for non-smokers. Key elements that contribute to an 
acceptable initiating experience include the nicotine content in the products as well as the 
response that consumers have in terms of satisfaction and prevention of symptoms of 
withdrawal. One commercially successful development in e-liquid formulations is the use of 
nicotine salts, which reproduce a form of nicotine naturally available in the tobacco leaf and 
are able to provide higher levels of nicotine delivery. 
 myblu™ e-cigarette system 
myblu™ is a commercially available, two-piece, closed e-cigarette system comprised of a 
rechargeable 350 milliampere hour (mAh) battery and a disposable pod. The pods connect 
directly and contain the mouthpiece, heating element, are pre-filled with e-liquid, and are 
compatible only with myblu™ batteries. During use, a consumer inhales through the 
mouthpiece and a sensor in the battery detects the change in air pressure which activates the 
heating element. The e-liquid heats to an aerosol which the consumer inhales. 
The battery is charged with a micro-USB charger and produces a typical output of 3.7 Volt 
(V) (maximum 3.9 V). Under typical usage conditions, the pods would contain 1.5 millilitre 
(mL) of e-liquid which lasts approximately 200 puffs, depending on individual use 
behaviours. Each e-liquid contains a mixture of glycerine, propylene glycol, nicotine freebase, 
and a proprietary blend of flavours.  
 Radiochemistry 
S-[Methyl-11C]nicotine is synthesised according to Figure 6.1-1. 
 
                 
 
Figure 6.1-1 
Synthetisation of S-[Methyl-11C]nicotine 
[11C]nicotine is purified from the crude product by separation on high-pressure liquid 
chromatography (HPLC) and obtained in ammonium formate (50 mM pH 3.5) and 
acetonitrile. The volume is reduced by evaporation to remove most of the acetonitrile and then 
the product solution is made alkaline by addition of Na2CO3 and passed through a C18 solid 
phase extraction column (SPE). The C18 SPE is washed with sterile water followed by 
flushing with nitrogen gas to remove ammonium formate and acetonitrile and most of the 
excess of water. The retained [11C]nicotine is eluted using the Nerudia nicotine solution to be 

 
 
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used in the inhalation device. The amount of radioactivity in the eluate is measured and the 
inhalation device is filled with appropriate volume corresponding to delivery of 25 
megabecquerel (MBq) by a 3-5 second inhalation. A volume of about 10-24 L is delivered to 
the subject. The volume administered is dependent on the yield of the 11C-labelling, that may 
vary between batches. 
The inhalation solution is quality controlled by: 
• Measurement of radioactivity 
• Determination of radiochemical purity 
• Determination of concentration of residual acetonitrile, to assure the level is below the 
IHC recommendations 
6.2 
Study rationale 
This study seeks to investigate the differences in airway deposition of nicotine from e-liquid 
vapour, between different e-liquid formulations. Two formulations are included in the study, 
one which uses freebase nicotine and one that uses nicotine salts (nicotine lactate). Both 
freebase nicotine and nicotine salts (nicotine lactate) are available in commercial products and 
previous clinical research carried out by Imperial Brands and by others in the industry has 
demonstrated differences in the pharmacokinetics (PK) between freebase and nicotine salts. 
This study aims to identify if the differences in PK responses between the formulations are 
linked to different sites of absorption of the nicotine. By radiolabelling and imaging, we aim 
to identify where the nicotine is within the respiratory tract which may give insight into the 
site of nicotine absorption. 
In addition to differences in PK, the formulations have different chemical properties. Nicotine 
has greater volatility in the freebase form compared to nicotine salts which has an impact on 
the sensorial effect when using the product, with greater throat irritation with freebase 
compared to nicotine salt formulations. This, in combination with literature on Nicorette 
Inhalator which demonstrated that gas phase nicotine mostly did not enter the lung, gives an 
indication that nicotine in the different formulations may reach different areas of the 
respiratory tract. 
6.3 
Risk/benefit assessment 
It may be considered problematic to expose research subjects to a nicotine delivery product. 
However, all research subjects are required to be daily cigarette users, so the participants are 
well acquainted with, and used to, the effects of nicotine. 
The investigational product (IP) myblu™ is currently commercially available on multiple 
markets. The amount of nicotine inhaled from a single puff of the IP is lower than in 
conventional tobacco-based cigarettes. This suggests that adverse effects from the nicotine 
exposure from the IP are less likely to occur among the research subjects. So far, no adverse 
effects have been reported associated with the use of the IP apart from known effects likely to 
be related to the nicotine exposure (such as salivation, nausea, and dyspepsia). 
Subjects below the age of 50 years are excluded from the study due to the radiation exposure. 
In addition, subjects with respiratory diseases or ongoing respiratory infections will be 
excluded from participation due to potential risks related to IP inhalation. 

 
 
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Risk associated with the e-cigarettes 
A risk assessment of the e-cigarette has been conducted as part of design control 
requirements. The identified hazards were mostly minor or moderate in severity and were 
mostly expected to occur only rarely. Potential hazards with a rating of “severe” or above 
included exposure to formaldehyde, nitrosamines or other contaminants, incorrect charging 
leading to some e-cigarettes overheating, and microbial infection from sharing devices. 
Chemical analysis of the condensate has confirmed very low levels of contaminants, and in 
the present study the e-cigarette will be used under supervision in a controlled environment. 
Therefore, the likelihood of any of these potential identified hazards occurring is considered 
to be extremely low. 
Risk associated with the arterial catheter 
There is a small risk for pain upon insertion, and also a few days after removal, of the arterial 
catheter. There is also a minor risk for numbness as well as impaired blood flow. Sufficient 
circulation is tested for with modified Allen test at screening. To minimise the risks with 
arterial catheters, the insertion is done by an anaesthesiologist, under hygienic conditions. 
Upon removal of the catheter, compression over the artery will be held for approximately 
10 minutes, after which a compressing bandage is applied. 
Risk associated with the radiation exposure 
Radioactively labelled nicotine will be given to the study subjects. The amount of 
radioactivity expected in the device is estimated to be about 200 MBq at the time of 
administration. Approximately up to 25 MBq is administered to the subject by two 3-5 second 
inhalations, as 1-2 consecutive puffs withdrawn from the device for each tracer 
administration. The effective dose of 50 MBq (2 inhalation administrations) of [11C]nicotine 
is not likely to exceed 0.2 millisievert (mSv). 
The effective dose in radiation protection and radiology is a measure of the cancer risk to a 
whole organism due to ionising radiation delivered non-uniformly to part(s) of its body. It 
takes into account both the type of radiation and the nature of each organ being irradiated. 
Additionally, the Ultra Low Dose Computed Tomography (ULDCT) used for attenuation 
correction and for anatomical orientation, will give a radiation dose of approximately 0.28 
mSv for the entire study. For subjects participating in Part B (Brain scan only) the dose from 
ULDCT is 0.05 mSv per investigation. 
The total radiation dose to the subjects participating in this study with two positron emission 
tomography/computerised tomography (PET/CT) investigations and two inhalations of [11C]-
labelled nicotine will thus be less than 1.0 mSv (0.5 mSv for part A and 0.3 mSv for part B). 
This dose falls within Category I that allows up to 1 mSv for subjects with an age above 
50 years. This dose limit relies on the guideline: “European commission Radiation protection 
99, Guidance on medical exposures in medical and biomedical research. 1998 Directorate-
General Environment, Nuclear Safety and Civil protection”. 
To our knowledge, there are no publications reporting effective dose due to [11C]nicotine 
inhalation or intravenous injection. The Swedish Radiation Safety Authority states an 
effective dose of 0.004 mSv/MBq for intravenous injection of neuroreceptor ligands, 
compared to 0.003 mSv/MBq for 11CO inhalation. Homogeneous distribution of a [11C]-
labelled tracer results in 0.003 mSv/MBq as well. Based on this data, the effective dose of 
50 MBq [11C]nicotine inhalation is not likely to exceed 0.004 mSv/MBq, or 0.2 mSv. 
Bergström et al, [1] reported 45% of administered nicotine vapour in the oral cavity at end of 
inhalation, and 8% at 45 minutes past inhalation. Assuming that 50% of the inhaled activity 
(25 MBq) remains on the mucosa of the oral cavity, with an effective half life of ten minutes, 

 
 
