Clinical trial NCT03509948
Conditions: Smoking Cessation
Sponsor: Achieve Life Sciences
Start date: 2018-04-27; Completion date: 2018-06-12
Enrollment: 13 participants; Allocation: RANDOMIZED; Intervention model: CROSSOVER

Brief summary:
This will be an open-label, randomised, 2-treatment period, single-dose crossover study to determine the comparative bioavailability and renal elimination following single-dose administration of 3.0 mg cytisine in healthy smokers under fed and fasted conditions.

Detailed description:

Arms: arm 1: Schedule A (6 participants):

* Period 1: cytisine (2 x 1.5 mg tablets) will be administered 30 minutes after the start of a high fat breakfast (fed state)
* Period 2: cytisine (2 x 1.5 mg tablets) will be administered after an overnight fast of at least 10 hours (fasting state) arm 2: Schedule B (6 participants):

* Period 1: cytisine (2 x 1.5 mg tablets) will be administered after an overnight fast of at least 10 hours (fasting state)
* Period 2: cytisine (2 x 1.5 mg tablets) will be administered 30 minutes after the start of a high fat breakfast (fed state)
Interventions: intervention 1: cytisine
Intervention descriptions: intervention 1: cytisine 1.5 mg film-coated tablets

Protocol documents:
Prot_000.pdf: 
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Protocol ACH-CYT-07, 03 April 2018, Version 2.0 
 
Cytisine 
 CONFIDENTIAL 
 
 
3 
SYNOPSIS  . 
NAME OF COMPANY: Achieve Life Sciences, Inc 
NAME OF INVESTIGATIONAL MEDICINAL PRODUCT (CYTISINE):  
Cytisine 1.5 mg film-coated tablet 
TITLE OF STUDY: A phase 1 open label, randomized, two-way crossover study in healthy 
smokers to investigate the effect of food on the bioavailability of cytisine in a new formulation 
PRINCIPAL INVESTIGATOR: 
 
STUDY CENTRE: Simbec Research Ltd (Simbec) 
     Merthyr Tydfil, CF48 4DR, UK 
CLINICAL PHASE: I 
OBJECTIVES: 
Primary:  
• To compare the bioavailability (Cmax and AUC0-∞) of cytisine in healthy smokers 
under fed and fasted conditions, following administration of 3 mg cytisine (2 x 
1.5 mg cytisine tablets). 
• To assess the extent of renal elimination of cytisine via measurement of urinary 
concentrations, following administration of 3 mg cytisine (2 x 1.5 mg cytisine 
tablets). 
Secondary:  
• To compare the AUC0-t, T½ and Tmax of cytisine in subjects under fed and fasted 
conditions, following administration of 3 mg cytisine (2 x 1.5 mg cytisine tablets). 
• To assess the safety and tolerability of cytisine in a new formulation at the 3 mg 
dose level under fed and fasted conditions. 
 
METHODOLOGY:  
This will be an open-label, randomised, 2- treatment period, single-dose crossover study to 
determine the comparative bioavailability and renal elimination of cytisine following single-
dose administration in healthy smokers under fed and fasted conditions.   
The study will be comprised of a pre-study screen, followed by 2 treatment periods (1 and 2) 
and a post-study follow-up.   
Screening (Day -28 to Day -1): Screening assessments will be carried out within 28 days 
before first administration of cytisine.   
Baseline (Day 0): Baseline assessments will be carried out one day before the first 
administration of cytisine.   
Treatment Periods (Day 1 to Day 5):  Eligible subjects will receive a single-dose of cytisine 
under fed and fasted conditions during 2 treatment periods over a 5-day time frame. Each 
treatment period will be approximately 2 days’ duration in which blood and urine samples will 
be collected over a 48 h period post cytisine dosing. This will require a total of 5 overnight 
stays in the Clinical Unit. The study will begin the afternoon before the first dosing on Day 0 
and the treatment period will start with the first dose on Day 1 and continue until 48 hours (h) 
after the second dosing on Day 5.   
Subjects will arrive at the Clinical Unit on Day 0 and will undergo an overnight fast of at least  


Protocol ACH-CYT-07, 03 April 2018, Version 2.0 
 
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10 h prior to first dosing on Day 1. Randomisation will occur on the morning of Day 1, Period 
1, and cytisine will be administered under fasted (after an overnight fast of at least 10 h) or fed 
(after a high fat breakfast) conditions.  Subjects will complete all 48 h postdose assessments 
prior to starting Period 2 and the second dosing on Day 3.   
Plasma and urine samples will be collected predose and up to 48 h postdose during each period 
(18 plasma samples and 11 urine samples per period) for the measurement of plasma and 
urinary cytisine concentrations. The 2 periods allow 48 hours between dose administrations. 
Safety will also be evaluated at specified times.   
Post Study: Post study telephone follow-up will be conducted 6 to 8 days after the second dose 
of cytisine. 
NUMBER OF SUBJECTS: 12 subjects will be randomised/enrolled to complete the study. 
MAIN INCLUSION CRITERIA: 
Healthy smokers (males or females who are non-pregnant/non-lactating) aged 18 - 55 years, 
with no allergy or sensitivity to cytisine or any of its excipients, who unless surgically sterile, 
abstaining from sexual intercourse or of non-child bearing potential, are willing to use an 
effective method of contraception from the first dose and for 3 months after the last dose of 
cytisine. 
CYTISINE ADMINISTRATION: 
Each subject will receive cytisine over 2 treatment periods under fed and fasted conditions in 
accordance with the randomisation schedule:   
Schedule A (6 subjects):  
• Period 1: Cytisine (2 x 1.5 mg tablets) will be administered 30 minutes after the start of 
a high fat breakfast (fed state). 
• Period 2: Cytisine (2 x 1.5 mg tablets) will be administered after an overnight fast of at 
least 10 hours (fasting state). 
Schedule B (6 subjects):  
• Period 1: Cytisine (2 x 1.5 mg tablets) will be administered after an overnight fast of at 
least 10 hours (fasting state). 
• Period 2: Cytisine (2 x 1.5 mg tablets) will be administered 30 minutes after the start of 
a high fat breakfast (fed state). 
240 mL water will be given with the dose. 
CRITERIA FOR EVALUATION: 
Study Endpoints:   
Plasma: The following PK endpoints will be derived from plasma cytisine concentration versus 
time data following administration of 3 mg cytisine (2 x 1.5 mg) under fed and fasted conditions: 
Maximum concentration (Cmax), time to Cmax (Tmax), elimination rate constant (kel), terminal 
elimination halflife (T1/2), area under the concentration versus time curve (AUC) from time of 
dosing to last measurable concentration (AUC0t), AUC extrapolated to infinity (AUC0inf) and 
AUC% extrapolated (residual area). 
Urine: Amount excreted in urine over time (Ae) and percent of cytisine excreted in urine 
(Ae%) will be calculated. 
Safety Endpoints: AEs, vital signs (supine systolic/diastolic pressure, pulse and oral 
temperature), 12 lead ECG, laboratory safety (haematology, biochemistry and urinalysis). 


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STATISTICAL METHODS:   
All statistical analysis will be performed using SAS® version 9.3. 
Demographic and Background Data: All demographic and background data will be listed. 
Demographic data will be summarised  (age, height, weight, BMI and race) by gender and 
overall. Subject disposition and analysis sets (safety, PK) will also be listed and summarised by 
frequency. 
Urine Excretion and PK:  All plasma and urine cytisine concentrations and derived PK data 
following administration of cytisine will be listed and the data summarised descriptively. 
Individual and mean plasma concentration time curves will also be plotted. In order to compare 
fed vs fasted conditions, following logarithmic transformation, Cmax, AUC0-∞ , AUC0-t and total 
Ae values will be subjected to an analysis of variance (ANOVA).  Point estimates and 90% 
confidence intervals (CI) will be constructed for the contrast between the two conditions. A 
comparison of Tmax will be performed using the Wilcoxon Signed-Rank test. In addition, the 
Hodges-Lehmann estimate of the median difference in Tmax and 95% confidence interval will be 
presented. 
Safety: All safety data will be listed.   
AEs will be coded using Medical Dictionary for Regulatory Activities (MedDRA) version 20.0 
or higher.  The incidence of treatment emergent AEs (TEAE) will be summarised by organ 
system, preferred term, severity and relationship to study drug.  Abnormal laboratory safety 
results will be listed separately.   
 
DURATION OF STUDY: Approximately 6 weeks for each individual (from screening to 
post-study follow-up). 


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TABLE OF CONTENTS 
SYNOPSIS  . ....................................................................................................................................3 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS ................................................11 
1. 
ETHICS ......................................................................................................................12 
1.1. 
Research Ethics Committee ........................................................................................12 
1.2. 
Subject Information and Consent ...............................................................................12 
1.3. 
Indemnity Arrangements ............................................................................................13 
2. 
INTRODUCTION AND BACKGROUND ...............................................................13 
2.1. 
Smoking and Health ...................................................................................................13 
2.2. 
Cytisine .......................................................................................................................13 
3. 
RATIONALE FOR THE STUDY ..............................................................................15 
4. 
STUDY OBJECTIVES ..............................................................................................15 
4.1. 
Primary Objective .......................................................................................................15 
4.2. 
Secondary Objectives .................................................................................................15 
5. 
INVESTIGATIONAL PLAN .....................................................................................15 
5.1. 
Study Design ...............................................................................................................15 
5.2. 
Discussion of Study Design ........................................................................................17 
6. 
SELECTION OF STUDY POPULATION ................................................................17 
6.1. 
Inclusion Criteria ........................................................................................................17 
6.1.1. 
To be Confirmed at Screening ....................................................................................17 
6.1.2. 
To be Re-Confirmed Prior to Dosing .........................................................................18 
6.2. 
Exclusion Criteria .......................................................................................................19 
6.2.1. 
To be Confirmed at Screening ....................................................................................19 
6.2.2. 
To be Re-Confirmed Prior to Dosing: ........................................................................19 
6.3. 
Removal of Subjects from Therapy or Assessment ....................................................19 
7. 
RESTRICTIONS PRIOR AND DURING THE STUDY ..........................................20 
7.1. 
Confinement ...............................................................................................................20 
7.2. 
Diet and Fluid Restrictions .........................................................................................20 
7.2.1. 
Meal Times .................................................................................................................20 
7.2.2. 
Fluid Intake .................................................................................................................21 
7.2.3. 
Smoking ......................................................................................................................21 
7.2.4. 
Alcohol Intake ............................................................................................................21 


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7.2.5. 
Caffeine .......................................................................................................................22 
7.2.6. 
Poppy and Sesame Seeds ............................................................................................22 
7.2.7. 
Grapefruit Juice and Other Restrictions .....................................................................22 
7.3. 
Other Life-Style Restrictions ......................................................................................22 
7.3.1. 
Strenuous Exercise ......................................................................................................22 
7.3.2. 
Blood Donation ...........................................................................................................22 
7.3.3. 
Contraception ..............................................................................................................22 
7.3.4. 
Sperm Donation ..........................................................................................................23 
8. 
INVESTIGATIONAL MEDICAL PRODUCT (CYTISINE) ...................................23 
8.1. 
Cytisine 1.5 mg Film-Coated Tablets .........................................................................23 
8.2. 
Receipt and Storage ....................................................................................................23 
8.3. 
Assembly and Certification ........................................................................................24 
8.4. 
Administration ............................................................................................................24 
8.5. 
Return/Destruction ......................................................................................................24 
8.6. 
Method of Assigning Subjects to Schedule A or Schedule B ....................................24 
8.7. 
Selection of Doses in the Study ..................................................................................25 
8.8. 
Selection and Timing of Dose for Each Subject .........................................................25 
8.9. 
Blinding ......................................................................................................................25 
9. 
PRIOR AND CONCOMITANT THERAPY .............................................................25 
10. 
TREATMENT COMPLIANCE .................................................................................25 
11. 
STUDY PROCEDURES ............................................................................................26 
11.1. 
Procedure Schedule ....................................................................................................26 
11.2. 
Detailed Description of Study Visits ..........................................................................28 
11.2.1. 
Screening Visit (Day -28 to Day -1) ...........................................................................28 
11.2.2. 
Day 0: Admission .......................................................................................................28 
11.2.3. 
Day 1 (Period 1) ..........................................................................................................28 
11.2.4. 
Day 2 (Period 1) ..........................................................................................................29 
11.2.5. 
Day 3 (Period 2) ..........................................................................................................29 
11.2.8. 
Days 6-8 post last dose: Post-Study Follow-up ..........................................................30 
12. 
DRUG CONCENTRATION MEASUREMENTS ....................................................31 
12.1. 
Pharmacokinetic Blood Sampling ..............................................................................31 
12.2. 
Pharmacokinetic Urine Collection ..............................................................................32 
12.3. 
Determination of Plasma and Urine Drug Concentrations .........................................32 


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12.4. 
Incurred Samples Reanalysis ......................................................................................33 
13. 
SAFETY ASSESSMENTS ........................................................................................33 
13.1. 
Subject’s Safety ..........................................................................................................33 
13.2. 
Adverse Events ...........................................................................................................34 
13.2.1. 
Definition of Adverse Event .......................................................................................34 
13.2.2. 
Recording of Adverse Events .....................................................................................35 
13.2.3. 
Grades of Adverse Event Severity ..............................................................................36 
13.2.4. 
Assessment of Causal Relationship ............................................................................36 
13.2.5. 
Reporting of Serious Adverse Events .........................................................................36 
13.2.6. 
Monitoring of Subjects with Adverse Events .............................................................38 
13.3. 
Pregnancy ...................................................................................................................38 
13.4. 
Laboratory ...................................................................................................................38 
13.4.1. 
Routine Laboratory Assessments ...............................................................................38 
13.4.2. 
Virology ......................................................................................................................39 
13.4.3. 
Drugs of Abuse and Alcohol ......................................................................................39 
13.5. 
Vital signs ...................................................................................................................39 
13.6. 
Physical Examination .................................................................................................39 
13.7. 
ECG Measurements ....................................................................................................39 
13.8. 
Concomitant Medication ............................................................................................40 
14. 
APPROPRIATENESS OF MEASUREMENTS ........................................................40 
15. 
STATISTICAL METHODS .......................................................................................40 
15.1. 
Primary Objectives and Analysis ................................................................................40 
15.2. 
Subject Populations for Analyses ...............................................................................41 
15.2.1. 
Safety Analysis Set .....................................................................................................41 
15.2.2. 
PK Analysis Set ..........................................................................................................41 
15.2.3. 
Urine Excretion Analysis Set ......................................................................................41 
15.3. 
Subjects Characteristics ..............................................................................................42 
15.4. 
Sample Size Calculation .............................................................................................42 
15.5. 
Plasma and Urine Concentrations ...............................................................................43 
15.6. 
Estimation of pharmacokinetic parameters ................................................................43 
15.7. 
Pharmacokinetic Analysis ..........................................................................................44 
15.8. 
Safety Variables and Analyses ...................................................................................44 
15.8.1. 
ECGs ...........................................................................................................................44 


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15.8.2. 
Vital Signs ..................................................................................................................44 
15.8.3. 
Adverse Events ...........................................................................................................44 
15.8.4. 
Safety Laboratory tests ...............................................................................................45 
15.9. 
Procedures for Deviations from Planned Analyses ....................................................45 
16. 
DOCUMENTATION .................................................................................................45 
16.1. 
Study File and Site Documents...................................................................................45 
16.2. 
Study Documents Supplied by the Sponsor ................................................................46 
16.3. 
Maintenance and Retention of Records ......................................................................46 
17. 
ADMINISTRATIVE PROCEDURES .......................................................................46 
17.1. 
Sponsor Responsibilities .............................................................................................46 
17.2. 
Investigator Responsibilities .......................................................................................47 
18. 
REGULATORY COMPLIANCE ..............................................................................47 
19. 
ETHICAL CONDUCT OF THE TRIAL / GOOD CLINICAL PRACTICE .............48 
20. 
PROTOCOL MODIFICATION/PREMATURE TERMINATION ...........................48 
21. 
POLICY FOR PUBLICATION AND DATA PRESENTATION .............................49 
INVESTIGATOR’S AGREEMENT .............................................................................................50 
REFERENCES ..............................................................................................................................51 
APPENDIX 1. NORMAL RANGES FOR VITAL SIGNS AND ECG PARAMETERS .........52 
 


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LIST OF TABLES 
Table 1: 
CYT-BIO-02-07 PK Results .......................................................................................14 
Table 2: 
Single Dose PK Results ..............................................................................................14 
Table 3: 
ACH-CYT-01 PK Results ..........................................................................................14 
Table 4: 
Cytisine Product Information .....................................................................................23 
Table 5: 
Schedule of Study Procedures ....................................................................................27 
Table 6: 
Blood Sampling for Pharmacokinetic Assessments During each Period ...................31 
Table 7: 
Urine Sampling for Pharmacokinetic Assessments During each Period ....................32 
Table 8: 
Adverse Event Severity ..............................................................................................36 
Table 9: 
Assessment of Causal Relationship ............................................................................36 
Table 10: Cytisine Pharmacokinetic Parameters ........................................................................43 
 
LIST OF FIGURES 
Figure 1: 
Study Design Overview ..............................................................................................16 
 


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LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS 
ABBREVIATION/TERM DEFINITION 
ADR 
Adverse Drug Reaction 
AE 
Adverse Event 
ANOVA 
Analysis of Variance 
ALT 
Alanine Aminotransferase 
AST 
Aspartate Aminotransferase 
AUC0-t 
AUC from Time Zero to Last Sampling Time with Quantifiable 
Concentrations 
AUC0-∞ 
AUC from Time Zero to Infinity 
%AUC 
Residual Area or Percentage of Extrapolated Part of AUC0-∞ 
BMI 
Body Mass Index 
Clast 
Last Quantifiable Concentration 
Cmax 
Maximum Observed Plasma Concentration 
CRF 
Case Report Form 
CV% 
Coefficient of Variation 
ECG 
Electrocardiogram 
EMA 
European Medicines Agency 
GCP 
Good Clinical Practice 
GMP 
Good Manufacturing Practice 
GMR 
Geometric Means Ratio` 
ICH 
International Conference on Harmonization 
IMP 
Investigational Medicinal Product (for this protocol indicates 
cytisine 1.5 mg film coated tablet) 
LC/MS/MS 
Liquid Chromatography-tandem Mass Spectrometry 
LLOQ 
Lower Limit of Quantification 
MedDRA 
Medical Dictionary for Regulatory Activities 
PK 
Pharmacokinetics 
RBC 
Red Blood Cell Count 
SAE 
Serious Adverse Event 
SAR 
Serious Adverse Reaction 
SD 
Standard Deviation 
SOC 
MedDRA System Organ Class 
Tmax 
Time to Maximum Observed Concentration 
T1/2 
Apparent Terminal Elimination Half-life 
UADR 
Unexpected Adverse Drug Reaction 
UAE 
Unexpected Adverse Event 
ULOQ 
Upper Limit of Quantification 
WBC 
White Blood Cell Count 
λz 
Apparent Terminal Elimination Rate Constant 


Protocol ACH-CYT-07, 03 April 2018, Version 2.0 
 
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1. 
ETHICS 
1.1. 
Research Ethics Committee  
This study protocol will be submitted to the Research Ethics Committee (REC) for review and 
approval.  The approval of the REC must be obtained before commencement of any study 
procedures. 
The favourable opinion is conditional upon the Sponsor registering the clinical trial in a publicly 
accessible database, within 6 weeks of the first participant recruited.   
All substantial protocol amendments must be approved by the REC responsible for the study.  
Non-substantial amendments will not require prior approval by the REC. 
If the study is stopped due to adverse events (AEs), it will not be recommenced without 
reference to the REC responsible for the study. 
The outcome of the study (e.g. completed) will be reported to the REC responsible for the study 
within 90 days of completion of the last subject’s final study procedures.  In the event of the 
study being prematurely terminated, a summary safety report will be submitted to the REC 
responsible for the study within 15 days. 
A summary of the clinical study report will be submitted to the REC responsible for the study 
within 1 year of completion of the last subject’s final study procedures. 
The REC will be informed that 
 is a commercial organisation and that the study is funded 
by Achieve Life Science. The subjects who take part in the clinical study will be paid for their 
inconvenience and have been informed that there will be no benefits gained by their 
participation. All potential conflicts of interest will be declared by the Investigators.  
1.2. 
Subject Information and Consent 
Potential subjects who volunteer for participation in the study will be informed of the aims, 
methods, anticipated benefits and potential hazards of the study and any possible discomfort it 
may entail.  Information will be given in both oral and written form and in the manner deemed 
appropriate by the Clinical Unit standard operating procedures (SOPs). Each subject will also be 
informed of his/her right to withdraw from the study at any time, for any reason. 
A written explanation (participant information sheet) and informed consent form will be 
provided and the subject will be allowed sufficient time to consider the study information. Prior 
to signing the informed consent form, the subject will be given an opportunity to discuss any 
issues concerning the study with an Investigator who has suitable knowledge of the study and 
will have all questions answered openly and honestly. 
If the subject is willing to participate in the study, the informed consent form will be signed and 
personally dated by the subject and the person taking consent. The subject will receive a copy of 
the informed consent form together with the participant information sheet and the original signed 
informed consent form will be retained with the study records at the Investigator site.  In 
addition, the actions and completion of the consenting process will be recorded in the subject’s 
medical record (i.e., source document). 


