Clinical trial NCT04756960
Conditions: Chronic Obstructive Pulmonary Disease (COPD) COPD
Keywords: Chronic Obstructive Pulmonary Disease COPD Healthy subjects Phase I CHF6001 Tanimilast Chiesi Farmaceutici S.p.A
Sponsor: Chiesi Farmaceutici S.p.A.
Start date: 2021-03-10; Completion date: 2021-04-29
Enrollment: 8 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
The objective of the study is to evaluate the bioavailability of CHF6001 after inhaled administration, to characterize the mass balance and route of elimination of CHF6001 along with its relevant metabolites, in healthy male subjects.

Detailed description:
This clinical trial is a single centre Phase I study, with a single dose, non-randomized, open-label, uncontrolled design. A total of 8 healthy male subjects were included in the study.

The aim was to assess the absolute bioavailability, the mass balance, and routes of elimination of CHF6001 (Tanimilast) in healthy male subjects, using \[14\^C\]-radiolabelled drug substance, administered as an intravenous (iv) infusion concomitantly with an inhaled (inh) non-radiolabelled dose of CHF6001.

Standard safety assessments were conducted during the study, including safety blood and urine laboratory tests, vital signs, physical examinations, ECGs, and assessment of any adverse events (AE). Blood, urine, and feces samples were collected for pharmacokinetic (PK) analyses.

Arms: arm 1: single dose of CHF6001 dry-powder inhaler (DPI) co-administered with an intravenous microdose of \[14\^C\]-labelled CHF6001
Interventions: intervention 1: CHF6001
Intervention descriptions: intervention 1: 4 inhalations of CHF6001 800 µg/20 mg NEXThaler® dry-powder inhaler (DPI), (total dose of 3200µg) co-administered with an intravenous microdose (18.5µg (18.5 kBq)) of \[14\^C\]-labelled CHF6001

Protocol documents:
Prot_000.pdf: 
CONFIDENTIAL     
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CLINICAL STUDY PROTOCOL 
STUDY CODE No.: CLI-06001AA1-02
EUDRACT No.: 2020-004201-31 
EVALUATION OF THE ABSOLUTE BIOAVAILABILITY AND MASS 
BALANCE OF CHF6001 FOLLOWING A SINGLE INHALED DOSE CO-
ADMINISTERED WITH AN INTRAVENOUS RADIOLABELLED 
MICROTRACER DOSE IN HEALTHY VOLUNTEERS
Version No.: 1.0
Date: 19 October 2020
The information contained in this document is confidential and will not be disclosed to others without written 
authorization from Chiesi Farmaceutici S.p.A., except to the extent necessary to obtain informed consent from those 
persons to whom the drug may be administered or for discussions with local regulatory authorities, Ethics 
Committee/Investigational Review Boards, or people participating in the conduct of the study.
Chiesi Farmaceutici S.p.A.
Via Palermo 26/A
43122 Parma - Italy
NCT no.: NCT04756960


Clinical Study Protocol
Clinical Study Code: CLI-06001AA1-02
Version No.: 1.0
EUDRACT No.: 2020-004201-31
Date: 19 October 2020
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GENERAL INFORMATION
SPONSOR:
Chiesi Farmaceutici S.p.A.* 
Via Palermo 26/A
43122 Parma - Italy
+ 39 0521 2791
*also reported as Chiesi thereafter 
SPONSOR MEDICAL EXPERT
(Clinical Research Physician)
, MD
Clinical Research Physician
Global Clinical Development
Tel.: 
            
Mobile: 
E-mail: 
Readily available in case of medical 
questions
INVESTIGATOR
(or Coordinating Investigator)
, MB ChB
 
MONITORING CRO
 
 
CENTRAL BIOANALYTICAL 
LABORATORY:
  
CENTRAL LABORATORY FOR 
RADIOANALYSIS
VERSION HISTORY
Version 
Date 
Change History
1.0
19 October 2020
First version


Clinical Study Protocol
Clinical Study Code: CLI-06001AA1-02
Version No.: 1.0
EUDRACT No.: 2020-004201-31
Date: 19 October 2020 
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PROTOCOL OUTLINE
Study title
Evaluation of the absolute bioavailability and mass balance of CHF6001 
following a single inhaled dose co-administered with an intravenous 
radiolabelled microtracer dose in healthy volunteers
Sponsor
Chiesi Farmaceutici S.p.A. - Via Palermo 26/A 43122 Parma - Italy
Name of the Product
CHF6001
Centre(s)
Single centre
Indication
Chronic Obstructive Pulmonary Disease (COPD) (study conducted in 
healthy subjects)
Study design
Open-label, uncontrolled, non-randomized, single dose
Study phase
Phase I
Objectives
The objectives of this study will be: 

To determine the absolute bioavailability of CHF6001 following a 
single inhaled dose co-administered with an intravenous 
microtracer dose

To characterize the mass balance and the elimination routes of 
CHF6001 after an intravenous microtracer dose

To characterize the relevant metabolites in plasma, urine and feces 

To assess the safety and tolerability of the study treatment
Treatment duration
One single dose treatment
Test product
dose/route/regimen
4 inhalations of CHF6001 800µg/20mg NEXThaler® DPI (total dose of 
3200µg) co-administered with an intravenous microdose (18.5µg and 
500nCi (18.5kBq)) of [14C]-labelled CHF6001.
Number of subjects
8 subjects 
Study population
Healthy male subjects
Inclusion/exclusion criteria
Inclusion criteria:
Subjects must meet all of the following inclusion criteria at screening, to 
be eligible for enrolment into the study:
1.
Subject’s written informed consent obtained prior to any study-
related procedure;
2.
Able to understand the study procedures, the risks involved and 
ability to be trained to use correctly the inhalers and to generate 
sufficient PIF (at least 40 L/min) using the In-Check device set as 
per NEXThaler® inhaler resistance and Placebo inhaler;
3.
Male subjects aged 30 to 55 years inclusive;
4.
Body mass index (BMI) within the range of 18 to 35 kg/m2 
inclusive;
5.
Non- or ex-smoker who smoked < 5 pack years (pack-years = the 
number of cigarette packs (20 cigarettes) or equivalent per day times 
the number of years) and who stopped smoking > 1 year prior to 
screening;
6.
Good physical and mental status, determined on the basis of the 
medical history and a general clinical examination, at screening and 
before treatment;
7.
Vital signs at screening within limits defined as: Diastolic Blood 
Pressure 40-90 mmHg and Systolic Blood Pressure 90-140 mmHg, 


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(three) measures performed in supine position after at least 5 
minutes of resting. The average from triplicate measurements must 
be within the defined range; 
Note: In case the values are outside the defined ranges, the 
assessment can be repeated once before treatment.
8.
12-lead digitised Electrocardiogram (12-lead ECG) in triplicate 
considered as normal (40 ≤ Heart rate ≤ 110 bpm, 120 ms ≤ PR ≤ 
210 ms, QRS ≤ 120 ms, QTcF ≤ 450 ms) at screening visit. The 
average from triplicate measurements must be within the defined 
range;  
Note: In case of values outside the defined ranges, the assessment 
can be repeated once before treatment.
9.
Lung function measurements within normal limits at screening 
(Normal values: FEV1/FVC > 0.70 and FEV1 >80% predicted);
Note: In case the values are outside the defines values, the 
assessment can be repeated once before treatment.
10. Regular bowel movements (averaging one or more bowel 
movements per day) at screening; 
11. Males fulfilling one of the following criteria: 
a.
Males with non-pregnant WOCBP partners: they and/or 
their partner of childbearing potential must be willing to 
use a highly effective birth control method in addition to 
the male condom from the signature of the informed 
consent and until 90 days after the follow-up visit or
b.
Males with pregnant WOCBP partner: they must be 
willing to use male contraception (condom) from the 
signature of the informed consent and until 90 days after 
the follow-up visit or
c.
Non-fertile male subjects (contraception is not required in 
this case) or
d.
Males with partner not of childbearing potential 
(contraception is not required in this case). 
Moreover, subjects must not donate sperm during the study and for 90 
days after the follow-up visit. For the definition of WOCBP and of 
fertile men and the list of highly effective birth control methods, refer 
to Appendix 2 (or section 4.1 of the CTFG guidance for more detailed 
information). 
Inclusion criteria 7, 8 and 9 could be checked again once before treatment.

If the criteria are met after the re-check, the subject can continue 
the study.

If the criteria are still not met, the subject will be discontinued and 
screen failed.
The inclusion criterion 6 will be re-checked at Day -1 

If the criterion is met, the subject can continue the study.

If the criterion is not met, the subject will be discontinued and 
screen failed.
Exclusion criteria:


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The presence of any of the following exclusion criteria at screening will 
exclude a subject from study enrolment:
1.
Participation in another clinical trial with an investigational drug in 
the 3 months or 5 half-lives of that investigational drug (whichever is 
longer) preceding the administration of the study drug; a longer and 
more appropriate time could be considered by the principal 
investigator based on the elimination half-life and/or long-term 
toxicity of the previous investigational drug;
2.
Clinically relevant and uncontrolled respiratory, cardiac, hepatic 
(including Gilbert syndrome), gastrointestinal, renal, endocrine, 
metabolic, neurologic, or psychiatric disorder that may interfere with 
successful completion of this protocol according to Investigator 
judgement;
3.
Clinically relevant abnormal laboratory values at screening suggesting 
an unknown disease and requiring further clinical investigation or 
which may impact the safety of the subject or the evaluation of the 
study results according to Investigator judgement;
Note: In case of abnormal laboratory values, that could indicate a 
temporary condition, the test can be repeated once before treatment.
4.
Subjects with history of breathing problems (e.g. history of asthma 
including childhood asthma);
5.
Positive to HIV1 or HIV2 serology at screening;
6.
Positive results from the Hepatitis serology which indicates acute or 
chronic Hepatitis B or Hepatitis C at screening (i.e. positive HB 
surface antigen (HBsAg), HB core antibody (IgM anti-HBc), HC 
antibody);
7.
Blood donation or blood loss (equal or more than 450 mL) less than 2 
months prior screening or prior to treatment;
8.
Positive urine test for cotinine at screening or before treatment;
9.
Documented history of alcohol abuse within 12 months prior to 
screening or a positive alcohol breath test at screening or before 
treatment;
10. Documented history of drug abuse within 12 months prior to screening 
or a positive urine drug screen at screening or before treatment;
11. Intake of non-permitted concomitant medications in the predefined 
period prior to screening or before treatment or the subject is expected 
to take non-permitted concomitant medications during the study (see 
“non-permitted concomitant medications” section);
12. Presence of any current infection, or previous infection that resolved 
less than 7 days prior to screening or before treatment;
13. Known intolerance and/or hypersensitivity to any of the excipients 
contained in the formulation used in the trial;
14. Unsuitable veins for repeated venipuncture;
15. Heavy caffeine drinker (> 5 caffeinated beverages e.g., coffee, tea, 
cola per day);
16. Abnormal haemoglobin level defines as <13mg/l for males and 
<11.3mg/l for female at screening;
17. Subjects using e-cigarettes within 6 months prior to screening;


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18. Subjects been involved in a study involving a 14C-labeled drug within 
the 12 months prior to enrollment;
19. Subjects with exposure to significant diagnostic or therapeutic 
radiation (eg, serial X-ray, computed tomography scan, barium meal) 
or current employment in a job requiring radiation exposure 
monitoring within 12 months prior to Day-1;
20. Documented COVID-19 diagnosis within the last 8 weeks or which 
has not resolved within 14 days prior to screening and before 
treatment.
The exclusion criterion 3, if met at the screening visit, can be checked 
again once before the treatment:

If the criterion is not met after re-check, the subject can continue 
the study.

If the criterion is still met, the subject will be discontinued and 
screen failed.
The following exclusion criteria 7, 8, 9, 10, 11, 12 and 20 will be re-
checked at Day -1 visit: 

If all criteria are not met, the subject can continue the study.

If at least one criterion is met, the subject will be discontinued and 
screen failed.
Study plan
After the screening visit, which will take place between 3 to 31 days prior 
to dosing, the eligible subjects will enter the study that comprises one 
single dose treatment period. 
During the treatment period, the subjects will come to the clinical site on 
Day -1. On Day 1, they will be administered with 4 inhalations of 
CHF6001 NEXThaler® 800µg followed by a 15 min intravenous infusion 
of the tracer (injection of 10 mL [14C]-CHF6001 1.85µg/mL) ending at the 
expected Tmax of 2 h for the inhaled dose, giving a total infused dose of 
18.5µg and 500nCi (18.5kBq). Inhaled drug administration will occur in 
fasting conditions (at least 10 hours fasting prior to administration) and the 
subjects will remain fasted until 2 hours post dose.
The subjects will remain at the clinical site under permanent supervision 
of the medical staff, until the morning of Day 11. They will be discharged 
after the 240h post-dose (i.v.) assessments on Day 11 are completed. 
A follow-up phone call (or visit, if necessary) will be performed from 7 to 
10 days after discharge or premature discontinuation to check and record 
concomitant medications and the status of any unresolved adverse events 
or any new adverse events
Most 
relevant 
allowed 
concomitant treatments
Occasional paracetamol (maximum 2 g per day with a maximum of 10 g 
per 14 days for mild conditions not meeting exclusion criteria).
Most 
relevant 
forbidden 
concomitant treatments

Intake of any enzyme-inducing drugs, enzyme-inhibiting drugs, 
biologic drugs or any drug known to have a well-defined potential 
for hepatotoxicity (e.g. isoniazide, nimesulide, ketoconazole) are 
not permitted within 3 months prior to screening and throughout 
the study period (including the follow up period)


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
Intake of any drug treatment, including prescribed or OTC 
medicines, vitamins, homeopathic remedies etc., with the 
exception of the allowed concomitant medications specified in the 
relative section are not permitted within 14 days prior to screening 
and throughout the study period including the follow up period
Pharmacokinetics
variables
Following intravenous (IV) administration: 

Plasma [14C]-CHF6001 AUC0-t iv, Cmax_iv, tmax_iv, AUC0-∞_iv, t1/2 iv, 
Vz, Vdss and systemic clearance (CL) 

Plasma [14C]-total AUC0-t iv, Cmax_iv, tmax_iv, AUC0-∞_iv, t1/2 iv

Urine and feces [14C]-CHF6001 and [14C]-total excreted fraction 
(Feu, Fef and Feu+f) 

Systemic blood clearance (CLblood) [14C]-CHF6001

Renal plasma and blood clearance (CLR and CLR_blood) [14C]-
CHF6001

Fraction of the relevant metabolites in plasma, and fraction of the 
relevant metabolites recovered in urine and feces (Fr_plasma, 
Fr_urine, Fr_feces, Fr_urine+feces)

Metabolized fraction (Femet) 

Blood to plasma ratio (Rb/p) [14C]-CHF6001

Blood to plasma ratio (Rbt/pt) [14C]-total

Hepatic extraction (Eh_blood) [14C]-CHF6001
Following inhaled administration: 

Plasma CHF6001 AUC(0-t) inh, Cmax_inh, tmax_inh, AUC(0-∞)_inh, t1/2_inh, 
absolute inhaled bioavailability (Finh)  
Measurement of PK 
variables

Blood samples for plasma non-radiolabeled CHF6001 to be 
collected at pre-dose (within 60 min from inhaled dosing) and at 
the following timepoints post inhaled dose: 15, 30, 60, 90 min, 2, 
3.75, 5.75, 7.75, 9.75, 11.75, 13.75, 25.75, 49.75, 73.75, 97.75, 
121.75, 145.75, 169.75, 193.75, 217.75 and 241.75 hours.

Blood samples for plasma [14C]-total, [14C]-CHF6001, [14C] of 
the relevant metabolites to be collected at pre-dose (within 60 min 
from inhaled dosing) and at the following timepoints relative to 
the start of the IV infusion: 5, 10, 15 [end of  infusion], 20, 25, 30, 
45, 60 min, 2, 4, 6, 8, 10, 12, 24, 48, 72, 96, 120, 144, 168, 192, 
216 and 240 hours. 

Blood samples for whole blood [14C]-total and [14C]-CHF6001 at 
pre-dose (within 60 min from inhaled dosing) and at 20 min after 
the start of IV infusion.

Urine samples for assessments of [14C]-total, [14C]-CHF6001 and 
[14C] of the relevant metabolites to be collected at pre-dose from  
-12 hours to immediately before study drug inhaled administration 


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(-12-0h) and at the following time frames relative to the start of 
the IV infusion:  0-4h, 4-8h, 8-12h, 12-24h, 24-48h, 48-72h, 72-
96h, 96-120h, 120-144h, 144-168h, 168-192h, 192-216h and 216-
240h.

Fecal samples for assessments of [14C]-total, [14C]-CHF6001 and 
[14C] of the relevant metabolites to be collected at pre-dose from 
check-in (Day-1) to immediately before study drug inhaled 
administration (check-in -0h) and at the following time frames 
relative to the start of the IV infusion:  0-24h, 24-48h, 48-72h, 72-
96h, 96-120h, 120-144h, 144-168h, 168-192h, 192-216h and 216-
240h.
Characterization of the relevant metabolites
For metabolites in plasma accounting for ≥10% of total radioactivity 
recovered in this matrix, structural characterization may be performed, 
where possible.
For metabolites in urine and feces (total excreta) accounting for ≥10% of 
total administered radioactivity, structural characterization may be 
performed, where possible. 
Safety variables

Adverse Events (AEs) and Adverse Drug Reactions (ADRs).

Systolic (SBP) and Diastolic (DBP) Blood Pressure.

Heart rate (HR) from local 12-lead safety ECG. 

Clinical laboratory evaluations (chemistry, haematology and fasting 
glucose)  
Measurement of safety 
variables

Vital Signs: blood pressure will be evaluated after 5 min in supine 
position at pre-dose and at 2,5 h post-dose

Local 12-lead Safety ECG: HR will be evaluated at pre-dose and at 2,5 
hours post dose

Chemistry, haematology and fasting glucose at screening and Day 11
Sample size calculation
According to the exploratory nature of the study, no formal sample size 
calculation was performed. A total of 8 subjects will be included in the 
study to characterize the absolute bioavailability and the mass balance of 
CHF6001.
Statistical methods
Pharmacokinetic variables
All PK variables (except for tmax, tmax_inh and tmax_iv) for plasma, urine and 
feces [14C]-total, [14C]-CHF6001 and non-radiolabeled CHF6001 will be 
summarized by means of descriptive statistics including n (number of 
observed values), arithmetic mean, standard deviation (SD), coefficient of 
variation (CV), geometric mean, geometric CV, median, minimum and 
maximum. tmax, tmax_inh, tmax_iv will be summarized by using n, median, 
minimum and maximum.
Concentration (mass equivalent per mL or mass per mL)/time curves for 
plasma [14C]-total, [14C]-CHF6001, and non-radiolabeled CHF6001 will be 
presented in linear/linear and log/linear scale. Plots will be presented based 
on arithmetic means and by subject (all individual plasma concentration/time 
curves in one graph).


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EUDRACT No.: 2020-004201-31
Date: 19 October 2020 
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Bar charts based on arithmetic means showing the percentage of urine and 
feces excreted fraction for [14C]-total, and [14C]-CHF6001 in the different 
time intervals will be presented.
Bar charts based on arithmetic means showing percentages of the relevant 
metabolites recovered in plasma, urine and feces will be presented.
All individual concentration (mass equivalent per mL or mass per mL) data 
and PK parameters will be listed. In addition, a listing of the actual sampling 
times relative to drug inhalation will be provided. Concentrations (mass 
equivalent per mL or mass per mL) will be summarized by scheduled 
sampling time by using n, arithmetic mean, SD, CV, median, minimum and 
maximum.
Safety Variables

The number and percentage of subjects experiencing adverse events, 
adverse drug reactions, serious adverse events and adverse events 
leading to study withdrawal will be summarized.