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and that all beta radiation is absorbed in the top 1 mm layer of the mucosa, the dose to oral 
cavity mucosa is estimated to be around 60 milligray (mGy). These absorbed doses are far 
below levels resulting in any risks of deterministic effects (e.g., 6 Gy for skin rash), even 
though mucosa is probably more radiation sensitive than regular skin. In another PET study 
reporting inhalation of [11C]nicotine [2], far larger amounts of radioactivity were inhaled (170 
MBq per inhalation) without any adverse effect. 
Risk associated with the magnetic resonance imaging (MRI) 
Subjects who have any condition that is contraindicated with the MRI examination will be 
excluded for PART B. 
Incidental findings during the MRI investigation will be evaluated and documented by a 
radiologist at the imaging site according to local practice. A clinical statement based on the 
evaluation will be provided to the Principal Investigator, who will inform the subject about 
the findings and handle the subject’s medical care as per standard medical/clinical judgement. 
In order to protect subjects and equipment during the MRI examination all personal and 
medical metallic and magnetic items (including credit cards etc.) have to be left outside the 
MRI room. In extensive research to evaluate whether the magnetic fields and radio waves 
used during an MRI scan pose a risk to the human body, no evidence for any risks have been 
identified. MRI is thus considered one of the safest medical imaging procedures currently 
available. 
General risks 
Besides the risks related to the insertion of arterial catheters, MRI and radiation as described 
above, there may also be risks related to the medical devices used in the study e.g. clinical 
laboratory sampling materials. However, these are devices that are used in routine medical 
care and the risk associated with their use is considered low and ethically justifiable. Study 
specific evaluations and sampling procedures, like blood-pressure measurements using a 
blood pressure cuff, may cause transient discomfort but the risk is deemed to be low and 
ethically justifiable. 
Subjects will remain in the research clinic for at least 2 hours after the administration of the IP 
and will be closely monitored by medical staff. Visits at the clinic may be prolonged in case 
the Investigator finds it medically warranted for safety reasons. 
Overdosing is not likely to occur since all IP will be administered by site personnel under 
medical surveillance. In cases of accidental overdose, standard supportive measures should be 
adopted as required. For further information regarding overdosing, refer to Section 11.4.2.13. 
The Principal Investigator at the research clinic will ascertain that adequate facilities and 
procedures are available to handle unexpected safety situations should they occur during the 
study. 
The potential adverse effects of the study procedures, which are likely to be minor and/or 
clinically insignificant, are from a research ethics perspective counterbalanced by the 
potential positive effects of the novel nicotine delivery system as a reduced toxicity 
alternative to conventional cigarettes. As the nicotine disposition and deposition profile of a 
product is likely to be central to its acceptability among current tobacco users, it is reasonable 
to conduct formal clinical studies to assess these features in more detail.  
 
 

 
 
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7 
STUDY OBJECTIVES AND ENDPOINTS 
7.1 
Primary objectives (PART A only) 
 The primary objectives are: 
• To measure the amount of nicotine deposited in the lungs and the oral cavity during 
40 minutes after administration of 11C-labelled nicotine via the myblu™ e-cigarette 
system using PET  
 Primary endpoint (PART A only) 
The primary endpoints are: 
• Dynamic PET Digital Imaging and Communications in Medicine (DICOM) 
images over the lungs and oral cavity 0-40 minutes after administration 
• Radioactivity vs. time profiles for the lungs and oral cavity 0-40 minutes after 
administration 
7.2 
Secondary objectives (PART A only) 
The secondary objectives are: 
• To describe the single-dose pharmacokinetics of 11C-labelled nicotine, given via the 
myblu™ e-cigarette system, by measuring the radioactivity in arterial blood in discrete 
samples up to 30 minutes following administration 
• To evaluate safety and tolerability of 11C-labelled nicotine inhaled via the myblu™  
e-cigarette system 
 Secondary endpoints (PART A only) 
• Radioactivity vs. time profiles for arterial blood at 2, 4,6, 8, 10, 15, 20 and 30 minutes 
after administration of [11C]nicotine inhalation 
• Frequency, intensity and seriousness of adverse events (AEs) 
• Clinically significant changes in vital signs 
7.3 
Exploratory objectives (PART B only) 
The exploratory objective for PART B is: 
• To measure the amount of nicotine deposited in the brain during 30 minutes after 
administration of 11C-labelled nicotine via the myblu™ e-cigarette system using PET 
• To describe the single-dose pharmacokinetics of 11C-labelled nicotine, given via the 
myblu™ e-cigarette system, by measuring the radioactivity in arterial blood in discrete 
samples up to 30 minutes following administration 
• To evaluate safety and tolerability of 11C-labelled nicotine inhaled via the myblu™  
e-cigarette system 

 
 
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 Exploratory endpoints (PART B only) 
• Dynamic PET DICOM images over the brain 0-30 minutes after administration 
• Radioactivity vs. time profiles for arterial blood at 2, 4,6, 8, 10, 15, 20 and 30 minutes 
after administration of [11C]nicotine inhalation 
• Frequency, intensity and seriousness of AEs 
• Clinically significant changes in vital signs 
 
 

 
 
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PET assessments (including insertion of an arterial catheter), two PET assessment sessions 
with at least 2 hours between the sessions and safety follow-up (including removal of arterial 
catheter) after the second PET assessment. 
The subjects will have breakfast (optional) at the research clinic before the first PET-session. 
Lunch will be served at the research clinic between the two PET-sessions. 
The subjects will be carefully monitored by clinical staff during and after the PET-sessions. 
There is immediate access to equipment and qualified staff in case of emergency.  
A final End-of-Study phone call (Visit 3) will take place on Day 7 (±2) or after early 
withdrawal to follow up on safety (AEs and concomitant medications). 
Each subject is expected to participate in the study for approximately 7 days from the day of 
PET-sessions (Day 1, Visit 2) to the End-of Study Visit. The Screening Visit will be 
performed within 28 days prior to PET-scanning (Day -28 to Day -1). 
The schedule of events is shown in Table 8.1-1. 
Study assessments are described in Section 11. 

 
 
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 PART B: PET 3 
In PART B of the study, two different formulations of [11C]nicotine will each be given to 2 
subjects. The subjects will have one PET-assessments each (PET 3) after two inhalations of 
[11C]nicotine via the myblu™. 
PART B will be completed for 2 subjects on each formulation before a decision will be taken 
if 3 additional subjects should be added to PART B. The decision is based on whether the 
results from the first scans show significant, quantifiable uptake of radioactivity in the brain. 
If 3 additional subjects are included, they will be treated with the formulation that showed the 
best uptake profile. 
The IP will be given as inhalation (“puffs”) via the myblu™ e-cigarette system to obtain a 
tracer dose of up to 50 MBq. 
Subjects will come for 2 visits to the clinic, 1 MRI assessment visit (if not on the PET 
assessment day) and have 1 End-of Study follow-up telephone call (Visit 3). 
Screening (Visit 1) will take place from Day -28 to Day -1 and will include an eligibility 
check, review of health status and inhalation training, see Table 8.1-2 for details.  
At Visit 2, subjects will arrive to the clinic in the morning of Day 1 and will remain at the 
clinic until the afternoon. Day 1 will contain pre-PET safety assessment, preparation for the 
PET assessments (including insertion of an arterial catheter), one PET assessment session and 
safety follow-up (including removal of arterial catheter) after the PET assessment. 
The subjects will have breakfast (optional) at the research clinic before the PET-session. 
Lunch (optional) will be served at the research clinic after the PET session. 
The subjects will be carefully monitored by clinical staff during and after the PET-session. 
There is immediate access to equipment and qualified staff in case of emergency. 
A final End-of-Study phone call (Visit 3) will take place on Day 7 (±2) or after early 
withdrawal to follow up on safety (AEs and concomitant medications). 
During the study period, the subjects will also have an MRI-scan. The MRI can be performed 
at any suitable timepoint during the study after the screening visit until the End-of-Study-
visit. 
Each subject is expected to participate in the study for approximately 7 days from the day of 
PET-session (Day 1, Visit 2) to the End-of Study Visit. The Screening Visit will be performed 
within 28 days prior to PET-scanning (Day -28 to Day -1). 
The schedule of events is shown in Table 8.1-2. 
Study assessments are described in Section 11. 

 
 
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9 
STUDY POPULATION  
Prospective approval of protocol deviations to eligibility criteria, also known as protocol 
waivers or exemptions, is not permitted. 
9.1 
Recruitment 
The subjects will be recruited from CTC’s database of healthy volunteers and patients and 
from advertising in media (including social media). 
Subjects may not participate in more than one part of the study. 
9.2 
Screening and enrolment log 
Investigators must keep a record of all screened subjects even if they were not subsequently 
included in the study. This information is necessary to verify that subjects were selected 
without bias. The reason for screen failure should be stated for all subjects screened but not 
included. The reason for withdrawal should be stated for all subjects included but not 
completed. 
A screening number will be allocated to each subject in connection to the informed consent 
process at the Screening visit. The screening number is generated automatically in the 
electronic case report Form (eCRF). The screening number will allow identification of 
subjects irrespective of their possible eligibility for the study.  
Subjects included will be assigned a subject number (101, 102, 103 etc. for PART A and 201, 
202, 203 etc. for PART B). The first digit will correspond to the study part, and the 2nd and 
the 3rd digits correspond to the subject number. 
If a subject cannot receive the planned dose of IP within 28 days after screening (i.e., the time 
interval between signing informed consent until dose administration) the subject should be 
rescreened before proceeding in the study. 
9.3 
Number of subjects 
Approximately 25 subjects will be screened to achieve a total of 14-17 included and evaluated 
subjects. 
For replacements of subjects who discontinue from the study, see Section 9.8. 
9.4 
Inclusion criteria 
For inclusion in the study, subjects must fulfil the following criteria: 
1. Willing and able to give written informed consent for participation in the study. 
2. Male subject or female subject of non-child bearing potential, aged 50-65 years 
inclusive. 
Women of non-childbearing potential are defined as pre-menopausal females who are 
sterilised (tubal ligation or permanent bilateral occlusion of fallopian tubes); or post-
menopausal defined as 12 months of amenorrhea (in questionable cases a blood sample 

 
 