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13 
1.3. 
Indemnity Arrangements 
The Sponsor and 
carry insurance to pay compensation for injury, accident, ill health or 
death caused by participation in this study without regard to proof of negligence in accordance 
with the insurance and compensation in the event of injury in phase I clinical trials 2012, 
guidance issued by the ABPI, the BioIndustry Association and the Clinical Contract Research 
Association in consultation with the Department of Health and the National Research Ethics 
Service. 
2. 
INTRODUCTION AND BACKGROUND 
2.1. 
Smoking and Health 
Tobacco smoking contributes to some 5 million premature deaths each year worldwide.1 
Smoking is highly addictive, with more than 95% of unaided attempts at cessation failing to last 
6 months.2  Every year that a smoker delays quitting beyond the mid-30s, it has been estimated 
that the person loses 3 months of life expectancy.3  The World Health Organization’s Framework 
Convention on Tobacco Control identifies evidence-based approaches to promote smoking 
cessation, which include mass-media campaigns, tax increases on tobacco, and help for smokers 
wanting to stop.4 
2.2. 
Cytisine 
) containing the active substance cytisine has 
been licensed and marketed in Central and Eastern Europe for several decades by 
 
), but remains relatively unknown outside Eastern Europe despite requests for 
market licensing worldwide.5-7 
Cytisine’s mechanism of action has assisted basic pharmacologists in understanding the complex 
pharmacology of the various subtypes of the nicotinic acetylcholine receptor.8  These studies 
have shown that both nicotine and cytisine bind strongly and preferentially to pre-synaptic α4β2 
receptors9,10 that mediate the release of dopamine in the shell of the nucleus accumbens.  This 
receptor subtype has been implicated in the development and maintenance of nicotine 
dependence11 and was the primary target for the drug varenicline, which has proved effective in 
aiding smoking cessation.12 
Three studies have evaluated the pharmacokinetics of cytisine in man.  
Study CYT–BIO-02-07 was conducted according to the principles of Good Clinical Practice 
(GCP) and sponsored by 
.  The pharmacokinetic profile of cytisine in plasma was 
determined following administration of 1.5 mg cytisine (in the form of a 
 tablet) in 36 male 
healthy non-smoking volunteers.  All drug administrations were given in the fasted state.  The 
primary PK outcomes are summarised in (Table 1): 


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Table 1: 
CYT-BIO-02-07 PK Results 
Cmax 
15.55 ng/mL 
AUC0-t 
59.80 ng.h/mL 
AUC0-∞ 
73.63 ng.h/mL 
Tmax 
0.92 h  
(≈ 55 min) 
T½ 
4.09 h 
Cummulative cytisine levels excreted in the urine during the 24 hours post administration period 
indicated that approximately 64% of administered dose was excreted as parent compound 
(34.71% - 100.24%). 
A second study13 reported the pharmacokinetic profile of cytisine in seven healthy subjects (aged 
between 20 and 39 years) following administration of 3 mg cytisine . Subjects were eligible if 
they were current cigarette smokers at the time of the study and on average, the subjects smoked 
10.6 cigarettes per day. There was no restriction on diet or smoking during the study. The 
primary PK outcomes are summarised in (Table 2): 
Table 2: 
Single Dose PK Results 
Cmax 
27.76 ng/mL 
T½ 
4.8 h 
Tmax 
1-2 h 
VD 
115 L 
CL 
16.7 L/h 
Cummulative urinary excretion was not evaluated; however, cytisine was still detectable in the 
urine collected at 24 h for all subjects (mean concentration at 24 hr was 428.15 ng/mL).  
A third study (ACH-CYT-01) examined the pharmacokinetic profile of cytisine in healthy non-
smoking subjects following administration of 3 mg cytisine in the original marketed formulation 
during the fed and fasted states. The primary PK outcomes are summarized in (Table 3):  
Table 3: 
ACH-CYT-01 PK Results 
 
Fasted 
Fed 
Cmax 
22.9 ng/mL 
22.3 ng/mL 
AUC0-t 
166 ng.h/mL 
160 ng.h/mL 
AUC0-∞ 
173 ng.h/mL 
168 ng.h/mL 
Tmax 
0.75 h  
2.75 h 
T½ 
4.59 h 
4.77 h 
Fasted subjects received a single dose of 3 mg cytisine and approximately 82% of the dose was 
recovered from urine within 24 hours.  Final urinary concentrations in the study were measured 
in the 12-24 hour epoch and were non-zero for all subjects, indicating that renal excretion was 
still ongoing beyond 24 hours after cytisine administration.  However, based on the diminishing 
urinary concentrations over time, it can be concluded that excretion is likely to be complete 
within 48 hours. 


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3. 
RATIONALE FOR THE STUDY 
In the recent bioavailability study evaluating the original marketed formulation for cytisine, no 
food effect was observed for AUC0-∞ (or AUC0-t) with values considered bioequivalent as the 
90% CI for the difference between fed and fasted states fell within the range 80 -125%.  
Consistent with this, the amounts of cytisine recovered from urine within 24 h were similar 
between fed and fasted states, despite Cmax being lower in the fed state. In that study, it was 
concluded that there was no clinically significant food effect for cytisine and there were no 
safety concerns following administration of a single dose of 3 mg cytisine under fed or fasted 
conditions for healthy non-smokers.  
A revised formulation has been subsequently developed, eliminating lactose and introducing 
lower formic acid containing-excipients, to retard the formation of the breakdown substance 
N-formylcytisine. Initial studies indicate that the revised formulation has improved stability at 
24 months and that the shelf-life could be extended to 36 months based on the preliminary 
stability information. This study will evaluate if there is any food effect for cytisine using this 
new formulation in healthy smokers. 
4. 
STUDY OBJECTIVES 
4.1. 
Primary Objective 
1. To compare the bioavailability (Cmax and AUC0-∞) of cytisine in smokers under fed and 
fasted conditions, following administration of 3 mg cytisine (2 x 1.5 mg cytisine tablets). 
2. To assess the extent of renal elimination (Ae and Ae%) of cytisine via measurement of 
urinary concentrations, following administration of 3 mg cytisine (2 x 1.5 mg cytisine 
tablets). 
4.2. 
Secondary Objectives 
1. To compare the AUC0-t, T½ and Tmax of cytisine in subjects under fed and fasted 
conditions, following administration of 3 mg cytisine (2 x 1.5 mg cytisine tablets). 
2. To assess the safety and tolerability of cytisine in a new formulation at the 3 mg dose 
level under fed and fasted conditions. 
5. 
INVESTIGATIONAL PLAN 
5.1. 
Study Design 
This will be an open-label, randomised, 2-treament period, single-dose crossover Phase I study 
conducted in healthy smokers as shown below: 
 


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Figure 1: 
Study Design Overview  
 
The study will be comprised of a pre-study screen, followed by 2 treatment periods (1 and 2) and 
a post-study follow-up.   
Screening (Day -28 to Day -1): Screening assessments will be carried out within 28 days before 
first administration of cytisine.  
Baseline (Day 0): Baseline assessments will be carried out one day before the first 
administration of cytisine.   
Treatment Periods (Day 1 to Day 5):  Eligible subjects will receive a single-dose of cytisine 
under fed and fasted conditions during the 2 treatment periods.  
• Each treatment period will be approximately 2 days’ duration in which plasma and 
urine samples will be collected for 48 hrs.   
• Subjects will arrive at the Clinical Unit on Day 0 and will undergo an overnight fast 
of at least 10 h prior to dosing on Day 1. Randomisation will occur on the morning of 
Day 1, Period 1, at least 1 hour  prior to first dosing. 
• Cytisine will be administered under fasted (after an overnight fast of at least 10 h) or 
fed (after a high fat breakfast) conditions with the last plasma and urine sample 
occurring at 48 h postdose (before the second cytisine dosing on Day 3).  All subjects 
will stay overnight on Day 2 and undergo an overnight fast of at least 10 h prior to 
dosing on Day 3. 
• Pharmacokinetic (PK) samples (plasma and urine) will be collected predose and up to 
48 h postdose during each period for the measurement of plasma and urine cytisine 
concentrations. Safety will also be evaluated at specified times.   
• Final ECGs and blood draw for repeat haematology and chemistries will be obtained 
just prior to discharge on Day 5. 
• There will be at least 48 hours between dose administrations on Day 1 and Day 3. 
Post Study: Post study telephone follow-up will be conducted 6 to 8 days after the last dose of 
cytisine. If any adverse events are recorded at post study telephone assessment, arrangements 
 
R 
A 
N 
D 
O 
M 
I 
Z 
E 
Schedule B 
 
Initial dose of cytisine (3 mg) given after fasted state 
(Treatment Period 1) followed by second dose of cytisine (3 
mg) given after fed state (Treatment Period 2). 
Schedule A 
 
Initial dose of cytisine (3 mg) given after fed state 
(Treatment Period 1) followed by second dose of cytisine (3 
mg) given after fasted state (Treatment Period 2). 
 
S 
C 
R 
E 
E 
N 
 
 
F 
 
O 
 
L 
 
L 
 
O 
   W 
 
 
 
U 
 
P 


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will be made with the subject, so that they are followed-up appropriately and the final outcome 
determined.  
The end of study is defined as completion of safety follow-up on all subjects. The maximum 
duration of this study is estimated to be around 6 weeks (screening to last subject’s last visit, via 
telephone contact).   
5.2. 
Discussion of Study Design 
This study design minimizes the influence of inter-subject variability, which is assumed to be 
higher than the intra-subject variability, as each subject will be administered cytisine in both the 
fed and fasted states. As each subject serves as his own control, the precision of the comparison 
should be improved. 
The blood and urine samples for the determination of cytisine plasma and urine concentrations 
will be collected up to 48 hours after cytisine administration. Taking into account the known 
terminal elimination half-life of cytisine (about 4 hours), this sampling schedule is expected to 
provide a reliable estimate of the extent of exposure.  
The washout of at least 48 hours between dose administration will allow a complete elimination 
of cytisine between doses and is deemed sufficiently long to avoid any relevant carry-over effect. 
6. 
SELECTION OF STUDY POPULATION 
Approximately 12 subjects represented in the Urine/PK Analysis Set will be required to 
complete the study.   
The study is to be conducted in healthy subjects who are current smokers.  A healthy subject 
population with carefully considered inclusion/exclusion criteria will avoid the potential for 
interaction of cytisine with any underlying disease state or concomitant medication that might be 
necessary to administer to patients, while ensuring that subjects are fit and well enough for 
participation in the study.  Since smokers are the intended population for cytisine treatment, 
current smokers will be enrolled in the study. 
The following eligibility criteria are designed to select subjects for whom protocol treatment and 
procedures are considered appropriate.  All relevant medical and non-medical conditions should 
be taken into consideration when deciding whether this protocol is suitable for a particular 
subject. 
6.1. 
Inclusion Criteria 
6.1.1. 
To be Confirmed at Screening 
1. Subject is current cigarette smoker. 
2. Healthy males and females between 18 and 55 years of age. 
a. If a female subject of child bearing potential, a negative pregnancy test at screening 
and admission and willing to use an effective method of contraception (unless of non-
childbearing potential or where abstaining from sexual intercourse is in line with the 


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preferred and usual lifestyle of the subject) from first dose until 3 months after last 
dose of cytisine.  
b. If a female subject of non-child bearing potential, a negative pregnancy test at 
screening and admission.  For the purposes of this study, this is defined as the subject 
being amenorrheic for at least 12 consecutive months or at least 4 months post-
surgical sterilisation (including bilateral fallopian tube ligation or bilateral 
oophorectomy with or without hysterectomy).  Menopausal status will be confirmed 
by demonstrating at screening that levels of follicle stimulating hormone (FSH) fall 
within the respective pathology reference range.  In the event a subject's menopause 
status has been clearly established (for example, the subject indicates she has been 
amenorrheic for 10 years), but FSH levels are not consistent with a post-menopausal 
condition, determination of subject eligibility will be at Investigator's discretion 
following consultation with the Sponsor. 
c. If a male subject, willing to use an effective method of contraception (unless 
anatomically sterile or where abstaining from sexual intercourse is in line with the 
preferred and usual lifestyle of the subject) from first dose until 3 months after last 
dose of cytisine. 
3. Subject with a body mass index (BMI) of 23-28 kg/m2. BMI = body weight in kg / 
[height in m2]. 
4. Subject with no clinically significant abnormal serum biochemistry or haematology 
values within 28 days before the first dose of cytisine.   
5. Subject with negative urinary drugs of abuse screen, determined within 28 days before 
the first dose of cytisine (a positive result may be repeated at Investigator's discretion). 
6. Subject with negative human immunodeficiency virus (HIV), hepatitis B surface antigen 
(Hep B) and hepatitis C virus antibody (Hep C) results. 
7. Subject with no clinically significant abnormalities in 12-lead ECG determined after 
minimum of 5 minutes in supine position within 28 days before the first dose of cytisine. 
8. Subject with no clinically significant abnormalities in vital signs (systolic blood pressure 
between 90-140 mmHg, diastolic blood pressure (DBP) between 50 and 90 mmHg, and 
pulse rate (PR) between 40-100 bpm, measured on the dominant arm after minimum of 
5 minutes in supine position) determined within 28 days before first dose of cytisine. 
9. Subject must be available to complete the study (including post study follow-up) and 
comply with study restrictions. 
10. Subject must provide written informed consent to participate in the study. 
6.1.2. 
To be Re-Confirmed Prior to Dosing 
1. Subject continues to meet all screening inclusion criteria (before the cytisine dose). 
2. Subject has a negative urinary drugs of abuse screen (including alcohol). 
3. Female subject has a negative urine pregnancy test.  


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6.2. 
Exclusion Criteria 
6.2.1. 
To be Confirmed at Screening 
1. Known hypersensitivity/allergy reaction to varenicline, other cytisine-derivatives or any 
of the excipients in the 
 formulation 
). 
2. History of severe hypersensitivity reactions to any other drugs. 
3. History of any medical condition (e.g. gastrointestinal, renal or hepatic) or surgical 
condition (e.g. cholecystectomy, gastrectomy) that may affect drug pharmacokinetics 
(absorption, distribution, metabolism or excretion). 
4. Female subjects who are breast feeding. 
5. Difficulty in donating blood on either arm or known history. 
6. History of alcoholism or drug abuse within last 2 years. 
7. Use of prescription or non-prescription drugs, including vitamins, herbal and dietary 
supplements within 14 days (or 5 half-lives, whichever is longer) prior to the cytisine 
dose, unless in the opinion of the Investigator the medication will not interfere with the 
study procedures or compromise subject safety. 
8. Participated in any investigational drug clinical trial within the previous 3 months or a 
marketed drug trial within the previous 30 days prior to randomization on Day 1 of 
Period 1. 
9. Donation of 450 mL or more blood or had history of significant blood loss due to any 
reason or had plasmapheresis within 3 months before the cytisine dose. 
10. Any inability or difficulty in fasting. 
11. Inability to communicate well with Investigators (i.e., language problem, poor mental 
development or impaired cerebral function). 
12. Any other condition that the Principal Investigator considers making the subject 
unsuitable for this study. 
6.2.2. 
To be Re-Confirmed Prior to Dosing: 
1. Development of any exclusion criteria since screening. 
2. Use of prescription or non-prescription drugs, including vitamins, herbal and dietary 
supplements since screening, unless in the opinion of the Investigator the medication will 
not interfere with the study procedures or compromise subject safety. 
3. Participation in a clinical study since the screening visit. 
4. Donation of 450 mL or more blood since the screening visit. 
6.3. 
Removal of Subjects from Therapy or Assessment 
Each subject will be informed of their right to withdraw from the study at any time and for any 
reason. 


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An Investigator will withdraw a subject from the study at any time for any of the following 
reasons: 
• If a subject experiences a serious or intolerable AE, that prevents them from 
continuing. 
• If a subject incurs a significant protocol violation which impacts on their safety or the 
scientific integrity of the study (this will be discussed on a case-by-case basis with the 
Sponsor). 
• At the request of the Sponsor. 
• If it is considered that the subject’s health is compromised by remaining in the study 
or the subject is not sufficiently cooperative. 
• If a subject is lost to follow-up. 
The reasons for any subject withdrawal will be recorded on the study completion form of the 
case report form (CRF). 
If a subject is withdrawn or chooses to withdraw from the study for any reason, every possible 
effort will be made to perform the evaluations described for the post-study follow-up. The data 
collected from withdrawn subjects will be included in the study report. 
In the event of any abnormalities considered to be clinically significant, subjects will be followed 
up with appropriate medical management until values are considered to be clinically acceptable. 
Referral or collaborative care will be organised if considered necessary. 
Since approximately 12 subjects are to be included in the urine excretion/PK dataset to complete 
the study, subjects who withdraw from the study before receiving any study medication will be 
replaced.  Subjects who are withdrawn from the study due to significant drug-related AEs will 
not be replaced.  Replacement of all other subjects withdrawn from the study after receiving 
study medication will be decided on a case-by-case basis by the Principle Investigator (or 
delegate) and Sponsor. 
7. 
RESTRICTIONS PRIOR AND DURING THE STUDY 
To avoid possible negative effects on the measurement of plasma drug concentrations and/or on 
subject’s safety, several restrictions are to be adopted as outlined in the eligibility criteria and in 
the following sections.  
7.1. 
Confinement 
Each subject will complete 2 treatment periods (Period 1 and Period 2) in confinement. This 
period will require 5 nights stay (arrive at the unit in the afternoon of Day 0 and discharged 
48 hours post second cytisine dose on Day 5). 
7.2. 
Diet and Fluid Restrictions 
7.2.1. 
Meal Times 
Fasting: 


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• Subjects will fast overnight for at least 10 hours before dosing. 
• Lunch will be served: approximately 4 h post-dose. 
• Dinner will be served: approximately 8 h post-dose. 
• Snack will be served: approximately 12 h post-dose. 
Fed: 
• Subjects will fast overnight for at least 10 hours. 
• A high fat breakfast will be served at 30 min pre-dose (breakfast needs to be finished 
at least 5 minutes before dosing).   
• Lunch will be served: approximately 4 h post-dose. 
• Dinner will be served: approximately 8 h post-dose. 
• Snack will be served: approximately 12 h post-dose. 
The example of a high fat (approximately 50% of total caloric content of the meal) and high-
calorie (approximately 800-1000 calories) breakfast provided in the US Food and Drug 
Administration guidance document for food-effect bioavailability and fed bioequivalence studies 
(Guidance for Industry, U.S. Department of Health and Human Services Food and Drug 
Administration Center for Drug Evaluation and Research (CDER), December 2002) is 
approximately 150, 250 and 500-600 calories from protein, carbohydrate and fat respectively. 
Suggested content to be:  2 eggs fried in butter, 2 strips of bacon, 2 slices of toast, 4 ounces of 
hash brown potatoes and 8 ounces of whole milk.  Consumption of the standardized breakfast 
will be documented. 
7.2.2. 
Fluid Intake 
No fluids (apart from water taken with dose) are allowed from 1 h prior to dosing until 1 h 
afterwards.  Water is then allowed ad libitum. Decaffeinated tea and coffee as well as 
squash/cordial are allowed from 4 h post-dose. 
7.2.3. 
Smoking 
There will be minimal restrictions to smoking during days of confinement, such as all smoking 
must occur within designated area(s) only and subjects cannot smoke during active PK sampling 
period (i.e. subjects must have their last cigarette a minimum of 2 hours before dosing and can 
only start smoking after final PK sample). Subjects will go for a cigarette as a group and will 
need to have their last cigarette before 23:00 while they stay in the clinic. Clinic staff will review 
these guidelines with subjects during confinement and will monitor compliance. 
7.2.4. 
Alcohol Intake 
The consumption of alcohol will be limited to a maximum of 2 units per day from 7 days prior to 
the first dose of cytisine.  Alcohol will be avoided completely for at least 2 days prior to the first 
dose of cytisine and throughout the study period.  Any deviation outside this alcohol intake 
restriction will be assessed on a case-by-case basis at Investigator’s discretion (provided the 


