Repeat-Dose Pharmacokinetic and Pharmacodynamic Evaluation of Cytisine in Healthy Smokers
Phase: PHASE1
Conditions: Smoking Cessation
Sponsor: Achieve Life Sciences

3URWRFRO$&+&<76HSWHPEHU9HUVLRQ

&\WLVLQH
&21),'(17,$/

3URWRFRO$&+&<7
5HSHDW'RVH3KDUPDFRNLQHWLFDQG3KDUPDFRG\QDPLF
(YDOXDWLRQRI&\WLVLQHLQ+HDOWK\6PRNHUV
6HSWHPEHU
9HUVLRQ



&21),'(17,$/

$FKLHYH/LIH6FLHQFHV,QF
$FKLHYH/LIH6FLHQFHV7HFKQRORJLHV,QF
1RUWK&UHHN3DUNZD\6XLWH:%URDGZD\6XLWH
%RWKHOO:DVKLQJWRQ86$9DQFRXYHU%ULWLVK&ROXPELD&DQDGD9+%
7HO)D[7HO)D[








Protocol ACH-CYT-02, 21 September 2017, Version 2,0
C),tisine
CONF'IDENTIAI
SPONSOR SIGNATURE PAGE
PROTOCOL NUMBER
PROTOCOL TITLE
EudraCT NUMBER:
PLANNED STUDY
DOSES:
PROTOCOL VERSION
PROTOCOL DATtr
ACH-CYT.O2
Repeat-Dose Pharmacokinetic and Phzumacodynamic
Evaluation of Cytisine in Healthy Smokers
2017-003257-42
Mutiple doses of 1.5 mg cytisine and 3,0 mg cytisine
Version 2.0
2l September 2017
Approvals:
Anthony Clarke, PhD
Chief Scientific Officer
Achieve Life Sciences, Inc.
Daniel Cain
Senior Director, Clinical Itesear ch
Achieve Life Sciences, Inc.
\
)I SePt eo t1
Date
Srf 9c t+
el 20 t-1
Lf Sq{- '?.c17*
Date
/1 
\
L'J r it
Regulatory Affairs Consultant
Chief Medical Officer

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0 
 
Cytisine 
 CONFIDENTIAL 
 
 
3 
SYNOPSIS  . 
NAME OF COMPANY: Achieve Life Sciences, Inc. 
NAME OF INVESTIGATIONAL MEDICINAL PRODUCT (IMP):  
Cytisine 1.5 mg film-coated tablets 
TITLE OF STUDY: Repeat-Dose Pharmacokinetic and Pharmacodynamic Evaluation of 
Cytisine in Healthy Smokers 
PRINCIPAL INVESTIGATOR: Dr An
e Koch 
STUDY CENTRE: Simbec Research Ltd (Simbec) 
     Merthyr Tydfil, CF48 4DR, UK 
CLINICAL PHASE:  1/2 
OBJECTIVES: 
Primary:  
• To evaluate the pharmacokinetic (PK) parameters during repeat dosing of 1.5 mg or 
3.0 mg cytisine when administered as the commercial 25-day schedule. 
• To evaluate the pharmacodynamic (PD) effects (e.g. reduction in smoking) with repeat 
dosing of 1.5 mg or 3.0 mg cytisine when administered as the commercial 25-day 
schedule. 
Secondary:  
• To compare the PK parameters and tolerability for repeat dosing of 1.5 and 3.0 mg 
cytisine during the 25-day schedule in healthy smokers at 18-65 and >65 (elderly) years 
of age. 
• To assess the renal elimination of cytisine via measurement of urinary concentrations of 
cytisine during treatment on Day 1 and Day 25. 
• To evaluate for effects on QT/QTc interval prolongation and cardiac safety during 
treatment on Day 1 and Day 25 in healthy smokers at 18-65 and >65 (elderly) years of 
age. 
 
METHODOLOGY:  
This is an open-label, randomised, multi-dose study to evaluate the PK profile and PD effect of 
cytisine when administered at doses of 1.5 mg and 3.0 mg following the commercialised  
25-day schedule. 
NUMBER OF SUBJECTS: A total of 36 subjects, such that 24 subjects are 18-65 years and 12 
subjects are >65 years of age, will be randomised/enrolled to complete the study. An attempt will 
be made for 50% male and 50% female enrollment. 
MAIN INCLUSION CRITERIA: 
Male or female smokers (minimum 10 cigarettes per day) aged 18 – 65+ years, with no allergy or 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0 
 
Cytisine 
 CONFIDENTIAL 
 
 
4 
sensitivity to cytisine or any of its excipients, who are deemed healthy by meeting enrollment 
criteria and, unless surgically sterile, are willing to abstain from sexual intercourse or willing to 
use an effective method of contraception from the first dose and for 3 months after the last dose 
of cytisine (refered to as the IMP). 
IMP ADMINISTRATION: 
Each subject will receive the IMP (cytisine in the form of the commercial product T
®), 
(1.5 mg or 3.0 mg dose cytisine) over a 25-day period according to the current T
ex SmPC as 
follows: 
Days 
Regimen 
Total Daily Dose 
Approximate Interval 
 
1.5 mg Dose 
3.0 mg Dose 
 
1-3 
6 times daily 
9.0 mg 
18.0 mg 
2 hours 
4-12 
5 times daily 
7.5 mg 
15.0 mg 
2.5 hours 
13-16 
4 times daily 
6.0 mg 
12.0 mg 
3 hours 
17-20 
3 times daily 
4.5 mg 
9.0 mg 
4-5 hours 
21-24 
2 times daily 
3.0 mg  
6.0 mg 
6 hours 
25 
Once daily 
1.5 mg 
3.0 mg 
- 
This schedule will require a total of 9 overnight stays (evenings of Day -1, Days 1, 2, 3 and Days 
12, 16, 20, 24, and Day 25). 
CRITERIA FOR EVALUATION: 
PK:  
PK Endpoints:   
Plasma:  
Estimated Cmax values will be determined as follows: 
• On Day 1, an estimate of Cmax will be made after the first daily (morning) dose from 
venous blood samples collected immediately post dose at 00:30, 00:45, 01:00, 01:15, 
01:30, and 02:00 hours. For the last evening dose on Day 1, Day 2, and Day 3, an 
estimate of Cmax will be made from venous blood samples collected immediately post 
dose at 00:30, 01:00, 01:30, 02:00, 02:30, 03:00, 03:30, 04:00, and 05:00 hours. 
• On Day 12, 16, 20, and 24, an estimate of Cmax will be made after the last daily (evening) 
dose from venous blood samples collected immediately post dose at  01:00,  01:30,  
02:00, 02:30, 03:00, 03:30 and 04:00 hours. 
Estimated Cmin values will be made from venous blood samples collected prior to the first 
morning dose on Day 4, Day 13, Day 17, Day 21 and Day 25. 
On Day 25 Cmax, Tmax, t1/2 and AUC will be determined after the administration of the final dose 
of cytisine from venous blood samples collected at the following timepoints: 00:30, 00:45, 01:00, 
01:15, 01:30, 01:45, 02:00, 02:15, 02:30, 02:45, 03:00, 03:15, 03:30, 03:45, 04:00, 05:00, 06:00, 
08:00, 10:00, 12:00 and 24:00 hours post-dose. 
Total volume of blood drawn for the pharmacokinetic and clinical laboratory safety assessments 
will be up to approximately 328 mL per subject over a 26-day period. 
Urine: Amount excreted in urine over 24 hours (Ae) and percent of drug excreted in urine (Ae%) 
over 24 hours will be calculated on Day 1 and Day 25 of treatment. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0 
 
Cytisine 
 CONFIDENTIAL 
 
 
5 
Safety:  
Safety Endpoints: The number of subjects with adverse events, serious adverse events, and 
adverse events leading to discontinuation will be documented. Adverse events will be assessed 
by clinical observation and spontaneous reporting by subjects. In addition, number of subjects 
with clinically significant abnormalities in vital signs (supine systolic/diastolic pressure, pulse 
and oral temperature), 12 lead ECG (heart rate, PR interval, QRS width, QT interval and QT 
interval corrected using Fridericia’s (QTcF interval) formula), laboratory parameters 
(haematology, biochemistry and urinalysis) will be evaluated. 
STATISTICAL METHODS:   
All statistical analysis will be performed using SAS® version 9.3 or higher. 
Demographic and Background Data: All demographic and background data will be listed. 
Demographic data will be summarised descriptively (age, height, weight and BMI) by dose 
group, age group and overall. Subject disposition and analysis sets (safety, PK, and PD) will also 
be listed and summarised by frequency. 
PK:  Pharmacokinetic parameters of cytisine will be derived from the plasma  
concentration-time profiles, by using a non-compartmental approach with a ln-linear terminal 
phase assumption. Actual times of sampling will be used. 
The following pharmacokinetic parameters will be estimated: maximum observed plasma 
concentration (Cmax); minimum observed plasma concentration (Cmin); time of occurrence of 
Cmax (Tmax); area under the plasma concentration versus time curve (AUC) from time zero (pre-
dose) to the last sampling time with quantifiable concentrations (AUC0-t); AUC from time zero 
to infinity (AUC0-∞); residual area or percentage of extrapolated part of AUC0-∞ (%AUC); 
apparent terminal elimination rate constant (λz); and apparent elimination terminal half-life (t1/2).  
In addition, the amount of cytisine excreted in urine over time (Ae) and percent of drug excreted 
in urine (Ae%) will be derived from Day 1 and Day 25 24-hour urine collections. 
Safety: All safety data will be listed.  
AEs will be coded using Medical Dictionary for Regulatory Activities (MedDRA) version 20.0. 
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.  
QT/QTc:  Triplicate ECGs, extracted at pre-dose and 0.5h, 1h, 2h, 3h, 4h, 6h, 8h, 12h and 24h 
after the first dose of cytisine on Day 1 and Day 25 will be obtained from the Holter recordings 
will be listed and analysed. 
 
DURATION OF STUDY: 
Approximately 9 weeks for each individual (from screening to 
post-study follow-up). 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0 
 
Cytisine 
 CONFIDENTIAL 
 
 
6 
 
TABLE OF CONTENTS 
SYNOPSIS  . ....................................................................................................................................3 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS ................................................12 
1. 
ETHICS ......................................................................................................................14 
1.1. 
Research Ethics Committee ........................................................................................14 
1.2. 
Subject Information and Consent ...............................................................................14 
1.3. 
Indemnity Arrangements ............................................................................................15 
2. 
INTRODUCTION AND BACKGROUND ...............................................................15 
2.1. 
Smoking and Health ...................................................................................................15 
2.2. 
Cytisine .......................................................................................................................15 
2.3. 
Recent Cytisine Pharmacokinetic Studies ..................................................................16 
2.3.1. 
Study CYT-BIO-02-07 in Healthy Subjects ...............................................................16 
2.3.2. 
PK Study in Healthy Smoking Subjects published by the University of 
Auckland .....................................................................................................................18 
2.4. 
Safety Summary ..........................................................................................................20 
2.5. 
Recent Nonclinical Toxicology Studies: Estimation for Higher Human 
Equivalent Doses ........................................................................................................20 
3. 
RATIONALE FOR THE STUDY ..............................................................................22 
4. 
STUDY OBJECTIVES ..............................................................................................22 
4.1. 
Primary Objectives .....................................................................................................22 
4.2. 
Secondary Objectives .................................................................................................22 
5. 
INVESTIGATIONAL PLAN .....................................................................................22 
5.1. 
Study Design ...............................................................................................................22 
6. 
SELECTION OF STUDY POPULATION ................................................................25 
6.1. 
Inclusion Criteria ........................................................................................................25 
6.1.1. 
To be Confirmed at Screening ....................................................................................25 
6.1.2. 
To be Re-Confirmed Prior to Start of Dosing (Day -1) ..............................................27 
6.2. 
Exclusion Criteria .......................................................................................................27 
6.2.1. 
To be Confirmed at Screening ....................................................................................27 
6.2.2. 
To be Re-Confirmed Prior to Start of Dosing (Day -1): .............................................27 
6.3. 
Removal of Subjects from Therapy or Assessment ....................................................28 
7. 
RETRICTIONS PRIOR AND DURING THE STUDY ............................................28 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0 
 
Cytisine 
 CONFIDENTIAL 
 
 
7 
7.1. 
Confinement ...............................................................................................................29 
7.2. 
Smoking, Diet and Fluid Restrictions .........................................................................29 
7.2.1. 
Smoking ......................................................................................................................29 
7.2.2. 
Meal Times .................................................................................................................29 
7.2.3. 
Fluid Intake .................................................................................................................29 
7.2.4. 
Alcohol Intake ............................................................................................................29 
7.2.5. 
Caffeine .......................................................................................................................29 
7.2.6. 
Poppy and Sesame Seeds ............................................................................................29 
7.2.7. 
Grapefruit Juice and Other Restrictions .....................................................................30 
7.3. 
Other Life-Style Restrictions ......................................................................................30 
7.3.1. 
Blood Donation ...........................................................................................................30 
7.3.2. 
Contraception ..............................................................................................................30 
7.3.3. 
Sperm Donation ..........................................................................................................30 
8. 
INVESTIGATIONAL MEDICINAL PRODUCT (IMP) ..........................................31 
8.1. 
Cytisine 1.5 mg Film-Coated Tablets .........................................................................31 
8.2. 
Receipt and Storage ....................................................................................................31 
8.3. 
Assembly and Certification ........................................................................................32 
8.4. 
Administration ............................................................................................................32 
8.5. 
Return/Destruction ......................................................................................................32 
8.6. 
Method of Assigning Subjects to Group A or Group B .............................................32 
8.7. 
Selection of Doses in the Study ..................................................................................32 
8.8. 
Selection and Timing of Dose for Each Subject .........................................................32 
8.9. 
Dose Modifications for 3.0 mg Dose or Elderly Subjects (>65) ................................33 
8.10. 
Blinding ......................................................................................................................34 
9. 
PRIOR AND CONCOMITANT THERAPY .............................................................34 
10. 
TREATMENT COMPLIANCE .................................................................................35 
11. 
STUDY PROCEDURES ............................................................................................35 
11.1. 
Procedure Schedule ....................................................................................................36 
11.2. 
Detailed Description of Study Visits ..........................................................................38 
11.2.1. 
Screening Visit (Day -28 to Day -2) ...........................................................................38 
11.2.2. 
Day -1 .........................................................................................................................38 
11.2.3. 
Day 1 to Morning of Day 4 .........................................................................................39 
11.2.4. 
Day 4 through Day 11 .................................................................................................40 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0 
 