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with simultaneous detection of follicle stimulating hormone [FSH] 25-140 IE/L and 
oestradiol <183 pmol/l is confirmatory). 
3. BMI ≥ 18.0 and ≤ 30.0 kg/m2. 
4. Habitual daily cigarette smoker. 
9.5 
Exclusion criteria 
Subjects must not enter the study if any of the following exclusion criteria are fulfilled: 
1. History of any clinically significant disease or disorder which, in the opinion of the 
Investigator, may either put the subject at risk because of participation in the study, or 
influence the results or the subject’s ability to participate in the study. 
2. Any clinically significant illness, medical/surgical procedure or trauma within 4 weeks 
of the (first) administration of IP that would influence the results or the subject’s ability 
to participate in the study, as judged by the Investigator. 
3. Any clinically significant disease affecting the respiratory tract that would influence the 
results of the study or the subject’s ability to participate in the study, as judged by the 
Investigator. 
4. Malignancy within the past 5 years with the exception of in situ removal of basal cell 
carcinoma. 
5. Any planned major surgery within the duration of the study. 
6. Any positive result on screening for serum hepatitis B surface antigen, hepatitis C 
antibodies and HIV. 
7. Untreated hypertension and/or, after 10 minutes supine rest at the time of screening, any 
vital signs values outside the following ranges: 
• Systolic blood pressure <90 or >160 mmHg, or 
• Diastolic blood pressure <50 or >100 mmHg, or  
• Pulse <40 or >90 bpm 
8. Clinically significant abnormalities in the resting ECG at the time of screening that 
would influence the results of the study or the subject’s ability to participate in the 
study, as judged by the Investigator. 
9. Previous participation in a PET-study or other nuclide medical study. 
10. Risk of occupational exposure to significant ionising radiation. 
11. Previous participation in any clinical procedures involving significant exposure to 
radiation (exceptions are dental X-rays and common X-rays of chest or extremities). 
12. Negative results of the modified Allen test on both arms at the time of screening or pre-
arterial catheter insertion at Visit 2. 
13. Current ongoing upper respiratory tract infection with e.g. coughing at the day of the 
PET-assessment, as judged by the Investigator. 

 
 
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14. Regular use of any prescribed or non-prescribed medication for the treatment of 
respiratory tract disorders within 2 weeks prior to the (first) administration of IP, at the 
discretion of the Investigator. 
15. Regular use of any anticoagulantia medication within 2 weeks prior to the (first) 
administration of IP, at the discretion of the Investigator. 
16. Known allergy towards local anaesthetics of the type used in connection with arterial 
catheter insertion. 
17. Planned treatment or treatment with investigational drugs within 3 months prior to 
Visit 2. 
18. Positive screen for drugs of abuse or alcohol at screening or on admission to the 
research unit on Visit 2. 
19. History of alcohol abuse or excessive intake of alcohol, as judged by the Investigator. 
20. Presence or history of drug abuse and/or anabolic steroids, as judged by the 
Investigator. 
21. Investigator considers the subject unlikely to comply with study procedures, restrictions 
and requirements. 
22. For PART B only: Conditions contraindicating MRI, including, but not limited to, 
claustrophobia, metallic implants or internal electrical devices (e.g., cochlear implant, 
nerve stimulator, gastric pacemaker, bladder stimulator, pacemaker, defibrillator, 
artificial valves in heart, aneurysm clips, etc.), and permanent makeup or tattoos which 
in the Investigator’s opinion might jeopardise the subject’s safety or interfere with the 
imaging measurements. Subjects with metal fragments in the head or body that would 
present a risk during the MRI scanning procedure, or subjects who have worked with 
ferrous metals either as a vocation or hobby (for example, as a sheet metal worker, 
welder or machinist) in such a way that might have led to unknown indwelling metal 
fragments that could cause injury if moved in response to the magnetic field during 
MRI. 
9.6 
Restrictions during the study 
The subjects must be willing to comply to the following restrictions during the entire study 
duration i.e., from screening to the end-of-study visit. 
 General restrictions 
• Meals and Dietary Restrictions: Meals (breakfast and lunch) will be served at the 
research clinic during Visit 2. Lunch will be served between the two PET-sessions (for 
subjects participating in PART A). Water is allowed ad libitum at the clinic except 
during PET-sessions. 
• Nicotine: Smoking or use of nicotine-containing products, except the IP, is not allowed 
during the study from midnight the day before the PET assessment until the subject 
leaves the clinic after the (last) PET assessment. 
• Alcohol: Consumption of alcohol is not allowed within 48 hours prior to or during 
Visit 2. 
• Coffee: Consumption of up to 5 cups of coffee will be allowed during Visit 2.  

 
 
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• Xanthine or taurine containing products/beverages: Energy drinks (e.g. Red bull) are not 
allowed during Visit 2. 
• Blood donation: The subjects must not donate blood or plasma during the study until 
one month after Visit 2. 
• Participation in other clinical studies: Study subjects are not allowed to participate in 
any other interventional clinical study during the study period. 
 Prior and concomitant therapy 
Prohibited medication 
• Regular use of any prescribed or non-prescribed medication for the treatment of 
respiratory tract disorders within 2 weeks prior to the (first) administration of IP, at the 
discretion of the Investigator. 
• Anticoagulantia. 
Allowed medication 
• Medications considered necessary for treatment of chronic/stable conditions that are 
part of the subject’s medical history, as judged by the Investigator. 
• Other medications considered necessary for the subject´s safety and wellbeing may be 
given at the discretion of the Investigator during the study period. 
9.7 
Screen failures 
Screen failures are defined as subjects who consent to participate in the clinical study but are 
not subsequently included in the study. A minimal set of screen failure information is required 
to ensure transparent reporting of screen failure subjects. Minimal information includes 
documentation of signed and dated informed consent form (ICF) and reason(s) for screening 
failure. 
Re-screening can be performed once if any of the following were reasons for screening failure 
or non-randomisation (as judged by the Investigator): 
• Practical reasons.  
• Non-significant medical conditions (e.g. influenza, nasopharyngitis).  
For subjects who are re-screened, a new screening number will be assigned and a new, signed 
ICF will be collected. 
9.8 
Subject withdrawal 
 General withdrawal criteria 
Subjects are free to discontinue their participation in the study at any time and for whatever 
reason without affecting their right to an appropriate follow-up investigation or their future 
care. If possible, the reason for withdrawal of consent should be documented. 
Subjects may be discontinued from the study at any time at the discretion of the Investigator. 

 
 
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Reasons for discontinuation include: 
• Subject decision 
• Severe non-compliance to study protocol procedures, as judged by the Investigator 
and/or Sponsor 
• Subject is lost to follow-up. 
• Significant AEs posing a risk for the subject, as judged by the Investigator and/or 
Sponsor 
• Withdrawal of informed consent to the use of biological samples 
• Pregnancy 
• Death 
• Meeting of an exclusion criterion during the study, which, in the opinion of the 
Investigator, may pose a risk for the subject  
• Use of prohibited medication 
 Procedures for discontinuation of a subject from the study 
A subject who prematurely discontinues participation in the study will always be asked about 
the reason(s) for discontinuation and the presence of any AEs. If a subject withdraws consent, 
the Investigator must ask the subject if he/she is willing, as soon as possible, to be assessed 
according to the procedures scheduled for the end-of-study visit. Any ongoing AEs will be 
followed as described in Section 11.4.2.11. 
The primary reason for discontinuation/early withdrawal must be specified in the eCRF and 
final drug accountability must be performed. 
 Subject replacement 
Subjects who are prematurely withdrawn from the study before completing the intended PET-
assessments may be replaced. 
9.9 
Randomisation 
Not applicable. 
9.10 Blinding 
This is an open-label study, i.e. the Investigator, study staff and subjects will know the type of 
IP to be received. 
 
 

 
 
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10 TREATMENTS  
10.1 Identity of investigational products  
The IPs will be supplied as the myblu™ e-cigarette system with e-liquid pods: 
The myblu™ e-cigarette will be supplied together with a USB charger. 
E-liquid pods will be supplied as: 
- 
Pods for training purposes, containing 2.5% freebase nicotine formulation (no flavour) 
corresponding to Formulation 1. 
- 
Pods for training purposes, containing 2.5% nicotine salt formulation (no flavour) 
corresponding to Formulation 2.  
- 
Empty pods (for filling at the PET Centre with radiolabelled Formulation 1 and 
Formulation 2) 
Bulk liquid Formulation 1 and Bulk liquid Formulation 2, will be supplied, for radiolabelling 
and filling of empty e-liquid pods at the PET Centre. 
10.2 Manufacturing, packaging and labelling 
The IP is manufactured, packed and labelled by Nerudia Ltd, Liverpool, United Kingdom. 
Labelling of e-liquid pods filled with radioactive e-liquid formulation will be labelled 
individually by the PET Centre before each administration to an individual study subject. 
10.3 Conditions for storage 
The IP will be stored at room temperature away from direct sunlight and heat. 
Temperature logs will be kept for the area where the IP is stored. The temperature should be 
noted on a daily basis (working days only unless automatic temperature readings are 
available). 
10.4 Preparation and accountability 
Preparation of the inhalation device (preparation of radiolabelled formulations and filling of e 
liquid pods) before each PET-sessions will be done by trained personnel at the PET Centre.  
CTC/PET Centre will maintain a Storage and Accountability Log detailing the dates and 
quantities of study medication received, prepared for and used by each subject and IP returned 
or destroyed at the end of the study. Any discrepancies between prepared and returned IP 
must be explained and documented. Products deliberately and/or accidentally destroyed by 
the site or the subject must be accounted for. 
10.5 Treatment administration 
Instructions on the proper use of the myblu™ products will be provided prior to use. Product 
use in the study sessions will consist of 1-2 puffs of each 3-5 seconds, followed by an 
inhalation of the vapour into the lungs. 