Percentage of subjects with clinical relevant changes at 2,5 hrs post-
dose in SBP, DBP and HR (from 12-lead ECG) defined as
o
Change from pre-dose in DBP > 10 mmHg; 
o
Change from pre-dose in SBP > 20 mmHg;
o
Change from pre-dose in in HR > 20 bpm
          will be summarized.

For quantitative laboratory parameters (chemistry, haematology and 
fasting glucose), shift tables from screening with regards to normal 
range will be presented.


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LIST OF ABBREVIATIONS AND DEFINITION OF TERMS
ADR
Adverse Drug Reaction
AE
Adverse Event
Ae
Excreted amount
ALT
Alanine Aminotransferase
AMS
Accelerator Mass Spectrometry
AST
Aspartate Aminotransferase
ATC
Anatomical Therapeutic Chemical classification
AUC
Area Under the Curve
AUC0-t
Area Under the Curve, from 0 to the last quantifiable concentration
AUC0-∞
Area Under the Curve, from 0 to infinite 
BMI
Body Mass Index
BP
Blood Pressure
BTPS
Body Temperature and Pressure, Saturated 
CI 
Confidence Interval
CIOMS
Council for International Organizations of Medical Sciences
CL
Systemic Clearance
CLblood
Total body blood clearance
CLR
Renal plasma clearance
CLR_blood
Renal blood clearance
Cmax
Maximum of concentration 
Cmin
Minimum of concentration 
COPD
Chronic Obstructive Pulmonary Disease
CRO
Contract Research Organization
CTFG
Clinical Trial Facilitation Group
CV
Coefficient of Variation
DBP
Diastolic Blood Pressure
DPI
Dry-powder inhaler
DRR
Data Review Report
EC
Ethics Committee
ECG
Electrocardiogram 
eCRF
Electronic Case Report Form
EDTA
Ethylenediaminetetraacetic acid
Eh_blood
Hepatic extraction
EMA
European Medicines Agency
FDA
Food and Drug Administration
fe
Percent of the dose excreted over a sampling interval
Fe
Cumulative percent of dose excreted 
FEV1
Forced Expiratory Volume in the first second
Finh
Absolute Inhaled Bioavailability
Femet
Metabolized fraction
Fr
Fraction of the relevant metabolites
FSH
Follicle-stimulating hormone
FVC
Forced Vital Capacity


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GCP
Good Clinical Practice
GLP
Good Laboratory Practice 
GMP
Good Manufacturing Practice
GOLD
Global Initiative for Chronic Obstructive Lung Disease
ɣ-GT
Gamma-Glutamyl Transpeptidase
hAME
Human Absorption Metabolism and Excretion
Hb
Total haemoglobin
HB
Hepatitis B
Anti HBc
Hepatitis B core antibody
HBsAg
Hepatitis B surface Antigen
Hct
Haematocrit
HCV
Hepatitis C
HIV1/HIV2
Human Immunodeficiency Virus 1/Human Immunodeficiency Virus 2
HR
Heart Rate 
HV
Healthy Volunteers
ICF
Informed Consent Form
ICH
International Conference on Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use
ICS
Inhaled Corticosteroid 
IMP
Investigational Medicinal Product
IRB
Institutional Review Board
IV
Intravenous
K
Potassium
LABA
Long Acting β2-Agonist
LAMA
Long Acting Muscarinic Antagonist
LC+AMS
Liquid Chromatography-Accelerator Mass Spectrometry
LC-MS/MS
Liquid Chromatography Tandem Mass Spectrometry 
LLOQ
Lower Limit Of Quantification 
LRTI
Lower Respiratory Tract Infection
MedDRA
Medical Dictionary for Regulatory Activities
MTD
Maximum Tolerated Dose
Na
Sodium
OECD
Organisation for Economic Co-operation and Development
OTC
Over The Counter
PDE4
Phosphodiesterase-4
PIF
Peak Inspiratory Flow
PK
Pharmacokinetic
PLT
Platelets count
PR
Time Interval Between the P and R wave in the ECG
QRS
Time Interval Between the Q and R and S wave in the ECG
QTcF
Fridericia corrected Time Interval Between the Q and T wave in the ECG
RBC
Red Blood Cells
Rb/p
Whole blood to plasma ratio
SAAC
Short Acting Anticholinergics
SABA
Short Acting Beta2 Agonists
SAE
Serious Adverse Event


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SAP
Statistical Analysis Plan
SBP
Systolic Blood Pressure
SD
Standard Deviation
SOP
Standard Operating Procedure
SUSAR
Suspected Unexpected Serious Adverse Reaction
tmax 
Time to reach the Cmax
tmin 
Time to reach the Cmin 
t½
Elimination half-life
TEAE
Treatment Emergent Adverse Event
TP
Treatment Period
Vdss
Volume of distribution at steady-state
Vz
Volume of distribution during the terminal phase
WBC
White Blood Cells
WHO
World Health Organization
WOCBP
Women of childbearing potential 
WONCBP
Women of non-childbearing potential
Note: 
“Chiesi Farmaceutici S.p.A.” is also reported as “Chiesi” throughout the text.


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CONTENT
GENERAL INFORMATION......................................................................................................................................................2
PROTOCOL OUTLINE...............................................................................................................................................................3
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS.......................................................................................10
CONTENT.....................................................................................................................................................................................13
1.
INTRODUCTION .............................................................................................................................................................15
1.1 BACKGROUND INFORMATION ...............................................................................................................................................................15
1.2 STUDY RATIONALE...................................................................................................................................................................................16
1.3 RISK/BENEFIT ASSESSMENT...................................................................................................................................................................16
2.
STUDY OBJECTIVES.....................................................................................................................................................17
3.
STUDY DESIGN ...............................................................................................................................................................17
4.
SUBJECT SELECTION CRITERIA ...........................................................................................................................17
4.1 SUBJECT RECRUITMENT..........................................................................................................................................................................17
4.2 INCLUSION CRITERIA...............................................................................................................................................................................18
4.3 EXCLUSION CRITERIA..............................................................................................................................................................................19
4.4 SUBJECT WITHDRAWALS........................................................................................................................................................................20
5.
CONCOMITANT MEDICATIONS .............................................................................................................................21
5.1 PERMITTED CONCOMITANT MEDICATIONS.......................................................................................................................................21
5.2 NON-PERMITTED CONCOMITANT MEDICATIONS.............................................................................................................................21
6.
TREATMENT(S)...............................................................................................................................................................21
6.1 
 
 
................................................................................................................................................................21
6.2 DOSAGE AND ADMINISTRATION ..........................................................................................................................................................22
6.3 PACKAGING ................................................................................................................................................................................................24
6.4 LABELING....................................................................................................................................................................................................25
6.5 TREATMENT ALLOCATION......................................................................................................................................................................25
6.6 TREATMENT CODE ...................................................................................................................................................................................25
6.7 TREATMENT COMPLIANCE .....................................................................................................................................................................25
6.8 DRUG STORAGE.........................................................................................................................................................................................26
6.9 DRUG ACCOUNTABILITY........................................................................................................................................................................26
6.10 PROVISION OF ADDITIONAL CARE......................................................................................................................................................26
7.
STUDY PLAN ....................................................................................................................................................................27
7.1 STUDY SCHEDULE ....................................................................................................................................................................................27
7.2 INVESTIGATIONS .......................................................................................................................................................................................32
8.
PHARMACOKINETIC ASSESSMENTS...................................................................................................................37
9.
SAFETY ASSESSMENTS...............................................................................................................................................40
10.
ADVERSE EVENT REPORTING................................................................................................................................40
10.1 DEFINITIONS.............................................................................................................................................................................................40
10.2 EXPECTEDNESS .......................................................................................................................................................................................41
10.3 INTENSITY OF ADVERSE EVENT.........................................................................................................................................................42
10.4 CAUSALITY ASSESSMENT....................................................................................................................................................................42
10.5 ACTION TAKEN WITH THE STUDY DRUG DUE TO AN AE.............................................................................................................42
10.6 OTHER ACTIONS TAKEN........................................................................................................................................................................43
10.7 OUTCOME .................................................................................................................................................................................................43
10.8 RECORDING ADVERSE EVENTS..........................................................................................................................................................43
10.9 REPORTING SERIOUS ADVERSE EVENTS TO CHIESI.....................................................................................................................43
10.10 REPORTING SERIOUS ADVERSE EVENTS TO REGULATORY AUTHORITIES/ETHICS COMMITTEES/IRB.....................44


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10.11 GENERAL NOTES..................................................................................................................................................................................44
11.
DATA MANAGEMENT..................................................................................................................................................45
12.
STATISTICAL METHODS............................................................................................................................................45
12.1 SAMPLE SIZE............................................................................................................................................................................................45
12.2 POPULATIONS FOR ANALYSIS .............................................................................................................................................................46
12.3 STATISTICAL ANALYSIS........................................................................................................................................................................46
13.
ETHICS COMMITTEE/INSTITUTIONAL REVIEW BOARD APPROVAL..................................................48
14.
REGULATORY REQUIREMENTS ............................................................................................................................48
15.
INFORMED CONSENT..................................................................................................................................................48
16.
SOURCE DOCUMENTS/DATA ...................................................................................................................................48
16.1 RECORDING OF SOURCE DATA............................................................................................................................................................48
16.2 DIRECT ACCESS TO SOURCE DOCUMENT/DATA.............................................................................................................................49
17.
STUDY MONITORING ..................................................................................................................................................49
18.
QUALITY ASSURANCE................................................................................................................................................49
19.
INSURANCE AND INDEMNITY.................................................................................................................................49
20.
CONFIDENTIALITY ......................................................................................................................................................50
21.
PREMATURE TERMINATION OF THE STUDY..................................................................................................50
22.
CLINICAL STUDY REPORT .......................................................................................................................................50
23.
RECORD RETENTION..................................................................................................................................................50
24.
PUBLICATION OF RESULTS .....................................................................................................................................50
25.
REFERENCES...................................................................................................................................................................51
APPENDIX 1 - APPROVAL OF THE PROTOCOL BY CLINICAL INVESTIGATOR(S) ....................................53
APPENDIX 2 – RECOMMENDATIONS RELATED TO CONTRACEPTION AND PREGNANCY TESTING 
IN CLINICAL TRIALS .............................................................................................................................................................54
APPENDIX 3 – BREAKDOWN OF BLOOD SAMPLES COLLECTION ...................................................................55


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1.
INTRODUCTION
1.1 Background information
The pathogenesis and progression of chronic obstructive pulmonary disease (COPD) is, in part, due 
to chronic inflammation [1]. However, the nature and severity of inflammation in COPD varies, and 
pharmacological anti-inflammatory treatments are unlikely to be effective in all patients; a precision 
medicine approach is needed to selectively target patients to increase the chance of therapeutic 
success [2]. 
Phosphodiesterase-4 (PDE4) is an enzyme that mediates the breakdown of cyclic adenosine 
monophosphate (cAMP), with PDE4 inhibition having anti-inflammatory effects in a broad range of 
cell types. The orally administered PDE4 inhibitor roflumilast prevents exacerbations in patients with 
COPD [3], [4]. However, systemic exposure after roflumilast administration can cause side effects 
such as nausea, weight loss and gastrointestinal disturbance, which may limit its use in clinical 
practice [5].
CHF6001 is a novel inhaled PDE4 inhibitor [6], currently in clinical development that has been 
specifically designed as an extra-fine formulation to be delivered via inhalation and to have a low 
systemic exposure. This allows CHF6001 to reach therapeutic concentration in the target organ, the 
lung, yet reduces exposure in the systemic circulation thus limiting systemic adverse effects [7]. 
Safety, tolerability and the PK profile of CHF6001 were evaluated in healthy volunteers (HV) in two 
phase 1 trials in which doses up to 4800µg/day were administered up to 14 days. All doses of 
CHF6001 were safe and well tolerated, both after single and repeated administration. Neither apparent 
gastrointestinal intolerance nor cardiovascular safety signal was noted [8]. 
Clinical pharmacology, safety, tolerability and PK profile of CHF6001 were investigated in an 
allergen challenge model in mild asthmatic subjects [9] and in subjects with COPD [10] evaluating 
the anti-inflammatory effect of CHF6001 on sputum and blood biomarkers of inflammation. Doses 
up to 1200µg/day were administered in asthma subjects and up to 3200µg/day were administered to 
COPD subjects, for 28 days. CHF6001 did show a reduction (~30%) in late asthmatic response to 
allergen challenge in comparison with placebo. In COPD subjects, on maintenance treatment with 
triple therapy, CHF6001 significantly decreased a number of key biomarkers of airway inflammation 
in sputum and in blood [11].
This observed anti-inflammatory effect of CHF6001 translated in a consistent and relevant reduction 
of moderate or severe exacerbations of COPD in patients with chronic bronchitis, on maintenance 
therapy with a LABA, in a 6-month dose finding trial [12]. CHF6001 proved to be safe and well 
tolerated especially with regard gastro-intestinal side effects, a known PDE4 inhibitor ‘drug class’ 
effects.
The objective of the proposed study is to evaluate the bioavailability of CHF6001 after inhaled 
administration and to characterize the mass balance and route of elimination of CHF6001 in human 
along with its relevant metabolites. 
For detailed information on preclinical and clinical data please refer to the Investigator's Brochure 
[13]. 


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1.2 Study rationale
Human radiolabelled mass balance or absorption, distribution, metabolism and excretion (ADME) 
studies are required by regulatory agencies for the registration of new drug, and therefore, are an 
integral part of the drugs development programs.
 
 
 
 
 
 
This study along with data from the mass balance studies in toxicology species, provides essential 
information on the exposure of the parent compound and the metabolites of CHF6001.
It will be an open-label study in 8 healthy male participants aimed to assess the absolute 
bioavailability, the mass balance and routes of elimination of CHF6001 in humans, using [14C]-
radiolabelled drug substance administered as an intravenous (IV) infusion concomitantly to an 
inhaled non-radiolabelled dose. 
The study involves a small amount of radioactivity; therefore, only adult males are included in the 
trial.  It will be conducted as an open-label trial as the study measures are objective outcomes (eg, 
total radioactivity in select biological matrices). Conducting the study in healthy subjects will allow 
the evaluation of CHF6001 metabolism in the absence of concomitant medications. The dose, subject 
population, study duration, and sample collection timing are considered adequate to achieve the study 
objectives.
1.3 Risk/benefit assessment
A single inhaled dose of CHF6001 3200µg DPI will be administered to healthy subjects in an open-
label fashion. This dose (and higher inhaled doses up to 4800µg) had been assessed after repeated 
dosing (14 days) in healthy subjects and had proved to be safe and well tolerated [8]. 
A microdose was selected because CHF6001 has not previously been administered by IV infusion to 
humans, although testing has been conducted in pre-clinical studies [13] and in order to evaluate the 
mass balance and absolute inhaled bioavailability in the same setting, thus reducing the overall 
exposure of healthy volunteers to CHF6001. The radioactive dose administered in this trial is an 
acceptable dose for [14C]-labeled human drug metabolism studies of this type. It is expected that this 
dose will provide a sufficient radioactive signal for total radioactivity counting and quantitative 
radioprofiling of [14C]-CHF6001 in plasma and excreta with minimal radiation risk to subjects.
The volunteers will be followed at the clinical site and closely monitored throughout the study period.
This trial is a mass balance study, so no benefits are expected for the healthy volunteers. Considering 
the safety profile of the IMP, the minimal radiation risk to subjects, the measures in place to assure 
the patients’ safety and the expected scientific value, the overall risk/benefit assessment can be 
considered acceptable for the proposed trial. 
This trial will be conducted in compliance with the Declaration of Helsinki (1964 and amendments) 
current ICH E6 Good Clinical Practices and all other applicable laws and regulations.


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2.
STUDY OBJECTIVES

To determine the absolute bioavailability of CHF6001 following single inhaled dose co-
administered with an IV microtracer dose

To characterize the mass balance and routes of elimination of CHF6001 after an IV 
microtracer dose

To characterize the relevant metabolites in plasma, urine and feces 

To assess the safety and tolerability of the study treatment
3.
STUDY DESIGN 
This clinical trial is a single centre Phase I study, with a single dose, non-randomized, open-label, 
uncontrolled design. 
                                            
Screening 
At screening visit (31 to 3 days before first study treatment administration), subjects will be selected 
to enter in the study according to the eligibility criteria. The signed informed consent form will be 
obtained prior to any study related procedures.
Treatment Period 
During the treatment period, subjects will remain at the clinical site (all subjects will be treated at 
 site) from Day -1 (the day before the study drug administration) until the morning of Day 
11. Each subject will fast from 10 hours pre-dose until 2 hours post-dose (between 2 hours post dose 
and discharge, standard meals will be provided at standard times as per site practice). They will be 
discharged after the 240h post-dose (i.v.) assessments are completed. 
Follow-up
A follow-up phone call (or visit, if necessary) will be performed between 7 to 10 days after discharge 
or premature discontinuation to check and record concomitant medications and the status of any 
unresolved adverse events or occurrence of any new adverse events.
End of the trial
The end of the trial is defined at execution of the Follow-up call (or visit) of the last subject of the 
trial.
4.
SUBJECT SELECTION CRITERIA
4.1 Subject Recruitment
A total of 8 subjects will be included in the study.
Subjects will be screened by selection from the subjects’ database of the clinical centre or via 
advertisement. Study specific advertisement will be submitted to the Ethics Committee for approval, 
before using it in the study.


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4.2 Inclusion Criteria 
Subjects must meet all of the following inclusion criteria to be eligible for enrolment into the study:
1. Subject’s written informed consent obtained prior to any study-related procedure;
2. Able to understand the study procedures, the risks involved and ability to be trained to use 
correctly the inhalers and to generate sufficient PIF (at least 40 L/min) using the In-Check 
device set as per NEXThaler® inhaler resistance and Placebo inhaler;
3. Male subjects aged 30 to 55 years inclusive;
4. Body mass index (BMI) within the range of 18 to 35 kg/m2 inclusive;
5. Non- or ex-smoker who smoked < 5 pack years (pack-years = the number of cigarette packs 
(20 cigarettes) or equivalent per day times the number of years) and who stopped smoking > 1 
year prior to screening;
6. Good physical and mental status, determined on the basis of the medical history and a general 
clinical examination, at screening and before treatment;
7. Vital signs at screening within limits defined as: Diastolic Blood Pressure 40-90 mmHg and 
Systolic Blood Pressure 90-140 mmHg, (three) measures performed in supine position after at 
least 5 minutes of resting. The average from triplicate measurements must be within the defined 
range; 
Note: In case the values are outside the defined ranges, the assessment can be repeated once 
before treatment.
8. 12-lead digitised Electrocardiogram (12-lead ECG) in triplicate considered as normal (40 ≤ 
Heart rate ≤ 110 bpm, 120 ms ≤ PR ≤ 210 ms, QRS ≤ 120 ms, QTcF ≤ 450 ms) at screening 
visit. The average from triplicate measurements must be within the defined range;  
Note: In case the values are outside the defined ranges, the assessment can be repeated once 
before treatment.
9. Lung function measurements within normal limits at screening: FEV1 >80% predicted and 
FEV1/FVC > 0.70; 
          Note: In case the values are outside the defined ranges, the assessment can be repeated once      
before treatment.
10. Regular bowel movements (averaging one or more bowel movements per day) at screening; 
11. Males fulfilling one of the following criteria: 
a.
Males with non-pregnant WOCBP partners: they and/or their partner of childbearing 
potential must be willing to use a highly effective birth control method in addition to 
the male condom from the signature of the informed consent and until 90 days after 
the follow-up visit or
b.
Males with pregnant WOCBP partner: they must be willing to use male contraception 
(condom) from the signature of the informed consent and until 90 days after the 
follow-up visit or
c.
Non-fertile male subjects (contraception is not required in this case) or
d.
Males with partner not of childbearing potential (contraception is not required in this 
case). 
Moreover, subjects must not donate sperm during the study and for 90 days after the follow-up visit. 
For the definition of WOCBP and of fertile men and the list of highly effective birth control methods, 
refer to Appendix 2 (or section 4.1 of the CTFG guidance for more detailed information). 
Inclusion criteria 7, 8 and 9 could be checked again once before treatment.