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subject’s alcohol intake will not impact in the safety aspects and objectives of the study and the 
subject has a negative alcohol screen prior to dosing). 
7.2.5. 
Caffeine 
Food or drink containing caffeine, including coffee, tea, cola, energy drinks or chocolates will be 
avoided completely for 2 days prior to dosing and whilst the subjects are resident in the Clinical 
Unit (during each dosing period). 
7.2.6. 
Poppy and Sesame Seeds 
Subjects must not eat food containing poppy or sesame seeds for 3 days before drug of abuse 
testing, as consumption can lead to a positive opiate result in the drugs of abuse test. 
7.2.7. 
Grapefruit Juice and Other Restrictions  
No food or drink containing grapefruit, cranberry, or Seville oranges (including marmalade and 
fruit juices), and/or food or drink, sweets, candies or other confectionary containing liquorice 
will be allowed from 7 days before the first dose of cytisine until the study completion. 
Each subject will be questioned on the specific points at admission to the clinical unit (Day 0). If 
a subject admits a non-compliance with these restrictions, the Principal Investigator or attending 
Research Physician will decide whether or not the subject will be permitted to remain in the 
study. Non-compliance with these restrictions will be noted. 
7.3. 
Other Life-Style Restrictions 
7.3.1. 
Strenuous Exercise 
Strenuous exercise must be avoided completely from 3 days before the first dose of cytisine until 
the final study visit. 
7.3.2. 
Blood Donation 
Subjects will be advised that they should not donate blood for at least 3 months after the final 
study visit. 
7.3.3. 
Contraception 
To prevent pregnancy female subjects of childbearing potential and male subjects with female 
partners of child bearing potential must take adequate contraceptive precautions for the entire 
duration of study participation. Adequate contraceptive precautions include: 
1. Established use of oral, injected or implanted hormonal methods of contraception. 
2. Placement of an intrauterine device (IUD) or intrauterine system (IUS).  
3. Barrier methods of contraception: Condom or occlusive cap (diaphragm or cervical/vault 
caps) with spermicidal foam/gel/film/cream/suppository.  
4. Male sterilisation (with the appropriate post-vasectomy documentation of the absence of 
sperm in the ejaculate). [For female subjects on the study, the vasectomised male partner 
should be the sole partner for that subject]. 


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5. True abstinence, when this is in line with the preferred and usual lifestyle of the subject. 
[Periodic abstinence (e.g., calendar, ovulation, symptothermal, post-ovulation methods) 
and withdrawal are not acceptable methods of contraception]. 
The chosen contraception method(s) must be followed from the first dose until at least 3 months 
after the last dose of cytisine. 
7.3.4. 
Sperm Donation 
Subjects must not donate sperm from the first dose and for at least 3 months after the last dose of 
cytisine.  
8. 
INVESTIGATIONAL MEDICAL PRODUCT (CYTISINE) 
8.1. 
Cytisine 1.5 mg Film-Coated Tablets 
Cytisine will be supplied by the Sponsor. A Cytisine tablet is formulated as a compressed film-
coated tablet containing 1.5 mg Cytisine in each single tablet (See Table 4).  
Cytisine tablets are manufactured by 
) in accordance with cGMP. 
Transition to a revised formulation for 
, as presented below, has been approved in 
 
and is being submitted in other countries where 
 is marketing the product. This study 
will evaluate the revised formulation tablets. The reformulation has been implemented to 
improve long term stability.  
Each tablet is composed of cytisine (as the base), with well-established tablet-forming excipients 
 
 
Cytisine should be administered orally in a single dose application of 
 
taken with 
  
Table 4: 
Cytisine Product Information 
Name of the Product: 
 
Strengths: 
 
Dosage form: 
 
Route/mode of administration: 
Oral 
Dosing Schedule: 
 
Dosing instructions: 
 
Manufacturer/Marketing Authorization Holder: 
 
 
The documentation supplied will make it possible to retrace the composition and pharmaceutical 
quality of the product. 
8.2. 
Receipt and Storage 
The test cytisine will be supplied by the Sponsor as product in the commercial presentation. 


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The Sponsor must notify the Chief Investigator, or the Project Manager, prior to dispatch of the 
cytisine supplies, and of the anticipated date of their arrival.  Cytisine should arrive at the study 
site at least 7 days before the first dosing day.  The Sponsor shall address all supplies to: 
 
 
 
 
 
The cytisine will be stored under quarantine in a segregated, study-specific area, at or below 
25 oC in a secure, temperature-controlled pharmacy. 
 Qualified Person (QP) will review 
the shipping documentation. If acceptable, the supplies will subsequently be removed from 
quarantine and approved for use. 
8.3. 
Assembly and Certification 
Cytisine tablets will be assembled into unit doses by suitably trained 
 staff according to 
SOP SR-IMP 010 and SR-PHARM 017 as appropriate.  
Cytisine will be labelled as specified in Annex 13 of the European Commission (EC) guide to 
Good Manufacturing Practice (GMP). 
Packaged cytisine tablets will be certified by
 Qualified Person according to SOP SR-
IMP 030. 
8.4. 
Administration 
Each subject will receive cytisine under both fed and fasted conditions over 2 treatment periods in 
accordance with the randomisation schedule, which will assign them to a scheduled period for 
either fed or fasting sequencing (Schedule A or Schedule B): 
The dose to be administered is 3 mg Cytisine (2 x 1.5 mg tablets) 
8.5. 
Return/Destruction 
All used cytisine containers and unused cytisine will be held under quarantine in the 
 
Pharmacy pending return/destruction. Following the Sponsor’s approval, all remaining cytisine 
will be destroyed or returned to the Sponsor within 8 weeks of the last dose of cytisine.  
8.6. 
Method of Assigning Subjects to Schedule A or Schedule B 
Subjects will be randomized to treatment schedule according to a code produced by 
 
using the PROC PLAN procedure of SAS® version 9.3. Subjects will be numbered sequentially 
from 001 (i.e. 001, 002 etc.).  Replacement subjects will be assigned the same randomisation as 
the subject they are replacing, however, 100 will be added to the number (i.e., 101 would replace 
001 etc.). 


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8.7. 
Selection of Doses in the Study 
The dose selected for this study is 3 mg cytisine. The dose level was selected to allow adequate 
detection of cytisine in plasma samples for PK analysis. 
8.8. 
Selection and Timing of Dose for Each Subject 
Doses will be administered at approximately 9:00 a.m. All subjects will be fasting for at least 
10 h overnight prior to dosing in both treatment periods. Subjects assigned to be fasting prior to 
dosing will remain fasting until lunch and subjects assigned to be fed prior to dosing will start 
their high-fat breakfast 30 minutes prior to the dose. Lunch will be provided 4 h after dosing for 
all subjects. 
8.9. 
Blinding 
The study is open-label. However, as stated in the EMA guidelines on investigation of 
bioavailability and bioequivalence (CPMP/EWP/QWP/1401/98 Rev.1/Corr** London, 
20 January 2010), analysis of the samples should be conducted without information on treatment, 
therefore staff responsible for drug concentration and ECG analysis will be blinded to the 
treatment schedule. 
9. 
PRIOR AND CONCOMITANT THERAPY 
Prior Medication: Prescription or non-prescription drugs, including vitamins, herbal and dietary 
supplements should not be taken within 14 days (or 5 half-lives, whichever is longer) prior to the 
first dose of cytisine, unless in the opinion of the Investigator and Sponsor's Responsible 
Physician the medication will not interfere with the study procedures or would compromise 
subject safety. Prescription or non-prescription drugs, including vitamins, herbal and dietary 
supplements taken during the 14 days before the first dose of cytisine, and the reason for taking 
them, will be noted in the subject’s CRF.  
Inclusion of subjects who have taken prior medication will be reviewed on a case-by-case basis 
in relation to the safety aspects and objectives of this study.   
Concomitant Medication: Prescription or non-prescription drugs, including vitamins, herbal 
and dietary supplements should not be taken throughout the duration of the study, with the 
exception of paracetamol (which may be taken as an analgesic to a maximum of 2 g in 24 h 
(500 mg 4 times a day) and ibuprofen (which may be taken as an analgesic to a maximum of 
1.2 g in 24 h (400 mg 3 times a day).  
If intake of ANY prior or concomitant medication is necessary during the study, the daily dosage, 
duration and reasons for administration will be recorded on the subject’s CRF. 
Chewing gum will not be allowed during confinement periods. 
10. 
TREATMENT COMPLIANCE 
Each dose of cytisine will be taken under supervision and a hand and mouth check conducted.  
The exact dosing time for each subject will be recorded on the subject’s CRF. 


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11. 
STUDY PROCEDURES 
All subjects will be evaluated for inclusion in the study during the Screening Period. Subjects 
who are eligible for the study will be randomized according to the randomization schedule. 
Subjects will receive their study medications on Day 1 and Day 3 of the 5-day clinic stay 
followed by the required blood and urine samples. The subjects will be released from the clinical 
research facility after the last PK and urine sample is collected and safety assessments completed 
on Day 5. All subjects are required to complete a telephone follow-up on Day 6-8 days post last 
dose. 
11.1. 
Procedure Schedule 
Table 5 provides a summary of required study evaluations. Screening evaluations are to occur 
within a 28-day interval from initiation of screening evaluations to randomization on Day 1. 


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Table 5: 
Schedule of Study Procedures 
Days 
Screening 
Baseline 
Study Day (Period 1 and Period 2) 
Post-Study Telephone Call  
Days 
-28 to -1 
Day 0 
1  
2 
3 
4 
5 
6-8 days after last dose 
Written informed consent 
X 
 
 
 
 
 
 
 
Demographic 
data/height/weight 
X 
 
 
 
 
 
 
 
Randomization 
 
 
X  
(Pre-first dose in 
Period 1) 
 
 
 
 
 
Vital signs 
X 
X 
X1 
 
X1 
 
X 
 
Medical history 
X 
 
 
 
 
 
 
 
Medical history update 
 
X2 
 
 
 
 
 
 
Prior and concomitant 
medication 
X 
X 
X 
X 
X 
X 
X 
 
Physical examination 
X 
 
 
 
 
 
 
 
12-lead ECG 
X 
 
 
 
 
 
X3 
 
Pregnancy test for all 
females 
X 
(Serum) 
X 
(Urine) 
 
 
 
 
 
 
FSH (post-menopausal 
females only) 
X 
 
 
 
 
 
 
 
Haematology 
X 
 
 
 
 
 
X 
 
Biochemistry 
X 
 
 
 
 
 
X 
 
Urinalysis 
X 
 
 
 
 
 
X 
 
Drugs-of-abuse tests in 
urine 
X 
X 
 
 
 
 
 
 
Verification of eligibility 
criteria  
X 
X 
X 
 
 
 
 
 
Cytisine administration 
 
 
X 
 
X 
 
 
 
Blood collection for PK 
analysis 
 
 
X 
X 
X 
X 
X 
 
Urine collection for PK 
analysis 
 
 
X 
X 
X 
X 
X 
 
Adverse events monitoring 
 
X4 
X4 
X4 
X4 
X4 
X4 
X4 
1 Vital signs (supine blood pressure, pulse rate and oral temperature) will be recorded at pre-dose and again approximately 2 hours after administration on Day 
1 of each period. 
2 Clinically relevant changes will be reported as adverse events.  
3 Repeat 12-lead ECG and assess prior to discharge on Day 5. 
4 Adverse event recording to begin upon admissions on Day 0 of Period 1 to post study. 


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11.2. 
Detailed Description of Study Visits 
11.2.1. 
Screening Visit (Day -28 to Day -1) 
All subjects will complete the following screening procedures within 28 days of Day 1 in order 
to verify required inclusion and exclusion criteria.  
1. Written informed consent. 
2. Demographic data, including height and weight. 
3. Number of cigarettes smoked in past 24 hours. 
4. Medical history data. 
5. Prior and concomitant medication. 
6. Physical examination. 
7. Vital signs. 
8. 12-lead ECGs (triplicate). 
9. Clinical laboratory safety tests (biochemistry, haematology, urinalysis, virology, drugs of 
abuse and serum pregnancy test for females). 
10. Verification of eligibility criteria. 
11.2.2. 
Day 0: Admission 
The results of screening for each subject must be reviewed by the Principal Investigator prior to 
subject’s admission to assure eligibility criteria have been met.   
On Day 0, the subject will check into the research facility and upon arrival to the research 
facility, the following procedures are to be performed: 
1. Repeat urine testing for drugs of abuse and urine pregnancy test for female subjects.  
2. Verify that all subjects still satisfy the inclusion and exclusion criteria. 
All subjects will fast for minimum of 10 hours prior to dosing on Day 1. 
All subjects will remain in clinic overnight. 
11.2.3. 
Day 1 (Period 1) 
Administration of cytisine will occur on Day 1 for all subjects.  
After Day 1 pre-dose study procedures have been successfully completed and all eligibility 
criteria has been verified, subjects will be randomized onto Schedule A or Schedule B. 
The following study procedures are to be performed prior to dosing:  
1. Vital signs (supine blood pressure, pulse rate and oral temperature) will be recorded at 
pre-dose and again approximately 2 hours after administration. Any clinically relevant 
changes should be documented as adverse events. 
2. Prior and concomitant medication documented. 


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3. Pre-dose blood draw and urine for PK to be taken within 60 minutes prior to dosing. 
4. Administration of cytisine according to randomization: 
Schedule A (6 subjects):  
Cytisine (2 x 1.5 mg tablets) will be administered 30 minutes after the start of a high fat 
breakfast (fed state) with 240 mL (8 fluid ounces) water. No additional fluids are to be 
taken until 1 hour post-dosing 
Schedule B (6 subjects):  
Cytisine (2 x 1.5 mg tablets) will be administered after the overnight fast of at least 
10 hours and subject will continue fasting until lunch (fasting state). Dosing will be 
administered with 240 mL (8 fluid ounces) water. Except for fluids taken for dosing, no 
fluids will be allowed from 1 hour before dosing until 1 hour post-dosing. 
Blood and urine samples will be collected and processed as outlined in (Section 12.1) and 
(Section 12.2) prior to, and after administration of cytisine (time zero).   
No food should be allowed for at least 4 hours post-dose for either arm until lunch is served for 
all subjects. 
All subjects will remain in the clinic overnight. 
11.2.4. 
Day 2 (Period 1) 
Blood and urine samples for pharmacokinetics for Period 1 will be collected (Sections 12.1 and 
Section 12.2). 
1. Document concomitant medications and any adverse events. 
2. Collect plasma samples for PK analysis. 
3. Collect urine samples for PK analysis. 
4. Initiate fasting for Period 2 on the evening of Day 2. All subjects will fast for minimum 
of 10 hours prior to dosing on Day 3. 
All subjects will remain in clinic overnight. 
11.2.5. 
Day 3 (Period 2) 
The following study procedures are to be performed prior to dosing:  
1. Remaining blood and urine samples to complete Period 1 (through 48 h) 
pharmacokinetics will be collected (Sections 12.1 and Section 12.2). 
2. Pre-dose blood draw and urine for PK to be taken within 60 minutes prior to dosing. 
3. Vital signs (supine blood pressure, pulse rate and oral temperature) will be recorded at 
pre-dose and again approximately 2 hours after administration. Any clinically relevant 
changes should be documented as adverse events. 
4. Administration of cytisine according to randomization: 
 


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Schedule A (6 subjects):  
Cytisine (2 x 1.5 mg tablets) will be administered after the overnight fast of at least 
10 hours and subject will continue fasting until lunch (fasting state). Dosing will be 
administered with 240 mL (8 fluid ounces) water. Except for fluids taken for dosing, no 
fluids will be allowed from 1 hour before dosing until 1 hour post-dosing. 
Schedule B (6 subjects):  
Cytisine (2 x 1.5 mg tablets) will be administered 30 minutes after the start of a high fat 
breakfast (fed state) with 240 mL (8 fluid ounces) water. No additional fluids are to be 
taken until 1 hour post-dosing 
1. Document concomitant medications and any adverse events. 
2. Collect plasma sample for PK analysis 
3. Collect urine sample for PK analysis.  
All subjects will remain in the clinic overnight. 
11.2.6. 
Day 4 
Blood and urine samples for pharmacokinetics in Period 2 will be collected (Section 12.1). 
1. Document concomitant medications and any adverse events. 
2. Collect plasma samples for PK analysis 
3. Collect urine samples for PK analysis. 
All subjects will remain in the clinic overnight. 
11.2.7. 
Day 5 
Remaining blood and urine samples for pharmacokinetics in Period 2 will be collected (Sections 
12.1 and 12.2). 
1. Document concomitant medications and any adverse events. 
2. At 48 hours post-dosing, collect blood for haematology and biochemistry and collect 
urine for urinalysis.  
3. Obtain 12-lead ECG (in triplicate) similar to what was done at screen and review prior to 
discharge. 
Upon final review by the study physician, the subject will be released from the clinical research 
facility.  
11.2.8. 
Days 6-8 post last dose: Post-Study Follow-up 
A Post-Study Follow-up telephone call will occur 6-8 days after last dosing to document any 
changes in adverse event(s) observed at discharge as well as any new adverse events. If any new 
adverse events are recorded at this follow-up call, arrangements will be made with the subject to 
come into the clinic for assessment and then followed until outcome determined.  


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12. 
DRUG CONCENTRATION MEASUREMENTS 
12.1. 
Pharmacokinetic Blood Sampling 
18 venous blood samples (volume of 2.7 mL each, total volume of approximately 48.6 mL) will 
be collected into pre-cooled lithium heparin tubes during each period, according to the following 
schedule: 
Table 6: 
Blood Sampling for Pharmacokinetic Assessments During each Period   
Blood Sample Number 
Time from Dosing 
1 
Pre-dose 
(within 60 minutes  
prior to dosing) 
2 
00:15 
3 
00:20 
4 
00:30 
5 
00:45 
6 
01:00 
7 
01:30 
8 
02:00 
9 
02:30 
10 
03:00 
11 
04:00 
12 
06:00 
13 
08:00 
14 
12:00 
15 
14:00 
16 
16:00 
17 
24:00 
18 
48:00 
The pre-dose blood samples will be collected within 60 minutes prior to dosing.  The post-dose 
blood samples will be collected within +/-1 minute from the scheduled sampling time. The clock 
time will be recorded and reported for all blood draws and all subjects.  A deviation greater than 
+/- 2 minutes will be reported as a protocol deviation and its cause will be recorded.  
In case blood sampling for pharmacokinetics and other procedures coincide in time, blood draws 
will have priority unless other procedures are necessary for assuring subject’s safety.  
Immediately upon sampling, the blood sample for pharmacokinetic analysis will be identified 
with a bar-coded label bearing details of study number, subject number, study phase/period, 
sampling time point, sample type and a unique 9-digit identification number. The samples will be 
logged and then centrifuged at 1500 x g, 4°C for 10 minutes. Two equal aliquots of plasma will 
be transferred to polypropylene tubes labeled identically to the original blood sample (except 
sample type being plasma instead of whole blood) and logged when stored at approximately -
20°C, pending analysis.   
The blood samples will be placed on ice immediately and centrifuged within 60 minutes of being 
taken. 