 
 
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Inhalation training of study subjects, see 11.3.1. 
10.6 Continuation of treatment with Investigational Product 
This is an exploratory study in volunteers who will have no medical benefit from the 
treatment and thus there will be no treatment after end of study participation. 
10.7 Treatment compliance 
All IP will be administered at the research clinic under supervision of trained study personnel 
to ensure compliance. 
10.8 Return and destruction of investigational products 
Any unused study product and all empty containers will be destructed at the site upon 
confirmation from the Sponsor. The Monitor will perform final IP accountability 
reconciliation at the study end to verify that all unused IP is adequately destroyed and 
documented. 
11 STUDY ASSESSMENTS 
The study assessments are described in the sections below and the timing of these assessments 
are detailed in the schedule of events (PART A: Table 8.1-1 and PART B: Table 8.1-2) 
11.1 Recording of data 
The Principal Investigator will provide the Sponsor with all data produced during the study 
from the scheduled study assessments. He/she ensures the accuracy, completeness, legibility, 
and timeliness of the data reported to Sponsor in the eCRF and in all required reports. 
11.2 Demographics and other baseline characteristics 
 Informed consent 
Signed informed consent must be obtained before any screening procedures are initiated. The 
informed consent procedure is further described in Section 14.3. 
 Eligibility criteria 
Eligibility criteria should be checked during screening and verified before IP administration. 
The criteria are specified in Sections 9.4 and 9.5. 
 Demographic information 
The following demographic data will be recorded: gender, age, ethnicity and race. 

 
 
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 Weight and height 
Weight and height will be measured without shoes. BMI will be calculated with one decimal 
from the height and weight recorded. 
 Medical/surgical history  
Medical/surgical history will be obtained by subject interview in order to verify that the 
eligibility criteria are met. 
The medical/surgical history should include all relevant diseases and surgeries within 2 
months prior to screening as judged by the Investigator. 
 Prior and concomitant medication 
Prior medications taken within 2 weeks will be obtained by subject interview in order to 
verify that the eligibility criteria are met (see also Section 9.6.2). 
Medications are classified as prior if the stop date was before or on the day of the first dose 
administration (pre-dose) and as concomitant if ongoing on the day of the first dose 
administration, stopped after the first dose administration or started after the first dose 
administration. To distinguish between prior and concomitant medications on Day 1 in each 
part (i.e. the first IP administration), the start time of any newly introduced medication or the 
stop time of any previously ongoing medication must be recorded in the eCRF.  
Any use of concomitant medication from screening until the last end-of-study visit must be 
documented appropriately in the subject’s eCRF. Relevant information (i.e. name of 
medication, dose, unit, frequency, start and stop dates, reason for use) must be recorded. All 
changes in medication should be noted in the eCRF. 
 HIV and Hepatitis B/C 
Subjects will be tested for HIV, hepatitis B surface antigen and hepatitis C antibodies prior to 
inclusion into the study. Any positive result will exclude the subject from participating in the 
study. 
 Urine drug screen 
Urine will be screened for drugs of abuse at timepoints outlined in the schedule of events 
(Table 8.1-1 and Table 8.1-2) using the AlereTM Drug Screen Test Panel. 
 Alcohol breath test 
An alcohol breath test will be performed at timepoints outlined in the schedule of events 
(Table 8.1-1 and Table 8.1-2). 
 
 Baseline symptoms 
A baseline symptom is defined as an event that occurs between the subject’s signing of the ICF 
until the insertion of the arterial catheter (i.e. an event that occurs during the screening 

 
 
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period). Such events are not AEs and will be recorded as baseline symptoms in the Medical 
History Log in the eCRF. 
  Clinical laboratory profile 
Blood samples for analysis of clinical chemistry and haematology will be collected through 
venepuncture or an indwelling venous catheter and sent to the certified clinical chemistry 
laboratory at Uppsala University Hospital and analysed by routine analytical methods.  
The clinical laboratory parameters are defined in Table 11.2-1 and will be assessed at 
screening. 
Abnormal values assessed by the Investigator as clinically significant will be reported as 
baseline symptoms. If an abnormal value is associated with corresponding clinical signs or 
symptoms, the sign/symptom should be reported in the Medical History Log in the eCRF. 
Table 11.2-1 
Clinical laboratory parameters 
Category 
Parameter 
Clinical chemistry 
Alanine aminotransferase (ALT) 
Aspartate aminotransferase (AST) 
Creatinine 
Glucose 
Haematology 
Haematocrit 
Haemoglobin (Hb) 
Platelet count 
Red blood cell (RBC) count 
White blood cell (WBC) count 
FSH-test (at screening, 
postmenopausal females 
only in questionable cases) 
FSH 
Estradiol 
  
Physical examination 
A complete physical examination will include assessments of the head, eyes, ears, nose, 
throat, skin, thyroid, neurological, lungs, cardiovascular, abdomen (liver and spleen), lymph 
nodes and extremities. 
Physical examination will be judged as normal, abnormal, not clinically significant or 
abnormal, clinically significant. The assessment will be recorded in the eCRF. 
  
Resting 12-lead ECG 
Single 12-lead ECG will be recorded in supine position after 10 minutes of rest using an ECG 
machine. HR and PR, QRS, QT and QTcF intervals will be recorded. 
ECGs will be reviewed and interpreted on-site by the Investigator. 
Any abnormalities will be specified and documented as clinically significant or not clinically 
significant. 

 
 
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11.3 Assessments related to PET-endpoints  
 Inhalation training 
At the end of the screening visit subjects, who consented to study participation and are found 
eligible (awaiting results of clinical laboratory sampling), will perform inhalation training 
with the myblu™ e-cigarette. Training will be held by study personnel at the research site. 
Inhalation training will be performed on dummy (nicotine free) devices and once on a regular 
myblu™ e-cigarette (containing nicotine corresponding to the formulations used during PET). 
Training on a nicotine-containing device is considered necessary for the subjects for them to 
be used to the experience when inhaling in the PET-camera. 
Inhalation training on dummy devices (nicotine free) may be repeated in the morning of 
Visit 2 (before PET), if considered necessary by the study personnel or the subject. 
 Insertion/removal of arterial catheter 
After a positive modified Allen test, an intraarterial catheter will be inserted preferably in the 
arteria radialis of the non-dominant arm if possible, else in another artery chosen by the 
anaesthesiologist. Local anaesthetics e.g. 0.5 mL of Xylocain® (active substance lidocaine) 
10 mg/mL may be applied for the arterial cannulation at the discretion of the anaesthesiologist 
inserting the cannula. 
Positive modified Allen test: If the hand flushes within 5-15 seconds, this indicates that the 
ulnar artery has good blood flow; this normal flushing of the hand is considered to be a 
positive test. 
Negative modified Allen test: If the hand does not flush within 5-15 seconds, this indicates 
that ulnar circulation is inadequate or non-existent; in this situation, the radial artery 
supplying arterial blood to that hand should not be punctured. 
The arterial catheter will be removed by trained personnel at the research clinic after the (last) 
PET session is completed. When the catheter is withdrawn, pressure should be applied to the 
injection site for approximately 10 minutes until bleeding ceases. The catheter is checked to 
see that it is intact. 
 Assessment of blood radioactivity 
Discrete blood samples will be analysed using an in house developed well counter detector 
system cross-calibrated with the PET/CT scanner. 
Serial discrete arterial blood samples with a volume of approximately 5 mL (2 mL used for 
each analysis) will be collected in test tubes for the analysis of radioactivity in whole blood 
withdrawn at specified timepoints (Section 11.3.4 and 11.3.5). The time of sampling will be 
recorded in the middle of the sampling period by using a foot pedal linked to a computer and 
the test tubes will be labelled with barcodes for identification. Data will be stored directly in 
separate files, one for each PET/CT-scan session, including sample weight, number of counts 
and time of measurement. The data will be corrected for physical decay of 11C to the time for 
tracer inhalation. The samples will be destructed directly after measurement. 