If the criteria are met after the re-check, the subject can continue the study.


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
If the criteria are still not met, the subject will be discontinued and screen failed.
The inclusion criterion n. 6 will be re-checked at Day -1.

If the criterion is met, subject can continue the study.

If the criterion is not met, subject will be discontinued and screen failed.
4.3 Exclusion Criteria
The presence of any of the following will exclude a subject from study enrolment: 
1. Participation in another clinical trial with an investigational drug in the 3 months or 5 half-lives 
of that investigational drug (whichever is longer) preceding the administration of the study drug; 
a longer and more appropriate time could be considered by the principal investigator based on 
the elimination half-life and/or long-term toxicity of the previous investigational drug;
2. Clinically relevant and uncontrolled respiratory, cardiac, hepatic (including Gilbert syndrome), 
gastrointestinal, renal, endocrine, metabolic, neurologic, or psychiatric disorder that may 
interfere with successful completion of this protocol according to Investigator judgement;
3. Clinically relevant abnormal laboratory values at screening suggesting an unknown disease and 
requiring further clinical investigation or which may impact the safety of the subject or the 
evaluation of the study results according to Investigator judgement;
Note: In case of abnormal laboratory values, that could indicate a temporary condition, the test 
can be repeated once before treatment.
4. Subjects with history of breathing problems (e.g. history of asthma including childhood asthma);
5. Positive to HIV1 or HIV2 serology at screening;
6. Positive results from the Hepatitis serology which indicates acute or chronic Hepatitis B or 
Hepatitis C at screening (i.e. positive HB surface antigen (HBsAg), HB core antibody (IgM anti-
HBc), HC antibody;
7. Blood donation or blood loss (equal or more than 450 mL) less than 2 months prior screening or 
prior to treatment;
8. Positive urine test for cotinine at screening or before treatment;
9. Documented history of alcohol abuse within 12 months prior to screening or a positive alcohol 
breath test at screening or before treatment;
10. Documented history of drug abuse within 12 months prior to screening or a positive urine drug 
screen at screening or before treatment;
11. Intake of non-permitted concomitant medications in the predefined period prior to screening or 
before treatment) or the subject is expected to take non-permitted concomitant medications 
during the study (see “non-permitted concomitant medications” section);
12. Presence of any current infection, or previous infection that resolved less than 7 days prior to 
screening or before treatment;
13. Known intolerance and/or hypersensitivity to any of the excipients contained in the formulation 
used in the trial;
14. Unsuitable veins for repeated venipuncture;
15. Heavy caffeine drinker (> 5 caffeinated beverages e.g., coffee, tea, cola per day);
16. Abnormal haemoglobin level defines as <13mg/l for males and <11.3mg/l for female at 
screening;
17. Subjects using e-cigarettes within 6 months prior to screening;
18. Subjects been involved in a study involving a 14C-labeled drug within the 12 months prior to 
enrollment;


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19. Subjects with exposure to significant diagnostic or therapeutic radiation (eg, serial X-ray, 
computed tomography scan, barium meal) or current employment in a job requiring radiation 
exposure monitoring within 12 months prior to Day-1;
20. Documented COVID-19 diagnosis within the last 8 weeks or which has not resolved within 14 
days prior to screening and before treatment.
 
The exclusion criterion 3, if met at the screening visit, can be checked again once before the 
treatment:

If the criterion is not met after the re-check, the subject can continue the study.

If the criterion is still met, the subject will be discontinued and screen failed.
The following exclusion criteria 7, 8, 9, 10, 11, 12 and 20 will be re-checked at Day -1 visit: 

If all the criteria are not met, the subject can continue the study.

If at least one criterion is met, the subject will be discontinued and screen failed.
4.4 Subject Withdrawals
Subjects must be discontinued from the study for any of the following reasons:

An adverse event occurs that, in the opinion of the investigator, makes it unsafe for the 
subject to continue in the study. In this case, the appropriate measures will be taken.

The subject is lost to follow-up.

The subject withdraws consent.

The subject's safety is affected by violation of inclusion or exclusion criteria or use of not-
permitted concomitant medication.

The subject is unwilling or unable to adhere to the study requirements, i.e., non-
compliance.

The sponsor or the regulatory authorities or the Ethics Committee(s), for any reason, 
terminates the entire study, or terminates the study for this trial site or this particular 
subject. 
It is understood by all concerned that an excessive rate of withdrawals can render the study 
uninterpretable; therefore, unnecessary withdrawals of subjects should be avoided.
However, should a subject discontinue the study, all efforts will be made to complete and report the 
observations as thoroughly as possible.
In case of withdrawal, the Investigator must fill in the “Study Termination” page in the eCRF, 
reporting the main reason for withdrawal.   
In case actual drop-out rate is higher than expected, additional subjects may be enrolled in order to 
reach a suitable number (at least 6) evaluable subjects.
If a subject is withdrawn/drops-out of the study after receiving the test treatment, the subject study 
number and corresponding test treatments should not be reassigned to another subject.


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5.
CONCOMITANT MEDICATIONS
5.1 Permitted concomitant Medications
Occasional paracetamol (maximum 2 g per day with a maximum of 10 g per 14 days for mild 
conditions not meeting exclusion criteria).
5.2 Non-permitted concomitant Medications

Intake of any enzyme-inducing drugs, enzyme-inhibiting drugs, biologic drugs or any drug 
known to have a well-defined potential for hepatotoxicity (e.g. isoniazide, nimesulide, 
ketoconazole) are not permitted within 3 months prior to screening and throughout the study 
period (including the follow up period)

Intake of any drug treatment, including prescribed or OTC medicines, vitamins, homeopathic 
remedies etc., with the exception of the allowed concomitant medications specified in the relative 
section are not permitted within 14 days prior to screening and throughout the study period 
including the follow up period
In case of intake of any forbidden concomitant medication during the study the need for this subject 
to be withdrawn from the study will be carefully evaluated by the Investigator on the basis of the 
potential impact on the study outcome and/or safety evaluation in the subject's best interest.
6.
TREATMENT(S)
The non-radiolabelled IMP will be supplied to the clinical site under the responsibility of the Sponsor, 
who will also provide the Pharmacist/Investigator with appropriate certificates of analytical 
conformity and QP release. 
The radiolabelled Drug substance (DS) will be provided by 
, 
 will manufacture the 
Drug Product (DP) for IV infusion, it will be manufactured and Certified by a
 QP and supply 
straight to the clinic. 
The analytical and microbiological methods related to the radiolabelling procedure will be validated 
and reports will be generated.
6.1 
CHF6001 800µg/20mg NEXThaler® DPI

Active ingredient: CHF6001 


 

CHF6001 DPI NEXThaler® matched placebo (for Training kit)

Active ingredient: None

       

 
 



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[14C]-CHF6001 1,85µg/ml (50nCi/ml) solution for IV infusion

Active ingredient: [14C]-CHF6001 

Formulation vehicle:  ethanol and kolliphor®HS 15 in 0.9% w/v saline 

Appearance: clear and colourless solution in a glass vial
6.2 Dosage and Administration 
6.2.1 Selection of doses in the study
Inhaled dose
The highest CHF6001 dose planned to be tested in the Phase 3 program studies is 1600µg 
administered twice-daily corresponding to a total daily dose of 3200µg. 
 
 
 
 
 
 
 
 
 
 
 
 
 
CHF6001 is mainly metabolized to CHF5656 and CHF6095, which are detected in the circulation of 
COPD patients at concentrations approximately 20- and 50- fold lower, respectively, than the parent 
compound CHF6001. CHF5956, the major metabolite of CHF6001, was 2,000-fold less potent than 
the parent compound in an in vitro model of LPS-induced TNF-alpha production in human isolated 
peripheral blood mononuclear cells.
Therefore, this clinical trial was designed taking into consideration the linear and time-independent 
PK at concentrations found to be safe and well tolerated in healthy subjects and patients for the active 
parent compound CHF6001. For this reason, mass balance characterization of CHF6001 after single 
dose administration of 3200µg is deemed appropriate.
Intravenous dose 
The proposed intravenous dose of 18.5µg (given over a 15-minutes infusion) accounts for less than 
0.6% of the concomitant cold inhaled dose (3200μg). The overall dose (3218.5μg) is therefore 
expected to be well within the highest dose tested in humans (4800μg by inhalation). 
The radioactive dose will be approximately 500nCi. The radioactive burden is considered to pose 
negligible additional risk above the background cosmic radiation, so no dosimetry estimation is 
necessary.


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6.2.2 Dosage
One single dose administration of CHF6001 will be given to all subjects, as follows:
CHF6001 DPI Inhalation:

4 inhalations of CHF6001 800µg/20mg DPI NEXThaler® giving a total dose of 3200 μg 
CHF6001
[14C]-CHF6001 IV infusion:

Detailed information on the process for formulation preparation are in the pharmacy protocol. 
The IV formulation will be administered intravenously as a single dose over 15 minutes, 
ending at the expected Tmax of 2 h for the inhaled dose giving a total infused dose of 18.5µg 
and 500nCi (18.5kBq).
6.2.3 Administration
The treatment administration will take place at the clinical site in the morning of Day 1 of the 
treatment period, according to the instruction for use (provided in separate documents) and under the 
supervision of the investigator or his/her designee.
In the morning of Day 1, before treatment administration, restrictions related to the study drug 
administration will be applied, as reported in section 7.1.4.
6.2.3.1 DPI administration 
The study medication will be administered using the DPI device. 
For inhalation treatments, time 0 is defined as the moment when the first inhalation takes place. 
Subjects will take the medication in an upright position. Additionally, the subjects will be instructed 
to hold their breath for 10 secs following each inhalation and to wait 50 seconds before taking the 
next inhalation, in order to have 1 min between consecutive inhalations. A delay of ± 5 seconds will 
not be considered as a protocol deviation.  
The dose counter of the NEXThaler® device should be inspected before and after each inhalation. 
Detailed Instruction for use of the NEXThaler device for IMP use will be provided in a separate 
document.
Treatment evidence will be recorded in the eCRF and in the administration log, by the investigator or 
designee. Any issues occurred during the inhalations, evaluated in this sense by the investigator or 
designee, will be reported in the eCRF. An incorrect inhalation is defined as “a significant reduction 
of delivered dose that reaches the lungs based on investigator judgement”. Even if this is the case, no 
extra inhalations (i.e., more than the 4 inhalations required) will be administered.
In order to prevent any kind of contamination, the administration of the drug will take place in a 
ventilated room well separated from the blood sampling room. Subjects will remain in the room of 
administration only for the time strictly needed for the administration. Furthermore, during inhalation, 
subjects and assistant will wear special protective coats and gloves, which will be removed before 
entering the blood sampling room. The assistant administering the inhalations will not be allowed to 
participate in the blood sampling procedures on Day 1.


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6.2.3.2 IV administration 
[14C]-CHF6001 will be administered according to the pharmacy protocol to participants at the site 
by a 15 min intravenous infusion ending at the expected Tmax of 2 h for the inhaled dose 
administered. Administration will be documented in the source documents and reported in the 
eCRF.
6.2.4
Subject Training
6.2.4.1 Training with In-Check DIAL
At screening and on Day -1 and Day 1 pre-dose of the treatment Period, the In-Check DIAL will be 
used to train subjects to use the DPI inhaler correctly. In-Check DIAL is a PIF meter that simulates 
the internal resistance of the device, to enable the measurement of airflow as if the subject is using 
the device. The In-Check DIAL resistance selector will be aligned with the internal resistance of the 
NEXThaler® device. In order to ensure an optimum aerosol dispersion threshold, a PIF of at least 40 
L/min is required. The optimal PIF is 60 L/min. Subjects who will reach the required PIF at the 
NEXThaler® setting, will be considered able to generate sufficient PIF for a correct inhalation with 
the NEXThaler®.
Following three successful inhalations with the In-Check DIAL device, the subjects will be asked to 
continue using the same inspiratory flow during drug inhalation. Detailed instructions for use of In-
Check DIAL are reported inside the commercial package.
Subject will not be treated if the training is not completed with success. Training evidence will be 
recorded in the eCRF including the three “PIF” values.
6.2.4.2 Training with DPI placebo
At screening and on Day-1 and Day1 pre-dose of the treatment period, the correct use of the DPI 
device (NEXThaler®) will be explained to the subject according to the instruction for use (provided 
in separate documents). 
Subject will be trained using placebo inhalers identical to the devices used for the administration of 
study drug. Therefore, the training will be done with DPI device, repeating inhalation for 
approximately 4 times.
At screening, and on Day -1 of the treatment period, the subject will receive a new training kit. The 
training kit dispensed at Day -1 will be used to follow the training procedure at Day -1 and Day 1 
before the administration of the study treatment.
Subject will not be treated if the training is not completed with success. Training evidence will be 
recorded in the administration log and in the eCRF, by the Investigator or designee.
6.3 Packaging 
All investigational products will be prepared in accordance with Good Manufacturing Practices 
(GMP) as required by the current Good Clinical Practices (GCP). Chiesi Farmaceutici S.p.A. will 
supply the non-radiolabelled DPI material while 
 will supply the radiolabelled IV solution 
as described in the following sections. 
6.3.1 Training Kit
The training kits will be supplied to the Investigator as one DPI training box for each subject, 
containing one placebo DPI inhaler.


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Clinical Study Code: CLI-06001AA1-02
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EUDRACT No.: 2020-004201-31
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
Primary packaging: 1 labelled inhaler of CHF6001 Placebo NEXThaler® dry powder for 
inhalation 

Secondary packaging: 1 labelled pouch containing 1 labelled inhaler of CHF6001 Placebo 
NEXThaler®

Tertiary packaging: Box containing 1 pouch containing 1 labelled inhaler of CHF6001 
Placebo NEXThaler®
6.3.2 Treatment Kit DPI
Treatment kit will be supplied to the Investigator as one treatment kit for each subject. 

Primary packaging: 1 labelled inhaler of CHF6001 NEXThaler® dry powder for inhalation
 

Secondary packaging: 1 labelled pouch containing 1 labelled inhaler of CHF6001 
NEXThaler®

Tertiary packaging: Box containing 1 pouch containing 1 labelled inhaler of CHF6001 
NEXThaler®
6.3.3 Treatment Kit [14C]-CHF6001 IV infusion
The radiolabelled Drug substance (DS) will be provided by 
, 
 will manufacture the 
Drug Product (DP) for IV infusion supply straight to the clinic and the treatment kit will be composed 
by one glass vial containing the [14C]-CHF6001 IV formulation.
Further information can be found in the pharmacy protocol.
6.4 Labeling 
All labeling will be in local language and according to local law and regulatory requirements and will 
be compliant with Annex 13 to the Volume 4 of the GMP. 
6.5 Treatment allocation
All subjects will be administered one single dose (4 inhalations) of CHF6001 800µg/20mg DPI 
NEXThaler® according to a non-randomized, open-label design co-administered with an intravenous 
microtracer dose.
6.6 Treatment Code
No treatment code will be available since the study is performed according to a non-randomized 
design.
6.7 Treatment compliance
For each subject, the number of correct and incorrect inhalations taken during the treatment period 
will be recorded in the source documents and reported in the eCRF.


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Clinical Study Code: CLI-06001AA1-02
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6.8 Drug Storage 
The Pharmacist/Investigator or designee will be responsible for the safe storage of all medications 
assigned to this study, in a secure place with restricted access, and maintained within the appropriate 
ranges of temperature.
DPI training kits: initial storage of pouched inhalers “Requires no special conditions”. Once 
removed from the pouch, the inhalers for training should be kept at ambient temperature but not above 
30°C. At this temperature condition the residual shelf life will be six months (180 days). Therefore, 
the Pharmacist/Investigator at the site must write the use-by-date on the training kit labels once the 
inhalers are removed from the pouch. The use-by-date corresponds to the dispensing date plus 6 
months. Please note that the use-by-date must not exceed the total shelf life of the product.
DPI treatment kits: initial storage of pouched inhalers: “Requires no special conditions”. Once 
removed from the pouch, the inhalers for treatment should be kept at ambient temperature but not 
above 25°C. At this temperature condition the residual shelf life will be six weeks (42 days).
All treatment and training kits, must be used by the subject only at the clinical site under 
Investigator/Pharmacist supervision.
[14C]-CHF6001 IV infusion treatment kits: The solution for IV infusion will be stored between 2 
to 8°C. Further information can be found in the pharmacy protocol.
6.9 Drug Accountability
The Investigator, or the designated/authorized representative, is responsible for the management of 
all the study medications to be used for the study. Study medications that should be stored in a locked, 
secure storage facility with access limited to those individuals authorized to dispense the study 
medications.
An inventory will be maintained by the Investigator or pharmacist (or other designated individual), 
to include a signed account of all the study medication(s) received and dispensed during the trial. 
At the conclusion or termination of the study, the Investigator or the pharmacist shall conduct and 
document a final drug supply (used and unused) inventory. An explanation will be given for any 
discrepancies.
All the study medications supplied, used or unused, will be destroyed directly from the Investigator 
by the centre. In this case, a destruction certificate must be asked to the investigational centre and 
filed both at site and at Sponsor. Destruction will not occur until authorized by Chiesi.
6.10 Provision of additional care
N.A.


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Clinical Study Code: CLI-06001AA1-02
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7.
STUDY PLAN
7.1 Study Schedule
Treatment Period
Follow-up
Screening
Day -1
Day 1
Day 2-10
Day 11
Informed consent
X
Residential period
X
X
X
Discharge
X
Treatment intake
IMP administration
X1
Training
Training with In-Check Dial
X
X
X
Training with Placebo
X
X
X
Subject Health Evaluation
In/Ex Criteria
X
X
Medical History
X
Demographic data
X
Height and weight
X
Alcohol breath test
X
X
Physical examination
X
X
X2
Adverse Events recording
X
X
X
X
X
X
Restrictions
X
X
X
X
X
Concomitant Med
X
X
X
X
X
X
Safety assessment in blood
Clinical Chemistry
X
X2
Serology
X
Haematology
X
X2
Fasting glucose
X
X2
Generic assessments in urine
Urinalysis
X
Drug panel
X
X
Cotinine
X
X
Samples for PK evaluations a
Blood 
X
X
X
Urine
X
X
X
X


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Clinical Study Code: CLI-06001AA1-02
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EUDRACT No.: 2020-004201-31
Date: 19 October 2020 
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Treatment Period
Follow-up
Screening
Day -1
Day 1
Day 2-10
Day 11
Feces
X
X
X
X
Cardiac assessments b
Local ECG
X
X
Vital Signs - Blood Pressure
X
X
Pulmonary Assessment
Spirometry
X
Footnotes for Evaluation Schedule:
1.
There will be 2 types of administration:
-
4 inhalations of CHF6001 800µg/20mg DPI NEXThaler®  
-
Intravenous infusion for 15min of the tracer starting 1h45min after the inhaled dose (time of the first 
inhalation) and ending at the expected Tmax of 2 h for the inhaled dose 
2.
A full physical examination and collection of blood safety test will be performed at discharge. A follow-up visit will 
be done only if deemed necessary by the principal investigator.
a)
Pharmacokinetic samples will be collected as:

Blood samples for plasma non-radiolabeled CHF6001 to be collected at pre-dose (within 60 min from 
inhaled dosing) and at the following timepoints post inhaled dose: 15, 30, 60, 90 min, 2, 3.75, 5.75, 7.75, 
9.75, 11.75, 13.75, 25.75, 49.75, 73.75, 97.75, 121.75, 145.75, 169.75, 193.75, 217.75 and 241.75 hours.