Cytisine 
 CONFIDENTIAL 
 
 
8 
11.2.5. 
Day 12 to Morning of Day 13 .....................................................................................41 
11.2.6. 
Day 13 through Day 15 ...............................................................................................41 
11.2.7. 
Day 16 through Morning of Day 17 ...........................................................................42 
11.2.8. 
Day 17 through Day 19 ...............................................................................................42 
11.2.9. 
Day 20 through Morning of Day 21 ...........................................................................43 
11.2.10. 
Day 21 through Day 23 ...............................................................................................43 
11.2.11. 
Day 24 through Day 26 ...............................................................................................44 
11.2.12. 
Post Study Follow-up .................................................................................................45 
12. 
DRUG CONCENTRATION MEASUREMENTS ....................................................45 
12.1. 
Pharmacokinetic Blood Sampling ..............................................................................45 
12.2. 
Pharmacokinetic Urine Collection ..............................................................................49 
12.3. 
Determination of Plasma and Urine Drug Concentrations .........................................49 
12.4. 
Incurred Samples Reanalysis ......................................................................................49 
13. 
SAFETY ASSESSMENTS ........................................................................................50 
13.1. 
Subject’s Safety ..........................................................................................................50 
13.2. 
Adverse Events ...........................................................................................................50 
13.2.1. 
Definition of Adverse Event .......................................................................................50 
13.2.2. 
Recording of Adverse Events .....................................................................................52 
13.2.3. 
Grades of Adverse Event Severity ..............................................................................52 
13.2.4. 
Assessment of Causal Relationship ............................................................................53 
13.2.5. 
Reporting of Serious Adverse Events .........................................................................53 
13.2.6. 
Monitoring of Subjects with Adverse Events .............................................................54 
13.3. 
Pregnancy ...................................................................................................................55 
13.4. 
Laboratory ...................................................................................................................55 
13.4.1. 
Routine Laboratory Assessments ...............................................................................55 
13.4.2. 
Expired Air Carbon Monoxide (CO) ..........................................................................56 
13.4.3. 
Virology ......................................................................................................................56 
13.4.4. 
Drugs of Abuse and Alcohol ......................................................................................56 
13.5. 
Vital signs ...................................................................................................................56 
13.6. 
Physical Examination .................................................................................................56 
13.7. 
ECG Measurements ....................................................................................................56 
13.8. 
12-Lead ECG Holter ...................................................................................................57 
13.9. 
Subject Diary ..............................................................................................................57 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0 
 
Cytisine 
 CONFIDENTIAL 
 
 
9 
13.10. 
Concomitant Medication ............................................................................................57 
14. 
APPROPRIATENESS OF MEASUREMENTS ........................................................57 
15. 
STATISTICAL METHODS .......................................................................................58 
15.1. 
Primary Objectives and Analysis ................................................................................58 
15.2. 
Subject Population for Analyses .................................................................................58 
15.2.1. 
Safety Analysis Set .....................................................................................................58 
15.2.2. 
PK Analysis Set ..........................................................................................................58 
15.2.3. 
Pharmacodynamic (PD) Analysis Set .........................................................................59 
15.3. 
Subjects Characteristics ..............................................................................................59 
15.4. 
Sample Size Calculation .............................................................................................59 
15.5. 
Plasma and Urine Concentrations ...............................................................................59 
15.6. 
Estimation of pharmacokinetic parameters ................................................................60 
15.7. 
Pharmacokinetic Analysis ..........................................................................................60 
15.8. 
Pharmacodynamic Data ..............................................................................................61 
15.9. 
Safety Variables and Analyses ...................................................................................61 
15.9.1. 
Holter ECGs ................................................................................................................61 
15.9.2. 
12-Lead ECGs ............................................................................................................62 
15.9.3. 
Vital Signs ..................................................................................................................62 
15.9.4. 
Adverse Events ...........................................................................................................62 
15.9.5. 
Safety Laboratory tests ...............................................................................................63 
15.10. 
Procedures for Deviations from Planned Analyses ....................................................63 
16. 
DOCUMENTATION .................................................................................................63 
16.1. 
Study File and Site Documents...................................................................................63 
16.2. 
Study Documents Supplied by the Sponsor ................................................................64 
16.3. 
Maintenance and Retention of Records ......................................................................64 
17. 
ADMINISTRATIVE PROCEDURES .......................................................................64 
17.1. 
Sponsor Responsibilities .............................................................................................64 
17.2. 
Investigator Responsibilities .......................................................................................65 
18. 
REGULATORY COMPLIANCE ..............................................................................65 
19. 
ETHICAL CONDUCT OF THE TRIAL / GOOD CLINICAL PRACTICE .............66 
20. 
PROTOCOL MODIFICATION/PREMATURE TERMINATION ...........................66 
21. 
POLICY FOR PUBLICATION AND DATA PRESENTATION .............................67 
22. 
INVESTIGATOR’S AGREEMENT ..........................................................................68 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0 
 
Cytisine 
 CONFIDENTIAL 
 
 
10 
23. 
REFERENCES ...........................................................................................................69 
APPENDIX 1. NORMAL RANGES FOR VITAL SIGNS AND ECG PARAMETERS .........70 
APPENDIX 2. FAGERSTRÖM NICOTINE DEPENDENCE QUESTIONNAIRE .................71 
APPENDIX 3. SHORT FORM OF THE TOBACCO CRAVING QUESTIONNAIRE ...........72 
 

3URWRFRO$&+&<76HSWHPEHU9HUVLRQ

&\WLVLQH
&21),'(17,$/

/,672)7$%/(6
7DEOH 
0HDQVRI3KDUPDFRNLQHWLFV3DUDPHWHUVIRU&\WLVLQH
7DEOH 
0HDQ&\WLVLQH8ULQH&RQFHQWUDWLRQVDQG3KDUPDFRNLQHWLF3DUDPHWHUV
7DEOH 
&DOFXODWLRQRI0DUJLQRI6DIHW\EDVHGRQ5DWDQG'RJ7R[LFRORJ\6WXGLHV
7DEOH 
&\WLVLQH3URGXFW,QIRUPDWLRQ
7DEOH 
'RVH0RGLILFDWLRQIRUPJ'RVH*URXS
7DEOH 
6FKHGXOHRI6WXG\3URFHGXUHV
7DEOH 
%ORRG6DPSOLQJIRU3KDUPDFRNLQHWLF$VVHVVPHQWV
7DEOH 
$GYHUVH(YHQW6HYHULW\
7DEOH 
$VVHVVPHQWRI&DXVDO5HODWLRQVKLSWR&\WLVLQH
7DEOH 6XPPDU\RI%ORRG9ROXPHV
7DEOH &\WLVLQH3KDUPDFRNLQHWLF3DUDPHWHUV
/,672)),*85(6
)LJXUH 
0DUNHWHG'RVLQJPJWDEOHWDQG7LPLQJVFKHGXOHIRU7D
[
)LJXUH 
0HDQSODVPDFRQFHQWUDWLRQVQJP/RIF\WLVLQHRYHUKRXUV
)LJXUH 
6WXG\'HVLJQ2YHUYLHZ
)LJXUH 
'RVH7LPLQJDQG7RWDO'DLO\'RVH

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
12 
LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS 
ABBREVIATION/TERM DEFINITION 
ADR 
Adverse Drug Reaction 
AE 
Adverse Event 
Ae 
Amount excreted  
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 
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 
MHRA 
Healthcare products Regulatory Agency 
NCCIH 
National Center for Complementary and Integrative Health 
(branch of US NIH) 
PD 
Pharmacodynamic  
PK 
Pharmacokinetics 
RBC 
Red Blood Cell Count 
REC 
Research Ethics Committee 
SAE 
Serious Adverse Event 
SAR 
Serious Adverse Reaction 
SD 
Standard Deviation 
SmPC 
Summary of Product Characteristics  
SOC 
(MedDRA) System Organ Class 
SAP 
Statistical Analysis Plan  
SUSAR 
Suspected Unexpected Serious Adverse Reactions 
TEAE 
Treatment emergent Adverse Events 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
13 
ABBREVIATION/TERM DEFINITION 
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-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
14 
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 S
ec 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 may be no benefits gained by their 
participation. All potential conflicts of interest will be declared by the Principal Investigator and 
designees.  
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. 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 Principal Investigator or designee 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. The original signed 
informed consent form will be retained with the study records at the Investigator site. In addition, 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
15 
the actions and completion of the consenting process will be recorded in the subject’s medical 
record (i.e., source document). 
1.3. 
Indemnity Arrangements 
The Sponsor and Si
c 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 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 
(-)-Cytisine, a plant-based alkaloid isolated from seeds of Cytisus laburnum (Golden rain acacia), 
has been used as a smoking cessation drug since the 1960’s in Eastern and Central Europe, 
marketed as 
 Despite its widespread use, cytisine has not 
been clinically developed outside of S
a’s Eastern and Central European territories. 
Achieve Life Sciences has acquired the license to cytisine outside of Sop
a’s territories to 
pursue clinical development and market approvals. There is widespread support for this strategy 
from leaders in the field.7-9 
So
ma’s focus on clinical development of cytisine was in its traditional markets in Eastern 
and Central European countries where the initial clinical studies were conducted and published, 
but not in English.5 Because most of the clinical development trials leading to market approval in 
these countries occurred decades ago, the trials were not performed according to today’s standard 
Good Clinical Practice (GCP) guidelines. Cytisine has been more recently evaluated in newer 
clinical trials, including additional pharmacokinetic (PK) studies and two large randomised 
Phase 3 clinical trials that were conducted according to GCP with more than 2,000 participants. 
The overall objectives in these more recent trials were to confirm the PK, efficacy, and safety of 
cytisine according to current clinical development standards. This clinical trial is being 
conducted to assess the pharmacokinetic parameters in far greater detail than has been reported 
for the commercial dose schedule as well as assessing a higher dose, as recommended by FDA 
for a future Phase 3 trial.  
As a smoking cessation treatment using cytisine, the designated number of 1.5 mg cytisine 
tablets to achieve a particular dose or schedule are taken orally with approximately 240 mL of 

3URWRFRO$&+&<76HSWHPEHU9HUVLRQ
&21),'(17,$/
&\WLVLQH


ZDWHU7KHPDUNHWHGGRVLQJPJWDEOHWDQGWLPLQJVFKHGXOHIRU
DUHVKRZQLQ
)LJXUH
)LJXUH
0DUNHWHG'RVLQJPJWDEOHWDQG7LPLQJVFKHGXOHIRU


5HFHQW&\WLVLQH3KDUPDFRNLQHWLF6WXGLHV

6WXG\&<7%,2LQ+HDOWK\6XEMHFWV
7KH3KDVH&<7%,2VWXG\HQUROOHGKHDOWK\PDOHVXEMHFWVZKRUHFHLYHGDVLQJOHRUDO
GRVHRIPJF\WLVLQHXQGHUIDVWLQJFRQGLWLRQV%ORRGVDPSOLQJDQGTXDQWLWDWLYHXULQH
FROOHFWLRQIRUWKHGHWHUPLQDWLRQRIF\WLVLQHLQSODVPDDQGXULQHZHUHSHUIRUPHGDWUHJXODU
LQWHUYDOVGXULQJWKHWUHDWPHQWSHULRG3K\VLFDOH[DPLQDWLRQVZHUHSHUIRUPHGE\WKH3ULQFLSDO
,QYHVWLJDWRURUGHVLJQHHDWWKHSUHVWXG\VFUHHQLQJDIWHUDGPLVVLRQWRWKHFOLQLFDOFHQWHUDQG
GXULQJWKHHYHQLQJEHIRUHGUXJDGPLQLVWUDWLRQWRWKHSRVWVWXG\H[DPLQDWLRQMXVWSULRUWR
GLVFKDUJHIURPWKHFOLQLF9LWDOVLJQVEORRGSUHVVXUHKHDUWUDWHDQGWHPSHUDWXUHZHUHDOVR
SHUIRUPHGDIWHUDGPLVVLRQWRWKHFOLQLFDOFHQWHUGXULQJWKHHYHQLQJEHIRUHGUXJDGPLQLVWUDWLRQ
DQGZLWKLQPLQEHIRUHGUXJDGPLQLVWUDWLRQSUHGRVHDQGDQGKDIWHUGUXJ
DGPLQLVWUDWLRQGXULQJWKH7UHDWPHQW3HULRG
%ORRGVDPSOLQJIRUGHWHUPLQDWLRQRIF\WLVLQHFRQFHQWUDWLRQVZDVFDUULHGRXWLQWKHWUHDWPHQW
SHULRGZLWKLQPLQEHIRUHSUHGRVHDQGDWDQGPLQDQGFRQWLQXHGDW
DQGKDIWHUDGPLQLVWUDWLRQRIPJ
F\WLVLQHRQH
WDEOHW8ULQHZDVFROOHFWHGEHIRUHEDVHOLQHDQGTXDQWLWDWLYHO\GXULQJWKH
LQWHUYDOVDQGKDIWHUDGPLQLVWUDWLRQRIVWXG\GUXJ$WWKHHQGRIHDFK
FROOHFWLRQLQWHUYDOLQGLYLGXDOXULQHSRUWLRQVZHUHSRROHGDQGDOLTXRWVZHUHZLWKGUDZQIURPHDFK
XULQHSRRO&UHDWLQLQHFRQFHQWUDWLRQVZHUHPHDVXUHGIRUHDFKXULQHSRRO3ODVPDDQGXULQHOHYHOV
RIF\WLVLQHZHUHGHWHUPLQHGXVLQJYDOLGDWHG/&0606DQDO\WLFDOPHWKRGV3KDUPDFRNLQHWLF
SDUDPHWHUVRIF\WLVLQHLQSODVPDWKDWZHUHHVWLPDWHGLQFOXGHG$8&ODVW$8&LQI&PD[7PD[DQG
7ò
5HVXOWVVKRZHGWKDWQRPHDVXUDEOHSODVPDOHYHOVRIF\WLVLQHZHUHREVHUYHGPLQDIWHUGUXJ
DGPLQLVWUDWLRQ$IWHUPLQDOHYHOH[FHHGLQJWKHOLPLWRITXDQWLWDWLRQ/24ZDVQRWHGLQRI
VXEMHFWVDIWHUPLQDOHYHOH[FHHGLQJWKH/24ZDVQRWHGLQRI