 
 
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 PET/CT (PART A) 
Administration/inhalation and subject preparation 
The subject will be moved from the research site to the PET-Centre approximately 30 minutes 
before the PET-scan. 
The subject is positioned in the PET-CT camera and a ULDCT scan is performed to correct 
for attenuation of the photons and to outline the anatomy. In supine position the subject inhale 
from the device, without changing the head or body position. The PET-CT operator takes the 
inhalation device and the PET-scan begins. 
The inhalation device is measured before and after inhalation to determine total amount of 
[11C]nicotine delivered to the subject. The amount of [11C]nicotine lost when the subject 
exhales cannot be determined. 
PET-protocol 
PET-scan 1: The subject inhales (1-2 puffs) for about 3-5 seconds to receive about 10-24 L 
solution corresponding to approximately 25 MBq [11C]nicotine. A dynamic 40 min PET scan 
is performed with the oral cavity/throat and large bronchi in field-of-view (20 cm). Arterial 
blood is collected as discrete samples of 5 mL each, taken at 2, 4, 6, 8, 10, 15, 20 and 30 
minutes to determine radioactivity in whole blood. 
PET-scan 2: The subject inhales (1-2 puffs) for about 3-5 seconds to receive about 10-24 L 
solution corresponding to approximately 25 MBq [11C]nicotine. A dynamic 40 min PET scan 
is performed with part of the lung in field-of-view (20 cm). Arterial blood is collected as 
discrete samples of 5 mL each, taken at 2, 4, 6, 8, 10, 15, 20 and 30 minutes to determine 
radioactivity in whole blood. 
For both lung and oral cavity scans, the dynamic PET data is acquired in list mode, and will 
initially be reconstructed into 6 x 10s, 3 x 20s, 2 x 30s, 2 x 60s, 2 x 150s and 6 x 300s frames. 
The scanner will be started 30 seconds before the tracer administration. The data collected 30 
seconds before inhalation will not be reconstructed. The registration of data is depending on 
the resulting images, the frame definition may be optimised, taking the inhalation protocol 
into account. The static scans over lung and oral cavity will be acquired as single frame scans 
of 600s. 
 PET/MR (PART B) 
PET scan 3: The subject inhales (1-2 puffs) for about 3-5 seconds to receive about 12 L 
solution corresponding to approximately 50 MBq [11C]nicotine. A dynamic PET scan is 
performed over the brain 0-30 min. Arterial blood is collected as discrete samples of 5 mL 
each, taken at 2, 4, 6, 8, 10, 15, 20 and 30 minutes to determine radioactivity in whole blood. 
Optional – extended brain uptake study 
If the results from PET-scan 3 show significant, quantifiable, uptake of radioactivity in the 
brain, the study is extended to include 3 more subjects, to a total of 5 individuals receiving the 
formulation that shows the best uptake. 
For the brain scans, the dynamic PET data is acquired in list mode, and will initially be 
reconstructed into 6 x 10s, 3 x 20s, 2 x 30s, 2 x 60s, 2 x 150s and 4x 300s frames. The scanner 
will be started 30 seconds before the tracer administration. The data collected 30s before 
inhalation will not be reconstructed. The registration of data is depending on the resulting 
images, the frame definition may be optimised, taking the inhalation protocol into account. 

 
 
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11.3.5.1 MR-scan for anatomical localisation of [11C]nicotine uptake: 
All subjects participating in PART B will perform an MRI at any suitable timepoint during 
the study period (after screening until end-of-study- visit). With MRI, a standard structural 
T1-weigthed volume is acquired to be used for anatomical localisation of [11C]nicotine uptake 
in combination with PET scans acquired. 
11.4 Assessments related to secondary endpoints 
 PET endpoints 
Refer to Section 11.3. 
 Adverse events 
The Principal Investigator is responsible for ensuring that all medical staff involved in the 
study is familiar with the content of this section and the content of the CTC standard 
operating procedures (SOPs) regarding handling of AEs. 
11.4.2.1 Definition of adverse event  
An AE is defined as any untoward medical occurrence in a subject administered an IP and 
which does not necessarily have a causal relationship with this treatment. An AE can 
therefore be any unfavourable and unintended sign (including clinically significant abnormal 
values from relevant tests, such as clinical safety laboratory tests, ECGs, vital signs), 
symptom, or disease temporally associated with the use of an IP, whether or not considered 
related to the IP. 
11.4.2.2 Definition of serious adverse event 
An SAE is any AE which: 
• results in death 
• is life-threatening (this refers to a reaction in which the subject was at risk of death 
at the time of the reaction; it does not refer to a reaction that hypothetically might 
had led to death if the reaction was more severe) 
• requires in-patient hospitalisation or prolongation of existing hospitalisation 
• results in persistent or significant disability/incapacity 
• is a congenital anomaly/birth defect 
• is an important medical event (IME) (this refers to a reaction that may not be 
immediately life-threatening or result in death or hospitalisation, but may jeopardise 
the subject or may require intervention to prevent any of the other outcomes defined 
above) 
Examples of IMEs are intensive treatment in an emergency room for allergic bronchospasm 
or blood dyscrasias, convulsions that do not result in hospitalisation, development of drug 
dependency, and drug abuse. 
Planned hospitalisations or surgical interventions for a condition that existed before the 
subject signed the ICF and that did not change in intensity are not SAEs. 

 
 
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If there is any doubt as to whether an AE meets the definition of an SAE, a conservative 
viewpoint must be taken, and the AE must be reported as an SAE. 
11.4.2.3 Time period and frequency for collecting adverse events 
All AEs (including SAEs) will be collected from the insertion of the arterial catheter until the 
end-of-study visit. 
Any AE with start date on the day IP administration must be recorded with start time. 
At the end-of-study visit, information on new AEs or SAEs, if any, and stop dates for ongoing 
events must be recorded as applicable. 
Investigators are not obligated to actively seek AEs or SAEs after conclusion of the study 
participation. However, if the Investigator learns of any SAE, including a death, at any time 
after a participant has been discharged from the study, and he/she considers the event to be 
reasonably related to the study intervention or study participation, the Investigator must 
promptly notify the Sponsor. 
11.4.2.4 Assessment of intensity 
The grading of the intensity of AEs will follow the common terminology criteria for adverse 
events (CTCAE) v5.0 [3]. Grade refers to the severity of the AE. The CTCAE displays Grades 
1 through 5 with unique clinical descriptions of severity for each AE based on this general 
guideline. 
The Investigator must assess the intensity of an AE using the following definitions, and record 
it on the AE Log in the eCRF: 
Grade 1 
Mild; asymptomatic or mild symptoms; clinical or diagnostic observations 
only; intervention not indicated. 
Grade 2 
Moderate; minimal, local or non-invasive intervention indicated; limiting age-
appropriate instrumental activities of daily living (ADL)*. 
Grade 3 
Severe or medically significant but not immediately life-threatening; 
hospitalisation or prolongation of hospitalisation indicated; disabling; limiting 
self- care ADL**. 
Grade 4 
Life-threatening consequences; urgent intervention indicated. 
Grade 5 
Death related to AE. 
*Instrumental ADL refer to preparing meals, shopping for groceries or clothes, using the 
telephone, managing money, etc. 
**Self- care ADL refer to bathing, dressing and undressing, feeding self, using the toilet, 
taking medications, and not bedridden. 
11.4.2.5  
Assessment of causal relationship 
The Investigator must assess the causal relationship between an AE and the IP using the 
definitions below and record it the AE Log of the eCRF: 
Probable 
The event has a strong temporal relationship to the IP or recurs on re-
challenge, and another aetiology is unlikely or significantly less likely. 

 
 
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Possible 
The event has a suggestive temporal relationship to the IP, and an alternative 
aetiology is equally or less likely. 
Unlikely 
The event has no temporal relationship to the IP or is due to 
underlying/concurrent illness or effect of another drug (that is, there is no 
causal relationship between the IP and the event). 
An AE is considered causally related to the use of the IP when the causality 
assessment is probable or possible. 
In addition, the Investigator will assess the causal relationship between an AE and the study 
procedures (e.g. injection site reaction, events in association with arterial catheter insertion, 
other arterial catheter associated AE). 
11.4.2.6 Assessment of outcome 
The Investigator must assess the outcome of an AE using the definitions below and record 
it on the AE Log of the eCRF: 
Recovered/resolved 
The subject has recovered completely, and no symptoms remain. 
Recovering/resolving  The subject’s condition is improving, but symptoms still remain. 
Recovered/resolved 
with sequelae 
The subject has recovered, but some symptoms remain (for 
example, the subject had a stroke and is functioning normally but 
has some motor impairment). 
Not recovered/not 
resolved 
The subject’s condition has not improved and the symptoms are 
unchanged (for example, an atrial fibrillation has become chronic). 
Fatal 
 
Unknown 
 
11.4.2.7 Reporting of action taken with study treatment 
The Investigator must report the action taken with study treatment using the definitions 
below and record it on the AE Log of the eCRF: 
Dose increased   
Dose not changed 
Dose rate reduced 
Dose reduced 
Drug interrupted 
Drug withdrawn 
Not applicable 
Unknown 
11.4.2.8 Collecting adverse events 
AEs identified using any of the following methods will be recorded: 
• AEs spontaneously reported by the subject 

 
 
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• AEs observed by the Investigator or medical personnel 
• AEs elicited based on non-leading questions from the Investigator or medical 
personnel 
11.4.2.9 Recording adverse events 
AEs must be recorded in the AE Log of the eCRF. The Investigator must provide 
information on the AE, preferably with a diagnosis or at least with signs and symptoms; 
start and stop dates, start and stop time; intensity; causal relationship to IP or study 
procedures; action taken, and outcome. 
If the AE is serious, this must be indicated in the eCRF. 
AEs, including out-of-range clinically significant clinical safety laboratory values, must be 
recorded individually, except when considered manifestations of the same medical condition 
or disease state; in such cases, they must be recorded under a single diagnosis. 
Malfunctions/incidents with the e-cigarette system will be reported to the Sponsor on a 
separate Product Form. 
11.4.2.10  Reporting of serious adverse events  
SAE reporting should be performed by the Investigator within 24 hours of awareness via the 
eCRF. All available information regarding the SAE should be entered in the AE Log for the 
specific subject. By saving the event as “serious” in the eCRF and once the Investigator has 
signed-off of the event, an e-mail alert is automatically sent to predefined recipients to 
highlight that an SAE has been registered. The same information is automatically sent to 
sae@ctc-ab.se. 
The SAE report is reviewed by a designated person at CTC’s Pharmacovigilance (CTC PV) 
department to ensure that the report is valid and correct. For fatal or life-threatening SAEs 
where important or relevant information is missing, immediate follow-up is undertaken and 
queries to the site are raised. Investigators or other site personnel should inform CTC PV of 
any follow-up information on a previously reported SAE immediately but no later than the 
end of the next business day of when he or she becomes aware of it. 
If the SAE report in the eCRF is updated, a new e-mail alert will be sent. 
If any additional documentation is required (e.g. autopsy report), CTC PV will request this 
information from the study site.  
In case the eCRF cannot be accessed, the SAE should be reported by manual completion of 
the paper SAE Form, provided in the Investigator Site File (ISF). The completed, signed and 
dated paper SAE Form should, within 24 hours, be scanned and e-mailed to:  
 
 
 
A copy of the paper SAE form must also be e-mailed to CTC at: sae@ctc-ab.se. 
The study site should notify the site Monitor via phone or e-mail about the submission of 
the SAE report. As soon as the site personnel have access to the eCRF, the SAE should be 
reported electronically as well. 