Blood samples for plasma [14C]-total, [14C]-CHF6001, [14C] of the relevant metabolites to be collected at 
pre-dose (within 60 min from inhaled dosing) and at the following timepoints relative to the start of the 
IV infusion: 5, 10, 15 [end of  infusion], 20, 25, 30, 45, 60 min, 2, 4, 6, 8, 10, 12, 24, 48, 72, 96, 120, 
144, 168, 192, 216 and 240 hours. 

Blood samples for whole blood [14C]-total and [14C]-CHF6001 at pre-dose (within 60 min from inhaled 
dosing) and 20 min after start of infusion

Urine samples for assessments of [14C]-total, [14C]-CHF6001 and [14C] of the relevant metabolites to be 
collected at pre-dose from -12 hours to immediately before study drug inhaled administration (-12-0h) 
and at the following time frames relative to the start of the IV infusion: 0-4h, 4-8h, 8-12h, 12-24h, 24-
48h, 48-72h, 72-96h, 96-120h, 120-144h, 144-168h, 168-192h, 192-216h and 216-240h.

Fecal samples for assessments of [14C]-total, [14C]-CHF6001 and [14C] of the relevant metabolites to be 
collected at pre-dose from check-in (Day -1) to immediately before study drug inhaled administration 
(check-in -0h) and at the following time frames relative to the start of the IV infusion: 0-24h, 24-48h, 48-
72h, 72-96h, 96-120h, 120-144h, 144-168h, 168-192h, 192-216h and 216-240h.
b)
Cardiac assessments will be performed at Screening and on Day 1 the treatment period as:

Local 12-lead ECG: A triplicate ECG will be done for assessing eligibility at screening and a single ECG 
will be performed for general safety at predose and at 2,5 hours post dose (inhaled administration).

Vital Signs - BP: BP measurement will be performed in triplicate at the following time points: at 
Screening and, for general safety, at predose and at 2,5 hours post dose (inhaled administration). 
7.1.1 Screening 
A Screening visit will be carried out from 3 to 31 days prior to the first administration of the study 
drug, in order to identify eligible consenting subjects for the study.
The following procedures will take place:


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
Collection of the signed written INFORMED CONSENT by potential eligible subjects, after 
the study has been fully explained by the investigator or designee. The investigator or his/her 
designee should provide the participants ample time and opportunity to inquire about details 
of the trial and to decide whether or not to participate in the trial;

SUBJECT SELECTION: inclusion/exclusion criteria will be checked 

SUBJECT TRAINING: subjects will be trained on a proper use of the inhaler with the In-
Check Dial and the DPI Nexthaler® placebo.

MEDICAL/SURGICAL HISTORY AND CONCOMITANT DISEASES: subject health 
history will be recorded.

DEMOGRAPHY DATA COLLECTION: demographic data including age, gender and race, 
will be recorded.

HEIGHT AND WEIGHT: will be recorded.

ALCOHOL BREATH TEST: will be performed

PHYSICAL EXAMINATION: a comprehensive physical examination will be performed.

ADVERSE EVENTS RECORDING: AEs occurred since the signature of the informed 
consent will be recorded. In case of any clinically significant abnormality revealed during the 
physical examination or screening procedures, it will be recorded in the subject’s medical 
history, unless its start date/time is after the informed consent signature date/time and it is not 
due to a pre-existing condition. In the latter case, it will be recorded as an adverse event.

STUDY RESTRICTIONS: study restrictions criteria will be checked.

PRIOR and CONCOMITANT MEDICATIONS CHECK: Any enzyme-inducing drugs, 
enzyme-inhibiting drugs, biologic drugs or any drug known to have a well-defined potential 
for hepatotoxicity (e.g. isoniazide, nimesulide, ketoconazole) taken in the last 3 months and 
any drug treatment, including prescribed or OTC medicines, vitamins, homeopathic 
remedies taken in the last 2 weeks will be recorded.  

BLOOD TESTING: blood samples collection for safety evaluations (clinical chemistry, 
haematology, serology, fasting glucose), according to section 7.2.2. Subjects have to be in 
fasting condition from at least 10 hours before blood samples are collected.

URINE TESTING: urine samples collection for urinalysis, drug panel and cotinine test will 
occur according to section 7.2.3.

LOCAL SAFETY ECG: Triplicate 12-lead ECG will be used to measure heart rate (HR) and 
will occur according to section 7.2.5.

BLOOD PRESSURE: measurement of Systolic and Diastolic Blood Pressure (SBP and DBP) 
will occur according to section 7.2.4.

SPIROMETRY: Lung function (FEV1 (L), FEV1 % of predicted normal value, FVC (L), FVC 
% of predicted normal value, FEV1/FVC) will be measured according to section 7.2.6.
7.1.2 Treatment Period
The subject will arrive at the clinical site on Day -1 and will stay overnight until the accomplishment 
of the 240 hours post-dose (i.v.) study procedures. All subjects will be discharged on Day 11.


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
On Day -1, subjects will be hospitalised and will undergo the planned assessments and checks 
to confirm eligibility)

On Day 1 of treatment period, subjects will undergo pre-dose assessments, then will receive 
the treatment and will start post-dose assessments.

From Day 2 until Day 11, subjects will continue the post-dose assessments.  Subjects will be 
instructed to continue to follow study restrictions (see section 7.1.4). They will then be 
discharged soon after the 240h post-dose (i.v.) assessment. 
The following procedures and assessments will take place during this period:

SUBJECT TRAINING with devices: subjects will be trained on a proper use of the inhaler 
with In-Check Dial (Day -1/Day 1) and with the DPI placebo training kit (Day -1 and Day 1 
pre-dose)

SUBJECT SELECTION (Day -1): Inclusion criterion 6 and Exclusion criteria 7, 8, 9, 10, 11, 
12 and 20, will be re-checked as requested in section 4.2 and 4.3. Subjects not fulfilling the 
mentioned criteria will be Screen Failed

ALCOHOL BREATH TEST (Day -1): will be performed 

PHYSICAL EXAMINATION (Day 1): a full physical examination will be performed and 
any new revealed clinically significant abnormality since the screening visit will be recorded 
as an AE. Furthermore, a full physical examination will be performed at discharge.

ADVERSE EVENTS RECORDING (Day -1 to Day 11): Any new AE/SAE occurred since 
the screening visit will be checked and recorded in the eCRF. The status of ongoing AEs will 
be checked and updated in the eCRF when applicable. Moreover, during this period in case 
of any new clinically significant abnormality revealed during the examination or procedures, 
it will be recorded as an adverse event. 

STUDY RESTRICTIONS (Day -1 to Day 11): study restrictions criteria will be checked.

CONCOMITANT MEDICATIONS CHECK (Day -1 to Day 11): concomitant medications 
taken by the subject in the last 14 days and during the treatment period will be checked and 
recorded in the eCRF.

BLOOD TESTING (at discharge): blood samples collection for safety evaluations (clinical 
chemistry, haematology, fasting glucose), according to section 7.2.2. Subjects have to be in 
fasting condition from at least 10 hours before blood samples are collected.

URINE TESTING (Day -1): urine samples collection for drug panel and cotinine test will 
occur according to section 7.2.3.

IMP ADMINISTRATION (Day 1)

PK INVESTIGATION: Collection of blood (Day 1 to Day 11), urine and fecal samples (Day 
-1 to Day 11) for PK assessment will occur according to section 7.2.1.

LOCAL SAFETY ECG: (Day 1) A single 12-lead ECG measurement will be performed on 
Day 1, at predose and at 2,5 hours post dose.

BLOOD PRESSURE (Day 1): measurement of Systolic and Diastolic Blood Pressure (SBP, 
DBP) will occur according to section 7.2.4.


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Clinical Study Code: CLI-06001AA1-02
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EUDRACT No.: 2020-004201-31
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7.1.3 Follow-up 
A follow-up call will be performed from 7 to 10 days after the discharge to perform the following:

CONCOMITANT MEDICATIONS CHECK: concomitant medications taken to treat AEs by 
the subjects since the discharge will be checked and recorded in eCRF.

ADVERSE EVENTS RECORDING: the status of unresolved AEs/SAEs at the discharge will 
be checked and any new AEs/SAEs occurred after the discharge will be recorded in the eCRF.
This safety follow-up call shall also be performed in case of premature discontinuation (7-10 days 
from the last administration intake).
If needed, according to the investigator's opinion, a follow-up visit can be performed.
7.1.4 Study restrictions and standardization
1. The following restriction must be applied during the study and must be checked at clinical 
unit from starting of Screening to discharge:
No smoking or intake of substances containing nicotine: within the entire study period.
2. The following restriction must be applied prior to the screening:
No strenuous activities: within 24h prior to the start of screening until the end of the 
visit.
3. The following restrictions must be applied during the treatment period (pre- and post-dose 
refer to the inhaled dose):
No food intake: within 10h pre-dose until 2h post-dose of study treatment drug. If 
needed, participants may consume prunes or prune juice to facilitate stool samples
No fluid intake: within 1h pre-dose and 1h post-dose of study treatment drug. 
Afterwards, in order to maintain adequate hydration, subjects should take at least 240 
mL of water every 2h for the following 6h
No strenuous activities: within 24h pre-dose until Day 11
No intake of beverages or food containing alcohol: within 48h pre-dose until discharge 
on Day 11
No intake of beverages or food containing xanthine (coffee, tea, chocolate, cola), or 
grapefruit (neither fruit nor juice): within 48h pre-dose until discharge on Day 11. 
Decaffeinated products are permitted except during fasting periods
No intake of food containing poppy seeds: within 24h pre-dose until Day 11
4. The following restriction must be applied to prior to the screening and discharge:

No food intake: within 10h prior to the blood collection for the lab test.
If these restrictions are not respected, the visit can be rescheduled once and all the information 
regarding restriction(s) not respected, will be recorded in the eCRF. If the restrictions are again not 
respected, the visit will be performed anyway and all the information regarding restriction not 
respected will be recorded in the eCRF.


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EUDRACT No.: 2020-004201-31
Date: 19 October 2020 
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7.2 Investigations
In case of coinciding investigations, the time point for blood collection for PK evaluation takes 
priority over any other scheduled study activities, e.g. blood pressure and local safety ECG. Where 
other activities are scheduled together with the PK blood collection, these will be performed in a 
sequence allowing for blood sampling exactly at the scheduled time point. The exact time of each 
activity will be recorded in the eCRF.
7.2.1 Sample collection for PK evaluation and samples preparation
The laboratory analysis will be carried out following the GCP and the applicable principles of Good 
Laboratory Practice (GLP) regulations of the Organisation for Economic Co-operation and 
Development (OECD).
The analytical procedures will be described in a separate analytical protocol. Validation data and 
details of the analytical procedures will be gathered in an analytical report that will be included in the 
clinical study report.
The breakdown of blood samples collection is reported in Appendix 3.
7.2.1.1 
Determination of CHF6001 and related analytes in human plasma
Blood samples for the determination of plasma total [14C], [14C]-CHF6001, metabolite profiling (if 
required) and non-radiolabeled CHF6001, will be collected and processed according to an established 
laboratory manual at pre-dose and in the 0-240h interval after dosing, as follows: 
•
22 blood samples of 4 mL for plasma non-radiolabeled CHF6001 will be collected at pre-dose 
(within 60 min from inhaled dosing) and at the following timepoints post inhaled dose: 15, 30, 60, 90 
min, 2, 3.75, 5.75, 7.75, 9.75, 11.75, 13.75, 25.75, 49.75, 73.75, 97.75, 121.75, 145.75, 169.75, 
193.75, 217.75 and 241.75 hours; 
•
25 blood samples of 5 mL for plasma total [14C] and [14C]-CHF6001 will be collected at pre-
dose (within 60 min from inhaled dosing) and at the following timepoints relative to the start of the 
IV infusion: 5, 10, 15 [end of infusion], 20, 25, 30, 45, 60 min, 2, 4, 6, 8, 10, 12, 24, 48, 72, 96, 120, 
144, 168, 192, 216 and 240 hours; 
•
25 blood samples of 10 mL for plasma [14C] of the relevant metabolites will be collected at 
pre-dose (within 60 min from inhaled dosing) and at the following timepoints relative to the start of 
the IV infusion: 5, 10, 15 [end of  infusion], 20, 25, 30, 45, 60 min, 2, 4, 6, 8, 10, 12, 24, 48, 72, 96, 
120, 144, 168, 192, 216 and 240 hours; 
•
2 blood samples of 2 mL for whole blood total [14C] and [14C]-CHF6001 will be collected at 
pre-dose (within 60 min from inhaled dosing) and 20 min after the start of infusion (for a total of 4 
samples).
The processing, storage and shipment of samples are detailed in the laboratory manual document.
Day
Clock time 
(exemplary)
Time 
after 
inhaled 
administration
Time after start 
of 
i.v. 
administration
Time deviation 
allowed
PK 
timepoint 
inhaled 
administration
PK 
timepoint 
i.v. 
administration
1
08:00
Pre-dose 
Within 60 minutes 
from inhalation
X1
X1


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Clinical Study Code: CLI-06001AA1-02
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Date: 19 October 2020 
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09:00
Start of inhaled 
administration
09:15
15 min 
± 2 min
X1
09:30
30 min 
± 2 min
X1
10:00
60 min 
± 5 min
X1
10:30
90 min 
± 5 min
X1
10:45
Start of infusion 
(SOI)
10:50
SOI + 5 min
± 2 min
X1
10:55
SOI + 10 min 
± 2 min
X1
11:00 
2 h 
SOI + 15 min 
(end of infusion)
± 2 min
X1
X1
11:05
SOI + 20 min 
± 2 min
X
11:10
SOI + 25 min 
± 2 min
X
11:15
SOI + 30 min 
± 2 min
X
11:30
SOI + 45 min 
± 5 min
X
11:45
SOI + 60 min 
± 5 min
X
12:45
3.75 h 
SOI + 2 h 
± 5 min
X
X
14:45
5.75 h 
SOI + 4 h 
± 10 min
X
X
16:45
7.75 h 
SOI + 6 h 
± 10 min
X
X
18:45
9.75 h 
SOI + 8 h 
± 10 min
X
X
20:45
11.75 h 
SOI + 10 h 
± 20 min
X
X
22:45
13.75 h 
SOI + 12 h 
± 20 min
X
X
2
10:45
25.75 h 
SOI + 24 h 
± 20 min
X
X
3
10:45
49.75 h 
SOI + 48 h 
± 1 h
X
X
4
10:45
73.75 h 
SOI + 72 h 
± 1 h
X
X
5
10:45
97.75 h 
SOI + 96 h 
± 1 h
X
X
6
10:45
121.75 h 
SOI + 120 h 
± 1 h
X
X
7
10:45
145.75 h 
SOI + 144 h 
± 1 h
X
X
8
10:45
169.75 h 
SOI + 168 h 
± 1 h
X
X
9
10:45
193.75 h 
SOI + 192 h 
± 1 h
X
X
10
10:45
217.75 h 
SOI + 216 h 
± 1 h
X
X
11
10:45
241.75 h 
SOI + 240 h 
± 1 h
X
X
SOI: Start Of Infusion
1: Fasted condition
Longer time deviations will be evaluated on a case by case basis during the Data Review Meeting. 
Metabolite profiling: pooling across timepoints for each subject will be done. This will be followed 


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by chromatography (LC+AMS) and radio-peaks will be detected and quantified. For metabolites in 
plasma accounting for ≥10% of total radioactivity recovered in this matrix structural characterization 
may be performed, where possible.
7.2.1.2 Determination of CHF6001 and related analytes in human urine
Urine samples for total [14C], metabolite profiling (if required) and [14C]-CHF6001 assessment will 
be collected at pre-dose from -12 hours to immediately before study drug inhaled administration (-
12-0h) and at the following time frames relative to the start of the IV from pre-dose infusion: 0-4h, 
4-8h, 8-12h, 12-24h, 24-48h, 48-72h, 72-96h, 96-120h, 120-144h, 144-168h, 168-192h, 192-216h 
and 216-240h, for a total of 14 urine collection intervals. 
Subjects will be asked to empty the bladder before both dosing and before the end of each collection 
interval. All urine voided during the collection intervals will be retained. Access to the toilet will be 
controlled by the site staff.  
Collection and processing are described in the laboratory manual document. 
The following time deviations from theoretical post-dose times related to the end of the collection 
interval will be allowed: 
For 4 and 8 hours post-dose
± 10 min
For 12 and 24 hours post-dose
± 20 min
For >24 hours post-dose
± 30 min
Longer time deviations will be evaluated on a case by case basis during the Data Review meeting.
Metabolite profiling: pooling across timepoints and subjects will be done. This will be followed by 
chromatography (LC+AMS) and radio-peaks will be detected and quantified. For metabolites in 
urine and feces (total excreta) accounting for ≥10% of total administered radioactivity structural 
characterization may be performed, where possible.
7.2.1.3 Determination of CHF6001 and related analytes in human feces
Fecal samples for total [14C], metabolite profiling (if required) and [14C]-CHF6001 assessment will 
be collected at pre-dose from check-in (Day -1) to immediately before study drug inhaled 
administration (check-in -0h) and at the following time frames relative to the start of the IV infusion: 
0-24h, 24-48h, 48-72h, 72-96h, 96-120h, 120-144h, 144-168h, 168-192h, 192-216h and 216-240h, 
for a total of 11 fecal collection intervals. 
The feaces samples will be pooled across the defined time windows. The time of each void will be 
recorded and then assigned to the appropriate collection interval. Collection and processing are 
described in the laboratory manual document. 
Metabolite profiling: pooling across timepoints and subjects will be done. This will be followed by 
chromatography (LC+AMS) and radio-peaks will be detected and quantified. For metabolites in urine 
and feces (total excreta) accounting for ≥10% of total administered radioactivity structural 
characterization may be performed, where possible.


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7.2.2 Blood collection for safety evaluation
The following blood samples will be collected at Screening and at discharge:

3.5 mL of blood sample for serum clinical chemistry testing (creatinine, urea, aspartate 
aminotransferase (AST), alanine aminotransferase (ALT), Gamma-glutamyl transpeptidase (γ-
GT), total bilirubin, alkaline phosphatase, sodium (Na), potassium (K)) and fasting glucose.

3.5 mL of blood sample for serum serology testing (HIV1, HIV2, Hepatitis B and Hepatitis C) 
will be collected only at Screening.