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12.2. 
Pharmacokinetic Urine Collection 
Eleven urine samples will be collected at the following time points during each period: 
Table 7: 
Urine Sampling for Pharmacokinetic Assessments During each Period   
Urine Sample Number 
Time from Dosing 
1 
Pre-dose 
(Pre-dose urine samples will 
be collected within 30 
minutes prior to dosing) 
2 
00:00-02:00* 
3 
02:00-04:00 
4 
04:00-08:00 
5 
08:00-12:00 
6 
12:00-24:00 
7 
24:00-26:00 
8 
26:00-28:00 
9 
28:00-32:00 
10 
32:00-36:00 
11 
36:00-48:00 
*Each urine collection must contain all urine excreted within the timeframes as indicated. 
The urine collection containers will be identified with a bar coded label bearing details of the 
study number, subject number, study period number, sampling time-point, sample type and a 
unique 9 digit identification number. 
The urine collection containers will be kept chilled (at approximately 4°C) during collections.  
After the termination of each fractional collection the volume excreted will be recorded and 2 x 
10 mL aliquots retained for determination of cytisine concentrations.  The aliquots will be 
labelled identically to the original collection container and will be stored at -20°C pending 
analysis. 
12.3. 
Determination of Plasma and Urine Drug Concentrations 
Cytisine concentrations (ng/mL) in plasma and urine will be determined using a validated liquid 
chromatography tandem mass spectrometry method and will be conducted in accordance with 
Good Laboratory Practice regulations and guidelines. Methods validation must be completed 
prior to start of the analysis. 
The sample analysis will only begin after the last sample of a study subject is collected. All 
samples from each subject will be analyzed in the same analytical batch.  In each run, standard 
and quality control samples will be distributed throughout the batch containing study samples. 
Samples with concentrations above the ULOQ will be diluted and re-assayed.  
Full details, including the lower (LLOQ) and upper (ULOQ) limits of quantification range,  will 
be provided in a Bioanalytical Phase Plan written by 
 
Plasma and urine samples of withdrawn subjects should also be analysed for drug concentrations 
and pharmacokinetic parameters (providing withdrawal was not due to withdrawal of consent).  


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Unacceptable values due to analytical reasons will be handled in accordance with the 
Bioanalytical Phase Plan.   
Details of the methodologies used and the results obtained, including details of re-measured 
samples, will be given in the Analytical Study Report (ASR). 
12.4. 
Incurred Samples Reanalysis 
In order to establish the reproducibility of the assay, 10% of the first 1000 samples and 5% of the 
number of samples exceeding 1000 samples will be reanalyzed. 
Both the original and replicate values will be presented in the Analytical Study Report with the 
percent difference between the two values. The original value will be the one used for 
pharmacokinetic analysis. 
13. 
SAFETY ASSESSMENTS 
13.1. 
Subject’s Safety 
Only healthy subjects are eligible for the study.  The healthy status will be determined by 
meeting the inclusion and exclusion criteria. Subjects’ safety will be monitored during the study.  
Adverse events will be monitored and recorded throughout the study.  
Prior to the first cytisine dosing only, the Principal Investigator or Research Physician will assess 
the clinical significance of results of laboratory investigations for haematology and serum 
chemistry values outside the defined normal ranges, as provided by the laboratory. All changes 
from screening that are observed on Day 2 during the study and meet clinically-significant 
definitions for laboratory adverse events will be recorded. 
In addition to the planned times, any safety procedures can be performed at any time considered 
necessary by the Principal Investigator or attending Research Physician.  
On each cytisine dosing day, at least one physician (Principal Investigator or Research 
Physician) will physically remain at the clinical site for investigational product administration 
and until 4 hours post-dose, and will remain available on call at all times during the entire period 
of the study.  
In case any clinically significant abnormality is observed at pre-dose, Principal Investigator or 
attending Research Physician will decide whether the subject will proceed to investigational 
product administration or will be withdrawn from the study. 
Safety will be assessed by consideration of all adverse events reported by or elicited from the 
subject and abnormalities detected on haematology, serum chemistry tests, urinalysis, and 
12-lead ECG. Worsening of other preexisting medical conditions and any changes to 
concomitant medications/treatments will also be taken into account in this evaluation.  
All adverse events (serious and non-serious) beginning on Day 1 (prior to dosing) through post-
study follow-up telephone call will be recorded in the subject’s CRF.  


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13.2. 
Adverse Events 
13.2.1. 
Definition of Adverse Event 
The definitions are adopted in accordance with the Directive 2001/20/EC, ICH-E2A and the 
European Commission Detailed Guidance 2011/C 172/01. 
An Adverse Event (AE) is defined as any untoward medical occurrence in a clinical trial subject 
administered a medicinal product and which does not necessarily have a causal relationship with 
the treatment. 
An AE can therefore be any unfavorable and unintended sign (including any clinical significant 
abnormal laboratory finding), symptom, or disease temporally associated with the use of an 
investigational medicinal product, whether or not considered related to the investigational 
medicinal product. 
An Adverse Drug Reaction (ADR) means all untoward and unintended responses to a 
medicinal product related to any dose administered.  The phrase ‘response to a medicinal 
product’ means that a causal relationship has at least a reasonable possibility, i.e. the relationship 
cannot be ruled out and is judged by the Investigator as at least possible (see definition below). 
An Unexpected Adverse Drug Reaction (UADR)/Unexpected Adverse Event (UAE) means 
an adverse reaction/event, the nature or severity of which is not consistent with the applicable 
product information, namely in the Investigator Brochure for an unauthorized investigational 
product or in the SmPC for an authorized product. 
The expected/unexpected status should be evaluated and assessed, by the Sponsor, based on the 
reference safety information available since expectedness in Pharmacovigilance refers strictly to 
the information listed or mentioned in the applicable reference safety information and not to 
events that might be anticipated from knowledge of the pharmacological properties of a 
substance or because it was foreseeable due to the health status (e.g., age, medical history) of the 
study subjects. 
A Serious Adverse Event (SAE) or Serious Adverse Reaction (SAR) is defined as an AE that 
results in any of the following: 
• Results in death  
• Is life-threatening  
• Requires hospitalization or prolongs existing inpatient’s hospitalization 
• Results in persistent or significant disability or incapacity 
• Results in a congenital abnormality or birth defect 
• Is an important medical event which requires medical intervention to prevent any of 
the above outcomes 
SUSARs: AEs which meet all of the following criteria: 
• Serious. 


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• Unexpected (i.e., is not consistent with the applicable product information e.g. 
Investigator's brochure for an unapproved cytisine or SmPC for an authorised 
product). 
• There is at least a reasonable possibility that there is a causal relationship between the 
event and the medicinal product.  
will be classified as suspected unexpected serious adverse reactions (SUSARs). 
Important medical events are those which may not be immediately life-threatening, but may 
jeopardize the subject and may require intervention to prevent one of the other serious outcomes 
listed above. Examples of such events are intensive treatment in an emergency room or at home 
for allergic bronchospasm, or blood dyscrasias or convulsions that do not result in 
hospitalization. 
The term “life-threatening” in the definition of “serious” refers to an event in which the subject 
was at risk of death at the time of the event; it does not refer to an event which hypothetically 
might have caused death if it were more severe. For example, drug-induced hepatitis that 
resolves without evidence of hepatic failure would not be considered life threatening even 
though drug-induced hepatitis can be fatal. 
Inpatient hospitalization or prolongation of existing hospitalization means that hospital inpatient 
admission and/or prolongation of hospital stay were required for treatment of AE or occurred as 
a consequence of the event. It does not refer to pre-planned elective hospital admission for 
treatment of a pre-existing condition that has not significantly worsened, or to diagnostic 
procedures. 
13.2.2. 
Recording of Adverse Events 
All of the following details will be recorded in the subject’s CRF for each AE: 
• Full description of AE. 
• Date and time of onset. 
• Date and time of resolution. 
• Severity of event, to be assessed by an Investigator in accordance with the definitions 
below. 
• Relationship to cytisine to be assessed by an Investigator in accordance with the 
definitions below. 
• Action taken (if any). 
• Outcome and details of any further follow-up. 
Adverse events documented in the CRF without a stop date should be reviewed at the end of 
each treatment period until final resolution or until it is medically justifiable to stop further 
follow up (e.g. a chronic condition has been reached.)  Documentation of adverse events should 
be updated as necessary. 


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13.2.3. 
Grades of Adverse Event Severity 
The following grades will be used by an Investigator to describe the severity of all AEs 
(including clinically-significant laboratory AEs) as shown in Table 8.  Only 1 severity grade will 
be used for each AE (e.g. mild - moderate is not acceptable). 
Table 8: 
Adverse Event Severity 
Severity of AE 
Definition 
Mild 
No interference with activity 
Moderate 
Some interference with activity requiring no or minimal  
medical intervention 
Severe 
Prevents daily activity and requires medical intervention 
If an adverse event has multiple aspects, the aspect in the highest intensity will be graded. It is 
emphasized that the term severe is a measure of intensity; thus a severe AE is not necessarily 
serious. For example, itching for several days may be rated as severe; however, may not be 
clinically serious.  
13.2.4. 
Assessment of Causal Relationship 
The causal relationship between an adverse event and cytisine will be determined and 
documented by the responsible Investigator, or designee, according to best medical judgment as 
shown in (Table 9). 
Table 9: 
Assessment of Causal Relationship 
Category 
Description 
Not Related 
The event is definitely not associated with cytisine.  
Unlikely 
The event was most probably produced by other factors such as the subject’s clinical 
state, therapeutic intervention or concomitant therapy, and does not follow a known 
response pattern to cytisine 
Possible 
The event follows a reasonable temporal sequence from the time of cytisine 
administration, and/or follows a known response pattern to the investigational 
product, but could have been produced by other factors such as the subject’s clinical 
state, therapeutic intervention or concomitant therapy. 
Probable 
The event follows a reasonable temporal sequence from the time of cytisine 
administration, and/or follows a known response pattern to the product, and could not 
have been produced by other factors such as the subject’s clinical state, therapeutic 
intervention or concomitant therapy. 
Definite 
The event follows a reasonable temporal sequence from the time of cytisine 
administration, and/or follows a known response pattern to the product, and could not 
have been produced by other factors such as the subject’s clinical state, therapeutic 
intervention or concomitant therapy, and either occurs immediately following 
investigational product administration, or improves on stopping the investigational 
product. 
13.2.5. 
Reporting of Serious Adverse Events 
Any new SAE that occurs during a treatment period should be recorded and reported 
immediately. All SAEs including those that are ongoing at the end of study treatment will be 
followed until each event resolves or is assessed as chronic.  


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In order to satisfy regulatory requirements, any Serious Adverse Event, whether deemed study 
drug-related or not, must be reported to the Sponsor or designee as soon as possible after the 
Investigator or coordinator has become aware of its occurrence.  SAE form completion and 
reporting must not be delayed even if all of the information is not available at the time of the 
initial contact. 
SAEs must be reported within 24 h of knowledge of the event by submitting an initial SAE 
report via email or fax to 
: 
Email: 
  
 
 will notify the Sponsor and Sponsor's Responsible Physician of the 
SAE via email within 24 h of receipt of the initial SAE report. 
Additional information (follow-up) about any SAE unavailable at the initial reporting should be 
forwarded by the site within 24 hours of the information becoming available.  
The following information should be provided to accurately and completely record the event: 
• Investigator name and center number 
• Subject number 
• Subject initials, if permitted 
• Subject demographics 
• Clinical event 
− description 
− date of onset 
− severity 
− treatment 
− relationship to study drug (causality) 
− action taken regarding study drug 
• If the AE resulted in death: 
− cause of death (whether or not the death was related to study drug) 
− autopsy findings (if available) 
• Medical history case report form (copy) 
• Concomitant medication case report form (copy) 
• Any relevant reports (laboratory, discharge, x-ray, etc.)  
Subjects who have had an SAE during the treatment period must be followed clinically until all 
parameters (including laboratory) have either resolved or been assessed as chronic.  


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SUSARs should be reported to the REC, MHRA and Sponsor in accordance with applicable 
regulatory requirements for expedited reporting.  It is 
 responsibility to report SUSARs 
to the REC and MHRA.  It is the Sponsor’s responsibility to report any SUSAR to the FDA. 
13.2.6. 
Monitoring of Subjects with Adverse Events 
In the event of any abnormalities considered to be clinically significant by the investigating 
physician, subjects will be followed up with appropriate medical management until: 
• It has resolved/returned to normal or baseline. 
• The event has stabilised at a level acceptable to the Investigator and is not considered 
to be clinically significant. 
13.3. 
Pregnancy 
Serum pregnancy test will be performed for all females regardless of post-menopausal or 
sterilised status at screening.  Pregnancy tests will be performed by 
  
Serum pregnancy tests will be performed using the Roche Cobas c6000 analyser series 
comprising of c501 and e601 modules.     
13.4. 
Laboratory 
13.4.1. 
Routine Laboratory Assessments 
Routine laboratory safety  samples will be analysed by
 at screening 
and again on Day 5 for each subject. Printed laboratory reports will include normal reference 
ranges.  A decision regarding whether the result outside the reference range is of clinical 
significance or not shall be made by an Investigator and the report will be annotated accordingly.  
Clinically significant abnormalities at screening or occurring during the study will be recorded 
on the AE page. The reference ranges for laboratory parameters will also be filed in the 
Investigator site file. 
Haematology: Haemoglobin (HGB), haematocrit (HCT), mean cell volume (MCV), mean cell 
haemoglobin (MCH), mean cell haemoglobin concentration (MCHC), red blood cells (RBC), 
white blood cells (WBC), neutrophils (NEUT), lymphocytes (LYMP), monocytes (MONO), 
eosinophils (EOS), basophils (BASO) and platelets (PLT).  
Biochemistry: Total protein (TP), albumin (ALB), total bilirubin (BIL-T), alanine transaminase 
(ALT), aspartate transaminase (AST), gamma glutamyl transferase (GGT), alkaline phosphatase 
(ALKP), glucose (GLU), sodium (NA), potassium (K), calcium (CA), bicarbonate (HCO3), 
creatinine (CREA) and urea. 
At screening, biochemistry samples will be collected into a z-gel collection tube.  The volume 
will allow sufficient serum for virology and pregnancy test screening.  At all other visits, 
biochemistry safety samples will be collected into a 4.5 mL plain Sarstedt Monovette® collection 
tube.  Biochemistry samples will be analysed using the Roche Cobas c6000 analyser series 
comprising of c501 and e601 modules. 
Urinalysis: pH, specific gravity, protein, blood, glucose, leucocytes, ketones, urobilinogen, 
bilirubin & nitrites.  In the event that the ‘dipstick’ is positive, RBCs, WBCs, epithelial cells, 


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crystals, bacteria and casts will be examined microscopically.  The microscopy will be 
performed at 
 or referred to a local specialist hospital laboratory for analysis. 
A mid-stream urine sample will be collected into a 20 mL Sterilin tube. Urinalysis will be 
performed using the Roche Cobas c6500 analyser series comprising of u601 and u701. 
13.4.2. 
Virology 
Virology (Hep B, Hep C and HIV) will be performed at screening.  
Virology will be analysed by 
 using the Roche Cobas c6000 
analyser series comprising of c501 and e601 modules. 
13.4.3. 
Drugs of Abuse and Alcohol 
Urine alcohol and drugs of abuse screen: Cannabinoids, amphetamines, cocaine, 
benzodiazepines, opiates and cotinine. 
A mid-stream urine sample will be collected into a 20 mL Sterilin tube.  At time-points when 
both urinalysis and drugs of abuse / alcohol screening is required, all testing will be performed 
from a single 20 mL sample. 
Drug of abuse and alcohol samples will be analysed by 
 using the 
Roche Cobas c6000 analyser series comprising of c501 and e601 modules.Vital signs  
13.5. 
Vital signs 
Systolic/diastolic blood pressure and pulse rate and oral temperature.   
Measurements will be recorded in the supine position after 5 min supine.  Blood pressure, pulse 
and temperature will be measured by the DINAMAP* Compact Vital Signs Monitor (Model TS) 
or equivalent.  Normal ranges for vital signs are presented in Appendix 1. 
13.6. 
Physical Examination 
A physical examination will be performed by an Investigator.  The examination will include 
general appearance, head, ears, eyes, nose, throat, neck, skin, cardiovascular system, respiratory 
system, gastrointestinal system, central nervous system, lymph nodes and musculoskeletal. An 
Investigator can examine other body systems if required, at their discretion. 
13.7. 
ECG Measurements 
Screening and discharge ECGs will be conducted using a 12-lead ECG.  Heart rate, PR interval, 
QRS width, QT interval, and QT interval corrected using Fridericia’s formula (QTcF) will be 
documented.  
12-lead ECG recordings will be made using a MAC 5500 or equivalent.  Each ECG trace should 
be labelled with the study number, subject number, subject initials and date of birth. An 
Investigator will provide an interpretation of each tracing.  Clinically significant abnormalities 
will be recorded on the AE page. Normal ranges for 12-lead ECG parameters are presented in 
Appendix 1. 


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13.8. 
Concomitant Medication 
All prior and concomitant medications taken during the study will be recorded in the subject’s 
CRF. 
14. 
APPROPRIATENESS OF MEASUREMENTS 
All measurements performed in the study are standard measurements. 
The total volume of blood to be collected from each subject during the study (approximately 
112 mL) is considered acceptable.   
Table 8: 
Summary of Blood Volume 
Procedure 
Visit 
No. Samples 
Blood Volume 
per Sample 
(mL) 
No. Treatment 
Periods 
Blood Volume 
(mL) 
Biochemistry  
Screeninga 
Study Day 5  
1 
2 
4.5 
4.5 
1 
4.5 
4.5 
Haematology  
Screening 
Study Day 5 
1 
2 
2.7 
2.7 
1 
2.7 
2.7 
PK Drug Conc. 
Measurement 
Treatment 
Period 
18 
2.7 
2 
97.2 
Total Blood Volume 111.6 
a From the screening biochemistry blood samples, the serum pregnancy test and virology screen will be conducted.  
15. 
STATISTICAL METHODS 
15.1. 
Primary Objectives and Analysis 
All data recorded during the course of the study or derived during the programming phase will be 
presented in individual data listings. 
Unless specified otherwise, continuous variables will be summarised with the following 
descriptive statistics: n (number of observations), arithmetic mean, standard deviation (SD), 
minimum value, median and maximum values. For PK summaries, the geometric mean and 
coefficient of variation (%CV) will also be used to summarise the data. Categorical data will be 
summarised with frequencies and percentages.  
Unless specified otherwise, the minimum and maximum statistics will be presented with the 
same precision as the original data.  The mean, median and geometric mean will be presented to 
one more decimal place than the original data. The SD will be presented to two more decimal 
places than the original data. Unscheduled measurements will be listed in the individual data 
listings.  With the exception of baseline, unscheduled measurements will be excluded from the 
descriptive statistical analysis.  In case an unscheduled measurement is performed immediately 
after the scheduled measurement due to an error in the previous measurement, the unscheduled 
measurement will be included in the analysis and the original erroneous measurement will be 
excluded. 
For subject characteristics and safety analyses, missing data will not be replaced; descriptive 
statistics and statistical analysis will be performed on the basis of the available data only.  