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
17 
subjects. At 10 h after drug administration, the cytisine plasma levels exceeded LOQ in all 
subjects. Mean values and other calculated PK parameters are presented in Table 1 (extracted 
from the Study CYT-BIO-02-07 report). 
Table 1: 
Means of Pharmacokinetics Parameters for Cytisine 
 
Cmax 
ng/mL 
Clast 
ng/mL 
AUCinf 
ng.h/mL 
AUClast 
ng.h/mL 
AUCres 
ng.h/mL 
AUC % 
Ex-inf 
% 
T½ 
h 
Ka 
l/h 
λz 
1/h 
MRT 
h 
Tmax 
h 
Mean 
SD 
SEM 
Median 
15.55 
6.55 
1.09 
14.97 
2.24 
0.81 
0.13 
1.92 
73.63 
21.27 
3.55 
70.72 
59.80 
16.77 
2.79 
58.58 
13.83 
7.36 
1.23 
11.66 
18.33 
6.46 
1.08 
17.89 
4.09 
0.82 
0.14 
4.00 
2.81 
1.30 
0.38 
0.95 
0.1758 
0.0336 
0.0056 
0.1735 
6.10 
1.23 
0.21 
6.02 
0.92 
0.40 
0.07 
0.83 
Min 
Max 
5.37 
36.67 
0.88 
4.40 
38.37 
121.92 
32.49 
103.47 
4.45 
40.33 
7.20 
35.38 
2.74 
6.36 
3.01 
4.42 
0.1090 
0.2531 
4.30 
9.65 
0.33 
1.75 
CV% 
Geo Mean 
42 
14.39 
36 
2.10 
29 
70.67 
28 
57.53 
53 
12.19 
35 
15.25 
20 
4.02 
46 
2.49 
19 
0.1726 
20 
5.99 
44 
0.84 
CI 95% 
Lower 
Upper 
KS p value 
 
13.33 
17.76 
0.65 
 
1.97 
2.52 
0.22 
 
66.43 
80.83 
0.64 
 
54.13 
65.48 
0.90 
 
 
16.15 
20.52 
0.61 
 
3.81 
4.37 
0.89 
 
1.98 
3.64 
0.282 
 
0.1644 
0.1872 
0.9837 
 
5.68 
6.52 
0.55 
 
0.78 
1.06 
0.12 
Mean Cmax (maximal plasma concentration) for the 36 subjects was 15.55 ± 6.55 ng/mL with a 
mean Tmax (time to maximal plasma concentration) of 0.92 h ± 0.40, ranging from 0.33-1.75 h. 
The mean AUClast (area under the concentration-time curves over 0-t period) was 59.80 h.ng/mL 
± 16.77 and the AUCinf (area under the concentration-time curve from 0 to infinity) was 
73.63 h.ng/mL ± 21.27. The mean AUCres was less than 20% of AUCinf. The mean T½ (plasma 
elimination half-life) was 4.09 h ± 0.82 with a range from 2.74-6.36 h and MRT (mean resistance 
time) was 6.10 ± 1.23 h. 
Mean urine concentration of cytisine in urine samples are presented in Table 2 (extracted from 
the Study CYT-BIO-02-07 report). 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
18 
Table 2: 
Mean Cytisine Urine Concentrations and Pharmacokinetic Parameters.  
 
Cytisine urinary concentrations (µg) 
Aea 
% 
recovery 
0 – 3 h 
3 – 6 h 
6 – 9 h 
9 – 12 h 
12 – 24 h 
Mean 
SD 
SEM 
Median 
329.02 
102.42 
17.07 
336.98 
250.58 
98.05 
16.34 
235.98 
154.25 
73.07 
12.18 
134.31 
105.28 
43.99 
7.33 
94.76 
119.62 
56.34 
9.39 
119.93 
958.75 
238.67 
39.78 
896.22 
63.92 
15.91 
2.65 
59.75 
Min 
Max 
70.72 
519.35 
14.73 
469.20 
11.14 
357.90 
20.95 
227.97 
24.80 
250.63 
24.89 
929.79 
34.71 
100.24 
CV% 
Geo Mean 
31.13 
308.24 
39.13 
221.92 
47.37 
135.49 
41.78 
96.07 
47.10 
105.64 
24.89 
929.79 
24.89 
61.99 
CI 95% 
Lower 
Upper 
 
294.36 
363.67 
 
217.41 
283.76 
 
129.53 
178.98 
 
90.40 
120.16 
 
100.56 
138.68 
 
877.99 
1039.50 
 
58.53 
69.30 
aAe: Amount excreted 
Mean cumulated values of cytisine excreted during a 24 h period evaluated as percent of 
administered dose was 958.75 ± 238.67 μg. The results indicated that, during a 24 h period, 
around 64% of the oral dose was excreted in the urine as parent compound. 
In summary, there were marked inter-individual differences observed in plasma concentrations 
as well as in all PK parameters after cytisine administration (e.g., CV exceeding 30% was noted 
for Tmax, Cmax, and AUC). The mean half-life for cytisine was 4.09 h ± 0.82. Cumulative 
amounts of cytisine were excreted in the urine within 24 h indicating that cytisine is most likely 
readily absorbed from the gastrointestinal tract after oral administration and may not undergo 
metabolic processes, since approximately 64% of the administered dose was excreted as 
unchanged cytisine within 24 h. More details are in the Investigator’s Brochure. 
2.3.2. 
PK Study in Healthy Smoking Subjects published by the University of Auckland 
A pilot PK study was conducted at the University of Auckland in New Zealand and published by 
Soo Hee Jeong10 et al in 2014. The study developed an analytical LC-MS assay for the detection 
of cytisine in plasma and urine following a single 3 mg oral dose. The LC-MS method in this 
study complied with limits set by US FDA guidelines including accuracy, precision, specificity, 
and linearity. The PK characteristics of cytisine were evaluated in seven healthy smokers after a 
single 3 mg oral cytisine dose (i.e. double the dose used in Study CYT-BIO-02-07) over a 24 h 
period. Subjects enrolled smoked an average of 10.6 cigarettes per day and had been smoking for 
an average of 9.5 years with only two subjects having previously made an attempt to quit 
smoking. 
Ten blood samples were collected in heparinised tubes at the following times from each subject: 
0 (just before dosing), 0.25, 0.5, 1, 2, 3, 4, 6, 8, and 24 h after cytisine administration. Blood 
pressure, heart rate and respiratory rate were also measured at 0 (just before dosing), 2, 4, 8, and 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
19 
24 h after drug administration. After providing a blood sample at 8 h, subjects went home and 
returned the next morning, 24 h post-dosing. Subjects also provided spot urine samples. Non-
linear mixed effects modelling (NONMEM) was used for modelling and estimating population 
pharmacokinetic parameters (CL, Vd). A single compartment model was fit to the data. 
After a single 3 mg oral dose, cytisine was absorbed into the bloodstream, with cytisine 
detectable in plasma as early as 15 min after dosing (Figure 2). Peak plasma concentrations were 
typically observed at 2 h after administration. In two subjects, the peak plasma concentration was 
observed at 1 h post-dose, suggesting that the peak plasma concentration may have been 
achieved between 1 and 2 h for all subjects. This is broadly consistent with the estimate of 0.92 h 
in the Study CYT-BIO-02-07. Since the blood sampling in this study had a less frequent 
sampling schedule of 0.25, 0.5, 1, and 2 h, this study was less able to accurately estimate a Tmax 
of approximately an hour. The mean Cmax was 27.76 ng/mL. Following the peak plasma 
concentration, cytisine concentrations declined in a monophasic manner and the T½ of cytisine 
was calculated to be 4.8 h.  
 
Figure 2: 
Mean plasma concentrations (ng/mL) of cytisine over 24 hours  
 
Mean plasma concentrations (ng/mL) of cytisine over 24 hours following a single 3 mg oral 
dose. Values are shown as mean ± SEM (n=7). 
The values for Vd and CL were estimated to be 115 L and 16.7 L/h, respectively with standard 
error values 0.003.  
Cytisine was still detectable in the urine collected at 24 h for all subjects (mean 24 h 
concentration was 428.15 ng/mL). Cytisine was detected in urine as unchanged cytisine and no 
metabolites were detected in plasma or in urine in any of the subjects in the study. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
20 
In summary, the logarithmic plot of mean cytisine concentration versus time after Cmax showed a 
single elimination phase (i.e., no distinction was made between the distribution and elimination 
phase of the drug). This indicated that single dose PK of cytisine may be described using a 
single-compartment model. Since the apparent volume of distribution for cytisine estimated for 
the population in the study (115 L) was more than three times that of the blood compartment 
(35 L), this suggested that cytisine may distribute to other compartments very rapidly. Plasma 
cytisine concentrations declined with an average elimination half-life of 4.8 h. Thus, in this pilot 
study, the half-life for 3 mg cytisine in smokers (4.8 h) was the same for 1.5 mg cytisine in 
healthy non-smokers (4.09 h ± 0.82). This study is described in more detail in the Investigator’s 
Brochure. 
2.4. 
Safety Summary 
Given that cytisine (T
x) has been marketed in Central and Eastern Europe, including four 
Member States of the European Union (EU), for several decades, there is considerable clinical 
experience with this product. Over 20 million subjects have been treated, and over 10,000 
subjects have taken part in clinical trials, the majority of which were conducted several years 
ago. S
ma, the manufacturer of T
x, has been filing Periodic Safety Update Reports 
(PSURs) since 2005. The cumulative denominator in PSURs is now over 15 million exposures.  
Since cytisine has a structural similarity to nicotine and is a partial agonist of nicotine 
acetylcholine receptors, the possible pharmacodynamic undesirable effects are manifested 
mainly as nicotine effects. In 6 placebo-controlled clinical trials (N=2844) 1389 subjects 
received cytisine. The most commonly reported adverse events in the cytisine group were effects 
on the gastro-intestinal system: abdominal pain upper, nausea, dyspepsia, dry mouth, vomiting, 
constipation and diarrhea. Nervous system and psychiatric disorders were also common, most 
frequently headache and dizziness, as well as somnolence and insomnia. However, the statistical 
analysis did not reveal any significant difference in the nervous system adverse events between 
the cytisine and placebo groups (p=0.12). 
Please reference the cytisine Summary of Product Characteristics (SmPC) for additional human 
safety information. 
2.5. 
Recent Nonclinical Toxicology Studies: Estimation for Higher 
Human Equivalent Doses 
A number of non-clinical studies with cytisine were performed more than 25 years ago and, thus, 
pre-dated current GLP standards. The US National Center for Complementary and Integrative 
Health (NCCIH) has sponsored new non-clinical GLP studies that have included 28-day repeat 
dosing for toxicology assessments in rats and dogs to current GLP guidance.  
Daily oral dose administration of cytisine in Sprague Dawley rats at 5 or 20 mg/kg/day for 28 
consecutive days resulted in clinical signs, such as drooling, in a dose dependent manner. Other 
clinical signs were only seen in the 20 mg/kg group including ruffled fur, hypoactivity, shoveling 
behaviors, and ataxia. Decreases in food consumption and body weight gain were observed in 
animals treated with 20 mg/kg of cytisine. No cytisine-related macroscopic and microscopic 
findings were observed. The no observed adverse effect level (NOAEL) of cytisine is considered 
to be approximately 5 mg/kg/day when given orally for 28 day consecutive days in rats. This 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
21 
would calculate to an estimated Human Equivalent Dose (HED) of 0.3 mg/kg (i.e. 21 mg for a 
70 kg human) when cytisine is administered for 28 consecutive days. 
Daily oral dose administration of cytisine via capsules in Beagle dogs at 0.5, 1.0, or 2 mg/kg/day 
for 28 consecutive days resulted in adverse effects. The maximum tolerated dose (MTD) was 
considered to be greater than 1 mg/kg/day but less than 2 mg/kg/day. Although emesis, drooling 
and diarrhea were observed in some animals treated with 0.5 mg/kg cytisine, the severities were 
mostly slight and not dose-limiting clinically; therefore, the NOAEL was slightly less or at 
approximately 0.5 mg/kg/day following 28 days oral administration. This gives an estimated 
HED of 0.28 mg/kg (i.e. 20 mg for a 70 kg human) when cytisine is administered for 28 
consecutive days. 
Based on the findings in both the 28-day rat and 28-day dog toxicology studies, the established 
commercial dose of 1.5 mg X 6 doses/day (maximum total exposure of 9 mg/day for the first 
3 days of treatment) is approximately half that of the estimated HED of ~20 mg/day for humans. 
The planned higher 3 mg/dose for this study yields a maximum daily dose exposure of 18 mg for 
the first 3 days of treatment. This again is below the calculated HED based on all of the studies 
as shown in Table 3.  
Table 3: 
Calculation of Margin of Safety based on Rat and Dog Toxicology Studies 
NOAEL 
Cmax 
(ng/mL) 
AUClast 
(ng*h/mL) 
HEDa 
(mg/kg/day) 
HEDa 
(mg/day) 
Human 
commercial 
Doseb 
Margin of 
Safety based 
on BSA 
7-day rat study:  
5 mg/kg  
383.5 
2926 
0.8 
56 
9  mg/day  
~6X 
7-day dog study:  
2 mg/kg 
317.5 
708.5 
1.1 
77 
 9  mg/day 
~8X 
28-day rat study: 
2 mg/kg 
292.6 
1071.7 
0.3 
21 
 9  mg/day 
~2X 
28-day dog study: 
0.5 mg/kg 
TBD 
TBD 
0.28 
20 
 9  mg/day  
~2X 
a HED = Human Equivalent Dose 
b Human commercial dose is 9 mg/day for the first 3 days in the 25-day schedule per commercial use. 
Notes: Cmax and AUC are determined from the NOAEL data.  
HED (mg/kg/day) = rat dose divided by 6.2 and dog dose divided by 1.8. 
HED (mg/day) = HED dose (mg/kg/day) x 70 kg. 
In conclusion, these margins of safety, based on HED calculations in newly conducted GLP 
toxicology studies support evaluating the pharmacokinetic parameters and pharmacodynamic 
effects of a higher 3 mg dose (maximum of 18 mg for the first 3 days) compared to the 
commercial dose of 1.5 mg (maximum of 9 mg for the first 3 days). In addition, these data are 
relevant for a future Phase 3 trial design.  