4 mL of blood sample for haematology testing (red blood cells count (RBC), white blood cells 
count (WBC) and differential (neutrophils, eosinophils, lymphocytes, monocytes) (% and 
values), total haemoglobin (Hb), haematocrit (Hct), platelets count (PLT)).
Below indications will be followed for safety evaluation in blood at screening:

As for Exclusion criteria no. 3, in case clinically relevant abnormal laboratory values are 
observed, suggesting a temporary condition and requiring further clinical investigation, 
subject will be asked to return to clinic once before treatment, to perform again the testing 
measurement. If at that time no abnormalities are observed, subject can be included in the 
study, otherwise subject will be discharged and recorded as Screening Failure.
Blood collection and sample preparation will be performed according to procedures provided by the 
local laboratory which will be in charge to transmit the results and the laboratory normal ranges for 
entry in the eCRF.
7.2.3 Urine collection for safety evaluation
The following urine sample will be collected at Screening:

urine sample for urinalysis – quantitative (proteins) and urinalysis – qualitative (ketones, 
microscopic examination of the sediment)

urine sample for drug test (cannabinoids, opiates, cocaine, benzodiazepines, amphetamines, 
barbiturates)

urine sample for cotinine test
The following urine samples will be collected on Day-1 of the Treatment Period:

urine sample for drug test (cannabinoids, opiates, cocaine, benzodiazepines, amphetamines, 
barbiturates)

urine sample for cotinine test
Urine collection and sample preparation will be performed according to procedures provided by the 
local laboratory which will be in charge to transmit the results to the Investigator for entry in the 
eCRF.
7.2.4 Blood Pressure 
At Screening and on Day 1 of the treatment period (before blood sampling) Systolic and Diastolic 
Blood Pressure (SBP, DBP) will be measured in triplicate, within 5 minutes, at the following time 
points: pre-dose and at 2,5 hours after inhaled administration, for safety reasons. Subjects should 


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be resting in a quiet supervised setting with minimal stimulation (e.g. no television, loud music, 
computer games) and lay in a supine position for 5 minutes (whenever possible) before each 
measurement. No time deviations will be recorded for BP. Below indications will be followed for 
Blood Pressure measurement at Screening:
As for Inclusion criterion no. 7, in case the values are out of the defined ranges (average of 
the triplicate parameters out of normal range i.e. Diastolic Blood Pressure 40-90 mmHg, 
Systolic Blood Pressure 90-140 mmHg), subject will repeat the Blood Pressure measurement. 
If the repeated values (the average of the triplicate parameters) are within the allowed ranges, 
subject can be included in the study, otherwise subject will be discharged and recorded as 
Screening Failure.
The single values and the average of triplicate Blood Pressure measurements will be reported in 
eCRF, and the average will be used to assess the inclusion criterion at screening.
7.2.5 Local Safety 12-Lead ECG
A local triplicate 12-lead digitalized Electrocardiogram (12-lead ECG) recording will be done and 
considered for safety purpose at Screening (before Blood pressure and blood sampling) to check the 
the subject eligibility (according to exclusion criterion) and a local single 12-lead ECG will be 
performed on Day 1 of the study period at predose and at 2,5 hours after inhaled administration. No 
time deviations will be recorded for ECG.
Subjects should be resting in a quiet supervised setting with minimal stimulation (e.g. no television, 
loud music, computer games) and lay in a supine position for 5 minutes (whenever possible) before 
each measurement. 
Below indications will be followed for Local ECG measurement at Screening:

As for Inclusion Criterion no. 8, in case abnormalities are observed with Local ECG (40 ≤ 
Heart rate ≤ 110 bpm, 120 ms ≤ PR ≤ 210 ms, QRS ≤ 120 ms, QTcF ≤ 450 ms for males) 
subject will perform again the ECG measurement. If at that time the average of the triplicate 
ECG parameters is within the allowed ranges, subject can be included in the study, otherwise 
subject will be discharged and recorded as Screening Failure. 
Below indications will be followed for local ECG measurement during the treatment period:

A single 12-lead ECG measurement will be performed on Day 1, at predose and at 2 hours 
post dose (in resting conditions as much as possible)
The values of all ECG parameters and the overall ECG evaluation will be reported in eCRF, at 
Screening and the average will be used to assess the inclusion criterion.
7.2.6 Lung function evaluation
The lung function measurements will be assessed locally at Screening.
Spirometry will be done according to the recommendation of the Official Statement of the European 
Respiratory Society and American Thoracic Society [17], [18]. Lung function measurements will be 
done with subjects either standing or sitting with the nose clipped after at least 10 minutes rest. Values 
will be corrected for BTPS conditions. 


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The following parameters will be recorded:

Forced Expiratory Volume in the 1st second (FEV1, L)

Forced Vital Capacity (FVC, L)

FEV1 % of predicted normal value

FVC % of predicted normal value

FEV1/FVC ratio
Predicted values of FEV1 will be calculated according to formulas reported by Quanjer et al [19]. 
For FEV1 and FVC, the highest value from three technically satisfactory attempts will be recorded 
in the eCRF (irrespective of the curve they come from). The chosen value should not exceed the next 
one by more than 150 mL. If the difference is larger, up to 8 measurements will be made and the 
largest value be reported. In order to be considered as technically satisfactory attempts, measurements 
should be free from cough and false-start. FEV1/FVC should be taken from the highest FEV1 and 
FVC values (even if not coming from the same curve). 
Below indications will be followed for lung function measurement at screening:

As for Inclusion Criterion no. 9, in case the criterion is not met, subject will repeat the test. If 
at that time the FEV1/FVC >0.70 and FEV1 >80% predicted, subject can be included in the 
study, otherwise subject will be discharged and recorded as Screening Failure. 
The FEV1/FVC values will be provided as ratio. For eligibility purposes, values will be considered 
as acceptable if FEV1/FVC >0.70. 
8.
PHARMACOKINETIC ASSESSMENTS
Main objective of this study is to assess the absolute bioavailability, the mass balance and routes of 
elimination of CHF6001, using [14C]-radiolabelled drug substance administered as an intravenous 
(IV) infusion concomitantly to an inhaled non-radiolabelled dose, in healthy volunteers.
Therefore, the following variables will be derived:
Following intravenous (IV) administration: 

Plasma [14C]-CHF6001, AUC0-t iv, Cmax_iv, tmax_iv, AUC0-∞_iv, t1/2 iv, Vz, Vdss and systemic 
clearance (CL) 

Plasma [14C]-total AUC0-t iv, Cmax_iv, tmax_iv, AUC0-∞_iv, t1/2 iv

Urine and feces [14C]-CHF6001and [14C]-total excreted fraction (Feu, Fef and Feu+f) 

Systemic blood clearance (CLblood) [14C]-CHF6001

Renal plasma and blood clearance (CLR and CLR_blood) [14C]-CHF6001

Fraction of the relevant metabolites in plasma, and fraction of the relevant metabolites 
recovered in urine and feces (Fr_plasma, Fr_urine, Fr_feces, Fr_urine+feces)

Metabolized fraction (Femet) 

Blood to plasma ratio (Rb/p) [14C]-CHF6001

Blood to plasma ratio (Rbt/pt) [14C]-total

Hepatic extraction (Eh_blood) [14C]-CHF6001
Following inhaled administration: 


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
Plasma CHF6001 AUC(0-t)_inh, Cmax_inh, tmax_inh, AUC(0-∞)_inh, t1/2_inh, absolute inhaled 
bioavailability (Finh)  
The pharmacokinetic analysis will be performed using WinNonlin Phoenix 6.2 or later (Pharsight 
Corporation, Palo Alto, CA, USA).  Actual elapsed time from dosing will be used to estimate all 
individual PK parameters.  Standard non-compartmental methods will be used for the calculation of 
the following parameters from the individual plasma drug concentrations versus time profile. 
The following PK parameters will be calculated, whenever possible, for each subject based on the 
whole blood concentrations of [14C]-CHF6001, plasma concentrations of CHF6001, plasma [14C]-
CHF6001 and plasma [14C]-total radioactivity:

AUC0-t: the area under the concentration-time curve from 0 to the last quantifiable 
concentration will be computed using the linear trapezoidal rule with linear interpolation;

AUC0-∞: the area under the concentration-time curve extrapolated to infinity will be calculated 
as the sum of AUC0-t and a residual part extrapolated to infinite time. The residual area from 
the last concentration data point to infinite time is calculated using the approximation:
Where λz is the first order terminal rate constant, estimated by linear regression of the terminal 
log-linear segment of the plasma log-concentration curve, and Ct is the last quantifiable 
concentration data point. In order to determine λz by log-linear regression, at least 3 points 
will be used. The quality of log-linear regression and the reliability of extrapolated PK 
parameters should be demonstrated by an adjusted coefficient of determination R2 of at least 
0.85;

Cmax and tmax: the value and time of the maximum concentration will be obtained directly from 
the experimental data without interpolation;

t½: the terminal half-life, associated with negative slope (-λz), calculated as 0.693/λz from the 
individual drug concentrations versus time profiles.

DoseIV: is the actual IV dose calculated as [weight of full syringe (g) minus syringe weight 
after dose (g)] multiplied by nominal concentration (μg/g)

Finh: inhaled absolute bioavailability based on AUC(0-∞) for CHF6001 calculated as Finh = 
(AUC0-∞_inh x Doseiv)/ (AUC0-∞_iv x Doseinh). If AUC0-∞ is not calculable AUC0-t will be used.  

Rb/p: whole blood to plasma ratio is determined on the [14C]-CHF6001 whole blood and plasma 
concentration (Cb and Cp) 20 min after the start of infusion, calculated as Rb/p=Cb/Cp

Rbt/pt: whole blood to plasma ratio is determined on the total-[14C] whole blood and plasma 
concentration (Cbt and Cpt) 20 min after the start of infusion, calculated as Rbt/pt=Cbt/Cpt

CL: the total body plasma clearance following IV administration, calculated only for [14C]-
CHF6001 as DoseIV/AUC0-∞

CLblood: the total body blood clearance following IV administration, calculated only for [14C]-
CHF6001 as CLblood =CL/Rb/p 

Vdss: Volume of distribution at steady-state will be calculated only for [14C]-CHF6001 as 
DoseIV x AUMC0-∞ / (AUC0-∞)2 
z
C
t
C
t
t
t




d


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
Vz: the volume of distribution during the terminal phase, calculated only for [14C]-CHF6001 
as DoseIV/(AUC0-∞ x λz)

Fr_plasma: fraction of the relevant metabolites in plasma calculated as 100 x AUC [14C]-
metabolite/ AUC [14C]-total radioactivity 
The following PK parameters will be calculated, whenever possible, for each subject based on the 
concentrations of urine [14C]-CHF6001 and urine [14C]-total radioactivity:

Aeu: amount excreted in urine (expressed in weight) over sampling interval calculated as: Ae 
= Cur x Vur, where Cur is the concentration in urine and Vur is the volume of urine

Cumulative Aeu: cumulative amount excreted in urine, calculated as the sum of the amount 
excreted in urine for each collection period

feu: percent of the dose excreted in urine over a sampling interval, where feu = 100 x 
(Aeu/DoseIV)

Feu: cumulative percent of dose excreted in urine, calculated as the sum of the percent of dose 
excreted in urine for each collection period

CLR: renal plasma clearance calculated only for [14C]-CHF6001, if plasma AUC0-∞ is 
calculable then CLR = Cumulative Aeu/AUC0-∞, if Plasma AUC0-∞ is not calculable then CLR= 
Cumulative Aeu/AUC0-t. 

CLR_blood: renal blood clearance calculated only for [14C]-CHF6001, where CLR_blood =CLR/Rb/p 

Fr_urine: fraction of the relevant metabolites recovered in urine calculated as amount 
recovered (Aru)/DoseIV x 100
The following PK parameters will be calculated, whenever possible, for each subject based on plasma 
[14C]-CHF6001 and urine [14C]-CHF6001 radioactivity:

Eh_blood: hepatic extraction calculated from intravenous systemic and renal blood clearance 
(CLblood = CL/Rb/p) and CLR_blood =CLR/Rb/p, respectively and human hepatic blood flow (QH; 
assumed to be 87 l/h [20]); Eh_blood = (CLblood – CLR_blood)/QH
The following PK parameters will be calculated, whenever possible, for each subject based on the 
concentrations of feces [14C]-CHF6001 and feces [14C]-total radioactivity:

Aef: amount excreted in the feces over sampling interval as Aef = (concentration in faecal 
collected sample x collected sample weight) for each faecal collection interval

Cumulative Aef: cumulative amount excreted in feces, calculated as the sum of the amount 
excreted in feces for each collection period

fef: percent of the dose excreted in feces over a sampling interval, where fef = 100 (Aef/doseIV)

Fef: cumulative percent of dose excreted in feces, calculated as the sum of the percent of dose 
excreted in feces for each collection period

Fr_feces: fraction of the relevant metabolites recovered in feces calculated as amount 
recovered (Arf)/DoseIV x 100
The following PK parameters will be calculated, whenever possible, for each subject based on the 
concentrations of urine and feces [14C]-CHF6001 and urine and feces [14C]-total radioactivity:


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
Cumulative Aeu+f: cumulative amount excreted in urine and feces, calculated as Cumulative 
Aeu + Cumulative Aef 

Fetot: cumulative percent of dose excreted in urine and feces, calculated as Feu + Fef

Fr_urine+feces: fraction of the relevant metabolites recovered in urine and feces calculated as 
Fr_urine + Fr_feces

Femet: Metabolized fraction calculated as Femet = [cumulative Aeu+f (14C-total) – cumulative 
Aeu+f ([14C]-CHF6001)]/ cumulative Aeu+f (14C-total) 
9.
SAFETY ASSESSMENTS
The safety profile will be assessed through the monitoring of the following:

Adverse Events (AEs) and Adverse Drug Reactions (ADRs)

Vital signs (Systolic and Diastolic Blood Pressure). Vital signs will be evaluated pre-dose 
and at 2,5 hours post-dose 

Heart Rate (HR) from local 12-lead safety ECG will be performed at pre-dose and at 2,5 
hours post-dose

Clinical laboratory evaluations: Blood chemistry, haematology and fasting glucose 
collected at Screening and Discharge
10.
ADVERSE EVENT REPORTING
10.1 Definitions
An Adverse Event is “any untoward medical occurrence in a patient or clinical trial subject 
administered a medicinal product and which does not necessarily have a causal relationship with this 
treatment”.
An adverse event can therefore be any unfavorable and unintended sign (including abnormal 
laboratory finding), symptom, or disease temporally associated with the use of a medicinal product, 
whether or not considered related to the medicinal product.
An Adverse Drug Reaction is an “untoward and unintended responses to an investigational 
medicinal product related to any dose administered”.
All adverse events judged by either the reporting Investigator or the Sponsor as having a reasonable 
causal relationship to a medicinal product qualify as adverse reactions. The expression “reasonable 
causal relationship” means to convey in general that there are facts (evidence) or arguments meant to 
suggest a causal relationship.
The definition covers also medication errors and uses outside what is foreseen in the protocol, 
including misuse and abuse of the product.
A Serious Adverse Event (SAE)/Serious Adverse Drug Reaction is any untoward medical 
occurrence or effect that at any dose falls in one or more of the following categories:
-
Results in death
Death is not an adverse event but an outcome. It is the cause of death that should be regarded as the 
adverse event. The only exception to this rule is “sudden death” where no cause has been established; 
in this latter instance, “sudden death” should be regarded as the adverse event and “fatal” as its reason 
for being serious.


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-
Is life-threatening
Life-threatening refers to an event in which the subject was at risk of death at the time of the event 
(e.g., aplastic anaemia, acute renal failure, and anaphylaxis). The term does not refer to an event 
which hypothetically might have caused death if it were more severe.
-
Requires hospitalisation or prolongation of existing hospitalisation
Hospitalization refers to a situation whereby an AE is associated with unplanned formal overnight 
admission into hospital, usually for purpose of investigating and/or treating the AE. Hospitalization 
for the treatment of a medical condition that occurs on an “elective” or “scheduled” basis or for a pre-
existing condition that did not worsen during the study should not necessarily be regarded as a AE. 
Complications that occur during the hospitalisation are AEs. If a complication prolongs 
hospitalisation, the event is an SAE. Emergency room visits that do not result in a formal admission 
into hospital should be evaluated for one of the other seriousness criteria (e.g., life-threatening; 
persistent or significant disability or incapacity; medically significant).
-
Results in persistent or significant disability or incapacity.
The term significant disability should be viewed as any situation whereby an AE has a clinically 
important effect on the subject’s physical or psychological well-being to the extent that the subject is 
unable to function normally.
-
     Is a congenital anomaly or birth defect
-
     Is a medically significant adverse event
This criterion allows for any situations in which important adverse events/reactions that are not 
immediately life-threatening or do not result in death or hospitalisation may jeopardise the subject’s 
health or may require intervention to prevent one of the above outcomes.
Examples of such events are: intensive treatment in an emergency room or at home for allergic 
bronchospasm; blood dyscrasias or convulsions that do not result in hospitalisation; or development 
of drug dependency or drug abuse.
Medical and scientific judgment should be exercised in deciding whether an event is serious because 
medically significant.
Any suspected transmission via a medicinal product of an infectious agent is also considered a serious 
adverse reaction.
A Non-Serious Adverse Event/Non-Serious Adverse Drug Reaction is an adverse event or adverse 
drug reaction that does not meet the criteria listed above for a serious adverse event/serious adverse 
drug reaction.
10.2 Expectedness
An expected adverse reaction is an adverse reaction, the nature or severity of which is consistent with 
the applicable reference safety information (included in the Investigator’s Brochure for CHF6001), 
otherwise it is considered unexpected. 
Reports which add significant information on specificity or severity of a known, already documented 
serious adverse drug reaction constitute unexpected events. For example, an event more specific or 
more severe than described in the reference safety information would be considered as “unexpected”. 


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Examples of such events are: (a) acute renal failure as a labelled ADR with a subsequent new report 
of interstitial nephritis and (b) hepatitis with a first report of fulminant hepatitis.
10.3 Intensity of Adverse Event
Each Adverse Event must be rated on a 3-point scale of increasing intensity:
Mild: The event causes a minor discomfort or does not interfere with daily activity of the subject or 
does not lead to either modification of test treatment dosage or establishment of a correcting 
treatment.
Moderate: The event perturbs the usual activity of the subject and is of a sufficient severity to make 
the subject uncomfortable. The event leads to a diminution of dosage of the test treatment, or a 
temporary interruption of its administration or to the establishment of a correcting treatment.
Severe: The event prevents any usual routine activity of the subject and causes severe discomfort. It 
may be of such severity to cause the definitive interruption of test treatment.
10.4 Causality Assessment
The following “binary” decision choice will be used by the Investigator to describe the causality 
assessment:

Reasonable possibility of a relatedness

No reasonable possibility of relatedness
The expression “reasonable possibility of relatedness” is meant to convey, in general, that there are 
facts (evidence) or arguments meant to suggest a causal relationship.
The Investigator will be asked to consider the following before reaching a decision on causality 
assessment:
Time relationship between study drug intake and event’s onset;
Dechallenge (did the event abate after stopping drug?);
Rechallenge (did the event reappear after reintroduction?);
Medical history;
Study treatment(s);
Mechanism of action of the study drug;
Class effects;
Other treatments-concomitant or previous;
Withdrawal of study treatment(s);
Lack of efficacy/worsening of existing condition;
Erroneous treatment with study medication (or concomitant);
Protocol related process.
10.5 Action taken with the study drug due to an AE
Dose not changed
Drug permanently withdrawn
Drug temporarily interrupted
Unknown
Not applicable


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10.6  Other actions taken
Specific therapy/Medication
Concomitant Procedure
10.7 Outcome
Each Adverse Event must be rated by choosing among:
Recovered/resolved
Recovering/resolving
Not recovered/not resolved
Recovered with sequelae/resolved with sequelae
Fatal
Unknown
10.8 Recording Adverse Events
All Adverse Events occurring during the course of the study must be documented in the Adverse 
Event page of the electronic Case Report Form (eCRF). Moreover, if the Adverse Event is serious, 
the Serious Adverse Event Form must also be completed. 
It is responsibility of the Investigator to collect all adverse events (both serious and non-serious) 
derived by spontaneous, unsolicited reports of subjects, by observation and by routine open 
questionings.
The recording period for Adverse Events is the period starting from the Informed Consent signature 
until the subject’s study participation ends.
Clinically significant abnormalities detected at Screening not due to a pre-existing condition or 
clinically significant changes at the following visits in the medical opinion of the investigator must 
be reported as adverse events in the eCRF.
If a clinically significant abnormal laboratory finding or other abnormal assessment meets the 
definition of an AE, then the AE eCRF page must be completed, as appropriate. A diagnosis, if 
known, or clinical signs and symptoms if diagnosis is unknown, rather than the clinically significant 
abnormal laboratory finding, should be reported on AE eCRF page. If no diagnosis is known and 
clinical signs and symptoms are not present, then the abnormal finding should be recorded.
For pharmacovigilance purposes, all SAEs should be followed-up in order to elucidate as completely 
and practically as possible their nature and/or causality until resolution of all queries, clinical recovery 
is complete, laboratory results have returned to normal, stable condition is reached or until the subject 
is lost to follow-up. Follow-up may therefore continue after the subject has left the study. In this case, 
the follow-up will continue with no timelines for related SAEs, while for unrelated SAEs the type 
and extent of follow-up undertaken will be determined for each individual case and will depend upon 
the nature (e.g. events with poor prognosis or which do not resolve), severity and medical significance 
of the event. 
10.9 Reporting Serious Adverse Events to Chiesi
The Investigator must report all Serious Adverse Events to the 
 Safety Contact listed below 
within 24 hours of awareness. The information must be sent by providing the completed paper Serious 
Adverse Event form. At a later date, the 
 Safety Contact will report all information to Chiesi 
Global Pharmacovigilance, the Clinical Project Manager and the Clinical Research Physician. 