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Subjects discontinued will be included in the descriptive statistics if they have received the 
investigational product and their examinations were performed at the same scheduled time as 
other subjects.  All data recorded on discontinued subjects will be listed. 
15.2. 
Subject Populations for Analyses 
15.2.1. 
Safety Analysis Set 
All subjects who received at least one dose of cytisine will constitute the Safety Set.  
15.2.2. 
PK Analysis Set 
• Subjects will be allocated to the PK Analysis Set on a per treatment basis. The PK 
Analysis Set for a treatment will include all subjects who receive the one dose of 
cytisine, do not violate the protocol in a way that may invalidate or bias the results 
(major protocol deviation) and have sufficient plasma concentration-time profiles 
complying with the following criteria: 
• Do not have an occurrence of vomiting or diarrhoea which renders the plasma 
concentration profile unreliable (e.g. if vomiting occurs at or before 2 times median 
Tmax);  
• Do not use a concomitant medication which renders the plasma concentration profile 
unreliable;  
• Do not have a pre-dose plasma concentration that is greater than 5% of the 
corresponding Cmax; 
• Have at least one evaluable plasma concentration that is preceded by a lower 
evaluable concentration and followed by a lower evaluable concentration for the 
calculation of Cmax, Tmax and AUCs (i.e. at least 3 evaluable concentrations in total). 
In order to be included in the statistical comparisons, a subject is required to be included in the 
PK Analysis Set for both treatments. 
15.2.3. 
Urine Excretion Analysis Set 
Subjects will be allocated to the Urine Excretion Analysis Set on a per treatment basis. The 
Urine Excretion Analysis Set for a treatment will include all subjects who receive the one dose of 
cytisine, do not violate the protocol in a way that may invalidate or bias the results (major 
protocol deviation) and comply with the following criteria: 
• Do not have any missing urine excretion collections (i.e. missing urine in a collection 
period specified in Section 12.2) which renders analysis of overall excretion 
unreliable; 
• Do not have an occurrence of vomiting or diarrhoea which renders the urine excretion 
profile unreliable (e.g. if vomiting occurs at or before 2 times median Tmax derived 
from plasma data);  
• Do not use a concomitant medication which renders the urine excretion profile 
unreliable.  


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In order to be included in the statistical comparisons,  a subject is required to be included in the 
Urine Excretion Analysis Set for both treatments. 
 
15.3. 
Subjects Characteristics 
Demographic data will be summarised (age, height, weight, BMI and race) by gender and 
overall. Separate summaries of  demographics,  will be provided for both safety and PK/Urine 
excretion analysis sets if applicable. Subject disposition and analysis sets (safety, PK) will also 
be listed and summarised by frequency. 
The protocol violations will be listed per subject, describing the nature of the violation. Subjects 
failing to complete the study (as well as the date and reasons for discontinuation) will be 
displayed. Time may be added if relevant to the nature of the violation. 
Medical history will be referred in accordance with the Medical Dictionary for Regulatory 
Activities (MedDRA), version 20.0 or higher. Medications will be mentioned according to the 
Anatomical Therapeutic Chemical (ATC) classification system using WHO Drug Dictionary. 
15.4. 
Sample Size Calculation 
A total of 12 subjects will be randomized to the study, with approximately 6 subjects per 
treatment schedule arm.  
Subject numbers are based on the following guidance: In study ACH-CYT-01, geometic mean 
AUC0-∞ values in the fed and fasted states were 168 ng.h/mL and 173 ng.h/mL respectively 
(yielding a geometric mean ratio of 0.97) with an intra subject CV% of approximately 6%. 
Geometric mean Cmax values in the fed and fasted states were 22.3 ng/mL and 29.8 ng/mL 
respectively (yielding a geometric mean ratio of 0.75) with an intra subject CV% of 
approximately 23%. Although this study is not a formal comparison of bioequivalence the 
assumptions used for a typical bioequivalence study have been used in order to estimate an 
appropriate sample size. Based on the assumption that similar results will be observed for this 
study, a sample size of 4 subjects would be sufficient to detect a 20% difference in AUC0-∞ 
between the treatments with a power of 80%, an alpha of 5% and a fed/fasted ratio of 1. 
Therefore, 12 subjects, which is the minimum number of subjects recommended for this type of 
trial, should be sufficient to compare the bioavailability with respect to AUC in the fed and 
fasted states. Given that the Cmax geometric mean ratio observed in the previous study did not fall 
within the bioequivalence range, likely due to the slower rate of absorption in the fed state, the 
sample size has been based on AUC0-∞.   
Subjects who withdraw from the study before receiving any study medication will be replaced.  
Subjects who are withdrawn from the study due to significant drug-related AEs will not be 
replaced.  Replacement of all other subjects withdrawn from the study after receiving study 
medication will be decided on a -case-by-case basis by the Principle Investigator (or delegate) 
and Sponsor.   


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15.5. 
Plasma and Urine Concentrations 
Descriptive statistics to summarize plasma and urine concentrations at each time point will be 
presented.   
Individual drug plasma concentrations versus time profiles will be plotted per subject on both a 
linear and a semi-logarithmic scale. For plotting of individual data in linear scale, concentrations 
below the LLOQ will be replaced by zero. For plotting of individual data in semi-logarithmic 
scale, concentrations below the LLOQ will be set to missing.  Graphical presentation of 
individual data will be based on actual blood sampling times. 
Arithmetic mean drug plasma concentration versus time profiles will be presented on both a 
linear and a semi-logarithmic scale.   
The post-dose blood samples will be collected within ±2 minutes from the scheduled sampling 
time. A deviation greater than ±2 minutes will be reported as a protocol deviation.   
15.6. 
Estimation of pharmacokinetic parameters 
Plasma: The following pharmacokinetic parameters for cytisine will be derived by standard non-
compartmental methods from the concentration versus time profiles: 
Table 10: 
Cytisine Pharmacokinetic Parameters 
Parameter 
Description 
Cmax 
Maximum observed plasma concentration post dose, directly obtained from the observed 
concentration versus time profile. 
Tmax 
Time of occurrence of maximum observed plasma concentration. 
AUC0-t 
Area under the plasma concentration versus time curve (AUC) from time zero to the last 
sampling time at which concentrations were at or above the LLOQ, calculated by the 
linear-up/log-down trapezoidal rule. 
AUC0-∞ 
Total AUC from time zero to infinity, calculated as AUC0-∞ = AUC0-t + (Clast/λz), where Clast 
is the last measurable plasma concentration and λz the apparent terminal elimination rate 
constant. 
%AUC 
Residual area or percentage of extrapolated part for the calculation of AUC0-∞, calculated as 
100*[1-(AUC0-t/AUC0-∞)] 
λz 
Apparent terminal elimination rate constant. 
T1/2 
Apparent terminal elimination half-life, calculated from ln 2/λz. 
No values of λz, AUC0-∞, %AUC and T1/2 will be reported for cases where λz cannot be reliably 
determined. 
Pharmacokinetic parameters will be estimated from the plasma concentration versus time 
profiles for all subjects in the safety analysis set.  Actual sampling times will be used for the 
pharmacokinetic analysis. All calculations will be performed using raw data.  
The terminal elimination rate constant (λz) will be determined by plotting the concentration data 
versus time on a semi-logarithmic scale.  The parameter will be estimated by linear least square 
regression analysis, using the last three (or more) non-zero concentrations.  The upper and the 
lower timepoints, as well as the number of timepoints, used for λz estimation will be reported.  
AUC0-t will be calculated by the linear-up/log-down trapezoidal method. 


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Individual pharmacokinetic parameters and descriptive statistics for the pharmacokinetic 
parameters obtained for each treatment will be presented.  If no reliable pharmacokinetic 
parameter could be determined for more than 1/3 of the subjects, only the minimum and 
maximum values will be presented for that parameter and all other descriptive statistics will be 
omitted.  Missing pharmacokinetic parameter data will not be imputed.  
Urine: The urine PK parameters amount excreted in urine over time (Ae) and percent of drug 
excreted in urine (Ae%) will be derived from the urine concentration-time data for all subjects in 
the safety analysis set. Derived urine PK parameters will be listed and summarised by treatment.   
15.7. 
Pharmacokinetic Analysis 
Following logarithmic transformation, Cmax, AUC0-t, AUC0-∞ and Total Ae values will be 
subjected to an analysis of variance (ANOVA) including fixed effects for sequence, period, 
treatment and subject nested within sequence. Point estimates and 90% confidence intervals (CI) 
will be constructed for the contrasts between fed and fasted states using the residual mean square 
error obtained from the ANOVA.  The point and interval estimates will be back-transformed to 
give estimates of the ratios of the geometric least squares means (LSmean) and corresponding 
90% CI. In addition, estimated geometric means will be presented. 
A comparison of Tmax will be performed using the Wilcoxon Signed-Rank test. In addition, the 
Hodges-Lehmann estimate of the median difference in Tmax and 95% CI will be presented. 
15.8. 
Safety Variables and Analyses  
All safety data will be listed using the safety analysis set. 
15.8.1. 
ECGs 
12-lead ECG parameters will be listed with any out of normal range values flagged (“H” -high or 
“L” -low) as described in the Statistical Analysis Plan (SAP). Abnormalities in ECGs will be 
classified in terms of clinical significance. Clinically significant abnormalities will be reported as 
AEs. 
15.8.2. 
Vital Signs 
Vital signs parameters will be listed with any out of normal range values flagged (“H” -high or 
“L” -low) as described in the Statistical Analysis Plan (SAP).   
Abnormalities in vital signs will be classified in terms of clinical significance. Clinically 
significant abnormalities will be reported as AEs. 
15.8.3. 
Adverse Events 
AEs will be tabulated and summarised according to the MedDRA version 20.0 or higher, and 
classified by system organ class (SOC) and preferred term (PT). The following information 
recorded or computed is used for the description of the adverse events: reported Term; SOC and 
PT by MedDRA coding; onset date, onset time, offset date and offset time; seriousness; severity 
(intensity); relationship (causality); action taken; outcome. 


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Treatment-emergent adverse events (TEAEs) are defined as AEs not present prior to first 
administration of investigational product, or AEs present before first administration of 
investigational product that worsen after the subject receives the first dose of investigational 
product. TEAEs that occur after administration of investigational product in a given period are 
assigned to the treatment administered in that period. A separate listing of SAEs will be 
presented, if applicable. 
Frequencies of TEAEs will be presented by SOC and PT, severity and relationship, by 
Fed/Fasting regimens and overall. 
15.8.4. 
Safety Laboratory tests 
Biochemistry, haematology and urinalysis parameters will be listed with any out of normal/alert 
range values flagged.  Laboratory test results which are out of normal range will also be 
presented separately along with normal reference ranges. 
Abnormalities in clinical laboratory tests will be classified in terms of clinical significance. 
Clinically significant abnormalities will be reported as AEs. 
15.9. 
Procedures for Deviations from Planned Analyses 
The planned statistical analyses for this study will be described in detail in a SAP. Modifications 
or additions to the analyses described above will be described in the SAP. Any decisions to 
deviate from the planned analyses described in the protocol and SAP will be documented in the 
clinical study report. The clinical study report will also provide a detailed explanation for any 
deviations from the planned analyses. 
16. 
DOCUMENTATION  
16.1. 
Study File and Site Documents 
Prior to the initiation of the study, the following items must be received by the Sponsor from the 
site: 
• Confidential Disclosure Agreement  
• Signed protocol, and amendment(s) page(s)  
• The Principal Investigator’s curriculum vitae and where required current medical 
license 
• Signed Clinical Study Agreement 
• Signed Financial Disclosure Form from the relevant site personnel 
• Signed Data Protection Form (for EU member states only) 
• Competent/Regulatory Authority written approval (if applicable) 
• EC written approval for the protocol, amendment(s), Informed Consent Form, Subject 
Information Sheet (if applicable), advertisements (if applicable) 


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• EC Membership list or an official statement from the EC/IRB stating the EC/IRB is in 
compliance with the EU Directive on Good Clinical Practice (GCP) 
• Local laboratory certification and normal ranges 
16.2. 
Study Documents Supplied by the Sponsor 
The Sponsor will supply the Investigator with the following items: 
• Current version of the Investigator’s Brochure  
• Current version of study protocol 
16.3. 
Maintenance and Retention of Records 
It is the responsibility of the Investigator to maintain a comprehensive and centralized filing 
system of all relevant documentation. 
• Investigators will be instructed to retain all study records required by the Sponsor and 
regulatory authorities in a secure and safe facility with limited access for one of the 
following time periods based on notification from the Sponsor: 
− For a period of at least 2 years from the last marketing approval worldwide or for 
at least 15 years, whichever is the greater 
− Or a period of at least two years after discontinuation of clinical development of 
the investigational product as confirmed by the Sponsor 
− For a longer period if required by local regulations 
The Investigator will be instructed to consult with the Sponsor before disposal of any study 
records and to provide written notification to the Sponsor of any change in the location, 
disposition, or custody of the study files. 
17. 
ADMINISTRATIVE PROCEDURES 
17.1. 
Sponsor Responsibilities 
The study will be monitored by representatives of the Sponsor and/or designated contract 
research organizations (CROs). Routine monitoring visits will be conducted to: 
a. Assure compliance with the study protocol. 
b. Verify that the research facilities, including laboratories and equipment, are adequate 
to safely and properly conduct the study. 
c. Verify that the investigational product is stored properly and under the proper 
conditions, is in sufficient supply, and that receipt, use, and return of investigational 
product at the study sites are controlled and documented adequately. 
d. Verify that written informed consent was obtained before any protocol-specific 
screening procedures are performed solely for the purpose of determining eligibility 
for the clinical study and/or prior to the provision of study medication. 
e. Review the subject CRFs and source documents to ensure that reported study data are 
accurate, complete, and verifiable from source documents. 


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f. Ensure that adequate records of clinical trial supplies are maintained. 
g. Verify that the Investigator and study site personnel are adequately qualified 
throughout the study. 
h. Verify that the safety information and amendments are submitted to the 
IRBs/ECs/REBs. 
17.2. 
Investigator Responsibilities 
All requested study data must be entered on the CRFs for the study. An explanation should be 
provided for all missing data. Correction of data on a CRF will be made with identification of the 
individual making the correction and date of the correction. Only individuals who are identified 
on the Delegation of Responsibility Form(s) may correct data on the CRF. For those subjects 
who withdraw before completion of their specified treatment regimen, all available efficacy and 
safety data must be entered in the CRF. The reason for withdrawal must be specified. Incomplete 
or inconsistent data on the CRFs will result in data queries that will be returned to the 
Investigator for resolution. 
The Investigator must maintain adequate and accurate source documents upon which CRFs for 
each subject are based. The source documents are to be separate and distinct from the CRFs, 
except for cases in which the Sponsor has predetermined that direct entry into specified pages of 
the subject’s CRF is appropriate. The documents to be maintained must include, but are not 
limited to, detailed notes on: 
a. The medical history prior to participation in the study 
b. The basic identifying information, such as demographics, that link the subject’s 
source documents with the CRFs 
c. The results of all diagnostic tests performed, diagnoses made, therapy provided, and 
any other data on the condition of the subject 
d. The subject’s exposure to study treatment 
e. All AEs 
f. The subject’s exposure to any concomitant therapy, including dates of administration 
g. All relevant observations and data on the condition of the subject throughout the 
study 
h. The oral and written communication with the subject regarding the study treatment, 
including the risks and benefits of the study. The date of informed consent must be 
recorded in the source documentation. 
18. 
REGULATORY COMPLIANCE 
Quality Assurance representatives from the Sponsor or their delegate, the MHRA, and all other 
regulatory agencies as required will be allowed to periodically visit the Investigators to discuss 
the conduct of the trial and, upon request, to inspect the records of the trial. These reviews are 
necessary to ensure that the study is conducted according to standards consistent with the ICH 
GCP Guideline. 
The Investigator agrees to discuss and correct, if necessary, any problems or deficiencies that are 
found during the course of these reviews. 


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19. 
ETHICAL CONDUCT OF THE TRIAL / GOOD CLINICAL 
PRACTICE 
This trial will be conducted in accordance with the Declaration of Helsinki, as well as the ICH 
Guidelines on GCP, the US Code of Federal Regulations, the Food and Drugs Act (Health 
Canada), and local requirements regarding IRB/EC/REB committee review, informed consent, 
and other statutes or regulations regarding the protection of the rights and welfare of human 
subjects participating in biomedical research.  The Chief Investigator shall be responsible for 
ensuring that the clinical study is performed in accordance with the following: 
• Declaration of Helsinki (Brazil, 2013). 
• Association of the British Pharmaceutical Industry (ABPI) Guidelines for Phase 1 
Trials (2012). 
• International Conference on Harmonisation (ICH) Harmonised Tripartite Guideline for 
Good Clinical Practice (GCP). 
• The Medicines for Human Use (Clinical Trials) Regulations 2004 (Statutory 
Instrument 2004 No. 1031) and subsequent amendments. 
• Applicable local standard operating procedures (SOPs). 
As this clinical study will be conducted in the United Kingdom, it has been registered in the 
EudraCT database and a Clinical Trials Authorisation (CTA) will be obtained from the 
Medicines and Healthcare products Regulatory Agency (MHRA) prior to the start of the study in 
accordance with Part 3, Regulation 12 of the United Kingdom (UK) Statutory Instrument.  In 
addition, this study protocol will be included in an Investigational New Drug (IND) application 
with the US FDA for cytisine development. 
20. 
PROTOCOL MODIFICATION/PREMATURE TERMINATION 
All protocol amendments must be written and approved by the Sponsor. Each IRB, EC, or REB 
will review and approve amendments prior to their implementation in the study. IRB, EC, or 
REB approval need not be obtained prior to removal of an immediate hazard to subjects. 
The Sponsor may terminate the protocol early if safety or other issues occur. 
The study may be suspended or terminated by either the Principal Investigator or the Sponsor 
after mutual consultation at any time for scientific and safety reasons.  Furthermore, the study 
may also be terminated prematurely by the Sponsor for important corporate reasons, or due to 
instruction of the Regulatory Authorities due to safety reasons.  
Following a decision of temporary suspension or discontinuation, it is a responsibility of the 
Principal Investigator to inform the study subjects, Ethics Committee and Regulatory Agency, 
stating the reasons for premature termination. 
 shall be responsible for expedited 
reporting and/or notification to MHRA/EMA. The Sponsor shall be responsible for expediting 
reporting and/or notification to other regulatory authorities, as applicable.  


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21. 
POLICY FOR PUBLICATION AND DATA PRESENTATION 
The Sponsor encourages the scientific publication of data from clinical research trials. However, 
Investigators may not present or publish partial or complete study results individually. The 
Principal Investigators and the Sponsor may propose appropriate scientific manuscripts or 
abstracts from the study data. Any manuscript or abstract proposed by the Investigators must be 
reviewed to ensure accuracy of data represented and commented upon in writing by the Sponsor 
prior to submission for publication. Investigators agree to consider the comments of the Sponsor 
in good faith and the Sponsor agrees in good faith not to impose limitations on access to the 
complete study data or unreasonable or inappropriate restrictions on publication of the study 
results. In case of publication, confidentiality of the study volunteers will be maintained. 


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INVESTIGATOR’S AGREEMENT  
Protocol No. ACH-CYT-07 
I have carefully read the foregoing protocol including all appendices and agree that it contains all 
the necessary information for conducting the study safely. 
I will conduct this study in strict accordance with this protocol and according to the current GCP 
guidelines and will attempt to complete the study within the time designated. 
I will provide copies of the protocol and all other information submitted by the Sponsor relating 
to pre-clinical and prior clinical experience to all personnel for whom I am responsible that 
participate in the study. I will discuss this information with them to assure that they are 
adequately informed regarding the drug and conduct of the study. 
I agree to keep records on all subject information (case report forms, shipment and drug return 
forms and all other information collected during the study) in accordance with the current GCP 
and local regulations. 
 