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
22 
3. 
RATIONALE FOR THE STUDY 
Although cytisine (T
x) has been marketed in Central and Eastern Europe using a standardised 
25-day treatment schedule at the manufactured 1.5 mg dose level, there have been no recent 
pharmacokinetic evaluations of repeat dosing at 1.5 mg or 3.0 mg in adult smokers, including the 
elderly (>65 years of age). Therefore, this study will evaluate the detailed pharmacokinetics of 
1.5 mg or 3.0 mg cytisine in adult smokers aged 18-65+ years as administered per the current 
T
ex labeling (25-day commercial schedule). In addition, assessment for any potential effect of 
cytisine on QT/QTc interval prolongation will be explored and analysed in relationship to the 
cytisine pharmacokinetic profile as well as general safety assessments for the 3 mg dose.  
4. 
STUDY OBJECTIVES 
4.1. 
Primary Objectives 
1. To evaluate the pharmacokinetic (PK) parameters during repeat dosing of 1.5 mg or 
3.0 mg cytisine when administered as the commercial 25-day schedule. 
2. To evaluate the pharmacodynamic (PD) effects (e.g. reduction in smoking) with repeat 
dosing of 1.5 mg or 3.0 mg cytisine when administered as the commercial 25-day 
schedule. 
4.2. 
Secondary Objectives 
1. To compare the PK parameters and tolerability for repeat dosing of 1.5 mg and 3.0 mg 
cytisine during the 25-day schedule in healthy smokers at 18-65 and >65 (elderly) years 
of age. 
2. To assess the renal elimination of cytisine via measurement of urinary concentrations of 
cytisine during treatment on Day 1 and Day 25.  
3. To evaluate for effects on QT/QTc interval prolongation and cardiac safety during 
treatment on Day 1 and Day 25 in healthy smokers at 18-65 and >65 (elderly) years of 
age. 
5. 
INVESTIGATIONAL PLAN 
5.1. 
Study Design 
This will be an open-label, randomised, multiple-dose PK and PD study conducted in adult 
smokers, 18-65+ years of age as shown in Figure 3: 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
23 
Figure 3: 
Study Design Overview  
 
 
The study will be comprised of a pre-study screen, followed by 25 days of treatment 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 (Section 11.2.1). Eligible subjects will then be asked to return on 
the evening prior to Day 1 (i.e. Day -1) for repeat of key entry testing, baseline questionnaires 
and provide overview of the risks of smoking and quit advice according to World Health 
Organizaion (WHO) guidelines11 (Section 11.2.2). Randomisation to treatment group will occur 
on the morning of Day 1 followed by initiation of study treatment. 
Treatment Period (Day 1 to Day 25):  Eligible subjects will take cytisine as outlined in the 
 product insert (SmPC). NOTE:  Group A subjects take 1 tablet at each timepoint 
(1.5 mg), Group B subjects take 2 tablets at each timepoint (3.0 mg). 
Figure 4: 
Dose Timing and Total Daily Dose 
Days 
Regimen 
Total Daily Dose 
Approximate Interval 
 
 
1.5 mg Dose 
3.0 mg Dose 
 
1-3 
6 times daily 
9.0 mg 
18.0 mg 
2 hours 
4-12 
5 times daily 
7.5 mg 
15.0 mg 
2.5 hours 
13-16 
4 times daily 
6.0 mg 
12.0 mg 
3 hours 
17-20 
3 times daily 
4.5 mg 
9.0 mg 
4-5 hours 
21-24 
2 times daily 
3.0 mg  
6.0 mg 
6 hours 
25 
Once daily 
1.5 mg 
3.0 mg 
- 
 
 
R 
A 
N 
D 
O 
M 
I 
S 
E 
Group B 
Multiple doses of 3.0 mg cytisine administered per 25-day 
schedule 
   12 subjects age 18-65 
   6 subjects age >65 
Group A 
Multiple doses of 1.5 mg cytisine administered per 25-day 
schedule 
   12 subjects age 18-65 
   6 subjects age >65 
 
S 
C 
R 
E 
E 
N 
 
 
F 
 
O 
 
L 
 
L 
 
O 
   W 
 
 
 
U 
 
P 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
24 
A total of 9 overnight stays in the clinic will be required over the 26-day study period. Subject 
entry into the study may be staggered in groups in order to facilitate study management activities 
but all subjects passing screening are to provide clinic visits as follows: 
• Day -1 and Days 1-3 (4 overnight stays in clinic):  Arrive at study clinic in the 
afternoon of Day -1, repeat select screening testing and verification of 
inclusion/exclusion criteria. Provide risks of smoking and quit advice according to 
WHO guidelines that evening.11  All subjects will fast for minimum of 10 hours prior 
to first dose on Day 1. Randomisation to treatment Group assignment will occur on 
the morning of Day 1, just prior to starting treatment. On the morning of Day 1, 
obtain baseline blood and urine and CO level, initiate Holter monitoring, receive 
study drug packets, begin treatment and provide required blood and urine samples. 
Subjects may have breakfast after first dose and continue with a standard meal 
schedule. On Days 1-3, various safety assessments and blood sample collections will 
occur. On the morning of Day 4 complete vital signs, ECG, blood for haematology 
and biochemistry testing, urine for cotinine test, expired CO, completion of tobacco 
craving questionnaire, safety assessment and collection of the pre-dose PK blood 
draw. Subject will then take first Day 4 dose followed by risks of smoking and quit 
advice, review of diary requirements, reminder of next clinic visit date and will be 
discharged. 
• Day 4-11:  Subjects to maintain dosing as per schedule. 
• Day 12-13 (1 overnight stay in clinic):  Subjects return to clinic prior to last evening 
dose on Day 12 for pre-dose PK blood collection. Blood collections occur post final 
dose on Day 12 for PK. On the morning of Day 13 complete vital signs, ECG, blood 
for haematology and biochemistry testing, urine for cotinine test, expired CO, 
completion of tobacco craving questionnaire, safety assessment, and collection of the 
pre-dose PK blood draw. Subject will then take first Day 13 dose followed by risks of 
smoking and quit advice, review of diary requirements, reminder of next clinic visit 
date and will be discharged. 
•  Day 13-15:  Subjects maintain dosing as per schedule. 
• Day 16-17 (1 overnight stay in clinic):  Subjects return to clinic prior to last evening 
dose on Day 16 for pre-dose PK blood collection. Blood collections occur post final 
dose on Day 16 for PK. On the morning of Day 17 complete vital signs, ECG, blood 
for haematology and biochemistry testing, urine for cotinine test, expired CO, 
completion of tobacco craving questionnaire, safety assessment, and collection of the 
pre-dose PK blood draw. Subject will then take first Day 17 dose followed by risks of 
smoking and quit advice, review of diary requirements, reminder of next clinic visit 
date and will be discharged. 
• Day 17-19:  Subjects maintain dosing as per schedule.  
• Day 20-21 (1 overnight stay in clinic):  Subjects return to clinic prior to last evening 
dose on Day 20 for pre-dose PK blood collection. Blood collections occur post final 
dose on Day 20 for PK. On the morning of Day 21 complete vital signs, ECG, blood 
for haematology and biochemistry testing, urine for urinalysis and cotinine test, 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
25 
expired CO, completion of tobacco craving questionnaire, safety assessment, and 
collection of the pre-dose PK blood draw. Subject will then take first Day 21 dose 
followed by risks of smoking and quit advice, review of diary requirements, reminder 
of next clinic visit date and will be discharged. 
• Day 21-23:  Subjects maintain dosing as per schedule. 
• Day 24-26 (2 overnight stays in clinic):  Subjects return to clinic prior to last evening 
dose on Day 24 for pre-dose PK blood collection. Blood collections occur post final 
dose on Day 24 for PK. All subjects will have breakfast 30 minutes prior to final dose 
on Day 25. Extensive PK blood sampling, urine collections and Holter monitoring 
occur pre and post final dose on Day 25, with final draw 24 hours post dose (Day 26). 
Final discharge from clinic on Day 26 will occur after 24 hour blood draw, vital 
signs, ECG, haematology and biochemistry testing, serum pregnancy test for female 
subjects, urine for urinalysis and cotinine test, expired CO, completion of tobacco 
craving questionnaire, safety assessment, risks of smoking and quit advice, review of 
diary requirements, and reminder of post study followup visit date. 
• Post Study: A post study follow-up visit will be conducted 6 to 8 days after last dose 
of cytisine. If any adverse events are recorded at post study follow up assessment, 
arrangements will be made with the subject, so that they are followed-up 
appropriately and the final outcome determined. Smoking history will be reviewed 
and final review of risks of smoking and quit advice provided. 
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 9 weeks (screening to last subject’s last visit).  
6. 
SELECTION OF STUDY POPULATION 
A total of 36 subjects will be required to complete the study. Of the 36 subjects, 24 are to be 
18-65 years and 12 are to be  >65 years of age. An attempt will be made for 50% male and 50% 
female enrollment. 
The study is to be conducted in adult smokers and therefore participants may receive therapeutic 
benefit from taking part. Outside of being active smokers, a healthy subject population with 
carefully considered inclusion/exclusion criteria will ensure that subjects are fit and well enough 
for participation 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. Regular moderate cigarette smokers (minimum 10 cigarettes per day) who want to stop 
smoking. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
26 
2. Urine cotinine >500 ng/mL. 
3. Expired air carbon monoxide (CO) > 11 parts per million (no cigarette 1 hour before 
test). 
4. Healthy males and females 18-65+ 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 
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. 
5. Subject with no clinically significant abnormal serum biochemistry, haematology and 
urine examination values within 28 days before the first dose of cytisine.  
6. Subject with negative urinary drugs of abuse screen, determined within 28 days before 
the first dose of cytisine (a positive alcohol result may be repeated at Investigator's 
discretion). 
7. Subject with negative human immunodeficiency virus (HIV), hepatitis B surface antigen 
(Hep B) and hepatitis C virus antibody (Hep C) results. 
8. 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. 
9. Subject with no clinically significant abnormalities in vital signs (systolic blood pressure 
between 90-150 mmHg (age 18-65) and 90-160 mmHg (age >65), diastolic blood 
pressure (DBP) between 50 and 90 mmHg, and pulse rate (PR) between 40-110 bpm, 
measured on the dominant arm after minimum of 5 minutes in supine position) 
determined within 28 days before first dose of cytisine.  
10. Subject must be available to complete the study (including in-clinic stays and post study 
follow-up) and comply with study restrictions. 
11. Subject must provide written informed consent to participate in the study. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
27 
6.1.2. 
To be Re-Confirmed Prior to Start of Dosing (Day -1) 
1. Subject continues to meet all screening inclusion criteria (before the first dose only). 
2. Subject has a negative urinary drugs of abuse screen (including alcohol). 
3. Female subject has a negative pregnancy test.  
6.2. 
Exclusion Criteria 
6.2.1. 
To be Confirmed at Screening 
1. Treatment with smoking cessation medications (bupropion, varenicline, any nicotine 
replacement therapy) within 8 weeks of first dose of cytisine. 
2. Use of other forms of nicotine (e-cigarettes, smokeless tobacco) within 8 weeks of first 
dose of cytisine or are planning to use these products during study. 
3. Known hypersensitivity/allergy reaction to varenicline, other cytisine-derivatives or any 
of the excipients in the 
 formulation. 
4. History of severe hypersensitivity reactions to any other drugs. 
5. Current treatment with antihypertensive medicinal products, statins, tuberculostatics, 
cholinomimetics or anticholinesterase medicinal products. 
6. 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). 
7. Female subjects who are breast feeding. 
8. Difficulty in donating blood on either arm or known history. 
9. History of alcoholism or drug abuse within last 2 years. 
10. Use of non-prescription drugs, including vitamins, herbal and dietary supplements within 
14 days (or 5 half-lives, whichever is longer) prior to the first dose of cytisine, unless in 
the opinion of the Principal Investigator the medication will not interfere with the study 
procedures or compromise subject safety. 
11. Participated in any investigational drug clinical trial within the previous 3 months or a 
marketed drug trial within the previous 30 days prior to randomisation on Day 1. 
12. 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 first dose of cytisine. 
13. Inability to communicate well with Principal Investigator or designees (i.e., language 
problem, poor mental development or impaired cerebral function). 
14. Any other condition that the Principal Investigator considers making the subject 
unsuitable for this study. 
6.2.2. 
To be Re-Confirmed Prior to Start of Dosing (Day -1): 
1. Development of any exclusion criteria since screening. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
28 
2. Use of non-prescription drugs, including vitamins, herbal and dietary supplements since 
screening, unless in the opinion of the Principal Investigator or designee 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. 
The Principal 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 36 subjects are required 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 Principal Investigator (or designee) and Sponsor. 
7. 
RETRICTIONS 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.  