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Name and Title
Telephone no.
Fax no.
E-mail
 Safety Contact     Not applicable
 
Chiesi Safety Contact
 
 
Reporting of SAEs from the investigator site is from the time of subject’s signature of informed 
consent and until the subject’s study participation ends. After this date, even if no active 
monitoring of subjects is required, SAEs occurring to a subject should be reported if the 
investigator becomes aware of them. 
Up to the closure of the site, SAE reports should be reported to the 
 Safety Contact. 
New serious adverse events occurring after the site is closed should be reported directly to the 
Chiesi Safety Contact.
10.10 Reporting Serious Adverse Events to Regulatory Authorities/Ethics Committees/IRB
All SUSARs, which occur with the investigational medicinal products within or outside the concerned 
clinical trial, if required, will be reported in compliance with the timelines and standards for reporting 
SUSARs set out in the EU Directive 2001/20/EC [Directive 2001/20/EC of the European parliament 
and of the council of 4/April/2001] and linked guidance [European Commission, Enterprise and 
Industry Directorate General: Detailed guidance on the collection, verification and presentation of 
adverse event/reaction reports arising from clinical trials on medicinal products for human use, latest 
version]. The EMA and the concerned national health authority (if applicable) will be informed 
through Eudravigilance, or according to local requirements (as applicable) while the Ethics 
Committees and the investigators by CIOMS I form or by periodic line- listings produced by Chiesi 
Global Pharmacovigilance.
With regard to regulations in force for Pharmacovigilance, the Investigator must fulfill his/her 
obligation according to the law in force in his country. 
10.11 General Notes
In case of death, a comprehensive narrative report of the case should be prepared by the 
Investigator and sent to the 
 Safety Contact together with the Serious Adverse Event 
form, retaining a copy on site.
If an autopsy is performed, copy of autopsy report should be actively sought by the Investigator 
and sent to the 
 Safety Contact as soon as available, retaining a copy on site.
All documents provided by the Investigator or site staff to the 
 Safety Contact must 
be carefully checked for respect of confidentiality. All personal subject’s data must be 
redacted.
All source documents provided by the Investigator or site staff to the 
 Safety Contact 
must be carefully checked for respect of confidentiality. All personal subjects’ data must be 
redacted.
In case of pregnancy occurred to the subject’s partner, the subject participating to the study 
should not be discontinued from the study. The pregnancy will be followed with due diligence 
until the outcome is known and till the age of one year to detect any congenital anomaly or birth 
defect. 
The pregnancy must be reported by the investigator within 24 hours by fax/e-mail/via Monitor 
to the 
 Safety Contact using the paper Pregnancy Report Form. The 
 Safety 
Contact will inform Chiesi of the pregnancy within one working day of being notified. 


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The first two pages of the Pregnancy Report Form should be completed by the investigator with 
all the available information and sent to the 
 Safety Contact. The third page will be 
completed as soon as the investigator has knowledge of the pregnancy outcome, together with a 
follow-up of the first two pages, if necessary (e.g. an update in the medications received during 
pregnancy by the mother). If it meets the criteria for immediate classification of a SAE (e.g. 
spontaneous or therapeutic abortion, stillbirth, neonatal death, congenital anomaly, birth defect) 
the Investigator should follow the procedure for reporting SAEs. 
Any Adverse Drug Reaction (ADR) occurring with any marketed non-investigational medicinal 
product and/or concomitant medication during the study must be reported by the Investigator 
to his/her concerned Health Authority according to the applicable laws. The Investigator is also 
recommended to report all adverse drug reactions to the relevant Marketing Authorisation 
Holders of the involved medicinal products. Additionally, also conditions of use outside the 
marketing authorisation of the medicinal products (i.e. off-label, overdose, misuse, abuse and 
medication errors) or from occupational exposure, as well as cases of suspected drug 
interaction, pregnancy, breast-feeding exposure and lack of efficacy should be reported.
11.
DATA MANAGEMENT
An electronic CRF (eCRF) will be filled-in by the Investigator and/or his/her representative designee. 
Front-end edit checks will run at the time of data collection and back-end edit checks will be used by 
the Data Manager to check for discrepancies and to ensure consistency and completeness of the data. 
Medical history, adverse events and concomitant procedures will be coded using the MedDRA 
dictionary; medications will be coded using the WHO Drug dictionary and Anatomical Therapeutic 
Chemical classification (ATC) up to level 5.
External data (PK concentrations) will be processed centrally, sent to the designated CRO and 
reconciled with the corresponding information recorded in the CRF as part of cleaning activities.
Access to electronic systems used for data collection will be granted to the study personnel only after 
appropriate training.
After the completion of data collection and cleaning, a review meeting will be held to determine the 
occurrence of any protocol violation and to define the subject populations for the analysis. Once the 
database has been declared to be complete and accurate, it will be locked, and the planned statistical 
analysis will be performed. 
If the database is unlocked after the initial lock, the process must be carefully controlled and 
documented; updates to the study data must be authorized by Chiesi.
At the study conclusion, a complete copy of the study data will be created for archival purposes at 
Chiesi. The investigators will receive copies of the subject data for retention at the investigational 
sites.
12.
STATISTICAL METHODS
12.1  Sample Size
According to the exploratory nature of the study, no formal sample size calculation was performed. 
A total of 8 subjects will be included in the study to characterize the absolute bioavailability and the 
mass balance of CHF6001.


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12.2 Populations for analysis

Safety population: all enrolled subjects who start study treatment inhalation.

PK population: all subjects from the safety population excluding subjects without any valid 
PK measurement or with major protocol deviations significantly affecting PK, for example: 
incorrect inhalation, change in subject condition (cold), failure in delivery of the device, use of 
non-permitted medications. Exact definition of major protocol deviations concerning PK will 
be discussed by the clinical team and described in a specific document before bioanalytical data 
are disclosed. Subjects will not be excluded on the basis of statistical analysis or for 
pharmacokinetic reasons, except in the cases defined in the EMA "Guideline on the 
Investigation of Bioequivalence" [21]. 
PK variables will be analysed in the PK population. The Safety population will be used in the 
analysis of all safety variables.
12.3 Statistical analysis
A detailed statistical analysis plan will be described in the Statistical Analysis Plan. The plan might 
be reviewed and updated as a result of the review of the data and will be finalized before locking the 
database. 
12.3.1 Descriptive Statistics 
All PK variables (except for tmax, tmax_inh and tmax_iv) will be summarized by means of descriptive 
statistics including n (number of observed values), arithmetic mean, standard deviation (SD), 
coefficient of variation (CV), geometric mean, geometric CV, median, minimum and maximum. tmax, 
tmax_inh, tmax_iv will be summarized by using n, median, minimum and maximum.
General descriptive statistics for other continuous variables will include the n (number of observed 
values), the mean, the standard deviation, the median, the minimum, and the maximum values. 
For categorical variables, the number and percent of subjects with a specific level of the variable will 
be presented.
12.3.2 Missing data
Details on dealing with missing data, along with the handling of possible outliers, will be described 
in the SAP. 
12.3.3 Subject demographics and baseline characteristics
Demographics and baseline variables will be summarized using descriptive statistics for the Safety 
population and PK (if different from safety population). 
 
12.3.4 PK variables
The following PK variables will be summarized by means of descriptive statistics as described in 
Section 12.3.1:

Plasma [14C]-CHF6001, AUC0-t iv, Cmax_iv, tmax_iv, AUC0-∞_iv, t1/2 iv, Vz, Vdss and CL

Plasma [14C]-total AUC0-t iv, Cmax_iv, tmax_iv, AUC0-∞_iv, t1/2 iv

Plasma CHF6001 AUC(0-t)_inh, Cmax_inh, tmax_inh, AUC(0-∞)_inh, t1/2_inh, Finh


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
Urine and feces [14C]-CHF6001and [14C]-total excreted fraction (Feu, Fef and Feu+f) 

[14C]-CHF6001 CLblood, CLR and CLR_blood

Blood to plasma ratio (Rb/p) [14C]-CHF6001

Blood to plasma ratio (Rbt/pt) [14C]-total

Femet

Metabolites Fr_plasma, Fr_urine, Fr_feces, Fr_urine+feces

[14C]-CHF6001 Eh_blood
Concentration (mass equivalent in mL or mass in mL)/time curves for plasma total [14C], [14C]-
CHF6001 and non-radiolabeled CHF6001 will be presented in linear/linear and log/linear scale. 
Plots will be presented based on arithmetic means and by subject (all individual plasma 
concentration/time curves in one graph).
Bar charts based on arithmetic means showing the percentage of urine and feces excreted fraction for 
total [14C] and [14C]-CHF6001 in the different time intervals will be presented.
Bar charts based on arithmetic means showing percenatages of the relevant metabolites recovered in 
plasma, urine and feces will be presented.
All individual concentration data (mass equivalent per mL or mass per mL) and PK parameters will 
be listed. In addition, a listing of the actual sampling times relative to drug inhalation will be provided. 
Concentrations will be summarized by scheduled sampling time by using n, arithmetic mean, SD, 
CV, median, minimum and maximum.
12.3.5 Safety variables
Adverse Events
All adverse events starting on or after the time of first study drug inhalation will be classified as 
treatment emergent adverse event (TEAE). Any adverse events started after the informed consent 
signature and before the time of first study drug inhalation will be classified as pre-treatment adverse 
event.
The number of subjects who experienced at least one TEAE, drug-related TEAE, serious TEAE and 
TEAE leading to study discontinuation will be summarized. Summaries will be presented overall 
(number and percentage of subjects having at least one event, total number of events) and by system 
organ class and preferred term (number and percentage of subjects having at least one occurrence of 
that event). 
All adverse events will be listed.
Pre-treatment adverse event will be listed only.
Vital signs
Percentage of subjects with clinical relevant changes from baseline to 2,5 hrs post-dose in SBP and 
DBP defined as

Change from pre-dose in DBP > 10 mmHg; 

Change from pre-dose in SBP > 20 mmHg;
  will be summarized.
12-lead safety ECG


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Percentage of subjects with clinical relevant changes from baseline to 2,5 hrs post-dose in HR defined 
as

Change from pre-dose in HR> 20 bpm
  will be summarized.
Laboratory data
For quantitative laboratory parameters (chemistry, haematology and fasting glucose). Shift tables 
from screening with reference to normal ranges will be summarized. 
All laboratory data will be listed with abnormal values flagged.
12.3.6 Interim analysis
Not Applicable.
13.
ETHICS COMMITTEE/INSTITUTIONAL REVIEW BOARD APPROVAL 
The study proposal will be submitted to the Ethics Committee in accordance with the local 
requirements. 
The EC shall give its opinion in writing -clearly identifying the study number, study title and informed 
consent form approved-, before the clinical trial commences.
A copy of all communications with the EC will be provided to the Sponsor.  
The Investigator should provide written reports to the EC annually or more frequently if requested on 
any changes significantly affecting the conduct of the trial and/or increasing risk to the subjects 
(according to the requirements of each country).
14.
 REGULATORY REQUIREMENTS
The study will be notified to the Health Authorities (or authorized by) according to the local legal 
requirements.  
Selection of the subjects will not start before the approval of the Ethics Committee has been obtained 
and the study notified to Health Authorities (or authorized by).
15.
INFORMED CONSENT 
Informed consent must be written in a language understandable to the subjects. It is the responsibility 
of the Investigator to obtain written consent from each subject prior to any study related procedures 
taking place, by using the latest EC approved version of the document. 
Adequate time shall be given to the subject to enquiry the PI about any clarification needed and to 
consider his or her decision to participate to the trial. 
If the subject is unable to read, the informed consent will be obtained in the presence of an impartial 
witness, e.g., a person independent of the study who will read the informed consent form and the 
written information for the subject.
Consent must be documented by the subject’s dated signature. The signature confirms that the consent 
is based on information that has been understood. Moreover, the Investigator must sign and date the 
informed consent form.
Each subject’s signed informed consent must be kept on file by the Investigator. One copy must be 
given to the subject.
16.
SOURCE DOCUMENTS/DATA
16.1 Recording of source data


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Data collection is the responsibility of the clinical trial staff at the site under the supervision of the 
site investigator. The investigator is responsible for ensuring the accuracy, completeness, legibility, 
and timeliness of the data reported. All source documents should be completed in a neat, legible 
manner to ensure accurate interpretation of data.  
Data recorded in the eCRF derived from source documents should be consistent with the data 
recorded on the source documents. 
16.2 Direct access to source document/data
The Investigators or designated must permit trial-related monitoring, audits, Ethics Committee review 
or regulatory inspection, providing direct access to source data/documents.
17.
STUDY MONITORING
Monitoring will be performed by 
 who has been designated by Chiesi.
It is understood that the monitor(s) will contact and visit the Investigator/centre before the study, 
regularly throughout the study and after the study had been completed, and that they will be permitted 
to inspect the various study records: case reports form, Investigator study file and source data, 
provided that subject confidentiality is respected.
The purposes of these visits are:

to assess the progress of the study;

to review the compliance with the study protocol;

to discuss any emergent problem;

to check the eCRFs for accuracy and completeness;

to validate the contents of the eCRFs against the source documents;

to assess the status of drug storage, dispensing and retrieval;

Prior to each monitoring visit, the Investigator or staff will record all data generated since the 
last visit on the case report forms. The Investigator and/or study staff will be expected to be 
available for at least a portion of the monitoring visit to answer questions and to provide any 
missing information;

It is possible that the Investigator site may be audited by Sponsor personnel or regulatory 
national and/or international regulatory agencies during and after the study has been 
completed.
18.
QUALITY ASSURANCE
The R&D Quality Assurance Department of Chiesi may perform an audit at any time according to 
the Sponsor’s Standard Operating Procedures, in order to verify whether the study is being conducted 
in agreement with Good Clinical Practices and the protocol. 
19.
INSURANCE AND INDEMNITY 
Chiesi holds and will maintain an adequate insurance policy covering damages arising out of Chiesi’s 
sponsored clinical research studies.
Chiesi will indemnify the Investigator and hold him/her harmless for claims for damages arising out 
of the investigation, in excess of those covered by his/her own professional liability insurance, 
providing that the drug was administered under his/her or deputy’s supervision and in strict 
accordance with accepted medical practice and with the study protocol.
The Investigator must notify Chiesi immediately upon notice of any claims or lawsuits. 


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20.
CONFIDENTIALITY
All study documents are provided by the Sponsor in confidence to the Investigator and his/her 
appointed staff. None of this material may be disclosed to any party not directly involved in the study 
without written permission from Chiesi.
The Investigator must assure the subject’s anonymity will be maintained. The Investigator will keep 
a separate list with at least the initials, the subject’s study numbers, names, and addresses and 
telephone numbers. The Investigator will maintain this for the longest period of time allowed by 
his/her own institution and, in any case, until further communication from Chiesi.
21.
PREMATURE TERMINATION OF THE STUDY 
Both the Sponsor and the Investigator reserve the right to terminate the study at any time. Should this 
be necessary, the procedures for an early termination or temporary halt will be arranged after 
consultation by all involved parties.
The Sponsor should submit a written notification to the Regulatory Authority concerned and Ethics 
Committee providing the justification of premature ending or of the temporary halt.
22.
CLINICAL STUDY REPORT
The clinical study report, including the statistical and clinical evaluations, shall be prepared and sent 
to principal Investigator for agreement and signature.
At the end of the trial a summary of the clinical study report will be provided to the Ethics 
Committees, to the Competent Authority of the EU Member State and to Investigator.
23.
RECORD RETENTION
After completion of the study, all documents and data relating to the study will be kept in an orderly 
manner by the Investigator in a secure study file. 
Regulations require that essential documents must be retained for at least two years after the final 
marketing approval in an ICH region or until two years have elapsed since the formal interruption of 
the clinical development of the product under study.
It is the responsibility of the Sponsor to inform the Investigator of when these documents can be 
destroyed. The Investigator must contact Chiesi before destroying any trial-related documentation. In 
addition, all subjects’ medical records and other source documentation will be kept for the maximum 
time permitted by the institution.
24.
PUBLICATION OF RESULTS
Chiesi is entitled to publish and/or present any results of this study at scientific meetings, and to 
submit the clinical trial data to national and international Regulatory Authorities and, if they fall under 
the Chiesi commitments on Clinical Trial Transparency, to make them available on www.chiesi.com 
website.
Chiesi furthermore reserves the right to use such data for industrial purposes.
In the absence of a Study Steering Committee, Investigators will inform Chiesi before using the 
results of the study for publication or presentation, and agree to provide the Sponsor with a copy of 
the proposed presentation. Data from individual study sites must not be published separately.
Negative as well as positive results should be published or otherwise made publicly available 
according to the relevant regulatory requirements. 


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25.
REFERENCES
[1]
Global Strategy for the Diagnosis, Management and Prevention of COPD, Global Initiative 
for Chronic Obstructive Lung Disease (GOLD) 2019. http://goldcopd.org/gold-2019-global-
strategydiagnosis- management-prevention-copd
[2]
Singh D, Roche N, Halpin D, Agusti A, Wedzicha JA, Martinez FJ. Current controversies in 
the pharmacological treatment of chronic obstructive pulmonary disease. Am. J. Respir. Crit. 
Care Med. 2016;194:541–9. doi:10.1164/rccm.201606-1179PP
[3]
Calverley PMA, Rabe KF, Goehring U-M, Kristiansen S, Fabbri LM, Martinez FJ, et al. 
Roflumilast in symptomatic chronic obstructive pulmonary disease: two randomised clinical 
trials. Lancet 2009;374:685–94. doi:10.1016/S0140-6736(09)61255-1
[4]
Fabbri LM, Calverley PMA, Izquierdo-Alonso JL, Bundschuh DS, Brose M, Martinez FJ, et 
al. Roflumilast in moderate-to-severe chronic obstructive pulmonary disease treated with 
long-acting bronchodilators: two randomised clinical trials. Lancet 2009;374:695–703. 
doi:10.1016/S0140-6736(09)61252-6
[5]
Martinez FJ, Calverley PMA, Goehring U-M, Brose M, Fabbri LM, Rabe KF. Effect of 
roflumilast on exacerbations in patients with severe chronic obstructive pulmonary disease 
uncontrolled by combination therapy (REACT): a multicentre randomised controlled trial. 
Lancet Elsevier; 2015;385:857–66. doi:10.1016/S0140-6736(14)62410-7
[6]
Villetti G, Carnini C, Battipaglia L, Preynat L, Bolzoni PT, Bassani F, et al. CHF6001 II: A 
novel phosphodiesterase 4 inhibitor, suitable for topical pulmonary administration - in vivo 
preclinical pharmacology profile defines a potent anti-inflammatory compound with a wide 
therapeutic window. J. Pharmacol. Exp. Ther. 2015;352:568–78. doi:10.1124/jpet.114.220558
[7]
Moretto N, Caruso P, Bosco R, Marchini G, Pastore F, Armani E, et al. CHF6001 I: A novel 
highly potent and selective phosphodiesterase 4 inhibitor with robust anti-inflammatory 
activity and suitable for topical pulmonary administration. J. Pharmacol. Exp. Ther. 
2015;352:559–67. doi:10.1124/jpet.114.220541
[8]
F Mariotti, M Govoni, G Lucci, D Santoro, M-A Nandeuil. Safety, tolerability, and 
pharmacokinetics of single and repeat ascending doses of CHF6001, a novel inhaled 
phosphodiesterase-4 inhibitor: two randomized trials in healthy volunteers.  Int J COPD  
2018; 13: 3399-3410. doi: 10.2147/COPD.S174156
[9]
Singh D, Leaker B, Boyce M, Nandeuil MA, Collarini S, Mariotti F, Santoro D, Barnes PJ A 
novel inhaled phosphodiesterase 4 inhibitor (CHF6001) reduces the allergen challenge 
response in asthmatic patients. Pulm Pharmacol Ther. 2016 Oct;40:1-6. doi: 
10.1016/j.pupt.2016.06.011.  Epub 2016 Jun 29.
[10] D. Singh, M.A. Nandeuil, G. Lucci, F. Mariotti, D. Santoro, S. Smith, A. DeSoyza, T. 
Harrison, M. Albayaty, G. Poli, D. Acerbi, M.A. Nandeuil. Safety, Efficacy and 
Pharmacokinetic profile of CHF6001, a novel selective inhaled PDE4 inhibitor in patients 
with COPD.  Am J Respir Crit Care Med, 2016


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[11] D Singh, K M Beeh, B Colgan, O Kornmann, B Leaker, H Watz, G Lucci, S Geraci, A 
Emirova, M Govoni, MA Nandeuil. Effect of the inhaled PDE4 inhibitor CHF6001 on 
biomarkers of inflammation in COPD. Respiratory Research (2019) 20:180 
doi.org/10.1186/s12931-019-1142-7
[12] Singh D, M-A Nandeuil, C Pigeon-Francisco, A Emirova, D Santoro, S Biondaro, G Cohuet, 
M Govoni, S Petruzzelli. Efficacy and safety of CHF6001, a novel inhaled PDE4 inhibitor in 
COPD: the PIONEER dose finding study. Am J Respir Crit Care Med 2019;199: A4529.
[13] CHF6001 IB, version 2.0 dated 25-Sep-2020
[14]
 
 
[15]
 
 
[16]
 
[17] Miller MR, Hankinson J, Brusasco V et al., Standardisation of spirometry. Eur Respir J, 2005. 
26(2): p. 319-38 
[18] Brian L. Graham et al. Standardization of Spirometry 2019 Update: An Official ATS/ERS 
Technical Statement. American Journal of Respiratory and Critical Care Medicine Volume 
200 Number 8. October 15 2019
[19] Philip H. Quanjer et al., Multi-ethnic reference values for spirometry for the 3–95-yr age 
range: the global lung function 2012 equations - European Respiratory Journal  2012; 40: 
1324–1343
[20] Davies B and Morris T (1993) Physiological parameters in laboratory animals and humans. 
Pharm Res 10:1093–1095.
[21] Guideline on the Investigation of Bioequivalence. Doc. Ref.: CPMP/EWP/QWP/1401/98 
Rev. 1/ Corr  - EMA – 2010. 