 
 
 
Site Principal Investigator’s name 
 
Sponsor’s Representative's name 
 
 
 
Signature 
 
Signature 
 
 
 
Date (ddMmmYYYY) 
 
Date (ddMmmYYYY) 
 
 
 
Institution 
 


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REFERENCES 
1. 
WHO. WHO Report on the Global Tobacco Epidemic, 2008 The MPOWER package. 
Geneva2008. 
2. 
Hughes JR, Keely J, Naud S. Shape of the relapse curve and long-term abstinence among 
untreated smokers. Addiction (Abingdon, England) 2004;99:29-38. 
3. 
Doll R, Peto R, Boreham J, Sutherland I. Mortality in relation to smoking: 50 years' 
observations on male British doctors. BMJ (Clinical research ed) 2004;328:1519. 
4. 
WHO. WHO Framework Convention on Tobacco Control2003. 
5. 
Aveyard P, West R. Cytisine and the failure to market and regulate for human health. 
Thorax 2013;68:989. 
6. 
Prochaska JJ, Das S, Benowitz NL. Cytisine, the world's oldest smoking cessation aid. 
BMJ (Clinical research ed) 2013;347:f5198. 
7. 
Stapleton JA. The case for licensing cytosine now for smoking cessation is 
overwhelming. BMJ (Clinical research ed) 2013;347:f5736. 
8. 
Gopalakrishnan M, Monteggia LM, Anderson DJ, et al. Stable expression, pharmacologic 
properties and regulation of the human neuronal nicotinic acetylcholine alpha 4 beta 2 receptor. 
The Journal of pharmacology and experimental therapeutics 1996;276:289-97. 
9. 
Tutka P, Zatonski W. Cytisine for the treatment of nicotine addiction: from a molecule to 
therapeutic efficacy. Pharmacological reports : PR 2006;58:777-98. 
10. 
Etter JF, Lukas RJ, Benowitz NL, West R, Dresler CM. Cytisine for smoking cessation: a 
research agenda. Drug and alcohol dependence 2008;92:3-8. 
11. 
Gotti C, Guiducci S, Tedesco V, et al. Nicotinic acetylcholine receptors in the 
mesolimbic pathway: primary role of ventral tegmental area alpha6beta2* receptors in mediating 
systemic nicotine effects on dopamine release, locomotion, and reinforcement. The Journal of 
neuroscience : the official journal of the Society for Neuroscience 2010;30:5311-25. 
12. 
Hughes JR, Stead LF, Hartmann-Boyce J, Cahill K, Lancaster T. Antidepressants for 
smoking cessation. The Cochrane database of systematic reviews 2014:Cd000031. 
13. 
Jeong SH, Newcombe D, Sheridan J, Tingle M. Pharmacokinetics of cytisine, an alpha4 
beta2 nicotinic receptor partial agonist, in healthy smokers following a single dose. Drug testing 
and analysis 2015;7:475-82. 
 
 


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APPENDIX 1. NORMAL RANGES FOR VITAL SIGNS AND ECG 
PARAMETERS 
 
 


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SAP_001.pdf: 
Achieve Life Sciences, Inc.: ACH-CYT-07 
 
Statistical Analysis Plan
 
 
 
 
Version date: 10 July 2018  
 
 
 
 
 
 
 
Page 1 of 88 
 
 
 
 
 
Statistical Analysis Plan 
 
Sponsor Protocol Number: ACH-CYT-07 
 
A Phase 1 Open Label, Randomized, Two-Way Crossover Study in 
Healthy Smokers to Investigate the Effect of Food on the 
Bioavailability of Cytisine in a New Formulation 
 
 
 
Prepared for: 
Achieve Life Sciences, Inc. 
 
Prepared by: 
 
  
 
 
 
 
CONFIDENTIAL 
 
Version  
Author 
Final 1.0 
 
 
 




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Statistical Analysis Plan
 
 
 
 
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Page 3 of 88 
 
 
 
TABLE OF CONTENTS 
1 
INTRODUCTION ................................................................................................................................... 7 
1.1 
GENERAL ...............................................................................................................................................7 
1.2 
CHANGES FROM PROTOCOL .......................................................................................................7 
1.3 
CHANGES FROM PREVIOUS VERSIONS OF THE SAP ...........................................................7 
2 STUDY OBJECTIVES ......................................................................................................................... 7 
3 STUDY DESIGN ................................................................................................................................... 8 
3.1 
OVERVIEW ............................................................................................................................................8 
3.2 
INCLUSION AND EXCLUSION CRITERIA .................................................................................8 
3.3 
STUDY TREATMENT ..........................................................................................................................9 
3.4 
SCHEDULE OF STUDY PROCEDURES .........................................................................................9 
3.5 
SAMPLE SIZE CONSIDERATIONS ............................................................................................... 10 
3.6 
RANDOMISATION .......................................................................................................................... 10 
4 STUDY VARIABLES AND COVARIATES .................................................................................... 11 
4.1 
PRIMARY VARIABLES ...................................................................................................................... 11 
4.2 
SECONDARY VARIABLES .............................................................................................................. 11 
5 DEFINITIONS AND DERIVED VARIABLES ................................................................................. 12 
6 ANALYSIS SETS ................................................................................................................................ 12 
6.1 
SAFETY SET......................................................................................................................................... 12 
6.2 
PK SET .................................................................................................................................................. 12 
6.3 
URINE EXCRETION SET ................................................................................................................ 13 
7 SAFETY MONITORING .................................................................................................................... 13 
8 INTERIM ANALYSES ........................................................................................................................ 13 
9 DATA .................................................................................................................................................... 13 
9.1 
CRF DATA .......................................................................................................................................... 13 
9.1.1 
Laboratory Data ................................................................................................................................. 13 
9.1.2 
Pharmacokinetic Data ....................................................................................................................... 14 
9.2 
RANDOMISATION LIST ................................................................................................................. 14 
9.3 
PROGRAMMING AND DATA REVIEW .................................................................................... 14 
10 STATISTICAL METHODS ................................................................................................................ 15 
10.1 
GENERAL PRINCIPLES .................................................................................................................... 15 
10.2 
STRATIFICATION AND COVARIATE ADJUSTMENT .......................................................... 16 
10.3 
MISSING DATA ................................................................................................................................. 16 
10.4 
POOLING OF SITES ......................................................................................................................... 16 
10.5 
MULTIPLE COMPARISONS ........................................................................................................... 16 


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10.6 
SUBGROUP ANALYSES .................................................................................................................. 16 
10.7 
STATISTICAL ISSUES ....................................................................................................................... 16 
11 STATISTICAL OUTPUT ................................................................................................................... 16 
11.1 
SUBJECT DISPOSITION .................................................................................................................. 16 
11.2 
SUBJECT CHARACTERISTICS AT BASELINE ........................................................................... 17 
11.2.1 Demographic and Baseline Characteristics ................................................................................. 17 
11.3 
EFFICACY ANALYSES...................................................................................................................... 17 
11.4 
PK ANALYSES .................................................................................................................................... 17 
11.4.1 Plasma Concentration Data ............................................................................................................ 17 
11.4.2 Derived Plasma PK Parameters ...................................................................................................... 18 
11.4.3 Urine Concentration Data .............................................................................................................. 20 
11.4.4 Derived Urine PK Parameters ........................................................................................................ 20 
11.5 
PD ANALYSES .................................................................................................................................... 21 
11.6 
SAFETY ANALYSES .......................................................................................................................... 21 
11.6.1 Adverse Events ................................................................................................................................... 21 
11.6.2 Laboratory Data ................................................................................................................................. 22 
11.6.3 Vital Signs ............................................................................................................................................. 23 
11.6.4 Electrocardiogram ............................................................................................................................. 23 
11.7 
OTHER DATA.................................................................................................................................... 23 
11.7.1 Prior and Concomitant Medications ............................................................................................. 23 
11.7.2 All Other Data ................................................................................................................................... 23 
12 VALIDATION..................................................................................................................................... 24 
13 LITERATURE CITATIONS/REFERENCES ................................................................................. 24 
14 LIST OF TABLES, FIGURES AND LISTINGS.............................................................................. 25 
15 SHELLS FOR TABLES, FIGURES AND LISTINGS .................................................................. 30 
16 APPENDICES .................................................................................................................................... 88 
16.1 
NORMAL RANGES ........................................................................................................................... 88 
 
 
 
 


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GLOSSARY OF ABBREVIATIONS 
 
ADaM 
Analysis Data Model 
AE 
Adverse Event 
Ae 
Amount Excreted 
ANOVA 
Analysis of Variance 
AUC 
Area Under the Plasma Concentration Curve 
AUC0-∞ 
Area Under the Plasma Concentration-time Curve calculated from the time 
of dosing to infinity 
AUC0-t 
Area Under the Plasma Concentration-time Curve calculated from the time 
of dosing to the last measurable concentration 
BLQ 
Below Limit of Quantification 
BMI 
Body Mass Index 
CI 
Confidence Interval 
Cmax 
Maximum observed plasma concentration 
CRF 
Case Report Form 
CS 
Clinically Significant 
CSR 
Clinical Study Report 
CumAe 
Cumulative Amount Excreted 
CV 
Coefficient of Variation 
DBL 
Database Lock 
DMP 
Data Management Plan 
DRM 
Data Review Meeting 
ECG 
Electrocardiogram 
GM 
Geometric Mean 
ICH 
International Conference on Harmonisation 
IMP 
Investigational Medicinal Product 
LLOQ 
Lower Limit of Quantification 
LSMean 
Least Squares Mean 
MedDRA 
Medical Dictionary for Regulatory Activities 
mg 
Milligram 
mL 
Millilitre 
 
 


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NCS 
Not Clinically Significant 
PD 
Pharmacodynamic 
PK 
Pharmacokinetics 
PT 
Preferred Term 
QC 
Quality Control 
RBC 
Red Blood Cell  
SAP 
Statistical Analysis Plan 
SD 
Standard Deviation 
SDTM 
Study Data Tabulation Model 
SOC 
System Organ Class 
TEAE 
Treatment Emergent Adverse Event 
TFL 
Table/Figure/Listing 
Tmax 
Time from dosing to the maximum observed plasma concentration 
ULOQ 
Upper Limit of Quantification 
WBC 
White Blood Cell  
 
  
 
 


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1 INTRODUCTION 
1.1 
GENERAL  
This statistical analysis plan (SAP) describes the statistical methods to be used during the reporting and 
analysis of data collected under the ACH-CYT-07 protocol dated 3rd April 2018 and should be read in 
conjunction with the study protocol and case report form (CRF).   
 
This version of the plan has been developed using the protocol version 2.0 dated 3rd April 2018 and the 
annotated CRF.  Any further changes to the protocol or CRF will be reviewed for potential impact on 
the SAP, which will be amended if it is deemed necessary. 
 
Draft versions of the SAP will undergo review by the Statistical Reviewer, Project Manager, PK Analyst, 
Medical Writer, Principal Investigator and the Sponsor/Sponsor representative. The analysis plan will be 
finalised and approved by the Sponsor prior to Database Lock (DBL). 
 
 
1.2 
CHANGES FROM PROTOCOL 
• 
Vital signs summary included. 
• 
The definitions of the PK Set and Urine Excretion Set have been amended to align with the 
ACH-CYT-01 study. Subjects will be required to receive both doses of cytisine in order to be 
included in the PK/Urine Excretion Set rather than just one dose,  
• 
The PK parameters, CL/F and Vd/F, will also be presented in line with FDA guidance.  
 
 
1.3 
CHANGES FROM PREVIOUS VERSIONS OF THE SAP 
Not applicable. 
 
2 STUDY OBJECTIVES 
This study will evaluate if there is any food effect for cytisine in a new formulation and further define 
renal elimination of cytisine over 48 hours.  
 
Primary Objective: 
The primary objectives of this study are:  
1. To compare the bioavailability (Cmax and AUC0-∞) of cytisine in smokers under fed and fasted 
conditions, following administration of 3 mg cytisine (2 x 1.5 mg cytisine tablets). 
 
2. To assess the extent of renal elimination (Ae and Ae%) of cytisine via measurement of urinary 
concentrations, following administration of 3 mg cytisine (2 x 1.5 mg cytisine tablets). 
 
Secondary Objectives: 
The secondary objectives of this study are: 
1. To compare the AUC0-t, T½, and Tmax of cytisine in subjects under fed and fasted conditions, 
following administration of 3 mg cytisine (2 x 1.5 mg cytisine tablets). 


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2. To assess the safety and tolerability of cytisine in a new formulation at the 3 mg dose level 
under fed and fasted conditions. 
 
 
3 STUDY DESIGN 
3.1 
OVERVIEW 
This is an open-label, randomized, 2-period, single-dose crossover Phase I study conducted in 12 healthy 
smokers to determine the comparative bioavailability and renal elimination of cytisine following single-
dose administration under fed and fasted conditions.  
 
 
Figure 1: Study Design Overview 
 
 
 
The study is comprised of a pre-study screen, followed by 2 treatment periods (1 and 2) and a post-
study follow-up. Screening assessments will be carried out within 28 days prior to first administration of 
cytisine. Eligible subjects will receive a single dose of cytisine under fed and fasted conditions during 2 
treatment periods. Each treatment period will be of approximately 2 days duration in which blood and 
urine samples will be collected over 48 h period post-cytisine dosing. Subjects will arrive at the Clinical 
Unit on Day 0 and will undergo an overnight fast of at least 10 h prior to first dosing on Day 1. 
Randomisation will occur on the morning of Day 1, Period 1, and cytisine will be administered under 
fasted (after an overnight fast of at least 10 h) or fed (after a high fat breakfast) conditions. Subjects will 
complete all 48 h post-dose assessments prior to starting Period 2 and the second dosing day on Day 3. 
A Post-Study telephone follow-up will be conducted 6 to 8 days after the second dose of cytisine.  
 
 
3.2 INCLUSION AND EXCLUSION CRITERIA 
To be eligible for inclusion into this study, each subject must fulfil all inclusion criteria and not violate 
any exclusion criteria (for the protocol under which they are entered) during screening prior to 
randomisation. Details of the inclusion and exclusion criteria are presented within the protocol (sections 
6.1, 6.2). 
 
R 
A 
N 
D 
O 
M 
I 
Z 
E 
Schedule B 
 
Initial dose of cytisine (3 mg) given after fasted state (Treatment Period 1) 
followed by second dose of cytisine (3 mg) given after fed state (Treatment 
Period 2). 
 
Schedule A 
 
Initial dose of cytisine (3 mg) given after fed state (Treatment Period 1) 
followed by second dose of cytisine (3 mg) given after fasted state (Treatment 
Period 2). 
 
S 
C 
R 
E 
E 
N 
 
 
F 
 
O 
 
L 
 
L 
 
O 
   W 
 
 
 
U 
 
P 


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3.3 STUDY TREATMENT 
Each subject will receive the following over 2 treatment periods in accordance with the randomisation 
schedule: 
 
• 
3 mg Cytisine (2 x 1.5 mg film coated tablets (
 under fasted conditions; 
• 
3 mg Cytisine (2 x 1.5 mg film coated tablets (
)) under fed conditions. 
 
Cytisine should be administered orally in a single dose application of two 1.5 mg tablets taken with 240 
mL water. 
 
 
3.4 SCHEDULE OF STUDY PROCEDURES 
Days 
Screening 
Baseline 
Study Day (Period 1 and Period 2) 
Post-Study 
Telephone Call 
Days 
-28 to -1 
Day 0 
1 
2 
3 
4 
5 
6-8 days after last 
dose 
Written informed consent 
X 
 
 
 
 
 
 
 
Demographic data/height/weight 
X 
 
 
 
 
 
 
 
Randomization 
 
 
X (Pre-first dose 
in Period 1) 
 
 
 
 
 
Vital signs 
X 
X 
X1 
 
X1 
 
X 
 
Medical history 
X 
 
 
 
 
 
 
 
Medical history update 
 
X2 
 
 
 
 
 
 
Prior and concomitant 
medication 
X 
X 
X 
X 
X 
X 
X 
 
Physical examination 
X 
 
 
 
 
 
 
 
12-lead ECG 
X 
 
 
 
 
 
X3 
 
Pregnancy test for all females 
X (Serum) 
X (Urine) 
 
 
 
 
 
 
FSH (post-menopausal females 
only) 
X 
 
 
 
 
 
 
 
Haematology 
X 
 
 
 
 
 
X 
 
Biochemistry 
X 
 
 
 
 
 
X 
 
Urinalysis 
X 
 
 
 
 
 
X 
 
Drugs-of-abuse tests in urine 
X 
X 
 
 
 
 
 
 
Verification of eligibility criteria  
X 
X 
X 
 
 
 
 
 
Cytisine administration 
 
 
X 
 
X 
 
 
 
Blood collection for PK analysis 
 
 
X 
X 
X 
X 
X 
 
Urine collection for PK analysis 
 
 
X 
X 
X 
X 
X 
 
Adverse events monitoring 
 
X4 
X4 
X4 
X4 
X4 
X4 
X4 
1 
Vital signs (supine blood pressure, pulse rate and oral temperature) will be recorded at pre-dose and again 
approximately 2 hours after administration on Day 1 of each period. 
2 
Clinically relevant changes will be reported as adverse events.  
3 
Repeat 12-lead ECG and assess prior to discharge on Day 5. 
4 
Adverse event recording to begin upon admissions on Day 0 of Period 1 to post study. 
 
 


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3.5 SAMPLE SIZE CONSIDERATIONS 
A total of 12 subjects will be randomized to the study, with 6 subjects per treatment schedule arm.  
Subject numbers are based on the following guidance:  In study ACH-CYT-01, geometric mean AUC0-∞ 
values in the fed and fasted states were 168 ng.h/mL and 173 ng.h/mL respectively (yielding a geometric 
mean ratio of 0.97) with an intra subject CV% of approximately 6%. Geometric mean Cmax values in the 
fed and fasted states were 22.3 ng/mL and 29.8 ng/mL respectively (yielding a geometric mean ratio of 
0.75) with an intra subject CV% of approximately 23%. Although this study is not a formal comparison 
of bioequivalence the assumptions used for a typical bioequivalence study have been used in order to 
estimate an appropriate sample size. Based on the assumption that similar results will be observed for 
this study, a sample size of 4 subjects would be sufficient to detect a 20% difference in AUC0-∞ between 
the treatments with a power of 80%, an alpha of 5% and a fed/fasted ratio of 1. Therefore, 12 subjects, 
which is the minimum number of subjects recommended for this type of trial, should be sufficient to 
compare the bioavailability with respect to AUC in the fed and fasted states. Given that the Cmax 
geometric mean ratio observed in the previous study did not fall within the bioequivalence range, likely 
due to the slower rate of absorption in the fed state, the sample size has been based on AUC0-∞.  
Subjects who withdraw from the study before receiving any study medication will be replaced. Subjects 
who are withdrawn from the study due to significant drug-related AEs will not be replaced.  Replacement 
of all other subjects withdrawn from the study after receiving study medication will be decided on a 
case-by-case basis by the Principle Investigator (or delegate) and Sponsor.   
 
3.6 RANDOMISATION  
Subjects will be allocated to a treatment schedule (sequence) in accordance with a randomisation code 
produced by
 using the PROC PLAN procedure of SAS® version 9.3. Treatment schedules are as 
follows: 
Schedule A (6 subjects): 
• 
Period 1: Cytisine (2 x 1.5 mg tablets) will be administered 30 minutes after the start of a high 
fat breakfast (fed state) with 240 mL water.   
• 
Period 2: Cytisine (2 x 1.5 mg tablets) will be administered after the overnight fast of at least 10 
hours (fasting state with 240 mL water.   
Schedule B (6 subjects): 
• 
Period 1: Cytisine (2 x 1.5 mg tablets) will be administered after the overnight fast of at least 10 
hours (fasting state with 240 mL water.   
• 
Period 2: Cytisine (2 x 1.5 mg tablets) will be administered 30 minutes after the start of a high 
fat breakfast (fed state) with 240 mL water.   
Subjects will be numbered sequentially from 001 (i.e. 001, 002 etc.). Any replacement subjects will be 
assigned the same randomisation as the subjects they are replacing with 100 added to the subject number 
(i.e. 101 would replace 001 etc.).  
 
 


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4 STUDY VARIABLES AND COVARIATES 
4.1 
PRIMARY VARIABLES 
The primary plasma cytisine pharmacokinetic endpoints for this study are as follows: 
• 
Cmax 
Maximum observed plasma concentration post dose, obtained from the 
observed concentration versus time profile.  
• 
AUC0-∞ 
Total area under the plasma concentration-time curve (AUC) from zero 
to infinity, calculated as AUC0-∞ = AUC0-t + (Clast/λz), where Clast is the 
last measurable plasma concentration and λz the apparent terminal 
elimination rate constant. 
The primary urine cytisine pharmacokinetic endpoints for this study are as follows: 
• 
Ae 
Amount excreted in urine over time.  
• 
Ae% 
Percentage of drug excreted in urine.  
 