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
29 
7.1. 
Confinement 
Each subject will be required to provide a total of 9 overnight stays in the clinic over a 26-day 
period.  
7.2. 
Smoking, Diet and Fluid Restrictions 
7.2.1. 
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. 
7.2.2. 
Meal Times 
During days of confinement, subjects will be provided standard meals (breakfast, lunch, dinner, 
snacks) at specific times, concurrent with study treatment dosing when possible. Subjects to be 
fasting prior to the first Day 1 dose and will be fed breakfast 30 minutes prior to the last Day 25 
dose. 
7.2.3. 
Fluid Intake 
Prior to PK sampling only, 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. 
7.2.4. 
Alcohol Intake 
The consumption of alcohol will be limited to a maximum of 2 units (50 mL spirits, 500 mL beer 
or lager or 250 mL wine) 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 on Day 1 and again on 
Day 25. Alcohol will be avoided completely for 24 hours prior to each day of confinement for 
PK sampling. 
7.2.5. 
Caffeine 
Food or drink containing caffeine, including coffee, tea, cola, energy drinks or chocolates will be 
avoided completely for 24 hours prior to each day of confinement for PK sampling. 
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. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
30 
7.2.7. 
Grapefruit Juice and Other Restrictions  
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 
avoided completely for 24 hours prior to each day of confinement for PK sampling. 
Each subject will be questioned on the specific points at admission to the clinical unit as 
appropriate. If a subject admits a non-compliance with these restrictions, the Principal 
Investigator or designee 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. 
Blood Donation 
Subjects will be advised that they should not donate blood for at least 3 months after the final 
study visit. 
7.3.2. 
Contraception 
To prevent pregnancy female subjects of childbearing potential and male subjects with female 
partners of child bearing potential must take / agree to 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]. 
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. 
7.3.3. 
Sperm Donation 
Subjects must not donate sperm from the first dose and for at least 3 months after the last dose of 
cytisine.  

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
31 
8. 
INVESTIGATIONAL MEDICINAL PRODUCT (IMP) 
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 
 
  accordance with cGMP. 
Each tablet is composed of cytisine (as the base), with well-established tablet-forming exc
nts 
 
oxides). 
Cytisine should be administered orally according to the dose timing schedule (see Figure 4) with 
each dose to be either one (1) 1.5 mg
) 1.5 mg tablets taken with 240 m
ater 
(8 fl
es) and, when possible, with a meal or snack.  
Table 4: 
Cytisine Product Information 
Name of the Product: 
Cytisine 1.5 mg, film coated tablets (T
®) 
Strengths: 
g 
Dosage form: 
Comp
ablet 
Route/mode of administration: 
Oral 
Dosing Schedule: 
(See Figure 4) 
Dosing instructions: 
Depending of Treatment Group assignment, 
tablets to be take
ater for each dose 
Manufacturer/Marketing Authorisation Holder: 
, 
ria 
 
The documentation supplied will make it possible to retrace the composition and pharmaceutical 
quality of the product. 
8.2. 
Receipt and Storage 
Cytisine will be supplied by the Sponsor as commercial product in a format suitable for study 
specific relabeling. 
The Sponsor must notify the Principal Investigator, or the Project Manager, prior to dispatch of 
the IMP 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: 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
32 
Cytisine will be stored under quarantine in a segregated, study-specific area, at or below 25 oC in 
a secure, temperature-controlled pharmacy. Si
ec’s 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 will be supplied to study subjects in its commercial configuration, as manufactured by 
S
ma.  
Cytisine will be labelled as specified in Annex 13 (Manufacture of IMPs) of the European 
Commission (EC) guide to Good Manufacturing Practice (GMP). 
Finished IMP will be certified by S
c’s Qualified Person according to standard operating 
procedure #BD/324/13/29. 
8.4. 
Administration 
Each subject will receive specified allocations of cytisine periodically during the treatment 
period that will allow for correct dosing as assigned at randomisation  (1.5 mg or 3.0 mg dose 
level). Cytisine allocations will be documented and actual dosing (and time of dosing) will be 
documented by each subject in their diary.  
8.5. 
Return/Destruction 
All used IMP containers and unused IMP will be held under quarantine in the Si
c Pharmacy 
pending return/destruction. Following the Sponsor’s approval, all remaining IMP will be 
destroyed or returned to the Sponsor within 8 weeks of the last dose of IMP.  
8.6. 
Method of Assigning Subjects to Group A or Group B 
Subjects will be randomised to treatment dosing (1.5 mg or 3.0 mg) according to a code 
produced by Si
c using the PROC PLAN procedure of SAS® version 9.1.3 or higher. Subjects 
will be numbered sequentially from 001 (i.e. 001, 002 etc.). Subjects will be stratified for age 
(elderly > 65 years: yes vs no) to assure that each treatment dosing has 6 elderly subjects and 12 
non-elderly subjects. 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.). 
8.7. 
Selection of Doses in the Study 
The dose selected for this study is either 1.5 mg or 3.0 mg cytisine. 
8.8. 
Selection and Timing of Dose for Each Subject 
First dose will be administered at approximately 9.00 a.m. on Day 1. Additional doses will be as 
per the Dosing Schedule (see Figure 4). A target time will be provided for each dose in the diary 
and actual time of dose will be recorded. For Days where dosing is every 2 to 3 hours, dosing 
should be ± 10 min within the targeted time. For Days where dosing is every 4-6 hours, dosing 
should be ± 30 min within the targeted time. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
33 
8.9. 
Dose Modifications for 3.0 mg Dose or Elderly Subjects (>65) 
Dose modifications should not be necessary, unless a severe adverse event warrants holding or 
missing a dose as outlined below during the first 1 – 16 days in the 3 mg cytisine dosage group 
or at any time in elderly subjects.  
Cytisine at higher levels in toxicology studies illicited adverse reactions primarily on the gastro-
intestinal system such as vomiting or diarrhea. Based on the HED calculations in newly 
conducted GLP toxicology studies (see Section 2.5), the safety margin supports the 3 mg dose 
(maximum of 18 mg for the first 3 days). The commercial dose of 1.5 mg (maximum of 9 mg for 
the first 3 days) has been well tolerated with no or minimal gastrointestinal symptoms. However, 
if a subject has nausea that results in vomiting or has minimal diarrhea progressing to severe 
diarrhea, the Principal Investigator/Sponsor Medical Monitor can delay, hold or skip doses. 
Dosing should resume within 48 hours when the adverse event has subsided to a moderate grade 
(see Table 8) or minimal symptoms. Dosing should then resume on the same schedule without 
modification. If the adverse event has not subsided to at least a moderate grade within 48 hours, 
the subject should be withdrawn from further study treatment. 
For all subjects randomised to the 3.0 mg dosing schedule, if a severe adverse event reoccurs 
during the Day 1-16 dosing, then dosing should be again held until the adverse event has 
subsided to a moderate grade or minimal symptoms within a 48-hour period. Dosing should then 
be reduced as specified in Table 5. If the severe adverse event reoccurs after reducing the dose, 
the subject should be withdrawn from further study treatment. If the severe adverse event does 
not reoccur, the patient can continue the study treatment with dosing at the 1.5 mg level as 
shown in Table 5. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
34 
Table 5: 
Dose Modification for 3.0 mg Dose Group  
Days 
Regimen 
Total Daily Dose Dose Modifications After Recurrence 
1-3 
6 times daily 
18.0 mg 
9 mg total daily dose (Reduce to one 1.5 mg 
cytisine tablet 6 times daily and follow the 
remaining schedule as for the 1.5 mg dosing 
group). 
4-12 
5 times daily 
15.0 mg 
7.5 mg total daily dose (Reduce to one 1.5 mg 
cytisine tablet 5 times daily and follow the 
remaining schedule as for the 1.5 mg dosing 
group). 
13-16 
4 times daily 
12.0 mg 
6 mg total daily dose (Reduce to one 1.5 mg 
cytisine tablet 4 times daily and follow the 
remaining schedule as for the 1.5 mg dosing 
group). 
17-20 
3 times daily 
9.0 mg 
No dose modification 
21-24 
2 times daily 
6.0 mg 
No dose modification 
25 
Once daily 
3.0 mg 
No dose modification 
For elderly subjects in the 1.5 mg dosing schedule, if a severe adverse event reoccurs at any time, 
then dosing should be held until the adverse event has subsided to a moderate grade or minimal 
symptoms within a 48-hour period. Dosing should then be reduced by one dose per day. If the 
severity of the event then reoccurs, the subject should be withdrawn from further study 
treatment. 
8.10. 
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 all samples will be conducted without information on treatment. 
9. 
PRIOR AND CONCOMITANT THERAPY 
Prior Medication:  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 Principal Investigator and Sponsor's Medical Monitor the 
medication will not interfere with the study procedures or would compromise subject safety. 
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.  

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
35 
Inclusion of subjects who are on prescription medications or 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: New prescription or other non-prescription drugs, including smoking 
cessation medications, tuberculostatics, cholinomimetics or anticholinesterase medicinal 
products, statins or antihypertensive medicinal products, 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 taken while in the clinic will be taken under supervision and a hand and 
mouth check conducted. Subjects will be required to maintain a diary to document dosing times 
and provide medication packets for tablet accountability at each clinic visit. These will be 
reviewed by clinic staff during clinic visits. 
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 randomised according to the randomisation schedule on 
Day 1. Subjects will receive their initial allocation of study medication on Day 1. The subjects 
will be required to make a total of 9 overnight clinic stays over a 26-day period. All subjects are 
required to complete a follow-up visit 6-8 days post last dose. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
36 
 
11.1. 
Procedure Schedule 
Table 6 provides a summary of required study evaluations. Screening evaluations are to occur within a 28-day interval from initiation 
of screening evaluations to Day -1, just prior to randomisation on Day 1. 
Table 6: 
Schedule of Study Procedures 
 
 
Screening 
Study Day 
Post-Study  
Study Day 
-28 to -2 
-11 
1-41 
4-11 
12-132 
13-15 
16-172 
17-19 
20-212 
21-23 
24-263 
6-8 days after last 
dose 
Written informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
Number of cigarettes smoked in 
previous 24 hours 
X 
X 
 
 
 
 
 
 
 
 
 
 
Daily Smoking and Study 
Medication Diary4 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Demographic data 
X 
 
 
 
 
 
 
 
 
 
 
 
Randomisation 
 
 
X5 
 
 
 
 
 
 
 
 
 
Vital signs 
X 
X6 
X6 
 
X6 
 
X6 
 
X6 
 
X6 
 
Medical history 
X 
X7 
 
 
 
 
 
 
 
 
 
 
Prior and concomitant medication 
X 
X 
X 
 
X 
 
X 
 
X 
 
X 
X 
Physical examination 
X 
 
 
 
 
 
 
 
 
 
 
 
12-lead ECG 
X 
X 
X8 
 
X8 
 
X8 
 
X8 
 
X8 
 
Holter monitor for continuous ECG 
readings9 
 
 
X 
 
 
 
 
 
 
 
X 
 
Pregnancy test for all females 
X 
Serum 
X 
Urine 
 
 
 
 
 
 
 
 
X 
Serum 
 
FSH (females of non-childbearing 
potential only) 
X 
 
 
 
 
 
 
 
 
 
 
 
Haematology10 
X 
 
X 
 
X 
 
X 
 
X 
 
X 
 
Biochemistry10 
X 
 
X 
 
X 
 
X 
 
X 
 
X 
 
Virology (HIV, Hep-B, Hep-C) 
X 
 
 
 
 
 
 
 
 
 
 
 
Urinalysis 
X 
 
 
 
 
 
 
 
 
 
X 
 
Drugs-of-abuse tests in urine 
X 
X 
 
 
 
 
 
 
 
 
 
 
Fagerström nicotine dependence 
 
X 
 
 
 
 
 
 
 
 
X 
 
Urine cotinine11 
X 
 
X 
 
X 
 
X 
 
X 
 
X 
 
Expired air CO11 
X 
 
X 
 
X 
 
X 
 
X 
 
X 
 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
37 
Verification of eligibility criteria  
X 
X 
X 
 
 
 
 
 
 
 
 
 
Tobacco craving questionnaire 
 
X 
X 
 
X 
 
X 
 
X 
 
X 
 
Risks of smoking and quit advice12 
 
X 
X 
 
X 
 
X 
 
X 
 
X 
X 
IMP (cytisine) administration13 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
Plasma collection for PK analysis14 
 
 
X 
 
X 
 
X 
 
X 
 
X 
 
Urine collection for PK analysis15 
 
 
X 
 
 
 
 
 
 
 