Clinical Study Protocol – Appendix 1
Clinical Study Code: CLI-06001AA1-02
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APPENDIX 1 - Approval of the protocol by clinical investigator(s)
EVALUATION OF THE ABSOLUTE BIOAVAILABILITY AND MASS BALANCE 
OF CHF6001 FOLLOWING A SINGLE INHALED DOSE CO-ADMINISTERED 
WITH AN INTRAVENOUS RADIOLABELLED MICROTRACER DOSE IN 
HEALTHY VOLUNTEERS 
Product: 

Inhalation: CHF6001 800µg/20mg 

Infusion: [14C]-CHF6001 1,85µg/1ml 
Pharmaceutical Form: 

Inhalation: Dry power Inhaler 

Infusion: Solution 
Approval of Clinical Study Protocol by the Investigator: 
I have carefully read this protocol and I agree that it contains all the necessary information 
required to conduct the study and I agree to conduct it as described.
I understand that this trial will not be initiated without Ethics Committee/Institutional Review 
Board approvals and that the administrative requirements of the governing body of the 
institution will be fully complied with.
Informed written consent will be obtained from all participating subjects and appropriately 
documented, prior to their enrolment in the study. 
The undersigned agrees that the trial will be carried out in conformity with the Declaration of 
Helsinki (as applicable, with attention being drawn to Section concerning freely given consent), 
ICH E6 Good Clinical Practices and with all the other local laws and regulations relevant to the 
use of new and approved therapeutic agents in subjects.
Principal Investigator’s Name:  __________________,MB ChB              Centre No. : 
_____________________
Signature
     Date
Chiesi Farmaceutici S.p.A.
Via Palermo 26/A
43122 Parma - Italy


Clinical Study Protocol – Appendix 2
Clinical Study Code: CLI-06001AA1-02
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APPENDIX 2 – RECOMMENDATIONS RELATED TO 
CONTRACEPTION 
AND 
PREGNANCY 
TESTING 
IN 
CLINICAL TRIALS
Definition of women of childbearing potential and of fertile men
For the purpose of this document, a woman is considered of childbearing potential (WOCBP), i.e. fertile, following 
menarche and until becoming post-menopausal unless permanently sterile. 
Definition of women of non-childbearing potential 
For the purpose of this document, a woman is considered of non-childbearing potential (WNOCBP), i.e. sterile, 
when physiologically incapable of becoming pregnant (i.e. post-menopausal or permanently sterile).
-
A postmenopausal state is defined as no menses for 12 months without an alternative medical cause. A high 
follicle stimulating hormone (FSH) level in the postmenopausal range may be used to confirm a post-
menopausal state in women not using hormonal contraception or hormonal replacement therapy. However, 
in the absence of 12 months of amenorrhea, a single FSH measurement is insufficient.
-
Permanent sterilisation methods include hysterectomy, bilateral salpingectomy and bilateral oophorectomy.
Definition of fertile men
For the purpose of this document, a man is considered fertile after puberty unless permanently sterile by bilateral 
orchidectomy.
Birth control methods, which may be considered as highly effective
For the purpose of this guidance, methods that can achieve a failure rate of less than 1% per year when used 
consistently and correctly are considered as highly effective birth control methods. Such methods include:
• combined (estrogen   and   progestogen   containing) hormonal contraception 
associated with inhibition of ovulation:

oral

intravaginal

transdermal
• progestogen-only hormonal contraception associated with inhibition of ovulation:

oral

injectable

implantable
• intrauterine device (IUD)
• intrauterine hormone-releasing system (IUS)
• bilateral tubal occlusion
• vasectomised partner 1
• sexual abstinence 2
1 Vasectomised partner is a highly effective birth control method provided that partner is the sole sexual partner of 
the WOCBP trial participant and that the vasectomised partner has received medical assessment of the surgical 
success.
2 Sexual abstinence is considered a highly effective method only if defined as refraining from heterosexual 
intercourse during the entire period of risk associated with the study treatments. The reliability of sexual abstinence 
needs to be evaluated in relation to the duration of the clinical trial and the preferred and usual lifestyle of the 
subject.
Clinical Trial Facilitation Group guidance: Recommendations related to contraception and pregnancy 
testing in clinical trials. Final version 2014-09-15


Clinical Study Protocol – Appendix 3
Clinical Study Code: CLI-06001AA1-02
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APPENDIX 3 – BREAKDOWN OF BLOOD SAMPLES COLLECTION
Detailed Blood Collection Volumes for PK sampling:
TOTAL VOLUME TO BE COLLECTED PER SUBJECT
Clinical chemistry
 & Fasting glucose
Hematology 
Serology 
Whole Blood AMS
PK
PK-AMS 
Metabolite profiling 
Screening
1
1
1
Day -1
Day 1
4
12
15
15
Day 2
1
1
1
Day 3
1
1
1
Day 4
1
1
1
Day 5
1
1
1
Day 6
1
1
1
Day 7
1
1
1
Day 8
1
1
1
Day 9
1
1
1
Day 10
1
1
1
Day 11
1
1
1
1
1
Follow up
Total vol (mL) 
per sample
3.5
4
3.5
2
4
5
10
Tot vol (mL)
7
8
3.5
8
88
125
250
Total vol (mL) 
per subject
489.5

SAP_001.pdf: 
STUDY CODE No.: CLI-06001AA1-02
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STATISTICAL ANALYSIS PLAN
STUDY CODE No.: CLI-06001AA1-02 
EVALUATION OF THE ABSOLUTE BIOAVAILABILITY AND MASS BALANCE OF 
CHF6001 FOLLOWING A SINGLE INHALED DOSE CO-ADMINISTERED WITH AN 
INTRAVENOUS RADIOLABELLED MICROTRACER DOSE IN HEALTHY 
VOLUNTEERS 
Version No.:1.0  
Date: 15 March 2022 
Author(s): 
, Sr Statistician II, 
 
 Sr Biostatistician I, 
 
The information contained in this document is confidential and will not be disclosed to others without written 
authorization from Chiesi Farmaceutici S.p.A, with the exception of appropriate Institutional Review Board and 
Ethics Committees or duly authorized representatives of a national regulatory authority under the condition that 
they are requested to keep it confidential. 
NCT no.: NCT04756960


 
 
STUDY CODE No.: CLI-06001AA1-02 
SAP Version Final 1.0 (15March2022) 
 
 
 
 
CONFIDENTIAL 
 
 
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Contents 
LIST OF ABBREVIATIONS ................................................................................................. 4 
1 
INTRODUCTION............................................................................................................ 6 
2 
STUDY DESIGN .............................................................................................................. 6 
3 
STUDY OBJECTIVES .................................................................................................... 8 
4 
STUDY VARIABLES ...................................................................................................... 9 
4.1 
PHARMACOKINETIC VARIABLES ................................................................................. 9 
4.2 
SAFETY VARIABLES .................................................................................................... 9 
4.3 
OTHER VARIABLES ..................................................................................................... 9 
5 
SAMPLE SIZE ............................................................................................................... 10 
6 
ANALYSIS SETS........................................................................................................... 10 
6.1 
SAFETY POPULATION ................................................................................................ 10 
6.2 
PHARMACOKINETIC (PK) POPULATION ..................................................................... 10 
6.3 
OTHER SETS DEFINED FOR TABLES AND LISTINGS.................................................... 10 
7 
GENERAL CONSIDERATIONS FOR STATISTICAL ANALYSIS ...................... 10 
7.1 
STATISTICAL SIGNIFICANCE ...................................................................................... 10 
7.2 
MULTIPLICITY ........................................................................................................... 10 
7.3 
HANDLING OF MISSING DATA ................................................................................... 10 
7.3.1 Missing/Incomplete Dates: .................................................................................. 11 
7.4 
COVARIATES ............................................................................................................. 11 
7.5 
INTERIM ANALYSES .................................................................................................. 11 
7.6 
EXAMINATIONS OF SUBGROUPS ................................................................................ 12 
7.7 
DESCRIPTIVE STATISTICS .......................................................................................... 12 
7.8 
DEFINITIONS ............................................................................................................. 12 
7.8.1 Baseline and Change from Baseline .................................................................... 12 
7.8.2 Study Day ............................................................................................................. 12 
7.9 
EXCLUSION OF DATA FROM THE STATISTICAL ANALYSES ........................................ 12 
7.9.1 Exclusion of Data from All Statistical Analyses .................................................. 12 
7.9.2 Exclusion of Data from PK Analyses ................................................................... 12 
7.10 
LISTINGS ................................................................................................................... 13 
7.11 
CODING ..................................................................................................................... 13 
8 
STUDY POPULATION ................................................................................................ 13 
8.1 
DISPOSITION OF SUBJECTS AND DISCONTINUATIONS ................................................ 13 
8.1.1 Disposition of Subjects ......................................................................................... 13 
8.1.2 Discontinuation from the Study ........................................................................... 13 
8.1.3 Protocol Deviations and Analysis Sets ................................................................ 13 
8.2 
DEMOGRAPHIC AND BASELINE CHARACTERISTICS ................................................... 13 
8.2.1 Demographic Characteristics .............................................................................. 13 
8.2.2 Physical Characteristics ...................................................................................... 14 
8.2.3 Alcohol Breath Test.............................................................................................. 14 
8.2.4 Spirometry at Screening ....................................................................................... 14 
8.3 
MEDICAL HISTORY AND CONCOMITANT DISEASES ................................................... 14 
8.4 
MEDICATIONS ........................................................................................................... 14 
8.5 
PROCEDURES ............................................................................................................ 14 


 
 
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8.6 
COMPLIANCE ............................................................................................................ 15 
9 
PHARMACOKINETICS ANALYSES ........................................................................ 15 
9.1 
PHARMACOKINETIC ANALYSIS ................................................................................. 15 
9.2 
CRITERIA FOR THE CALCULATION OF APPARENT TERMINAL ELIMINATION RATE 
CONSTANT AND HALF-LIFE ................................................................................................... 18 
9.3 
CRITERIA FOR CALCULATION AND REPORTING OF AREA UNDER THE 
CONCENTRATION-TIME CURVE ............................................................................................. 19 
9.4 
CALCULATION OF [14C]-CHF6001 AND [14C]-TOTAL RADIOACTIVITY URINE 
PARAMETERS ........................................................................................................................ 19 
9.5 
CALCULATION OF [14C]-CHF6001 AND [14C]-TOTAL RADIOACTIVITY FECES 
PARAMETERS ........................................................................................................................ 20 
9.6 
CRITERIA FOR HANDLING BELOW THE LIMIT OF QUANTIFICATION OR MISSING 
CONCENTRATIONS FOR PHARMACOKINETIC ANALYSIS ........................................................ 20 
9.7 
TREATMENT OF OUTLIERS IN PHARMACOKINETIC ANALYSIS ................................... 21 
9.8 
PRESENTATION OF PHARMACOKINETIC DATA ........................................................... 21 
10 SAFETY ANALYSES ................................................................................................... 22 
10.1 
EXTENT OF EXPOSURE .............................................................................................. 22 
10.2 
ADVERSE EVENTS ..................................................................................................... 22 
10.3 
VITAL SIGNS ............................................................................................................. 24 
10.4 
12-LEAD SAFETY ELECTROCARDIOGRAM ................................................................. 24 
10.5 
LABORATORY DATA ................................................................................................. 24 
11 OTHER ANALYSES ..................................................................................................... 24 
12 CHANGES IN THE PLANNED ANALYSES FROM STUDY PROTOCOL ........ 24 
13 OUTPUT ......................................................................................................................... 25 
13.1 
SOFTWARE ................................................................................................................ 25 
13.2 
REPORTING CONVENTIONS ....................................................................................... 26 
13.2.1 
Treatment, Visit and Subgroup Descriptors .................................................... 26 
13.2.2 
Decimal places ................................................................................................. 26 
13.2.3 
Other reporting conventions ............................................................................ 26 
13.3 
FORMAT .................................................................................................................... 27 
13.4 
QUALITY CONTROL ................................................................................................... 27 
14 SAS CODE ...................................................................................................................... 28 
14.1 
TABLES THAT NEED DESCRIPTIVE STATISTICS ........................................................... 28 
14.2 
TABLES THAT NEED FREQUENCY COUNTS ................................................................. 28 
15 REFERENCES ............................................................................................................... 28 
16 LIST OF TABLES, LISTINGS AND FIGURES ........................................................ 29 
16.1 
TABLES ..................................................................................................................... 29 
16.2 
LISTINGS ................................................................................................................... 41 
16.3 
FIGURES .................................................................................................................... 51 
 


 
 
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List of Abbreviations 
 
ADR 
Adverse Drug Reaction 
AE 
Adverse Event 
ATC 
Anatomical Therapeutic Chemical classification 
AUC 
Area Under the Curve 
AUC0-∞ 
Area Under the Curve, from 0 to infinite 
AUC0-t 
Area Under the Curve, from 0 to the last quantifiable concentration 
BMI 
Body Mass Index 
BLQ 
Below Limit of Quantification 
CI 
Confidence Interval 
CL 
Systemic Clearance 
CLblood 
Total body blood clearance 
CLR 
Renal plasma clearance 
CLR_blood 
Renal blood clearance 
CP 
Clinical Pharmacologist  
Cmax 
Maximum of concentration 
Cmin 
Minimum of concentration 
CSR 
Clinical Study Report 
CV 
Coefficient of Variation 
DBP 
Diastolic Blood Pressure 
DPI 
Dry-Powder Inhaler 
DRM 
Data Review Meeting 
DRR 
Data Review Report 
ECG 
Electrocardiogram 
eCRF 
Electronic Case Report Form 
EMA 
European Medicines Agency 
Fe 
Cumulative percent of dose excreted 
Fr 
Fraction of the relevant metabolites 
HR 
Heart Rate 
IV 
Intravenous 
LLQ 
Limit of Quantification 
MedDRA 
Medical Dictionary for Regulatory Activities 
PK 
Pharmacokinetics 
PT 
Preferred Term 
Rb/p 
Whole blood to plasma ratio 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SBP 
Systolic Blood Pressure 
SD 
Standard Deviation 
SOC 
System Organ Class 
t½ 
Elimination half-life 
TEAE 
Treatment Emergent Adverse Event 
tmax 
Time to reach the Cmax 
tmin 
Time to reach the Cmin 
Vdss 
Volume of distribution at steady-state 
Vz 
Volume of distribution during the terminal phase 


 
 
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WHO 
World Health Organisation 
WHO-DD 
World Health Organisation Drug Dictionary 
 
 
VERSION HISTORY 
Version  
Date  
Change History 
1.0 
15March2022 
First version 
 
 
 
 
 


 
 
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1 Introduction 
This document presents the Statistical Analysis Plan (SAP) for Chiesi Farmaceutici S.p.A. 
protocol CLI-06001AA1-02: Evaluation of the absolute bioavailability and mass balance of 
CHF6001 following a single inhaled dose co-administered with an intravenous radiolabelled 
microtracer dose in healthy volunteers. 
This analysis plan is based on the final protocol (version 1.0, 19 October 2020) and the final 
electronic case report form (eCRF) (version 2.0, 02Mar2021).  
The SAP provides the description of the final analyses. In case of deviations from the SAP, 
explanations will be provided in the Clinical Study Report (CSR). 
 will perform the statistical analyses and is responsible for the production and quality 
control of all outputs described in this document. 
 
2  Study Design 
This is a phase I, open-label, uncontrolled, non-randomized, single dose, single centre study in 
8 healthy male subjects designed to evaluate absolute bioavailability and mass balance of 
CHF6001 following a single inhaled dose co-administered with an intravenous radiolabelled 
microtracer dose. 
The study consists of two periods (Screening and Treatment) and Follow-up call.  
Screening: 
At Screening visit (31 to 3 days before first study treatment administration), subjects will be 
selected to enter the study according to the eligibility criteria. Study related procedures will be 
performed upon obtaining a signed informed consent. 
Treatment Period: 
During treatment period, subjects will remain at the clinical site from Day -1 until the morning 
of Day 11 and will be administered a single dose of CHF6001 on Day 1. 4 inhalations of 
CHF6001 800µg/20mg NEXThaler® DPI (total dose of 3200µg) co-administered with an 
intravenous microdose (18.5µg and 500nCi (18.5kBq)) of [14C]-labelled CHF6001 will be 
administered. The subjects will be discharged after the 240h post-dose assessments are 
completed. 
Follow-up: 
A follow-up phone call (or visit, if necessary) will be performed between 7 to 10 days after 
discharge or premature discontinuation to check and record concomitant medications and the 
status of any unresolved adverse events or occurrence of any new adverse events (AEs). 
The end of the trial is defined at execution of the Follow-up call (or visit) of the last subject of 
the trial. 
The study plan and scheduled tests are summarised in the following flow-chart: 
 
 
 
 


 
 
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Table 1: Study Plan 
 


 
 
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3 Study Objectives 
The objectives of this study are: 
 To determine the absolute bioavailability of CHF6001 following single inhaled dose 
co-administered with an IV microtracer dose 
 To characterize the mass balance and routes of elimination of CHF6001 after an 
intravenous (IV) microtracer dose 
 To characterize the relevant metabolites in plasma, urine and feces 
 To assess the safety and tolerability of the study treatment 


 
 
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4 Study Variables 
4.1 Pharmacokinetic Variables 
Following are the pharmacokinetic (PK) variables of this study: 
 Plasma [14C]-CHF6001, AUC0-t iv, Cmax_iv, tmax_iv, AUC0-∞_iv, t1/2 iv, Vz, Vdss and CL 
 Plasma [14C]-total AUC0-t iv, Cmax_iv, tmax_iv, AUC0-∞_iv, t1/2 iv 
 Plasma CHF6001 AUC(0-t)_inh, Cmax_inh, tmax_inh, AUC(0-∞)_inh, t1/2_inh, Finh 
 Urine and feces [14C]-CHF6001and [14C]-total excreted fraction (Feu, Fef and Feu+f) 
 Systemic blood clearance [14C]-CHF6001 CLblood, renal plasma and blood clearance 
CLR and CLR_blood 
 Blood to plasma ratio (Rb/p) [14C]-CHF6001 
 Blood to plasma ratio (Rbt/pt) [14C]-total 
 Metabolized fraction (Femet) 
  Hepatic extraction [14C]-CHF6001 (Eh_blood) 
 
Characterization of the relevant metabolites 
For metabolites in plasma accounting for ≥10% of total radioactivity recovered in this matrix, 
structural characterization may be performed, where possible. For metabolites in urine and 
feces (total excreta) accounting for ≥10% of total administered radioactivity, structural 
characterization may be performed, where possible: 
 Fraction of the relevant metabolites in plasma (Fr_plasma), fraction of the relevant 
metabolites recovered in urine and feces (Fr_urine, Fr_feces, Fr_urine+feces) 
Where possible, these data will be reported in a separate report together with structural 
characterization. 
 