 
 
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The Sponsor or delegate will assume responsibility for reporting SAEs to the independent 
ethics committee (IEC) in accordance with local regulations. 
11.4.2.11 Treatment and follow-up of adverse events 
Subjects with AEs that occur during the study must be treated according to daily clinical 
practice at the discretion of the Investigator. 
AEs must be followed up until resolution or to the end-of-study visit, whichever comes first. 
At the end-of-study visit, information on new AEs, if any, and stop dates for previously 
reported AEs must be recorded (if known). AEs assessed as stable by the Investigator at the 
end-of-study visit will not have to be followed up until resolution. 
It is the responsibility of the Investigator to follow up on all SAEs until the subject has 
recovered, stabilised, or recovered with sequelae, and to report to the Sponsor all relevant new 
information using the same procedures and timelines as those for the initial report. Relevant 
information includes discharge summaries, autopsy reports, and medical consultation. 
11.4.2.12 Procedures in case of pregnancy 
In case of pregnancy or suspicion of possible pregnancy of any female subjects, the study 
treatment must be stopped immediately, and the subject discontinued from participation in the 
study. Pregnancy itself is not regarded as an AE unless there is a suspicion that the IP may 
have interfered with the effectiveness of the contraceptive medication. However, the outcome 
of all pregnancies (spontaneous miscarriage, elective termination, normal birth or congenital 
abnormality) must be followed up and documented even after the subject was discontinued 
from the study. 
All events of congenital abnormalities/birth defects are SAEs. Spontaneous miscarriages 
should also be reported and handled as AEs. All outcomes of pregnancy must be reported to 
the Sponsor and the Principal Investigator on the pregnancy outcomes report form. 
11.4.2.13 Treatment of overdose 
An overdose is a dose in excess of the dose specified for each cohort in this clinical study 
protocol (CSP).   
Over-dosing is not likely to occur in this study since all IP will be administered by site 
personnel under medical surveillance. In cases of accidental overdose, standard supportive 
measures should be adopted as required. 
An overdose should be documented as follows:  
• An overdose with associated AE is recorded as the AE diagnosis/symptoms in the AE 
Log of the eCRF.  
• 
An overdose without associated symptoms is only reported in the subject’s medical 
records. 

 
 
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 Vital signs 
Systolic and diastolic blood pressure and pulse will be measured in supine position after 
10 minutes of rest.  
Vital signs will be judged as normal, abnormal, not clinically significant or abnormal, 
clinically significant. The assessment will be recorded in the eCRF.  
Post-IP vital signs, where the change from the values measured at the screening visit is judged 
as “abnormal, clinically significant” by the Investigator will be reported as AEs. 
11.5 Appropriateness of measurements 
All methods used for demography and safety assessments are commonly used in standard 
medical care and in clinical studies. 
12 PROCEDURES FOR BIOLOGICAL SAMPLES 
12.1 Sample collection  
The sample collection procedure for radiation analysis during PET analysis is described in 
Section 11.2.12. 
Clinical laboratory samples are collected according to standard procedures. 
12.2 Volume of blood 
The estimated volume of blood collected during the study from each subject is presented in 
Table 12.2-1. 
Table 12.2-1 
Estimated blood volumes 
PART A 
Estimated number of 
sampling occasions 
Estimated 
volume per 
occasion 
(mL) 
Total (mL) 
Clinical chemistry, haematology, microbiology 
1 
20 
20 
Blood sampling PET 
2 
40 
80 
Total: 
 
 
100 
PART B 
Estimated number of 
sampling occasions 
Estimated 
volume per 
occasion 
(mL) 
Total (mL) 
Clinical chemistry, haematology, microbiology 
1 
20 
20 
Blood sampling PET 
1 
40 
40 
Total: 
 
 
60 

 
 
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12.3 Handling, storage and destruction of laboratory samples 
All biological samples will be registered in a biobank at CTC (893), if applicable. 
Any remains from radioactive samples drawn and analysed at the PET Centre or the safety 
laboratory samples will be disposed of immediately after analyses according to standard 
routines. 
12.4 Chain of custody of biological samples 
A full chain of custody is maintained for all samples throughout their lifecycle. 
CTC keeps full traceability of collected biological samples from the subjects while in storage 
at the research clinic until shipment and keeps documentation of receipt of arrival. 
The sample receiver (the analytical laboratory) keeps full traceability of the samples while in 
their storage and during use until used or disposed of. 
The Sponsor keeps oversight of the entire life cycle through internal procedures, monitoring 
of study sites and auditing of external laboratory providers. 
12.5 Withdrawal of informed consent for donated biological samples 
If a subject withdraws consent to the use of biological samples donated, the samples will be 
disposed of /destroyed, if not already analysed and documented. 
The Principal Investigator will ensure that: 
• Subject withdrawal of informed consent is notified immediately to Sponsor. 
• Biological samples from the subject, if stored at the research clinic, are immediately 
identified, disposed of/destroyed and the action is documented. 
The Sponsor has to ensure that the laboratory(ies) holding the samples is/are informed about 
the withdrawn consent immediately and that samples are disposed of/destroyed or returned to 
the research clinic and the action is documented. 
13 QUALITY MANAGEMENT, QUALITY ASSURANCE AND QUALITY 
CONTROL 
13.1 Quality management: critical process, system and data identification 
During CSP development, the Sponsor will identify those processes, systems (facilities, 
computerised systems) and data that are critical to ensure human subject protection and the 
reliability of trial results according to applicable SOPs and International Council for 
Harmonisation (ICH) E6 R2. 
Identified risks will be categorised separately from the CSP. 
13.2 Quality assurance and quality control 
The Sponsor is responsible for implementing and maintaining quality assurance (QA) and 
quality control (QC) systems with written SOPs with regards to management of identified 

 
 
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risks, CSP compliance, good clinical practice (GCP) compliance and applicable regulatory 
requirements. 
The Sponsor is responsible for securing agreements with involved subcontractors and to 
perform regular subcontractor oversight to ensure CSP compliance, GCP compliance and 
compliance with applicable regulatory requirements.  
The Sponsor is responsible for implementing a risk-based validated electronic data capture 
system and maintain SOPs for the whole life cycle of the system. 
QC should be applied to each stage of data handling to ensure that all data are reliable and 
have been processed correctly. 
The Sponsor has delegated the responsibilities outlined above to CTC whilst maintaining 
overall study oversight. 
14 ETHICAL AND REGULATORY REQUIREMENTS  
14.1 Ethical conduct of the study 
The study will be performed in accordance with ethical principles that have their origin in the 
Declaration of Helsinki [4] and are consistent with ICH/GCP E6 (R2), EU Clinical Trials 
Directive, and applicable local regulatory requirements. 
14.2 Ethics review 
The Principal Investigator is responsible for submission of the CSP, the subject information 
and ICF, any other written information to be provided to the subjects and any advertisements 
used for recruitment of subjects to applicable IEC for approval. 
Approval must be obtained in writing from the IEC before the first subject can be recruited. 
14.3 Subject information and consent 
It is the responsibility of the Investigator or an authorised associate to give each potential 
study subject adequate verbal and written information before any study specific assessments 
are performed. 
The information will include the objectives and the procedures of the study as well as any 
risks or inconvenience involved. It will be emphasised that participation in the study is 
voluntary and that the subject may withdraw from participation at any time and for any 
reason, without any prejudice. All subjects will be given the opportunity to ask questions 
about the study and will be given sufficient time to consider participation before signing the 
ICF. 
Before performing any study-related procedures the ICF must be signed and personally dated 
by the subject and by the Investigator. A copy of the subject information including the signed 
ICF will be provided to the subject. 
Documentation of the discussion and the date of informed consent must be recorded in the 
source documentation and in the eCRF. The subject information sheet and the signed ICF 
should be filed by the Investigator for possible future audits and/or inspections.  