X 
 
Adverse events monitoring16 
 
 
X 
 
X 
 
X 
 
X 
 
X 
X 
1 4 overnight stays in clinic, starting evening of Day -1. 
2 1 overnight say in clinic. 
3 2 overnight stays in clinic. 
4 Subject to record number of cigarettes smoked in past 24 hours on a daily basis throughout study, actual time of each medication dose and record any adverse 
events. Information to be reviewed and documented during clinic days. Subject to also maintain record of number of cigarettes smoked in each 24 hour period 
from Day 26 until final Post-Study Visit (see Section 13.9). 
5 Pre-First Dose on Day 1 
6  Vital signs (supine blood pressure, pulse rate and oral temperature) will be recorded at pre-dose and again prior to discharge from clinic. Vital signs will also be 
recorded 3 hours post last dose on Days 1, 2, 3, 12, 16, 20 and 24. 
7 Clinically relevant changes will be reported as adverse events.  
8 Obtain 12-lead ECG (triplicate) at any time if clinically indicated during clinic visits.  Repeat 12-lead ECG and assess prior to discharge. 
9  Attach ECG Holter monitor and begin recording prior to first dose on Day 1 and continue for 24 hours (discontinue just prior to first dose on Day 2.  Repeat 
again by starting Holter monitor prior to dose on Day 25 and record for 24 hours. 
10 Haematology and biochemistry testing to be completed prior to discharge on Days 4, 13, 17, 21, 26 
11 Urine cotinine and expired CO testing to be obtained at screen and prior to discharge on Days 4, 13, 17, 21, 26. 
12 Each subject given advice about the risks of smoking and quitting. This will take the form of a brief interview following WHO evidence-based 
recommendations on the treatment of tobacco dependence on Day -1 and reminders prior to discharge on Days 4, 13, 17, 21, 26 and on the final post-study visit. 
13IMP administration to follow 25-day dosing schedule (see Figure 4). 
14Plasma collection for PK anlaysis is outlined in Section 12.1. 
15 Urine to be collected for PK on Day 1 and Day 25 as follows:  pre-dose and 0-24 hours. 
16 Adverse event recording to begin upon admissions on Day -1 to post study. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
38 
11.2. 
Detailed Description of Study Visits 
11.2.1. 
Screening Visit (Day -28 to Day -2) 
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. Document actual number of cigarettes smoked in the past 24 hours. 
3. Expired CO. 
4. Demographic data. 
5. Medical history data. 
6. Prior and concomitant medication. 
7. Physical examination, including height and weight. 
8. Vital signs (supine blood pressure, pulse rate and oral temperature). 
9. 12-lead ECGs (triplicate). 
10. Clinical laboratory safety tests (biochemistry, haematology, urinalysis, urine cotinine, 
virology, drugs of abuse, serum pregnancy for females and FSH for females of non-
childbearing potential). 
11. Verification of eligibility criteria. 
11.2.2. 
Day -1 
All subjects will check into the research facility and upon arrival to the clinic, the following 
procedures will be performed: 
1. Vital signs (supine blood pressure, pulse rate and oral temperature). 
2. Update any medical history or concomitant medications. 
3. Repeat urine testing for drugs of abuse and urine pregnancy test for female subjects. 
4. Obtain 12-lead ECG (triplicate). 
5. Verification of eligibility criteria. 
6. Record number of cigarettes smoked in past 24 hours and instruct subject how to 
document cigarette smoking and dosing times in the daily smoking/medication diary (see 
Section 13.9). 
7. Subject to complete the Fagerström nicotine dependence questionnaire (see Appendix 2). 
8. Subject to complete tobacco craving questionnaire (see Appendix 3). 
9. Each subject given advice about the risks of smoking and quitting. This will take the form 
of a brief interview following WHO evidence-based recommendations on the treatment 
of tobacco dependence. 
10. Each subject must fast for minimum of 10 hours prior to first dose on Day 1.   

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
39 
 
11.2.3. 
Day 1 to Morning of Day 4 
The results of screening for each subject must be reviewed by the Principal Investigator or 
designee prior to randomisation to assure eligibility criteria have been met.  
On Day 1: 
1. Vital signs (supine blood pressure, pulse rate and oral temperature). 
2. Verify and document any baseline clinically relevant adverse events. 
3. Randomise subject to treatment group. Provide IMP and instructions for administration, 
reviewing dosing schedule and documentation requirements in the study diary. 
4. Attach ECG Holter monitor, checking to ensure recording has started. Note: Holter 
monitors to remain attached to subjects approximately 24 hours, discontinued just prior to 
first dose on Day 2. 
5. Obtain pre-dose blood sample prior to first dose and PK sampling after first dose 
(Section 12.1). 
6. First dose to be taken prior to breakfast (minimum fasting of 10 hours). Breakfast and 
standard meal schedule to begin after taking first dose. 
7. Obtain urine PK samples starting prior to first dose and continuing for 24 hours (Section 
Section 12.2). 
8. Subject to take 6 doses of cytisine as outlined in Figure 4. 
9. Obtain pre-last dose PK blood sample and additional PK sampling after last dose (Section 
Section 12.1). 
10. Record vital signs (supine blood pressure, pulse rate and oral temperature) at 3 hours 
after the last dose (Note: this is the estimated highest Cmax timepoint for the daily dosing). 
11. Obtain 12-lead ECG (triplicate) at any time if clinically indicated. 
12. Record any adverse events and update concomitant medications. 
13. The number of cigarettes smoked and actual dosing times will be recorded in the CRF.  
14. All subjects will remain in the clinic overnight. 
On Day 2: 
1. Discontinue Holter monitoring approximately 24 hours after initial dose taken on Day 1 
(just prior to first dose on Day 2). 
2. Subject to take 6 doses of cytisine as outlined in Figure 4. 
3. Obtain pre-last dose PK blood sample and additional PK sampling after last dose 
(Section 12.1). 
4. Record vital signs (supine blood pressure, pulse rate and oral temperature) at 3 hours 
after the last dose (Note: this is the estimated highest Cmax timepoint for the daily dosing). 
5. Obtain 12-lead ECG (triplicate) at any time if clinically indicated. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
40 
6. Record any adverse events and update concomitant medications. 
7. The number of cigarettes smoked and actual dosing times will be recorded in the CRF.  
8. All subjects will remain in the clinic overnight. 
On Day 3: 
1. Subject to take 6 doses of cytisine as outlined in Figure 4. 
2. Obtain pre-last dose PK blood sample and additional PK sampling after last dose 
(Section 12.1). 
3. Record vital signs (supine blood pressure, pulse rate and oral temperature) at 3 hours 
after the last dose (Note: this is the estimated highest Cmax timepoint for the daily dosing). 
4. Obtain 12-lead ECG (triplicate) at any time if clinically indicated. 
5. Record any adverse events and update concomitant medications. 
6. The number of cigarettes smoked and actual dosing times will be recorded in the CRF.  
7. All subjects will remain in the clinic overnight. 
On Day 4: 
1. Obtain pre-dose PK blood sample prior to Day 4 dose (Section 12.1). 
2. Vital signs (supine blood pressure, pulse rate and oral temperature). 
3. Obtain 12-lead ECG (triplicate). 
4. Obtain blood for haematology and biochemistry testing. 
5. Obtain urine for cotinine test and expired air CO prior to discharge. 
6. Subject to complete tobacco craving questionnaire (see Appendix 3). 
7. Record any adverse events and update concomitant medications. 
8. Subject to take first Day 4 dose. 
9. Subject given advice about the risks of smoking and quitting.  
10. Remind subject to record number of cigarettes smoked daily and actual dosing times in 
diary, discharge subject from clinic and schedule return for Day 12. 
11.2.4. 
Day 4 through Day 11 
1. Subject to maintain dosing schedule (5 times daily). 
2. Subject to record number of cigarettes smoked and actual dosing times daily in diary (see 
Section 13.9). 
3. Subject to maintain adverse event log. 
4. Subject to bring medication packets and diary to clinic on Day 12. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
41 
11.2.5. 
Day 12 to Morning of Day 13 
On Day 12: 
1. Subject will check into the research facility in the afternoon, just prior to the last Day 12 
dose. 
2. Obtain pre-dose PK blood sample prior to the last Day 12 dose. Blood samples will then 
be collected and processed as outlined in (Section 12.1) after the last Day 12 dose of 
cytisine. 
3. Record vital signs (supine blood pressure, pulse rate and oral temperature) at 3 hours 
after the last dose (Note: this is the estimated highest Cmax timepoint for the daily dosing). 
4. Obtain 12-lead ECG (triplicate) if clinically indicated. 
5. Record any adverse events and update concomitant medications. 
6. Review smoking/medication diary for completeness. The number of cigarettes smoked 
and actual dosing times will be recorded in the CRF. 
7. All subjects will remain in the clinic overnight. 
On Day 13: 
1. Obtain pre-dose PK blood sample prior to initial dose (Section 12.1). 
2. Vital signs (supine blood pressure, pulse rate and oral temperature). 
3. Obtain 12-lead ECG (triplicate). 
4. Obtain blood for haematology and biochemistry testing. 
5. Obtain urine for cotinine test and expired air CO prior to discharge.  
6. Subject to complete tobacco craving questionnaire (see Appendix 3). 
7. Record any adverse events and update concomitant medications. 
8. Subject to take first Day 13 dose in clinic. 
9. Subject given advice about the risks of smoking and quitting.  
10. Remind subject to record number of cigarettes smoked and actual dosing times daily in 
diary, discharge subject from clinic and schedule return for Day 16. 
11.2.6. 
Day 13 through Day 15 
1. Subject to maintain dosing schedule (4 times daily). 
2. Subject to record number of cigarettes smoked daily and actual dosing times in diary. 
3. Subject to maintain adverse event log. 
4. Subject to bring medication packets and diary to clinic on Day 16. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
42 
11.2.7. 
Day 16 through Morning of Day 17 
On Day 16: 
1. Subject will check into the research facility in the afternoon, just prior to the last Day 16 
dose. 
2. Obtain pre-dose PK blood sample prior to the last Day 16 dose. Blood samples will then 
be collected and processed as outlined in (Section 12.1) after the last Day 16 dose of 
cytisine. 
3. Record vital signs (supine blood pressure, pulse rate and oral temperature) at 3 hours 
after the last dose (Note: this is the estimated highest Cmax timepoint for the daily dosing). 
4. Obtain 12-lead ECG (triplicate) if clinically indicated. 
5. Record any adverse events and update concomitant medications. 
6. Review smoking/medication diary for completeness. The number of cigarettes smoked 
and actual dosing times will be recorded in the CRF. 
7. All subjects will remain in the clinic overnight. 
On Day 17: 
1. Obtain pre-dose PK blood sample prior to initial dose (Section 12.1). 
2. Vital signs (supine blood pressure, pulse rate and oral temperature). 
3. Obtain 12-lead ECG (triplicate). 
4. Obtain blood for haematology and biochemistry testing. 
5. Obtain urine for cotinine test and expired air CO prior to discharge.  
6. Subject to complete tobacco craving questionnaire (see Appendix 3). 
7. Record any adverse events and update concomitant medications. 
8. Subject to take first Day 17 dose in clinic. 
9. Subject given advice about the risks of smoking and quitting.  
10. Remind subject to record number of cigarettes smoked daily and actual dosing times in 
diary, discharge subject from clinic and schedule return for Day 20. 
11.2.8. 
Day 17 through Day 19 
1. Subject to maintain dosing schedule (3 times daily). 
2. Subject to record number of cigarettes smoked and actual dosing times daily in diary (see 
Section 13.9). 
3. Subject to maintain adverse event log. 
4. Subject to bring medication packets to clinic on Day 20. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
43 
11.2.9. 
Day 20 through Morning of Day 21 
On Day 20: 
1. Subject will check into the research facility in the afternoon, just prior to the last Day 20 
dose. 
2. Obtain pre-dose PK blood sample prior to the last Day 20 dose. Blood samples will then 
be collected and processed as outlined in (Section 12.1) after the last Day 20 dose of 
cytisine. 
3. Record vital signs (supine blood pressure, pulse rate and oral temperature) at 3 hours 
after the last dose (Note: this is the estimated highest Cmax timepoint for the daily dosing). 
4. Obtain 12-lead ECG (triplicate) if clinically indicated. 
5. Record any adverse events and update concomitant medications. 
6. Review smoking/medication diary for completeness. The number of cigarettes smoked 
and actual dosing times will be recorded in the CRF. 
7. All subjects will remain in the clinic overnight. 
On Day 21: 
1. Obtain pre-dose PK blood sample prior to initial dose (Section 12.1). 
2. Vital signs (supine blood pressure, pulse rate and oral temperature). 
3. Obtain 12-lead ECG (triplicate). 
4. Obtain blood for haematology and biochemistry testing. 
5. Obtain urine for cotinine test and expired air CO prior to discharge. 
6. Subject to complete tobacco craving questionnaire (see Appendix 3). 
7. Record any adverse events and update concomitant medications. 
8. Subject to take first Day 21 dose in clinic. 
9. Subject given advice about the risks of smoking and quitting.  
10. Remind subject to record number of cigarettes smoked daily and actual dosing times in 
diary, discharge subject from clinic and schedule return for Day 24. 
11.2.10. 
Day 21 through Day 23 
1. Subject to maintain dosing schedule (2 times daily). 
2. Subject to record number of cigarettes smoked and actual dosing times daily in diary. 
3. Subject to maintain adverse event log. 
4. Subject to bring medication packets and diary to clinic on Day 24. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
44 
11.2.11. 
Day 24 through Day 26 
On Day 24: 
1. Subject will check into the research facility in the afternoon, just prior to the last Day 24 
dose. 
2. Obtain pre-dose PK blood sample prior to the last Day 24 dose. Blood samples will then 
be collected and processed as outlined in (Section 12.1) after the last Day 24 dose of 
cytisine. 
3. Repeat vital signs (supine blood pressure, pulse rate and oral temperature) at 3 hours after 
the last dose (Note: this is the estimated highest Cmax timepoint for the daily dosing). 
4. Obtain 12-lead ECG (triplicate) if clinically indicated. 
5. Record any adverse events and update concomitant medications. 
6. The number of cigarettes smoked and actual dosing times will be recorded in the CRF. 
7. All subjects will remain in the clinic overnight. 
On Day 25: 
1. All subjects to have breakfast 30 minutes prior to final dose. 
2. Attach ECG Holter monitor, checking to ensure recording has started. NOTE:  Holter 
monitors to remain attached to subjects approximately 24 hours, discontinued on Day 26. 
3. Record Vital signs (supine blood pressure, pulse rate and oral temperature). 
4. Obtain pre-dose PK blood sample (Section 12.1). 
5. Begin urine collection for PK as outlined in (Section 12.2). 
6. Subject to take final Day 25 dose of cytisine (documenting time of dose in diary) after 
breakfast and begin extensive PK blood and urine sampling as outlined in (Section 12.1) 
throughout day and up through 24 hours post. 
7. Obtain 12-lead ECG (triplicate) if clinically indicated. 
8. Record any adverse events and update concomitant medications. 
9. The number of cigarettes smoked will be recorded in the CRF. 
10. All subjects will remain in the clinic overnight. 
On Day 26: 
1. Complete final PK blood draw and urine collection (Section 12.1). 
2. Obtain blood and urine for haematology, biochemistry, serum pregnancy test (females 
only) and urinalysis (Section 13.4.1). 
3. Vital signs (supine blood pressure, pulse rate and oral temperature). 
4. 12-lead ECGs (triplicate). 
5. Obtain urine for cotinine test and expired air CO prior to discharge. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
45 
6. Subject to complete tobacco craving questionnaire (see Appendix 3). 
7. Record any adverse events and update concomitant medications. 
8. Subject given advice about the risks of smoking and quitting.  
9. Remind subject to record number of cigarettes smoked daily in diary, discharge subject 
from clinic and schedule follow up visit (within 6-8 days). 
11.2.12. 
Post Study Follow-up 
A Post-Study Follow-up visit will occur 6-8 days after last dosing to document any changes in 
concomitant medication(s) and adverse event(s) observed/documented at discharge as well as 
any new concomitant medications or adverse events. If any new adverse events are recorded at 
this follow-up visit, arrangements will be made with the subject to come into the clinic for 
assessment and then followed until outcome determined. Subject’s diary will be reviewed and 
collected (Note: diary will be used as source document for smoking history, actual dosing times 
and any self-reported adverse events). Subject given advice about the risks of smoking and 
quitting.  
12. 
DRUG CONCENTRATION MEASUREMENTS 
12.1. 
Pharmacokinetic Blood Sampling 
A total of 95 venous blood samples will be collected over a 26-day period (approximate volume 
of 3.0 mL each, total volume of approximately 285 mL) into pre-cooled lithium heparin tubes, 
according to the following schedule: 
Table 7: 
Blood Sampling for Pharmacokinetic Assessments   
Test Period 
Blood Sample 
Number 
Time from Dosing 
Day 1 (first dose) 
 