4.2 SECONDARY VARIABLES 
The secondary plasma cytisine pharmacokinetic endpoints for this study are as follows: 
• 
Tmax 
Time of occurrence of maximum observed plasma concentration.  
• 
AUC0-t 
AUC from the time of zero to the last sampling time at which 
concentrations were at or above the lower limit of quantification (LLOQ), 
calculated by the linear-up/log-down trapezoidal rule. 
• 
%AUC 
Residual area or percentage of extrapolated part for the calculation of 
AUC0-∞, calculated as 100*[1-(AUC0-t/ AUC0-∞)].  
• 
λz 
Apparent terminal elimination rate constant.  
• 
t1/2 
Apparent terminal elimination half-life, calculated from ln 2/λz. 
• 
CL/F 
Apparent total body clearance. 
• 
Vd/F 
Apparent volume of distribution. 
 
The safety endpoints for this study are as follows: 
• 
Adverse events (AEs). 
• 
Laboratory safety data (biochemistry, haematology, urinalysis and microscopy). 
• 
Vital signs (supine systolic/diastolic blood pressure, pulse rate and oral temperature). 
• 
12-lead ECG (heart rate, PR interval, QRS width, QT interval and QT interval corrected using 
 
Fridericia’s formula (QTcF interval)). 
 
 


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5 DEFINITIONS AND DERIVED VARIABLES 
Study drug/IMP: Study drug/IMP means 3 mg cytisine (2 x 1.5 mg cytisine tablets). 
Treatment: Treatment means 3 mg cytisine in the fed or fasted state. 
Baseline: In general, baseline is defined by subject and by variable as the last non-missing value (including 
repeats) before the first dose of study drug. This is normally the pre-dose assessment on Day 1 but if 
this assessment is missing (or not planned) then the assessment at the Screening/Day 0 visit will be used 
instead if available. 
Study Day: Study day is the number of days since start of treatment where the date of first dose is 
counted as Day 1. 
Bleed Time Deviation: Actual time of blood sample – theoretical time of blood sample. 
Protocol Deviation: a deviation related to study inclusion or exclusion criteria, conduct of the trial, 
subject management or subject assessment. This refers to any change, divergence, or departure from 
the study design or procedures defined in the protocol. Deviations recorded by the Project Manager or 
detected by Data Management or by statistical programming checks will be identified and discussed at 
the Data Review Meeting (DRM) before Database Lock (DBL). All protocol deviations within the study 
database will be classified as either ‘Major’ or ‘Minor’ prior to DBL, details of which will be included 
within the Protocol Deviations listing. 
6 ANALYSIS SETS 
Membership of the analysis sets will be reviewed and agreed prior to DBL. These will be reviewed by 
the Sponsor, Study Statistician, PK analyst and Project Manager and signed off. If PK data is not available 
at the time of DBL, then subjects will be assumed to be included in the analysis set unless the PK data 
provide reason to exclude a subject, in which case this will be discussed with the Sponsor and 
documented within the Analysis Sets listing (Listing 16.2.3.1). 
 
6.1 
SAFETY SET 
All randomised subjects who receive at least one dose of cytisine will constitute the Safety Set. 
This analysis set will be used for baseline and safety summaries as well as for all study listings. 
 
6.2 PK SET 
 The PK Set will include all subjects who receive both doses of cytisine and have sufficient plasma 
concentration-time profiles complying with the following criteria: 
• 
Do not have an occurrence of vomiting or diarrhoea which renders the concentration profile 
unreliable (e.g. if vomiting occurs at or before 2 times median Tmax);  
• 
Do not use a concomitant medication which renders the concentration profile unreliable;  
• 
Do not have a pre-dose concentration that is greater than 5% of the corresponding Cmax; 


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• 
Have at least one evaluable concentration that is preceded by a lower evaluable concentration 
and followed by a lower evaluable concentration for the calculation of Cmax, Tmax and AUCs (i.e. 
at least 3 evaluable concentrations in total); 
• 
Do not violate the protocol (major protocol deviation) in a way that may invalidate or bias the 
results. 
 
6.3 URINE EXCRETION SET 
 The Urine Excretion Set will include all subjects who receive both doses of cytisine and have sufficient 
urine excretion profiles complying with the following criteria: 
• 
Do not have any missing urine excretion collections (i.e. missing urine in a collection period 
specified in section11.4.3) which renders analysis of overall excretion unreliable;  
• 
Do not have an occurrence of vomiting or diarrhoea which renders the urine excretion profile 
unreliable (e.g. if vomiting occurs at or before 2 times median Tmax derived from plasma data);  
• 
Do not use a concomitant medication which renders the concentration profile unreliable;  
• 
Do not violate the protocol (major protocol deviation) in a way that may invalidate or bias the 
results. 
 
 
7 SAFETY MONITORING 
No interim safety analyses are planned for this study. 
8 INTERIM ANALYSES 
No interim efficacy/PK analyses are planned for this study. 
9 DATA 
9.1 
CRF DATA 
 
CRF data will be provided by 
Data Management to the Statistics department as SAS datasets in 
 standard format. SDTM datasets will be derived from the raw database and ADaM from SDTM.  
Both SDTM and ADaM domains will be used for programming the outputs to be included in the Clinical 
Study Report (CSR). SDTM/ADaM programming will begin when populated 
 standard SAS 
datasets are available.  
9.1.1 Laboratory Data 
Transfers of safety laboratory data will be available from the Pathology laboratory and delivered to Data 
Management via electronic transfer and stored within the study database. Details of laboratory data are 
documented in the Data Management Plan (DMP). Populated test transfers will be received before 
programming can start. The following results will be included: 


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• 
Haematology: Haemoglobin, haematocrit, mean cell volume, mean cell haemoglobin, mean cell 
haemoglobin concentration, red blood cells, white blood cells, neutrophils, lymphocytes, 
monocytes, eosinophils, basophils and platelets. 
• 
Biochemistry: Total protein, albumin, total bilirubin, alanine transaminase, aspartate 
transaminase, gamma glutamyl transferase, alkaline phosphatase, glucose, sodium, potassium, 
calcium, bicarbonate, creatinine and urea. 
• 
Urinalysis: Glucose, specific gravity, protein, leucocytes, ketones, urobilinogen, bilirubin, 
nitrites, pH and blood. 
• 
Microscopy: In the event that the ‘dipstick’ is positive, RBCs, WBCs, epithelial cells, crystals, 
bacteria and casts will be examined microscopically. 
• 
Virology: Hep B, Hep C and HIV. 
• 
Drugs of Abuse and Alcohol: Urine alcohol and drugs of abuse screen: cannabinoids, 
amphetamines, cocaine, benzodiazepines, opiates and cotinine. 
• 
Pregnancy/post-menopausal assessment: serum/urine pregnancy test and follicle 
stimulating hormone.  
• 
Other parameters: Any further parameters that are taken that are not included as part of the 
above categories will be included in an ‘Other Laboratory Data’ listing. 
 
 
9.1.2 Pharmacokinetic Data 
Plasma and urine concentration data will be received as an Excel file from the Bioanalytical department 
via an electronic transfer and stored as a SAS dataset. This data will then be stored in the appropriate 
SDTM domain and subsequent ADaM domain which will be used to produce the file provided to the 
pharmacokinetic team in order to derive the PK parameters using Phoenix WinNonlin 6.4.  Derived PK 
parameters will be received by the Statistics department from the Project Manager/PK analyst in a 
SAS.xpt file in an agreed format and then stored as a SAS dataset and subsequently within SDTM/ADaM 
domains. 
 
9.2 RANDOMISATION LIST 
 
The randomisation list will be uploaded to a SAS dataset and incorporated into the relevant 
SDTM/ADaM domains. 
 
 
9.3 PROGRAMMING AND DATA REVIEW 
 
Programming of datasets, tables, figures and listings may be ongoing while study data management 
activities are in progress.  
Prior to DBL, a review of the clinical database (i.e. CRF data, laboratory data) in the form of Excel data 
listings will be conducted.  A Data Review Meeting (DRM) will be held to discuss the outcome of this 
review, any potential impact on the analyses, analysis sets and protocol deviations.  Once all data issues 
have been resolved and the analysis sets approved, the database will be locked.  The SDTM/ADaM 
datasets will be finalised and the final run of outputs and quality control (QC) will take place. 


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10 STATISTICAL METHODS 
10.1 GENERAL PRINCIPLES 
• 
All statistical methods will be based on the International Conference on Harmonisation (ICH) 
E9 document “Statistical Principles for Clinical Trials”. 
• 
All data collected will be presented within data listings and any unscheduled visits will be listed.  
• 
Data will be summarised by treatment and overall (data from both treatments pooled) where 
appropriate. The format of the summaries is defined in the shells at the end of this document. 
• 
In summary and analysis tables of continuous variables, standard descriptive statistics (N 
[number within population], n [number of observations included in analysis], mean, standard 
deviation [SD], median, minimum and maximum) will be presented. Least squares mean (LS 
mean) and 90/95% confidence interval (CI) will be presented in the statistical analysis outputs as 
appropriate. For PK summaries, geometric mean, and coefficient of variation (%CV) will also be 
used to summarise the data.  
• 
Unless otherwise specified, the minimum and maximum statistics will be presented in summary 
tables to the same number of decimal places as the original data.  The mean, median, LS mean, 
geometric mean and CI will be presented to one more decimal place than the original data. SD 
will be presented to two more decimal places than the original data. %CV will be presented to 
one decimal place. 
• 
In summary tables of categorical variables, the number of non-missing observations by category 
will be presented along with percentages. Unless otherwise specified, the denominator for each 
percentage will be the number of non-missing observations. All percentages will be presented 
to one decimal place. 
• 
All plots will use a linear time scale for the nominal times of the visits and will be labelled by 
time point. 
• 
Original values will be used in summary tables, unscheduled measurements will be listed only.  
However, where repeats of baseline values occur, the last assessment will be used within any 
summary tables and used to calculate change from baseline. In case an unscheduled measurement 
is performed immediately after the scheduled measurement due to an error in the original 
measurement, the unscheduled measurement will be included in the analysis and the original 
erroneous measurement will be excluded. 
• 
The date format for all output presentations will be as captured in the database. 
• 
All statistical analysis will be performed using SAS 9.3 or higher. 
• 
All hypothesis testing will be carried out at the 5% (2-sided) significance level unless stated 
otherwise.  
• 
P-values will be rounded to four decimal places.  P-values less than 0.0001 will be reported as 
<0.0001 in tables.   
• 
Generally, character values will be left aligned and numeric values will be decimally aligned. 
• 
If any of the assumptions underlying the formal statistical methods proposed are violated during 
the analysis of the final data, alternative statistical methods will be used and any changes 
documented in the statistical methods section of the clinical study report (CSR), including the 
rationale for use. 


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• 
For numeric data which includes non-numeric values (e.g. PK data reported as BLQ or 
laboratory results reported as < 10 or >100) the following principles will be applied when 
summarising the data: 
 BLQ will be replaced with a zero. 
 Results reported as <x or >x will be treated as x. 
 
10.2 STRATIFICATION AND COVARIATE ADJUSTMENT 
Not applicable. 
 
 
10.3 MISSING DATA 
Generally, no methods to impute missing data will be used.  However, for the purpose of calculating 
change from baseline, in the instance of a missing baseline result for Period 1 (generally the assessment 
at Day 1, Pre-Dose) the results obtained at the Screening/Day 0 visit will be used instead, if available. 
Missing Period 2 baseline results will not be replaced. 
 
In the instance of missing pharmacokinetic blood samples, the linear trapezoidal rule will be employed 
between the samples immediately before and after the missing sample for the AUC calculations. 
 
 
10.4 POOLING OF SITES 
Not applicable. 
 
10.5 MULTIPLE COMPARISONS 
Not applicable. 
 
10.6 SUBGROUP ANALYSES 
Not applicable. 
 
10.7 STATISTICAL ISSUES 
None. 
11 STATISTICAL OUTPUT 
General principles for layout of the statistical output are described in Section 10.1.  Layout and 
specifications are illustrated for each unique table and listing within the shells presented in Section 14.  
 
11.1 SUBJECT DISPOSITION 
The subject disposition table will summarise the following data for all randomised subjects: 


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• 
The number (%) of subjects within each analysis set; 
• 
The number (%) of subjects receiving each treatment; 
• 
The number (%) of subjects within each treatment schedule (sequence); 
• 
The number (%) of subjects who completed each study period, where completion is defined as 
having the 48 h sample collections on Day 3 (Period 1) or Day 5 (Period 2). 
• 
The number (%) of subjects who completed the study/withdrew from the study and the 
associated reasons for early study termination. A subject is deemed to have completed the study 
if they complete the Post-Study follow-up telephone contact.  
Screening and study completion/termination data will also be listed.  A listing of all subjects with protocol 
deviations will be presented including major/minor classification. A data listing presenting subject 
eligibility for each analysis set and the reason for exclusion from an analysis set will also be presented.  
 
11.2 SUBJECT CHARACTERISTICS AT BASELINE 
11.2.1 Demographic and Baseline Characteristics 
Demographic data will be listed (including informed consent information) and descriptive statistics for 
the continuous variables age, height, weight and BMI and frequencies for the categorical variable race 
will be tabulated by gender and overall.  
Demographic and baseline data will be listed and summarised using the Safety Set. Demographic data 
may also be summarised using the PK Set/Urine Excretion Set, if appropriate. 
 
11.3 EFFICACY ANALYSES 
Not applicable. 
 
11.4 PK ANALYSES 
The primary analysis sets for summaries of plasma and urine PK data will be the PK Set and Urine 
Excretion Set respectively. If the PK Set/Urine Excretion Set and the Safety Set are not equivalent, then 
all PK summaries described below will also be produced for the Safety Set.  
11.4.1 Plasma Concentration Data 
Plasma PK samples will be collected for measurement of cytisine at the following time points during each 
period: Pre-dose, 15 min, 20 min, 30 min, 45 min, 1h, 1.5h, 2h, 2.5h, 3h, 4h, 6h, 8h, 12h, 14h, 16h, 24h 
and 48h post-dose. 
 
Cytisine concentrations in plasma will be listed and summarised for each treatment using the descriptive 
statistics N, n, arithmetic mean, arithmetic standard deviation (SD), coefficient of variation (CV%), 
minimum, median and maximum.  
 
The individual subject plasma cytisine concentration profiles over time will be presented graphically using 
actual blood sampling times on a linear and semi-logarithmic scale for each subject with both treatments 


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presented on the same plot. Arithmetic mean plasma cytisine concentration profiles over time will be 
presented on linear and semi-logarithmic scales.  
 
For inclusion within the summary tables and linear scale plots, concentrations below the limit of 
quantification (BLQ) will be assigned a value of zero. Concentrations above the upper limit of 
quantification (ULOQ) will be obtained via dilution into the calibration range and are valid results. For 
inclusion within the semi-logarithmic scale plots, concentrations below the limit of quantification will be 
set to missing.  
 
Plasma concentration data listings and individual plots will be presented using the Safety Set. Plasma 
concentration data summaries and mean plots will be presented using the PK Set and also the Safety Set 
if it differs from the PK Set. All plasma concentration data included in listings and summaries will be 
presented to three significant figures. 
 
11.4.2 Derived Plasma PK Parameters 
The following derived pharmacokinetic parameters of cytisine in plasma will be determined using 
WinNonlin Phoenix 6.4 from the individual concentration versus time profiles using standard non-
compartmental methods:  
 
Pharmacokinetic 
Parameter 
Definition 
WinNonlin computation 
WinNonlin 
Parameter 
Name 
Cmax (ng/mL) 
Maximum observed plasma 
concentration post dose 
Maximum observed concentration, 
occurring at Tmax. If not unique, 
then the first maximum is used. 
Cmax 
Tmax (h) 
Time of occurrence of 
maximum observed plasma 
concentration 
Time of maximum observed 
concentration.  If the maximum 
observed concentration is not 
unique, then the first maximum is 
used. 
Tmax 
AUC0-t (h*ng/mL) 
The area under the plasma 
concentration-time curve 
(AUC) from 0 h to the last 
sampling time at which 
concentrations were at or 
above the LLOQ 
AUC measured from the 
concentration at time of dosing to 
the last measurable positive 
concentration.  The AUC is 
computed using the linear-up/log-
down trapezoidal method. 
AUClast 
AUC(0-inf) (h*ng/mL) 
The AUC from 0 h 
extrapolated to infinity 
AUC measured from the 
concentration at time of dosing 
extrapolated to infinity based on 
the last observed concentration.   
= 𝐴𝐴𝐴𝐴𝐴𝐴0−𝑡𝑡+ 𝐶𝐶𝑙𝑙𝑙𝑙𝑙𝑙𝑙𝑙𝑙𝑙𝑙𝑙𝑙𝑙
λ𝑧𝑧
 
AUCINF_obs 
AUC% extrapolated (%) 
Residual area. 
Percentage of AUC0-inf due to 
extrapolation from time of last 
measurable positive concentration 
to infinity. 
= 𝐴𝐴𝐴𝐴𝐴𝐴0−𝑖𝑖𝑖𝑖𝑖𝑖−𝐴𝐴𝑈𝑈𝑈𝑈0−𝑡𝑡
𝐴𝐴𝐴𝐴𝐴𝐴0−𝑖𝑖𝑖𝑖𝑖𝑖
∙100 
AUC_%Extrap_
obs 
𝜆𝜆z (1/h) 
Apparent terminal elimination 
rate constant. 
First order rate constant associated 
with the terminal (log-linear) 
Lambda_z 


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portion of the curve. Estimated by 
linear regression of time vs. log 
concentration; the regression 
analysis should contain data from at 
least 3 different time points in the 
terminal phase. 
t1/2 (h) 
Apparent terminal elimination 
half-life. 
= ln(2)/ λ𝑧𝑧 
HL_Lambda_z 
CL/F 
Apparent total body 
clearance.  
= 𝐷𝐷𝐷𝐷𝐷𝐷𝐷𝐷/𝐴𝐴𝐴𝐴𝐴𝐴0−𝑖𝑖𝑖𝑖𝑖𝑖 
Cl_F_obs 
Vd/F 
Apparent volume of 
distribution based on the 
terminal phase.  
=
𝐷𝐷𝐷𝐷𝐷𝐷𝐷𝐷
𝐾𝐾𝑒𝑒𝑒𝑒. 𝐴𝐴𝐴𝐴𝐴𝐴0−𝑖𝑖𝑖𝑖𝑖𝑖
 
Vz_F_obs 
 
The terminal elimination rate constant (λz) will be determined by plotting the concentration data versus 
time on a semi-logarithmic scale.  The parameter will be estimated by linear least square regression 
analysis, using the last three (or more) non-zero concentrations.  The upper and the lower time points, 
as well as the number of time points, used for λz estimation will be reported. Values of λz, AUC0-∞, 
%AUC and t½ will not be reported for cases where λz cannot be reliably determined. 
 
AUC0-t will be calculated using the linear-up/log-down trapezoidal method. 
 
For the calculation of derived pharmacokinetic parameters, concentrations below the limit of 
quantification (BLQ) will be assigned a value of zero. In case of a deviation from the theoretical time, the 
actual time of blood sample will be used in the calculation of the derived pharmacokinetic parameters. 
Should the actual time be unavailable but a blood sample was taken, the nominal time point will be 
assigned. 
 
Derived pharmacokinetic parameters will be listed and summarised for each treatment. The descriptive 
statistics presented will be N, n, arithmetic mean, arithmetic standard deviation (SD), coefficient of 
variation (CV%), minimum, median, maximum and geometric mean (with the exception of Tmax). 
 
If no reliable PK parameter can be determined for more than a third of the subjects, only n, minimum 
and maximum values will be presented for that parameter and all other descriptive statistics will be 
omitted. 
 
Plasma PK parameter data listings will be presented using the Safety Set. Plasma PK data summaries will 
be presented using the PK Set and also the Safety Set if they differ. All PK parameter data included in 
listings and summaries will be presented to three significant figures with the exception of the CV% which 
will be presented to one decimal place. 
 