 
 
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The final approved version of the subject information and ICF must not be changed without 
approval from the Sponsor and the applicable IEC.  
14.4 Subject data protection 
The ICF includes information that data will be recorded, collected and processed and may be 
transferred to European Economic Area (EEA) or non-EEA countries. In accordance with the 
European Union Data Protection Directive (95/46/EC) and General Data Protection 
Regulation (GDPR), the data will not identify any persons taking part in the study. 
The potential study subject should be informed that by signing the ICF he/she approves that 
authorised representatives from Sponsor and CTC, the concerned IEC have direct access to 
his/her medical records for verification of clinical study procedures. For further details on the 
subject information and ICF process, refer to Section 14.3. 
The subject has the right to request access to his/her personal data and the right to request 
rectification of any data that is not correct and/or complete in accordance with the European 
Union Data Protection Directive (95/46/EC) and the request will be raised to the Principal 
Investigator.  
The Investigator must file a Subject Identification List which includes sufficient information 
to link records, i.e. the eCRF and clinical records. This list should be preserved for possible 
future inspections/audits but must not be made available to the Sponsor except for monitoring 
or auditing purposes. 
Personal data that are collected in the study such as health information and ethnicity are 
considered as sensitive personal data. This data will be pseudoanonymised, i.e. personally 
identifiable information (PII) will be removed and replaced by a unique subject ID and will be 
processed by the Sponsor and other involved parties during the study. After the study end, 
only anonymised data, i.e. aggregated data sets, can be used. 
For this study, the Sponsor Nerudia Ltd is the data controller of all data processed during the 
study (e.g. Trial Master File [TMF], study reports) and CTC AB is the data processor. Any 
subcontractors used in the study, are also data processors. 
For data that are processed at the clinic(s) (e.g. medical records and ISF), CTC AB is the data 
controller. 
14.5 Changes to the approved clinical study protocol  
Any proposed change to the approved final CSP (including appendices) will be documented 
in a written and numbered clinical protocol amendment. All substantial amendments to the 
protocol must be approved by the appropriate IEC before implementation according to 
applicable regulations. 
14.6 Audits and inspections 
Authorised representatives of Sponsor, or an IEC may perform audits or inspections at the 
research clinic, including source data verification (SDV). The purpose of an audit or 
inspection is to systematically and independently examine all study-related activities and 
documents, to determine whether these activities were conducted, and data were recorded, 
analysed, and accurately reported according to the protocol, ICH-GCP guidelines and any 

 
 
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applicable regulatory requirements. The Investigator will contact the Sponsor immediately if 
contacted about an inspection at the centre. 
14.7 Insurance 
Subjects will be covered under Nerudia Ltd’s liability insurance policy. The certificate of 
insurance and an information leaflet containing essential information about the insurance 
coverage can be provided upon request. The participating subjects are also protected in 
accordance with national regulations, as applicable. CTC has a company insurance covering 
services performed by CTC. 
15 STUDY MANAGEMENT 
15.1 Training of study site personnel 
Before enrolment of the first study subject a Sponsor representative or delegate will perform a 
study initiation visit at the research clinic. The requirements of the CSP and related 
documents will be reviewed and discussed and the investigational staff will be trained in any 
study specific procedures and system(s) utilised. 
It is the responsibility of the Investigator to ensure that all personnel involved in the study are 
fully informed of all relevant aspects of the study and have a detailed knowledge of and 
training in the procedures that are to be executed by them. Any new information of relevance 
to the performance of this study must be forwarded to the staff involved in a timely manner. 
The Investigator will keep a list of all personnel involved in the study together with their 
function and study related duties delegated. A Curriculum Vitae will be available for all staff 
to whom study-specific duties are delegated.  
15.2 Clinical monitoring  
The Sponsor is responsible for securing agreement from all involved parties to ensure direct 
access to all study related sites, source data/documents, and reports for the purpose of 
monitoring and auditing by the Sponsor, and inspection by domestic and foreign regulatory 
authorities.  
As defined in the risk-based monitoring (RBM) plan, approved by the Sponsor and provided 
separately, the responsible Monitor will periodically visit the study site at times agreed upon 
by the Investigator and the Monitor. At the time of each monitoring visit, the role of the 
Monitor is (but not limited to) to:  
• provide information and support to the investigational team. 
• confirm that facilities and resources remain acceptable. 
• confirm that the investigational team is adhering to the CSP, applicable SOPs, 
guidelines, manuals and regulatory requirements.  
• verify that data are being accurately and timely recorded in the eCRFs and that IP 
accountability checks are being performed. 
• verify that data in the eCRF are consistent with the clinical records (SDV) in 
accordance with the RBM plan.  

 
 
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• verify that the correct informed consent procedure has been adhered to for participating 
subjects.  
• ensure that withdrawal of informed consent to the use of the subject’s biological 
samples will be reported and biological samples are identified and disposed 
of/destructed accordingly, and that this action is documented and reported to the 
subject. 
• verify that AEs are recorded and reported in a timely manner and according to the CSP. 
• raise and escalate any serious quality issues, serious GCP breach and any data privacy 
breach to the Sponsor.  
Centralised monitoring will also be performed continuously by study team members at CTC 
in accordance with the RBM plan.  
When the study has been completed and all queries have been resolved and the database has 
been locked, the Monitor will perform a close-out visit. 
15.3 Source data documents 
A separate Origin of Source Data List will be generated for each site before start of 
enrolment, specifying the location of the source of derived information appearing in the 
eCRF. This document must be signed by the Principal Investigator and the Monitor to confirm 
agreement before start of recruitment. 
Source documents are all documents used by the Investigator or hospital that relate to the 
subject’s medical history, that verifies the existence of the subject, the inclusion and exclusion 
criteria, and all records covering the subject’s participation in the trial. They include 
laboratory notes, memoranda, material dispensing records, subject files, etc. The eCRF may 
constitute source data if clearly defined in the Origin of Source Data List.  
The Investigator should guarantee access to source documents to the Monitor and the IECs, if 
required. 
15.4 Study agreements 
The Principal Investigator must comply with all the terms, conditions, and obligations of the 
Clinical Study Agreement for this study. 
Agreements between Sponsor and CTC must be in place before any study-related procedures 
can take place, or subjects be enrolled. 
15.5 Study timetable and end of study 
The study is expected to start in Q1 2020 and to be completed by Q2 2020. 
A subject is considered to have completed the study if he/she has completed all visits in the 
study including the last visit. 
The end of the study is defined as the date of the last visit of the last subject in the study. 

 
 
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15.6 Termination of the study 
The Sponsor reserves the right to discontinue the study at any time but intends only to 
exercise this right for valid scientific or administrative reasons. 
After such a decision, the PI must inform all participating subjects and perform relevant 
assessments, preferably according to the scheme for the final assessments. All delivered and 
unused IP and other study materials must be returned and all eCRFs completed as far as 
possible. 
15.7 Reporting and publication 
 Clinical study report  
A clinical study report (CSR), describing the conduct of the study, any statistical analyses 
performed and the results obtained, will be prepared by CTC and the PET Centre. The report 
will be reviewed and approved by, as a minimum, the Principal Investigator and the Sponsor. 
 Confidentiality and ownership of study data 
Any confidential information relating to the IP or the study, including any data and results 
from the study, will be the exclusive property of the Sponsor. The Investigator and any other 
persons involved in the study are responsible for protecting the confidentiality of this 
proprietary information belonging to the Sponsor. 
 Publication 
The results from this study may be submitted for publication at the discretion of the Sponsor. 
15.8 Archiving  
The Principal Investigator is responsible for maintaining essential documents, (as defined in 
ICH E6 GCP, Section 8) for 10 years after finalisation of the CSR. This includes any original 
source documents related to the study, the Subject Identification List (providing the sole link 
between named subject source records and anonymous eCRF data), the original signed ICFs 
and detailed records of disposition of IP. 
It is the responsibility of the Sponsor to inform the Investigator/institution as to when these 
documents no longer need to be retained. 
The Sponsor will archive the TMF in accordance with ICH E6 GCP, Section 8 and applicable 
regulatory requirements. 
The data from the eCRFs will be sent to the Sponsor and a copy will be sent to the clinic and 
filed in the Investigator Study File for archiving for 10 years after finalisation of the CSR. 
The completed eCRFs are the sole property of the Sponsor and should not be made available 
in any form to third parties, except for authorised representatives of appropriate 
Health/Regulatory Authorities, without written permission from the Sponsor. 