1 
Pre-Dose 
(within 60 minutes prior to dosing) 
2 
00:30 
3 
00:45 
4 
01:00 
5 
01:15 
6 
01:30 
7 
02:00 
Day 1 (last dose)  
 
 
 
8 
Pre-Last Dose 
9 
00:30 
10 
01:00 
11 
01:30 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
46 
Test Period 
Blood Sample 
Number 
Time from Dosing 
 
 
 
 
 
 
12 
02:00 
13 
02:30 
14 
03:00 
15 
03:30 
16 
04:00 
17 
05:00 
 
Day 2 (last dose) 
18 
Pre-Last Dose 
19 
00:30 
20 
01:00 
21 
01:30 
22 
02:00 
23 
02:30 
24 
03:00 
25 
03:30 
26 
04:00 
27 
05:00 
Day 3 (last dose) 
 
28 
Pre-Last Dose 
29 
00:30 
30 
01:00 
31 
01:30 
32 
02:00 
33 
02:30 
34 
03:00 
35 
03:30  
36 
04:00 
37 
05:00 
Day 4 (prior to first dose) 
38 
Pre-Dose 
Day 12 (last dose) 
 
39 
Pre-Last Dose 
40 
01:00 
41 
01:30 
42 
02:00 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
47 
Test Period 
Blood Sample 
Number 
Time from Dosing 
43 
02:30 
44 
03:00 
45 
03:30 
46 
04:00 
Day 13 (prior to first dose) 
47 
Pre-Dose 
 
Day 16 (last dose) 
 
48 
Pre-Last Dose 
49 
01:00 
50 
01:30 
51 
02:00 
52 
02:30 
53 
03:00 
54 
03:30 
55 
04:00 
Day 17 (prior to first dose) 
56 
Pre-Dose 
Day 20 (last dose) 
 
57 
Pre-Last Dose 
58 
01:00 
59 
01:30 
60 
02:00 
61 
02:30 
62 
03:00 
63 
03:30 
64 
04:00 
Day 21 (prior to first dose) 
65 
Pre-Dose 
Day 24 (last dose) 
 
66 
Pre-Last Dose 
67 
01:00 
68 
01:30 
69 
02:00 
70 
02:30 
71 
03:00 
72 
03:30 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
48 
Test Period 
Blood Sample 
Number 
Time from Dosing 
73 
04:00 
Day 25 
74 
Pre-dose 
75 
00:30 
76 
00:45 
77 
01:00 
78 
01:15 
79 
01:30 
80 
01:45 
81 
02:00 
82 
02:15 
83 
02:30 
84 
02:45 
85 
03:00 
86 
03:15 
87 
03:30 
88 
03:45 
89 
04:00 
90 
05:00 
91 
06:00 
92 
08:00 
93 
10:00 
94 
12:00 
Day 26 
95 
24:00 (post Day 25 dose) 
 
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 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
49 
logged, immediately placed on ice and then centrifuged within 60 minutes of draw 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.  
12.2. 
Pharmacokinetic Urine Collection 
Two urine samples will be collected during treatment on Day 1 and on Day 25 as follows: 
• Pre-dose (pre-dose urine samples will be taken upon waking up) 
• 0-24 hour 
The urine collection containers will be identified with a bar coded label bearing details of the 
study number, subject number, study period number (Day 1 or Day 25), 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 the 24 hour 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.  
The sample analysis will only begin after the last sample of a study subject is collected. All 
samples from each subject will be analysed 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 
esearch Ltd. 
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).  
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. 
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 reanalysed. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
50 
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 current smokers who are healthy by inclusion and exclusion criteria are eligible for the 
study. Subjects’ safety will be monitored during the study. Adverse events will be monitored and 
recorded throughout the study.  
Prior to the first IMP dosing only, the Principal Investigator or designee will assess the clinical 
significance of results of ECG readings and laboratory investigations for haematology and serum 
chemistry values outside the defined normal ranges, as provided by the laboratory. All changes 
from screening during the study and meet clinically-significant definitions for laboratory adverse 
events will be recorded as an adverse event. 
In addition to the planned times, any safety procedures can be performed at any time considered 
necessary by the Principal Investigator or designee.  
On each day where subjects are in the clinic, at least one physician (Principal Investigator or 
designee) will physically remain at the clinical site, 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, the Principal Investigator 
or designee 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 visit will be recorded in the subject’s CRF.  
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 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
51 
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 Principal 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 unauthorised investigational 
product or in the SmPC for an authorised 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 hospitalisation or prolongs existing inpatient’s hospitalisation 
• 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. 
• Unexpected (i.e., is not consistent with the applicable product information e.g. 
Investigator's brochure for an unapproved IMP 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 
jeopardise 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 
hospitalisation. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
52 
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 hospitalisation or prolongation of existing hospitalisation 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 the Principal Investigator or designee in 
accordance with the definitions below. 
• Relationship to IMP to be assessed by the Principal Investigator or designee 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. 
13.2.3. 
Grades of Adverse Event Severity 
The following grades will be used by the Principal Investigator or designee 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 or significantly interferes with daily activity and 
requires medical intervention 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
53 
If an adverse event has multiple aspects, the aspect in the highest intensity will be graded. It is 
emphasised 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 Principal Investigator or designee, according to best medical judgment as 
shown in Table 9.  
Table 9: 
Assessment of Causal Relationship to Cytisine 
Catagory 
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.  
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 
Principal Investigator or designee 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 
 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
54 
vices Ltd will notify the Sponsor and Sponsor's Medical Monitor 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: 
• Principal 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.  
SUSARs should be reported to the REC, Healthcare products Regulatory Agency (MHRA) and 
Sponsor in accordance with applicable regulatory requirements for expedited reporting. It is 
Si
ec’s 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 Principal Investigator or designee 
and is not considered to be clinically significant. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
55 
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 S
 L
 
Serum pregnancy tests will be performed using the Roche Cobas c6000 analyser series 
comprising of c501 and e601 modules.  
Pregnancies must be reported immediately and followed up to determine outcome, including 
spontaneous or voluntary termination, details of the birth, and the presence or absence of any 
birth defects, congenital abnormalities, or maternal and/or new-born complications. Pregnancy 
outcomes must be collected for females who took the IMP and the female partners of any males 
who took the IMP. Consent to report information regarding these pregnancy outcomes should be 
obtained from the mother.  
13.4. 
Laboratory 
13.4.1. 
Routine Laboratory Assessments 
Routine laboratory safety samples will be analysed by 
at screening 
and prior to discharge at each clinic visit 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 the Principal Investigator or designee 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).  
Samples will be collected into a 2.7 mL ethylenediaminetetraacetic acid (EDTA) Sarstedt 
Monovette® collection tube (or equivalent) and analysed using the Siemens Advia 2120® or 
Siemens Advia 120®. 
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 Sarstedt Monovette 4.7 mL z-gel 
collection tube (or equivalent). This volume allows 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 (or equivalent). Biochemistry samples will be analysed 
using the Roche Cobas c6000 analyser series comprising of c501 and e601 modules. 
Urinalysis: A mid-stream urine sample will be collected into a 20 mL Sterilin tube. Urinalysis 
will be performed using the Siemens Clinitek 500 analyser or the cobas U6500. Tests included in 
the Urinalysis screen are SG, pH, leukocytes, nitrites, protein, glucose, ketones, urobilinogen, 
bilirubin, blood.  

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
56 
 In the event that the ‘dipstick’ is positive for leukocytes, nitrites, protein and blood, RBCs, 
WBCs, epithelial cells, crystals, bacteria and casts will be examined microscopically. Samples of 
menstruating females may not be send for microscopy if blood is positive. The microscopy will 
be performed at 
 using the cobas u6500 701 module or referred to a local specialist 
hospital laboratory for analysis. Other positive parameters will be managed as appropriate.  
13.4.2. 
Expired Air Carbon Monoxide (CO) 
Expired CO will be obtained using the Bedfont Micro+ Smokerlyzer®. Levels for moderate 
smokers are defined as >11 ppm, compared to 0-6 ppm for non-smokers. 
13.4.3. 
Virology 
Virology (Hep B, Hep C and HIV) will be performed at screening.  
Virology will be analysed from the Sarstedt Monovette 4.7 mL z-gel biochemistry screen by 
, S
 using the Roche Cobas c6000 analyser series comprising of c501 
and e601 modules. 
13.4.4. 
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 the Principal Investigator or designee. 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. The Principal Investigator or designee can examine other 
body systems if required, at their discretion. 
13.7. 
ECG Measurements 
ECGs will be conducted using a 12-lead ECG at Screening, Day -1 and again prior to discharge 
from the clinic on Days 4, 13, 17, 21 and 26. Heart rate, PR interval, QRS width, QT interval, 
and QT interval corrected using Fridericia’s formula (QTcF) will be documented.  

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
57 
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. 
13.8. 
12-Lead ECG Holter 
12-lead ECG Holter recording will begin on Day 1 and Day 25, prior to dosing and continue 
until approximately 24 hours post dosing. 
13.9. 
Subject Diary 
All subjects will maintain a daily smoking/medication diary, starting on Day 1 and ending at the 
Post-Study Visit (6-8 days after the last dose). This will include all time spent in the clinic. 
Subjects are to record the number of cigarettes smoked in past 24 hours (daily consumption) and 
actual time they take medication. In addition, subjects are to record any adverse events 
experienced outside of clinic. Clinic personnel are to review diary when subject is in the clinic 
and document the number of cigarettes smoked and the actual time medication was taken.  
13.10. 
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 
328 mL) is considered acceptable. (See Table 10).  
Table 10: 
Summary of Blood Volumes 
Procedure 
Visit 
No. Samples 
Blood 
Volume per 
Sample (mL) 
Total Blood Volume 
(mL) 
Biochemistry  Screeninga 
Day 4, 13, 
17, 21, 26 
1 
5 
4.7 
4.5 
4.7 
22.5 
Haematology  Screening 
Day 4, 13, 
17, 21, 26 
1 
5 
2.7 
2.7 
2.7 
13.5 
PK Drug 
Conc. 
Measurement 
Treatment 
Period 
95 
3.0 
285 
Total Blood Volume 
328 
a From the screening biochemistry blood samples, the serum pregnancy test and virology screen will be conducted.  