11.4.2.1 Statistical Analysis of Plasma PK Parameters 
 
Following logarithmic transformation, Cmax, AUC0-t, and AUC0-∞ values will be subjected to an analysis of 
variance (ANOVA) including fixed effects for sequence (treatment schedule), period, treatment and 
subject nested within sequence.  Point estimates and 90% confidence intervals (CI) will be constructed 
for the contrasts between fed and fasted states using the residual mean square error obtained from the 
ANOVA.  The point and interval estimates will be back-transformed to give estimates of the ratios of 
the geometric least squares means (LSmean) and corresponding 90% CI.  In addition, estimated 
geometric means will be presented. 


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A comparison of Tmax will be performed using the Wilcoxon Signed-Rank test.  In addition, the Hodges-
Lehmann estimate of the median difference in Tmax and 95% CI will be presented.  
 
The statistical analysis of the plasma PK parameters will be performed using the PK Set and also the 
Safety Set if they differ. Geometric LSmeans will be presented to three significant figures and the ratios 
and 90% CIs will be presented to 2 decimal places. 
 
11.4.3 Urine Concentration Data 
Urine PK samples will be collected for measurement of cytisine at the following time intervals during 
each study period: Pre-dose, 0-2h, 2-4h, 4-8h, 8-12h, 12-24h, 24-26h, 26-28h, 28-32h, 32-36h, 36-48h 
post-dose. 
 
Cytisine concentrations in urine will be listed and summarised for each using the descriptive statistics 
N, n, arithmetic mean, arithmetic standard deviation (SD), coefficient of variation (CV%), minimum, 
median and maximum.  
 
Urine concentrations will be listed using the Safety Set and summarised using the Urine Excretion Set. 
The summary may also be presented using the Safety Set if it differs from the Urine Excretion Set. All 
urine concentration data included in listings and summaries will be presented to three significant figures. 
 
11.4.4 Derived Urine PK Parameters 
The derived urine pharmacokinetic parameters, Ae and Ae% (individual and cumulative), will be 
determined from the individual concentration versus time data using SAS version 9.3. 
 
The derived urine PK parameter Ae will be calculated as: urine volume * urine cytisine concentration. 
Ae% will be derived as: 100*Ae/Dose (where dose is expressed in ng). The cumulative parameters 
(CumAe, CumAe%) are the sum of the individual values calculated for each time interval from 0h to 48h. 
Total Ae0-48h is defined as the total amount excreted from 0 h to 48 h. 
 
For the calculation of derived urine pharmacokinetic parameters, concentrations below the limit of 
quantification will be assigned a value of zero.  
 
The derived urine parameters Ae, Ae%, CumAe and CumAe% for each time interval will be listed and 
summarised with the urine cytsisine concentration data,  
 
The derived urine pharmacokinetic parameters Total Ae0-48h and Total Ae0-48h% will be listed and 
summarised separately using the descriptive statistics N, n, arithmetic mean, arithmetic standard 
deviation (SD), coefficient of variation (CV%), minimum, median, maximum and geometric mean. 
 
Urine PK parameter data listings will be presented using the Safety Set. Urine PK data summaries will be 
presented using the Urine Excretion Set and also the Safety Set should they differ. Urine PK parameter 
data included in listings and summaries will be presented to three significant figures with the exception 
of Ae% and CumAe% which will be presented to one decimal place. 
 
 


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11.4.4.1 Statistical Analysis of Urine PK Parameters 
 
Following logarithmic transformation, Total Ae0-48h values will be subjected to an analysis of variance 
(ANOVA) including fixed effects for sequence (treatment schedule), period, treatment and subject 
nested within sequence.  Point estimates and 90% confidence intervals (CI) will be constructed for the 
contrasts between fed and fasted states using the residual mean square error obtained from the 
ANOVA.  The point and interval estimates will be back-transformed to give estimates of the ratios of 
the geometric least squares means (LSmean) and corresponding 90% CI.  In addition, estimated 
geometric means will be presented. 
 
The statistical analysis of the urine PK parameter will be performed using the Urine Excretion Set and 
also the Safety Set should they differ. Geometric LSmeans will be presented to three significant figures 
and the ratios and 90% CIs will be presented to 2 decimal places. 
 
 
11.5 PD ANALYSES 
Not applicable. 
 
11.6 SAFETY ANALYSES 
11.6.1 Adverse Events 
All AEs will be coded using the MedDRA dictionary using the version specified in the Data Management 
Plan (DMP).  
All AEs, including those which occurred prior to the first dose of study drug, will be listed.  Only 
treatment emergent adverse events (TEAEs), i.e., existing conditions that worsen or events that occur 
during the course of the study after administration of IMP, will be included within the summary tables. 
AEs occurring post-dose on Period 1 up to immediately prior to dosing on Period 2 will be assigned to 
the corresponding Period 1 treatment. Similarly, any AEs occurring post-dose on Period 2 up to Post-
Study will be assigned to the corresponding Period 2 treatment. TEAEs will be summarised by treatment 
and overall.   
An overall summary of AEs will be produced including the number of TEAEs; the number and % of 
subjects reporting at least 1 TEAE, serious TEAE (where SAE is reported as ‘Yes’), TEAE leading to 
withdrawal from study drug (Action recorded as ‘Study Drug Discontinued’); the number and % of 
subjects reporting TEAEs by severity and relationship to study drug.  A subject with multiple occurrences 
of any AE is counted only once at the maximum level of severity or the highest association to study drug. 
The number of TEAEs and the number and % of subjects reporting at least one TEAE will be tabulated 
by system organ class (SOC) and preferred term (PT).  A subject reporting multiple episodes of a 
particular AE within a treatment period will only contribute one count towards the corresponding SOC 
and preferred term. The number of TEAEs and the number and % of subjects reporting at least one 
TEAE will also be tabulated by preferred term (PT), with the PTs sorted in descending order of 
frequency.  A subject reporting multiple episodes of a particular AE within a treatment period will only 
contribute one count towards the corresponding PT. 


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In addition, the number and % of subjects reporting TEAEs will be tabulated by maximum severity and 
strongest relationship to study drug.  For the summary of TEAEs by severity, if a subject has multiple 
events occurring within the same SOC or preferred term the event with the highest severity will be 
counted.  Similarly, for TEAEs by relationship to study drug, if a subject has multiple events occurring 
within the same SOC or PT, the event with the highest association to study drug will be counted. 
Data will be listed by treatment.  For any adverse event taken prior to first administration of study drug, 
treatment will be described as ‘Prior to Treatment’.  
Where there are only partial dates/times recorded for adverse events, adverse events will be assigned 
to treatment unless it can be ruled out based on the partial information.  
The derived variables, ‘Time from Dose’ and ‘Duration’ will be presented where full date and time are 
present. If partial dates are present for any parameter required in the calculation, then the variable will 
not be populated. The following will be used to calculate the variables: 
Duration: (Date/Time of Resolution-Date/Time of Onset) + 1 minute; 
 
Time from Dose: (Date/Time of Onset-Date/Time of Start of Dose). 
 
The following will be presented in listing format within the data summaries: 
 
Serious Adverse Events – If there are none present, the listing will be produced stating: ‘No subjects 
experienced any serious adverse events’. 
 
Adverse Events which Led to Withdrawal of Study Drug – If there are none present, the listing will be 
produced stating: ‘No subjects experienced any adverse events that led to withdrawal of study drug’. 
 
Adverse event data will be summarised and listed using the Safety Set. 
11.6.2 Laboratory Data 
Routine safety laboratory tests will be carried out at Screening and Day 5.  
The safety laboratory parameters required for this study are listed in section 9.1.1. 
 
Safety laboratory data listings will be presented in two ways: 
• 
Out of range values - any values that fall outside of the normal/alert ranges (presented in listing 
format within the data summaries) 
• 
All safety laboratory data (including physician’s review (Normal, Abnormal-NCS, Abnormal-CS)) 
with any out of range values flagged (presented within the data listings).  
Microscopy, virology, urine drugs of abuse and alcohol screen, pregnancy tests and other parameters 
will also be listed. 
 
If there are no further parameters databased other than those specified in section 9.1.1 then the ‘Other 
Laboratory Data’ listings should display, ‘No other laboratory parameters to report’. 
 
Laboratory data will be listed using the Safety Set. 
 


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11.6.3 Vital Signs 
Vital signs will be taken at Screening; Day 0; Day 1 at pre-dose and 2h post-dose (Period 1); Day 3 at 
pre-dose and 2h post-dose (Period 2); Day 5. 
Vital signs parameters (supine systolic and diastolic blood pressure and pulse rate and oral temperature) 
will be listed with any out of normal range values (see Appendix 16.1) flagged (flag ‘H’ or ‘L’ appended 
to relevant result). Descriptive statistics (N, n, mean, SD, minimum, median and maximum) of absolute 
and change from pre-dose to 2 h post-dose will be tabulated by treatment. 
 
Vital signs data will be listed and summarised using the Safety Set. 
 
11.6.4 Electrocardiogram 
11.6.4.1 12-Lead ECG 
12-lead ECGs will be performed in triplicate at Screening and prior to discharge on Day 5. 
12-lead ECG parameters (heart rate, PR interval, QRS width, QT interval and QTcF interval) will be 
listed with any out of normal range values (see Appendix 16.1) flagged (flag ‘H’ or ‘L’ appended to relevant 
result). The ECG results will also be assessed as either ‘normal’ or abnormal’ with comments on 
abnormal results also presented. 
 
12-lead ECG data will be listed using the Safety Set. 
 
 
11.7 OTHER DATA 
11.7.1 Prior and Concomitant Medications 
All medications will be coded using the WHO Drug Dictionary (version as specified in the DMP) and 
listed using the ATC Level 4 class, Preferred Term and verbatim text. A medication will be regarded as 
prior if it stops prior to administration of first treatment and will be assigned a treatment phase of ‘Prior 
to treatment’. A medication which starts prior to first treatment and continues after dosing and will be 
assigned a treatment phase of ‘Prior and ongoing’. Otherwise, treatment phase will be described as the 
treatment the medication started on. Where there are only partial dates/times recorded for medications, 
medications will be assigned to all treatments that cannot be ruled out based on the partial information.  
Prior and concomitant medications will be listed using the Safety Set. 
11.7.2 All Other Data 
All data will be listed using the Safety Set, including the following: Tobacco and Alcohol Use, Visit Dates, 
Medical History, Menstrual and Obstetric History, Physical Examination, Inclusion/Exclusion Criteria 
Failures, Bag Search and Cigarette Collection, Eligibility Check, Dose Administration and Additional 
Notes. 
 
 


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Derivations within listings: 
PK Blood Sampling Time Deviations: Calculate sample time deviation as: actual time – theoretical time, 
display in minutes. 
Analysis Sets: Detail whether subject should be included in each of the analysis sets along with 
corresponding comments if not included. 
Inclusion/Exclusion Criteria: Only failures to be recorded.  If no failures display ‘All subjects passed 
inclusion/exclusion criteria’. 
Protocol Deviations: Major/minor classification to be assigned and confirmed by Sponsor. 
 
12 VALIDATION 
All tables, figures and listings will be subject to independent quality control and visual review. Unique 
tables will be independently programmed. Findings will be documented in an Output Summary file quality 
control form and actions taken will also be documented.  
13 LITERATURE CITATIONS/REFERENCES 
None. 
 


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14 LIST OF TABLES, FIGURES AND LISTINGS 
List of Tables and Figures Contained in Report Section 14 
 
14.1 
Disposition and Demographic Data 
14.1.1 Disposition Data 
Table 14.1.1.1 
Summary of Study Disposition 
Safety Set 
 
14.1.2 Demographic Data 
Table 14.1.2.1 
Summary of Demographic Data 
Safety Set 
Table 14.1.2.2 
Summary of Demographic Data - (if required) 
PK Set 
Table 14.1.2.3 
Summary of Demographic Data - (if required) 
Urine Excretion Set 
 
14.2 
Efficacy Data 
Not Applicable. 
 
14.3 
Safety Data 
14.3.1 Adverse Events 
Table 14.3.1.1 
Summary of Treatment Emergent Adverse Events 
Safety Set 
Table 14.3.1.2 
Summary of Treatment Emergent Adverse Events by System 
Organ Class and Preferred Term  
 
Safety Set 
Table 14.3.1.3 
Summary of Treatment Emergent Adverse Events by 
Preferred Term  
 
Safety Set 
Table 14.3.1.4 
Summary of Treatment Emergent Adverse Events by System 
Organ Class, Preferred Term and Severity 
Safety Set 
Table 14.3.1.5 
Summary of Treatment Emergent Adverse Events by System 
Organ Class, Preferred Term and Relationship 
Safety Set 
Table 14.3.1.6 
Serious Adverse Events 
Safety Set 
Table 14.3.1.7 
Adverse Events Leading to Withdrawal of Study Drug 
Safety Set 
 
 
14.3.2 Laboratory Safety  
Table 14.3.2.1 
Biochemistry Out of Range Data 
Safety Set 
Table 14.3.2.2 
Haematology Out of Range Data 
Safety Set 
Table 14.3.2.3 
Urinalysis Out of Range Data 
Safety Set 
 
 


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 14.3.3 Vital Signs  
Table 14.3.3.1  
Summary of Absolute and Change from Pre-Dose to 
Post-Dose Vital Signs Data  
Safety Set 
 
14.4 
Pharmacokinetics 
 
14.4.1 Concentration-Time Data   
Table 14.4.1.1 
Summary of Plasma Cytisine Concentration Data 
PK Set 
Table 14.4.1.2 
Summary of Urine Cytisine Concentration Data 
Urine Excretion Set 
Table 14.4.1.3 
Summary of Plasma Cytisine Concentration Data – (if required) 
Safety Set 
Table 14.4.1.4 
Summary of Urine Cytisine Concentration Data – (if required) 
Safety Set 
Figure 14.4.1.1 
Mean Plasma Cytisine Concentration-Time Curves on a 
Linear Scale 
PK Set 
Figure 14.4.1.2  
Mean Plasma Cytisine Concentration-Time Curves on a 
Semi-Logarithmic Scale 
PK Set 
Figure 14.4.1.3  
Mean Plasma Cytisine Concentration-Time Curves on a Linear 
Scale – (if required) 
Safety Set 
Figure 14.4.1.4 
Mean Plasma Cytisine Concentration-Time Curves on a Semi-
Logarithmic Scale – (if required) 
Safety Set 
14.4.2 Derived Pharmacokinetic Data 
Table 14.4.2.1 
Summary of Derived Plasma Cytisine Pharmacokinetic 
Parameters 
PK Set 
Table 14.4.2.2 
Summary of Derived Urine Cytisine Pharmacokinetic 
Parameters 
Urine Excretion Set 
Table 14.4.2.3 
Summary of Statistical Analysis of Cytisine Cmax and 
AUC 
PK Set 
Table 14.4.2.4 
Summary of Statistical Analysis of Cytisine Tmax 
PK Set 
Table 14.4.2.5 
Summary of Statistical Analysis of Cytisine Total 
Amount Excreted in Urine 
Urine Excretion Set 
Table 14.4.2.6 
Summary of Derived Plasma Cytisine Pharmacokinetic 
Parameters – (if required) 
Safety Set 
Table 14.4.2.7 
Summary of Derived Urine Cytisine Pharmacokinetic 
Parameters – (if required) 
Safety Set 
Table 14.4.2.8 
Summary of Statistical Analysis of Cytisine Cmax and AUC – 
(if required) 
Safety Set 
Table 14.4.2.9 
Summary of Statistical Analysis of Cytisine Tmax – (if required) Safety Set 
 
 
 


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14.5 
Pharmacodynamics 
Not applicable. 
 
14.6 
Other 
Not applicable. 
 
Subject Data: Listings Contained in Report Appendix 16.2 
 
 
 
  
  
Table 14.4.2.10 
Summary of Statistical Analysis of Cytisine Total Amount 
Excreted in Urine – (if required) 
Safety Set 
16.2.1 
Visit Dates, Dosing Information and Disposition 
Listing 16.2.1.1 
Visit Dates 
Safety Set 
Listing 16.2.1.2 
Dose Administration 
Safety Set 
Listing 16.2.1.3 
Subject Disposition 
Safety Set 
Listing 16.2.1.4  
Additional Notes 
Safety Set 
16.2.2 
Protocol Deviations  
Listing 16.2.2.1 
Protocol Deviations 
Safety Set 
16.2.3 
Analysis Sets 
Listing 16.2.3.1 
Analysis Sets 
Safety Set 
16.2.4 
Demographic Data and Other Baseline Characteristics  
Listing 16.2.4.1 
Demographic Data 
Safety Set 
Listing 16.2.4.2 
Medical History and Concurrent Conditions 
Safety Set 
Listing 16.2.4.3 
Virology Results 
Safety Set 
Listing 16.2.4.4 
Tobacco and Alcohol Use 
Safety Set 
Listing 16.2.4.5 
Drugs of Abuse Results 
Safety Set 
Listing 16.2.4.6 
Menstrual and Obstetric History 
Safety Set 
Listing 16.2.4.7 
Pregnancy Test Results 
Safety Set 
Listing 16.2.4.8 
Inclusion/Exclusion Criteria Failures 
Safety Set 
Listing16.2.4.9 
Eligibility Check 
Safety Set 
Listing 16.2.4.10 
Bag Search and Cigarette Collection 
Safety Set 


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Not applicable. 
 
 
 
 
 
 
 
 
 
16.2.5 
Drug Concentration Data and Pharmacokinetics 
Listing 16.2.5.1 
Plasma Cytisine Concentration Data 
Safety Set 
Listing 16.2.5.2 
Urine Cytisine Concentration Data 
Safety Set 
Listing 16.2.5.3 
Individual Derived Plasma and Urine Cytisine 
Pharmacokinetic Parameters 
Safety Set 
Figure 16.2.5.1 
Individual Plasma Cytisine Concentration-Time Curves 
on a Linear Scale 
Safety Set 
Figure 16.2.5.2 
Individual Plasma Cytisine Concentration-Time Curves 
on a Semi-Logarithmic Scale  
Safety Set 
16.2.6 
Efficacy  
16.2.7 
Adverse Events  
Listing 16.2.7.1 
Adverse Events  
Safety Set 
16.2.8 
Individual Laboratory Safety Measurements  
Listing 16.2.8.1 
Biochemistry Data 
Safety Set 
Listing 16.2.8.2 
Haematology Data 
Safety Set 
Listing 16.2.8.3 
Urinalysis Data 
Safety Set 
Listing 16.2.8.4 
Microscopy Data 
Safety Set 
Listing 16.2.8.5 
Other Laboratory Data 
Safety Set 
16.2.9 
Vital Signs 
Listing 16.2.9.1 
Vital Signs Data 
Safety Set 
16.2.10 
Physical Examination 
Listing 16.2.10.1 
Physical Examination Data 
Safety Set 
16.2.11 
ECG 
Listing 16.2.11.1 
12-Lead ECG Data 
Safety Set 


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Not applicable. 
 
 
16.2.12 
Prior and Concomitant Medication 
Listing 16.2.12.1 
Prior and Concomitant Medication Use 
Safety Set 
16.2.13 
Pharmacodynamics  


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15 SHELLS FOR TABLES, FIGURES AND LISTINGS 
 
  
 
  
 
 
 
 
                                                         
  
                                                    
 
                                                       
 
                                                     
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 


 
 
 
 
 
 
 
     
 
 
 
 
 
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x 
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Listing 16.2.4.10 
Bag 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
    
 
 
 
 


 
 
 
 
 
 
 
     
 
 
 
 
 
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16 APPENDICES 
16.1 NORMAL RANGES 
 
Vital Signs Normal Ranges: 
 
Parameter 
Normal Range 
Units 
Pulse Rate 
40-100 
Beats per minute (bpm) 
Systolic Blood Pressure 
90-140 
mmHg 
Diastolic Blood Pressure 
50-90 
mmHg 
Respiratory Rate 
12-18 
Breaths per minute 
Oral Temperature 
35.0-37.5 
Degrees Celsius (°C) 
Pulse Oximetry 
94-100 
% 
 
12-Lead ECG Normal Ranges: 
 
Parameter 
Normal Range 
Units 
Heart Rate 
40-100 
Beats per minute (bpm) 
PR Interval 
120-220 
mSec 
QRS Width 
70-120 
mSec 
QT Interval 
N/a 
N/a 
QTc Interval (=QTcF) (Fridericia’s) 
350-450 
mSec