 
 
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16 DATA MANAGEMENT 
The data management routines include procedures for handling of the eCRF, database set-up 
and management, data entry and verification, data validation, QC of the database, and 
documentation of the performed activities including information of discrepancies in the 
process. The database, data entry screens, and program will be designed in accordance with 
the CSP. 
Data validation/data cleaning procedures are designed to assure validity and accuracy of 
clinical data. These procedures consist of computerised online edit checks identifying e.g. 
data values that are outside the allowed range and SAS-programmed batch checks on data 
exports. All study-specific and standard data validation programming will be tested in a 
separate testing environment prior to use on production data. 
Detailed information on data management will be described in a study-specific Data 
Management Plan (DMP). 
16.1 The web-based eCRF 
Clinical data will be entered into a 21 CFR Part 11-compliant eCRF (Viedoc™) provided by 
PCG Solutions AB. The eCRF includes password protection and internal quality checks, such 
as automatic range checks, to identify data that appear inconsistent, incomplete, or inaccurate. 
Clinical data will be entered directly from the source documents or at bedside (if the eCRF 
data constitutes source data). Source data are to be defined at the site before inclusion of the 
first subject (Section 15.3). 
Authorised site personnel designated by the Investigator will complete data collection. 
Appropriate training and security measures will be completed with the Investigator and all 
authorised trial site personnel prior to the trial being initiated and any data being entered into 
the system for any study subject. 
16.2 The entering of data into the eCRF 
All entries, corrections, and alterations are to be made by the Investigator or designee. Neither 
the Monitor nor any other study team member besides site staff can enter data in the eCRF. 
All data should be entered in English. The eCRFs should be completed as soon as possible 
during or after the subject’s visit. The Investigator must verify that all data entries in the 
eCRFs are accurate and correct. If some assessments are not done, or if certain information is 
not available, not applicable or unknown, the Investigator or assigned clinical staff should 
record such information in the eCRF. The Investigator will be required to electronically sign 
off the clinical data. This will be performed by means of the Investigator’s unique User ID 
and password; date and time stamps will be added automatically at time of electronic 
signature. 
16.3 The query process 
The Monitor will review the eCRFs and evaluate them for completeness and consistency. 
Data in the eCRF will be compared with the respective source documents to ensure that there 
are no discrepancies for critical data as described in the RBM plan. All entries, corrections, 

 
 
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and alterations are to be made by the Investigator or designee. Neither the Monitor nor any 
other study team member besides site staff can enter data in the eCRF.  
If corrections are needed, queries will be raised within the eCRF, either as a result of built-in 
edit checks or manually raised by the monitor. An appropriate member of the site staff will 
answer the queries in the eCRF either by correcting the data or by entering a response to the 
query. The monitor will either approve the answer/correction or re-issue the query. 
16.4 Audit trail 
All entries in the eCRF will be fully recorded in a protected audit trail. Once clinical data 
have been saved, corrections to the data fields will be audit trailed, meaning that the reason 
for change, the name of the person who made the change, together with time and date will be 
logged. 
16.5 External data 
External data consists of data that are not recorded in the eCRF. Data may be received in 
electronic format or as a paper printout. Key variables are defined in order to uniquely 
identify each sample record. File and data formats are agreed with the external data provider. 
16.6 Medical coding 
Medical coding will be performed by trained personnel at CTC. AEs and medical/surgical 
history verbatim terms will be coded using the Medical Dictionary of Regulatory Activities 
(MedDRA; latest version available at eCRF finalisation) as specified in the DMP. Prior and 
concomitant medications will be coded according to the World Health Organisation (WHO) 
Anatomic Therapeutic Chemical (ATC) classification system. All coding will be approved by 
Sponsor prior to database lock. 
16.7 Database lock 
When all data have been entered and discrepancies solved, clean file will be declared, the 
database will be locked, and the data will be analysed.  
17 STATISTICAL METHODS AND DETERMINATION OF SAMPLE SIZE  
The principal features of the statistical analysis to be performed are described in this section. 
A more technical and detailed elaboration of the principal features will be presented in a 
separate Statistical Analysis Plan (SAP), which will be signed and approved prior to database 
lock. 
Analyses of the primary, secondary and exploratory endpoints will be performed by CTC and 
the PET Centre. 
17.1 General 
Continuous data will be presented in terms of evaluable and missing observations, arithmetic 
mean, standard deviation (SD), median, minimum and maximum value.  

 
 
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Categorical data will be presented as counts and percentages. When applicable, summary data 
will be presented by treatment, and by assessment time. Individual subject data will be listed 
by subject number, treatment, and, where applicable, by assessment time. 
All descriptive summaries and statistical analyses will be performed using SAS Version 9.4 or 
later (SAS Institute, Inc., Cary, NC). 
Baseline will be defined as the visit with last data collection point prior to the first 
administration of IP. 
No imputation of missing data will be performed. 
17.2 Determination of sample size 
No formal sample size calculation has been performed for this study. The proposed sample 
size is considered sufficient to provide adequate information for the study objectives. 
Approximately 25 subjects will be screened to achieve 14-17 included and evaluable subjects. 
17.3 Analysis data sets 
 Full analysis set 
The Full Analysis Set (FAS) will consist of all subjects who have been randomised and 
received at least one dose of IP and who has at least one post-baseline assessment data.  
 Safety analysis set 
The Safety Analysis Set will comprise all subjects who received at least one dose of the IP.  
 Per protocol set 
The Per Protocol Set (PPS) will consist of all subjects who have been randomised and 
completed the study without any major protocol deviations that are judged to compromise the 
analysis of the data. All protocol violations will be judged as major or minor prior to database 
lock. 
17.4 Description of study population 
 Demographics and baseline characteristics 
Descriptive statistics for demographics, weight and height will be presented by treatment and 
study part. 
 Medical/surgical history and prior/concomitant medication 
Medical/surgical history will be presented by system-organ-class (SOC) and preferred term 
(PT). Prior/concomitant medications will be presented by ATC level 1, 3 and 5. 
All data will be listed by subject. 

 
 
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 Treatment compliance 
The number of subjects treated with each formulation in each part will be listed. 
 Physical examination 
Clinically significant and non-clinically significant abnormal findings will be specified and 
presented by subject and summarised by treatment and study part. 
All data will be listed by subject. 
 12-lead ECG 
All ECGs will be categorised as ”normal”, ”abnormal, not clinically significant”, or 
”abnormal, clinically significant” (as judged by the Investigator) and summarised by 
treatment and study part using frequency tables.  
All data will be listed by subject. 
 Clinical laboratory analyses 
Clinical laboratory data will be summarised by treatment and study part. 
Abnormal, clinically significant values will be summarised separately if considered 
appropriate.  
All data will be listed by subject. 
17.5 Analysis of PET endpoints 
The PET Centre will be responsible for evaluation of PET data, and present time activity data 
on deposition during the dynamic scans in PET-scan 1 and 2. For the evaluation of 
[11C]nicotine kinetics in the lung, primary bronchi, oral cavity and airways, volumes of 
interest (VOIs) will be delineated, and time activity data generated utilising PET-image 
analysis software. The VOIs will be delineated based on the anatomical information from the 
low dose CT-images combined with the uptake seen in the PET images. Deposition in the 
lungs will be measured and presented for one large VOI covering the entire lung. 
Furthermore, a small VOI may be delineated to be representative for deep lung tissue, 
however this VOI will not include the entire deep lung tissue. Even though alveolar tissue 
cannot be identified in detail on the ULDCT or PET images, the highest density of alveolar 
tissue, combined with lowest density of bronchi and major blood vessels, will be found in the 
peripheral part of the lung and a small VOI will be delineated at a position representative for 
deep lung tissue. 
For PET-scan 3 acquired over the brain, the amount of nicotine distributed to the brain will be 
estimated. A whole brain VOI will be delineated on the T1-MR images overlaid on the brain 
PET-image. This part of the study is explorative and dependent on the time course of the 
[11C]nicotine uptake to the blood and the time activity uptake for brain nicotine may be 
difficult to measure with a sufficient signal-to-noise ratio. 
The uptake data will be stored in spreadsheet format and sent to study sponsor to enable 
further data analysis. Furthermore, and as an exploratory part of the PET-image analysis, 
other regions may be delineated and analysed, if visual inspection of overlaid PET and 
ULDCT images shows substantial uptake in other tissues or regions. 

 
 
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Input data for evaluation of [11C] uptake and distribution: 
• Amount of radioactivity administered from inhalator (inhalator measured before and 
after inhalation) 
• PET-images showing deposition of radioactivity in the lung 
• PET-images showing deposition of radioactivity in the oral cavity  
• Radioactivity measured in arterial blood.  
The relative amount of radioactivity that is deposited in the lung can be determined from the 
activity measured in the PET images and the known amount of activity in the inhalator before 
and after the inhalation. 
Blood radioactivity data will be collected processed and stored in excel spreadsheets. 
Time course of delivered nicotine as well as fraction that reach lung or is deposited in oral 
cavity, throat and large bronchi will be tabulated and sent to the study sponsor as excel files. 
The radioactivity measured continuously in arterial blood as well in the discrete blood 
samples will tabulated in excel sheets and sent to the study sponsor. 
17.6 Analysis of safety endpoints 
 Adverse events  
An overview of all AEs, including SAEs, intensity, relationship to IP, and deaths will be 
presented by SOC and PT. 
Incidence of AEs and SAEs will be summarised by SOC and PT by treatment and study part 
and overall. 
All AE data will be listed subject and include the verbatim term entered by the Investigator. 
 Vital signs 
Vital signs (systolic/diastolic blood pressure and pulse) will be summarised by treatment and 
study part. Data will be presented with absolute and percent change from baseline. 
All data will be listed by subject. 
 
 

 
 
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18 REFERENCES 
1. Bergström M, Nordberg A, Lunell E, Antoni G, Långström B. Regional deposition of 
inhaled 11C-nicotine vapor in the human airway as visualized by positron emission 
tomography. Clin Pharmacol Ther 57:309-317, 1995 
2. Berridge MS, Apana SM, Nagano KK, Berridge CE, Leisure GP, Boswell MV. Smoking 
produces rapid rise of [11C]nicotine in human brain. Phychopharmacology (Berl) 
298:383-394, 2010 
3. National Cancer Institute Cancer Therapy Evaluation Program. Common terminology 
criteria for adverse events, CTCAE v5.0 (2017). 
4. Declaration of Helsinki: https://www.wma.net/policies-post/wma-declaration-of-helsinki-
ethical-principles-for-medical-research-involving-human-subjects/