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
58 
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. 
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 Population for Analyses 
15.2.1. 
Safety Analysis Set 
All subjects who received at least one dose of cytisine will constitute the safety population.  
15.2.2. 
PK Analysis Set 
The PK Analysis Set will include subjects who have completed all cytisine dosing on Day 1-3, 
completed >90% cytisine dosing on Days 4-25, and comply with the following criteria: 
• Do not have an occurrence of vomiting or severe diarrhoea which renders the 
concentration profile unreliable;  
• Do not use a concomitant medication which renders the concentration profile 
unreliable;  
• 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; 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
59 
• Do not violate the protocol (major protocol violation) in a way that may invalidate or 
bias the PK results. 
15.2.3. 
Pharmacodynamic (PD) Analysis Set 
The PD Analysis Set will include all subjects in the PK Analysis Set who have an available 
baseline result and at least one on-treatment result with regards to urine cotinine, expired air CO 
or daily cigarette consumption and do not incur a major protocol violation in a way that may 
invalidate or bias the PD results. 
15.3. 
Subjects Characteristics 
Separate summaries of demographics will be provided for both safety and PK analysis sets if 
applicable. 
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 approximately 36 subjects will be randomised to the study, with approximately 18 
subjects per treatment schedule arm (12 age 18-65 and 6 age >65).  
No previous pharmacokinetic studies with cytisine have reported the effects of repeated 
administration, so no estimates of intra-subject variability of Cmax, Cmin or AUC are available. No 
specific hypothesis testing is planned. Post-hoc analyses may be performed for PK or PD effects 
that may appear significantly different between the dosage groups or age groups. In general the 
sample size of 36 should be sufficient to meet the exploratory objectives of the study. 
Subjects who withdraw from study at any time after randomisation will not be automatically 
replaced. Replacement based on the number and reason of withdrawals will be at discretion of 
the Sponsor, following discussion with the Principal Investigator.  
15.5. 
Plasma and Urine Concentrations 
Descriptive statistics will be provided to summarise plasma and urine concentrations at each time 
point. Concentrations below the LLOQ will be replaced by zero for calculation of descriptive 
statistics.  
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.  

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
60 
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 11: 
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 Cmax. 
Cmin  
Minimum observed plasma concentration prior to the last daily dose or prior to each 
scheduled dose change in both treatment group A and B. 
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 of cytisine will be derived from the plasma concentrations-time 
profiles, by using a non-compartmental approach with a ln-linear terminal phase assumption. 
Actual times of sampling will be used. 
Cmax will be calculated after the first dose and the last dose on Day 1, after the last dose on days 
2, 3, 12, 16, 20, 24 and after the morning dose on Day 25. 
Cmin will be derived from pre the first dose plasma values on days 4, 13, 17, 21 and 25.  
The t1/2 and AUC (AUC0-t and AUC0-∞) will be determined after the administration of the final 
dose of cytisine on Day 25. 
Derived plasma pharmacokinetic parameters will be listed and summarised by dose group 
(1.5 mg, 3.0 mg), and age (18-65, >65 and overall). 
Urine: The  amount excreted in urine over time (Ae) and percent of drug excreted in urine 
(Ae%) will be derived from Day 1 and Day 25 24 hour urine data for all subjects in the safety 
analysis set. Derived urine PK parameters will be listed and summarised by treatment dosing 
(1.5 mg, 3.0 mg), and age (18-65, >65 and overall).  
15.7. 
Pharmacokinetic Analysis 
Comparisons of Cmax and Cmin between schedule changes within a dose group will be performed 
using analysis of variance (ANOVA) on log-transformed data. Point estimates and 90% 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
61 
confidence intervals (CI) will be constructed for the contrasts between schedule changes which 
will be back-transformed to give estimates of the ratios of the geometric least squares means 
(LSmean) and corresponding 90% CI. This analysis will be performed by age group (18-65, >65 
and overall) within a dose group.  
Comparisons of the Day 25 Cmax, AUC0-t and AUC0-∞ values between dose groups will be 
performed using an ANOVA on log-transformed data. Point estimates and 90% confidence 
intervals (CI) will be constructed for the contrast between the 1.5 mg and 3 mg dose groups 
which will be back-transformed to provide an estimate of the ratio of the geometric LSmean and 
corresponding 90% CI. This analysis will be performed by age group (18-65, >65 and overall). 
15.8. 
Pharmacodynamic Data 
In order to evaluate any reduction in smoking, expired air monoxide (CO), urine cotinine results 
and the number of cigarettes smoked daily will be listed and descriptive statistics of absolute and 
change from baseline results will be tabulated by dose group and age group (18-65, >65 and 
overall). The mean change from baseline values will also be presented graphically. 
15.9. 
Safety Variables and Analyses  
All safety data will be listed using the safety analysis set. Safety summaries will include the 
number of subjects with adverse events, serious adverse events, and adverse events leading to 
discontinuation. Adverse events will be assessed by clinical observation and spontaneous 
reporting by subjects. In addition, number of subjects with clinically significant abnormalities in 
laboratory values will be summarized. 
15.9.1. 
Holter ECGs 
The following ECG parameters will be extracted in triplicate from Holter recordings on Day 1 
and Day 25 at 30 and 15 minutes prior to the first dose and 0.5 h, 1 h, 2 h, 3 h, 4 h, 6 h, 8 h, 12 h 
and 24 h post dose: 
• Heart Rate (HR) 
• PR intervals  
• QRS  
• QT  
• QTcF 
If a large change in heart rate is observed during the study, the individual QT correction method 
(QTcI) will be used as the primary QT correction method. 
The extracted ECG parameters will be listed with any out of normal range values flagged 
(“H” -high or “L” -low). The ECG results will assessed as either “normal” or “abnormal”. Any 
comments on abnormal results will also be provided. Descriptive statistics of absolute and 
change from baseline values at each time point will be tabulated by dose group and age group 
(18-65, >65 and overall).  
In addition, frequencies of QTcF data will be calculated according to the following categories: 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
62 
For absolute values: 
• QTcF ≤ 450 mSec 
• 450 < QTcF ≤ 480 mSec 
• 480 < QTcF ≤ 500 mSec 
• QTcF > 500 mSec. 
For change from baseline: 
• QTcF increase ≤ 30 mSec  
• 30 < QTcF increase ≤ 60 mSec  
• QTcF increase > 60 mSec. 
Abnormalities in ECGs will be classified in terms of clinical significance. Clinically significant 
abnormalities will be reported as AEs. The number and percentage of subjects developing new 
ECG abnormalities will be summarised. The presence of outliers will be explored for heart rate, 
PR, QRS, QT and QTcF. Details of this analysis will be described in the Statistical Analysis Plan 
(SAP). 
In order to explore the effects of cytisine on QTc prolongation, plasma concentrations will be 
plotted against change from baseline QTcF values. 
15.9.2. 
12-Lead 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 SAP. Descriptive statistics of absolute and change from baseline 
values at each time point will be tabulated by dose group and age group (18-65, >65 and overall). 
Abnormalities in ECGs will be classified in terms of clinical significance. Clinically significant 
abnormalities will be reported as AEs. 
15.9.3. 
Vital Signs 
Vital signs parameters will be listed with any out of normal range values flagged (“H” -high or 
“L” -low). Descriptive statistics of absolute and change from baseline values at each time point 
will be tabulated by dose group and age group (18-65, >65 and overall). 
Abnormalities in vital signs will be classified in terms of clinical significance. Clinically 
significant abnormalities will be reported as AEs. 
15.9.4. 
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, resolution date and resolution time; seriousness; 
severity (intensity); relationship (causality); action taken; outcome. 
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 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
63 
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 dose 
group, age group and overall. 
15.9.5. 
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. Descriptive statistics of absolute and change from 
baseline biochemistry and haematology values at each time point will be tabulated by dose group 
and age group (18-65, >65 and overall). 
Abnormalities in clinical laboratory tests will be classified in terms of clinical significance. 
Clinically significant abnormalities will be reported as AEs. 
15.10. 
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). 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
64 
• 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 centralised 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 organisations. Routine monitoring visits will be conducted to: 
1. Assure compliance with the study protocol. 
2. Verify that the research facilities, including laboratories and equipment, are adequate to 
safely and properly conduct the study. 
3. 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. 
4. 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. 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
65 
5. Review the subject CRFs and source documents to ensure that reported study data are 
accurate, complete, and verifiable from source documents. 
6. Ensure that adequate records of clinical trial supplies are maintained. 
7. Verify that the Investigator and study site personnel are adequately qualified throughout 
the study. 
8. 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: 
1. The medical history prior to participation in the study. 
2. The basic identifying information, such as demographics, that link the subject’s source 
documents with the CRFs. 
3. The results of all diagnostic tests performed, diagnoses made, therapy provided, and any 
other data on the condition of the subject. 
4. The subject’s exposure to study treatment. 
5. All AEs. 
6. The subject’s exposure to any concomitant therapy, including dates of administration. 
7. All relevant observations and data on the condition of the subject throughout the study. 
8. 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 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
66 
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. 
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, 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
67 
stating the reasons for premature termination. Si
ec 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.  
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.  

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
68 
22. 
INVESTIGATOR’S AGREEMENT  
 
Protocol No. ACH-CYT-02 
 
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 
 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
69 
23. 
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. 
Etter J. Cytisine for smoking cessation: A literature review and a meta-analysis. Archives 
of Internal Medicine 2006;166:1553-9. 
6. 
Cahill K, Lindson-Hawley N, Thomas KH, Fanshawe TR, Lancaster T. Nicotine receptor 
partial agonists for smoking cessation. The Cochrane database of systematic reviews 
2016:Cd006103. 
7. 
Rigotti NA. Cytisine--a tobacco treatment hiding in plain sight. The New England journal 
of medicine 2014;371:2429-30. 
8. 
Aveyard P, West R. Cytisine and the failure to market and regulate for human health. 
Thorax 2013;68:989. 
9. 
Walker N, Bullen C, Barnes J, et al. Getting cytisine licensed for use world-wide: a call 
to action. 2016;111:1895-8. 
10. 
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. 
11. 
Raw M, Anderson P, Batra A, et al. WHO Europe evidence based recommendations on 
the treatment of tobacco dependence. Tobacco control 2002;11:44-6. 
12. 
Heatherton TF, Kozlowski LT, Frecker RC, Fagerstrom KO. The Fagerstrom Test for 
Nicotine Dependence: a revision of the Fagerstrom Tolerance Questionnaire. British journal of 
addiction 1991;86:1119-27. 
13. 
Heishman SJ, Singleton EG, Pickworth WB. Reliability and validity of a Short Form of 
the Tobacco Craving Questionnaire. Nicotine & tobacco research : official journal of the Society 
for Research on Nicotine and Tobacco 2008;10:643-51. 
 
 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
70 
APPENDIX 1. NORMAL RANGES FOR VITAL SIGNS AND ECG 
PARAMETERS 
 
 
Vital Signs: 
 
PARAMETER 
Normal Range for 
ages 18 to 65 
Normal Range for 
ages > 65 
Units 
Pulse 
40 - 110 
40 - 110 
Beats per Minute 
(BPM) 
Systolic Blood 
Pressure 
90 - 150 
90 - 160 
mmHg 
Diastolic Blood 
Pressure 
50 - 90 
50 - 90 
mmHg 
Oral Temperature 
35 - 37.5 
35 - 37.5 
Degree Celsius 
 
ECG Parameters: 
 
PARAMETER 
Normal Range for all ages  
Units 
Heart Rate 
40 - 110 
Beats per Minute (BPM) 
PR Interval 
120 - 220 
msec 
QRS width 
70 - 120 
msec 
QT 
N/A - should be corrected for heart rate 
QTcB - males 
350 - 430 
msec 
QTcB - females 
350 - 450 
msec 
QTcF - males 
350 - 430 
msec 
QTcF - females 
350 - 450 
msec 
 
 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
71 
APPENDIX 2. FAGERSTRÖM NICOTINE DEPENDENCE 
QUESTIONNAIRE 
 
The Fagerström Nicotine Dependence Questionnaire (FTND) has been shown to be a reliable 
and valid measure of nicotine dependence12 and will be administered as outlined in this protocol. 
Analysis of results is outlined in the study Statistical Analysis Plan. 
 
Question 
Response 
Points* 
How soon after you wake up do you smoke your 
first cigarette? 
Within 5 minutes 
 
3 
6-30 minutes 
 
2 
31-60 minutes 
 
1 
After 60 minutes 
 
0 
 
 
Do you find it difficult to refrain from smoking in 
places where it is forbidden (for example in church, 
at the library, in a cinema, etc.)? 
Yes 
 
1 
No 
 
0 
 
 
Which cigarette would you hate most to give up? 
The first one in the 
morning 
 
1 
All others 
 
0 
 
 
How many cigarettes/day do you smoke? 
10 or less 
 
0 
11-20 
 
1 
21-30 
 
2 
31 or more 
 
3 
 
 
Do you smoke more frequently during the first 
hours after waking than during the rest of the day? 
Yes 
 
1 
No 
 
0 
 
 
Do you smoke if you are so ill that you are in bed 
most of the day? 
Yes 
 
1 
No 
 
0 
 
 
*Note: The actual questionnaire will not contain the points column, which is provided here to 
show how responses will be scored for analysis. 
 

Protocol ACH-CYT-02, 21 September, 2017, Version 2.0  
  CONFIDENTIAL 
 
Cytisine  
 
 
 
72 
APPENDIX 3. SHORT FORM OF THE TOBACCO CRAVING 
QUESTIONNAIRE 
 
The template below has been validated as a self-report instrument to measure tobacco craving13 
and will be administered to study subjects as outlined in this protocol. Analysis of results is 
outlined in the study Statistical Analysis Plan. 
 
 
Question 
Response 
 
Strongly Disagree....Strongly Agree 
Factor 1 (emotionality) 
I would be less irritable now if I could smoke. 
 
 
 
 
 
 
 
If I were smoking now I could think more clearly. 
 
 
 
 
 
 
 
I could control things better right now if I could smoke. 
 
 
 
 
 
 
 
Factor 2 (expectancy) 
I would enjoy a cigarette right now. 
 
 
 
 
 
 
 
A cigarette would taste good right now. 
 
 
 
 
 
 
 
Smoking a cigarette would be pleasant. 
 
 
 
 
 
 
 
Factor 3 (compulsivity) 
If I smoked right now, I would not be able to stop. 
 
 
 
 
 
 
 
I could not stop myself from smoking if I had some 
cigarettes here. 
 
 
 
 
 
 
 
Factor 4 (purposefulness) 
If I had a lit cigarette in my hand, I probably would 
smoke it. 
 
 
 
 
 
 
 
It would be hard to pass up the chance to smoke. 
 
 
 
 
 
 
 
I could not easily limit how much I smoked right now.