4.2 Safety Variables 
Following are the safety variables for this study: 
 AEs and ADRs 
 Vital signs parameters (SBP and DBP) 
 HR from 12-lead electrocardiogram (ECG) 
 Laboratory parameters (haematology, chemistry and fasting glucose) 
 
4.3 Other Variables 
No other variables defined. 
 


 
 
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5 Sample Size 
According to the exploratory nature of the study, no formal sample size calculation was 
performed. A total of 8 subjects will be included in the study to characterize the absolute 
bioavailability and the mass balance of CHF6001. 
 
6 Analysis Sets 
The definitions of the analysis sets are summarised below. A final agreement on the subjects 
be included in or excluded from each analysis set will be reached during the Data Review 
Meeting (DRM) before the database lock. Inclusions and exclusions from analysis sets will be 
fully documented in the Data Review Report (DRR).  
 
6.1 Safety Population 
Safety Population is defined as all enrolled subjects who start study treatment inhalation. 
The Safety population will be used in the analysis of all safety variables. 
 
6.2 Pharmacokinetic (PK) Population 
PK population consists of all subjects from the safety population excluding subjects without 
any valid PK measurement or with major protocol deviations significantly affecting PK, for 
example: incorrect inhalation, change in subject condition (cold), failure in delivery of the 
device, use of non-permitted medications. 
PK variables will be analysed in the PK population. 
 
6.3 Other Sets Defined for Tables and Listings 
For the purposes of tables and listings the following set is defined: 
 Enrolled Set: all subjects who provided informed consent for the study. 
 
7 General Considerations for Statistical Analysis 
7.1 Statistical Significance 
There is no hypothesis testing or confidence interval (CI) calculation performed hence 
statistical significance is not applicable. 
 
7.2 Multiplicity 
Since there is no hypothesis testing involved, multiplicity adjustment is not applicable. 
 
7.3 Handling of Missing Data 
The number of subjects with missing data will be presented under a “Missing” category. Unless 
otherwise stated, missing values will not be included in the denominator count when 
calculating percentages. 


 
 
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When quantitative variables are being summarised, only the non-missing values will be 
evaluated for calculating summary statistics. 
 
7.3.1 Missing/Incomplete Dates: 
Medications: 
In case of missing or incomplete dates not directly allowing allocation to any category of 
medications, a worst-case allocation will be done according to the available parts of the start 
and the stop dates. The medications will be allocated to the first category allowed by the 
available data, according to the following order: 
 Concomitant medication; 
 Post-Study medication; 
 Prior medication. 
Procedures: 
In case of missing or incomplete dates not directly allowing allocation to any category of 
procedure, a worst-case allocation will be done according to the available parts of the start and 
the stop dates. The procedure will be allocated to the first category allowed by the available 
data, according to the following order: 
 Concomitant procedure; 
 Post-Study procedure 
 Prior procedure. 
Adverse events: 
In case of missing or incomplete date/time not directly allowing allocation to any of the 
category of AEs, a worst-case allocation will be done according to the available parts of the 
start and the stop dates/times. The AE will be allocated to the first category allowed by the 
available data, according to the following order: 
 Treatment emergent;  
 Post-study; 
 Pre-treatment. 
Missing severity of adverse events: 
In case of missing severity, the severity will not be imputed and will be reported as “Missing”.  
Other critical missing data, if any, will be discussed during the review of the data. Decisions 
will be fully documented in the DRR. 
 
7.4 Covariates 
Not applicable. 
 
7.5 Interim Analyses 
No interim analysis will be performed. 
 


 
 
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7.6 Examinations of Subgroups 
No subgroup analysis will be performed. 
 
7.7 Descriptive Statistics 
PK variables (except for tmax, tmax_inh and tmax_iv) will be summarized by means of descriptive 
statistics including n (number of observed values), arithmetic mean, standard deviation (SD), 
coefficient of variation (CV), geometric mean, geometric CV, median, minimum and 
maximum. tmax, tmax_inh, tmax_iv will be summarized by using n, median, minimum and 
maximum. 
Descriptive statistics for quantitative variables will include n (the number of non-missing 
values), mean, standard deviation (SD), median, minimum and maximum values.  
Categorical variables will be summarised by using frequency count and percent distributions.  
 
7.8 Definitions 
7.8.1 Baseline and Change from Baseline 
Baseline values are defined as the last evaluation before the first treatment dose (Inhalation). 
For each of the above variable, change from baseline is defined at each post-dose as: 
Post-dose value – baseline value. 
 
7.8.2 Study Day 
The study day relative to the date of study medication intake will be calculated as: 
 date of event – date of first study drug inhalation + 1 (if date of event ≥ date of first 
study drug inhalation); 
or 
 date of event – date of first study drug inhalation (if date of event < date of first study 
drug inhalation). 
 
7.9 Exclusion of Data from the Statistical Analyses 
7.9.1 Exclusion of Data from All Statistical Analyses 
There will not be any exclusions of the collected data from the statistical analyses. 
 
7.9.2 Exclusion of Data from PK Analyses 
Subjects may be excluded from the PK population and hence from the PK analysis if they fall 
in the cases as defined in the European Medicines Agency (EMA) "Guideline on the 
Investigation of Bioequivalence”. 
All data will be reviewed during the DRM in order to make decisions of exclusion from PK 
analyses and documented in the DRR. 
If any data or subject is excluded from the tables or figures, a footnote will be added to notify 
the exclusions. 
 


 
 
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7.10 Listings 
All data collected in the eCRF will be presented in the listings. 
 
7.11 Coding 
Medical and surgical history, concomitant diseases, procedures and adverse events will be 
coded according to Medical Dictionary for Regulatory Activities (MedDRA) version 24.0 
Medications will be coded using the World Health Organization Drug Dictionary (WHO-DD) 
WHO Global B3 format Version -March 2021. 
 
8 Study Population 
8.1 Disposition of Subjects and Discontinuations 
8.1.1 Disposition of Subjects 
The number of subjects treated, who completed treatment period and who completed the 
follow-up will be summarized. All subjects will be included. 
 
8.1.2 Discontinuation from the Study 
The number and percentage of subjects who completed the study, withdrew from the study and 
the number and percentage of subjects with each reason for withdrawal from the study and 
from the treatment period will be presented on Enrolled Set. 
 
8.1.3 Protocol Deviations and Analysis Sets 
Deviations will be classified according to the following categories: 
 Violation of Inclusion Criterion; 
 Violation of Exclusion Criterion; 
 Treatment Administration Deviation; 
 Non-Permitted Medication; 
 Assessment Performed Outside the Allowed Time Window; 
 Visit Performed Outside the Allowed Time Window; 
 Study Procedure Deviation; 
Major and minor protocol deviations will be summarised using the Safety Set.  
The number of subjects included in the Safety and PK Sets will be summarised. 
All deviations, including significant GCP deviation, will be listed. 
 
8.2 Demographic and Baseline Characteristics 
8.2.1 Demographic Characteristics 
Demographic characteristics will be summarised. These will include age (years) and race.  
 


 
 
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8.2.2 Physical Characteristics 
Height (cm), weight (kg) and body mass index (BMI, kg/m2) will be summarized. 
Note:  
 BMI (kg/m²) will be calculated as: weight at the visit (kg) / height (m)2 at Visit 1. 
 
8.2.3 Alcohol Breath Test 
Results of alcohol breath test will only be listed. 
 
8.2.4 Spirometry at Screening 
Spirometry results at Screening will only be listed. 
 
8.3 Medical History and Concomitant Diseases 
Medical/surgical history and concomitant diseases will be summarised by MedDRA system 
organ class (SOC) and preferred term (PT) using Safety Set.  
Notes:  
 medical/surgical history is defined as conditions in the medical/surgical history and 
concomitant diseases eCRF form which are not ongoing at Day 1; 
 concomitant diseases are defined as conditions in the medical/surgical history and 
concomitant diseases eCRF form which are ongoing at Day 1. 
 
8.4 Medications 
Previous medications and concomitant medications will be summarised for the Safety Set 
through frequency distributions and percentages by Anatomical Main Group (1st level of the 
anatomical therapeutic chemical (ATC) classification), Therapeutic Subgroup (2nd level of the 
ATC classification), Chemical Subgroup (4th level of the ATC classification) and preferred 
name. Subjects with at least one prior/concomitant medications will also be summarized. Post-
study medications will only be presented in a listing. 
The medications will be classified according to the following rules: 
 previous medication: medication start date < date of first study drug inhalation and stop 
medication date ≤ date of first study drug inhalation; 
 concomitant medication: date of first study drug inhalation ≤ medication end date; 
 post-study medication: medication start date ≥ date of discharge. 
 
8.5 Procedures 
Previous procedures and concomitant procedures will be summarised for the Safety Set 
through frequency distributions and percentages by MedDRA SOC and PT. 
The procedures will be classified according to the following rules: 
 previous procedures: procedure start date < date of first study drug inhalation and end 
procedure date ≤ date of first study drug inhalation; 


 
 
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 concomitant procedures: date of first study drug inhalation ≤ procedure end date; 
 post-study procedures: procedure start date ≥ date of discharge. 
 
8.6 Compliance 
Treatment Compliance 
In general, treatment compliance will be evaluated on the basis of the information recorded in 
the eCRF. 
Number of correct and incorrect inhalations, number of interruptions during the infusion will 
be summarized using descriptive statistics on Safety population.  
In addition, all variables related to the inhalation and to the IV infusion will be listed. 
 
9 Pharmacokinetics Analyses 
9.1 Pharmacokinetic Analysis 
The concentration ratios of whole blood [14C]-total and [14C]-CHF6001 radioactivity to plasma 
[14C]-total and [14C]-CHF6001 radioactivity, individual and cumulative mass balance of [14C]-
total radioactivity (amount (Aeu, cumulative Aeu, Aef and cumulative Aeu) and percentage of 
dose (feu, Feu, fef and Fef) recovered as [14C]-total radioactivity in urine, feces and total excreta) 
will be calculated by radioanalysis laboratory (
).  
In addition, metabolites in plasma accounting for ≥10% of total radioactivity recovered in this 
matrix, structural characterization may be performed, where possible. For metabolites in urine 
and feces (total excreta) accounting for ≥10% of total administered radioactivity, structural 
characterization may be performed, where possible.  
If available, fraction of the relevant metabolites in plasma (Fr_plasma), fraction of the relevant 
metabolites recovered in urine and feces (Fr_urine, Fr_feces, Fr_urine+feces) will be reported 
in a separate report together with structural characterization performed by radioanalysis 
laboratory (
). 
 
Urine and Feces [14C]-CHF6001 and [14C]-total radioactivity 
Parameter 
Unitsa 
Definition 
Aeu 
mg 
amount excreted in urine (expressed in weight) over sampling 
interval 
Cumulative Aeu 
mg 
cumulative amount excreted in urine, calculated as the sum of 
the amount excreted in urine for each collection period 
feu 
% 
percent of the dose excreted in urine over a sampling interval  
Feu 
% 
cumulative percent of dose excreted in urine, calculated as the 
sum of the percent of dose excreted in urine for each collection 
period 
Aef 
mg 
amount excreted in the feces over sampling interval  
Cumulative Aef 
mg 
cumulative amount excreted in feces, calculated as the sum of 
the amount excreted in feces for each collection period 
fef 
% 
percent of the dose excreted in feces over a sampling interval  


 
 
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Fef 
% 
cumulative percent of dose excreted in feces, calculated as the 
sum of the percent of dose excreted in feces for each collection 
period 
CLR 
L/h 
renal plasma clearance (urine [14C]-CHF6001 only) 
CLR_blood 
L/h 
renal blood clearance where CLR_blood =CLR/Rb/p (urine 
[14C]-CHF6001 only) 
Eh blood 
 
hepatic extraction calculated from intravenous systemic and 
renal blood clearance (CLblood = CL/Rb/p) and CLR_blood 
=CLR/Rb/p, respectively and human hepatic blood flow (QH; 
assumed to be 87 l/h). Eh blood = (CLblood - CLR_blood)/ QH 
Cumulative Aeu+f 
mg 
cumulative amount excreted in urine and feces, calculated as 
Cumulative Aeu + Cumulative Aef 
Fetot 
% 
cumulative amount excreted in urine and feces, calculated as Feu 
+ Fef 
Femet 
 
metabolized fraction calculated as Femet = [cumulative Aeu+f 
(14C-total) – cumulative Aeu+f ([14C]-CHF6001)]/ cumulative 
Aeu+f (14C-total) 
a Units are based on concentration units (provided by the bioanalytical lab or preferred units for presentation of PK parameters) 
and dose units used in the study. 
 
 The following PK parameters will be determined by 
 where possible from the plasma 
concentrations of CHF6001 following inhaled administration using noncompartmental 
methods in validated software program Phoenix WinNonlin (Phoenix WNL, Certara, Version 
8.1 or higher): 
Parameter 
Unitsa 
Definition 
AUC(0-t)_inh 
h*xg/mL 
area under the concentration-time curve from 0 to the last 
quantifiable concentration (tlast)b 
AUC(0-∞)_inh 
h*xg/mL 
area under the concentration-time curve extrapolated to infinityc 
%AUCextrap 
% 
percentage of AUC due to extrapolation from the last 
quantifiable concentration to infinity 
Cmax_inh 
xg/mL 
the value of the maximum observed concentration 
tmax_inh 
h 
the time of the maximum observed concentration 
tlast_inh 
h 
the time of the last quantifiable concentration 
t1/2_inh 
h 
terminal half-life 
Finh 
 
inhaled absolute bioavailability based on AUC(0-∞)  
a Units are based on concentration units (provided by the bioanalytical lab or preferred units for presentation of PK parameters) 
and dose units used in the study.   
b Area under the concentration-time curve will be calculated using the linear trapezoidal rule with linear interpolation 
c Based on the last observed quantifiable concentration 
 
 


 
 
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The following PK parameters will be determined where possible from the plasma, whole blood, 
urine and feces [14C]-CHF6001 concentrations, plasma whole blood, urine and feces [14C]-total 
concentrations following intravenous (IV) infusion administration of [14C]-CHF6001 in each 
subject using noncompartmental methods in validated software program Phoenix WNL 
(Certara, Version 8.1 or higher): 
Plasma [14C]-CHF6001 and [14-C]-total radioactivity 
 
 
Parameter 
Unitsa 
Definition 
[14C]- CHF6001 
and [14C]- Total 
AUC(0-t)_iv 
h*xg/mL area under the concentration-time curve from 0 to 
the last quantifiable concentration (tlast)b 
[14C]- CHF6001 
and [14C]- Total 
AUC(0-∞)_iv 
h*xg/mL area 
under 
the 
concentration-time 
curve 
extrapolated to infinityc 
[14C]- CHF6001 
and [14C]- Total 
%AUCextrap 
% 
percentage of AUC due to extrapolation from the 
last quantifiable concentration to infinity 
[14C]- CHF6001 
and [14C]- Total 
Cmax_iv 
xg/mL 
the 
value 
of 
the 
maximum 
observed 
concentration 
[14C]- CHF6001 
and [14C]- Total 
tmax_iv 
h 
the time of the maximum observed concentration 
 
tlast_iv 
h 
the time of the last quantifiable concentration 
[14C]- CHF6001 
and [14-C]-Total 
t1/2_iv 
h 
terminal half-life 
[14C]- CHF6001 
CL 
L/h 
total body plasma clearance (plasma [14C]-
CHF6001) 
[14C]- CHF6001 
Vz 
L 
volume of distribution during the terminal phase 
(plasma [14C]-CHF6001) 
[14C]- CHF6001 
Vdss 
L 
volume of distribution at steady state (plasma 
[14C]-CHF6001) 
 
Plasma 
concentration of 
[14C]- 
CHF6001/Total 
Radioactivity 
Ratio  
NA 
ratio of plasma concentration of [14C]-CHF6001 
to plasma concentration of [14C]-total 
 
AUC(0-∞) Plasma 
[14C]- 
CHF6001/Total 
Radioactivity 
Ratio  
NA 
AUC(0-∞) of plasma [14C]-CHF6001 relative to 
AUC(0-∞) of plasma [14C]-total radioactivity  
 
Cmax 
Plasma 
[14C]- 
CHF6001/Total 
Radioactivity 
Ratio  
NA 
Cmax_iv of plasma [14C]-CHF6001 relative to 
Cmax_iv of plasma [14C]-total radioactivity 
a Units are based on concentration units (provided by the bioanalytical lab or preferred units for presentation of PK parameters) 
and dose units used in the study.  For total radioactivity, Cmax and AUC units will be presented as mass equivalents (ng Eq/g 
and h*ng Eq/g, respectively).  
b Area under the concentration-time curve will be calculated using the linear trapezoidal rule with linear interpolation 
c Based on the last observed quantifiable concentration 
 
 


 
 
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Blood [14C]-CHF6001 and [14-C]-total radioactivity 
 
 
Parameter 
Units 
Definition 
[14C]-
CHF6001 
Rb/p 
NA 
whole blood to plasma ratio is to be determined on the 
[14C]-CHF6001 
whole 
blood 
and 
plasma 
concentration (Cb and Cp) 20 minutes after the start of 
infusion, calculated as Rb/p =Cb/Cp 
[14C]-total 
Rbt/pt 
NA 
whole blood to plasma ratio is to be determined on the 
total-[14C] whole blood and plasma concentration (Cbt 
and Cpt) 20 minutes after the start of infusion, 
calculated as Rbt/pt =Cbt/Cpt 
Additional PK parameters may be determined where appropriate. 
Concentration ratios of plasma [14C]-CHF6001 to plasma [14C]-total radioactivity will be listed 
by time point and summarized. 
Pharmacokinetic analysis will be carried out using actual dose administered (mg) and actual 
blood sampling times post-dose. If an actual time is missing, the sample concentration result 
will be treated as missing unless there is scientific justification to include the result using the 
nominal time.  
The parameters Cmax_inh, Cmax_iv, tlast_inh, tlast_iv, tmax_inh and tmax_iv will be obtained directly from the 
concentration-time profiles. If Cmax occurs at more than 1 timepoint, tmax will be assigned to 
the first occurrence of Cmax. 
Inhaled absolute bioavailability (Finh) based on AUC(0-∞) will be calculated as follows: 
Finh = (AUC(0-∞)_inh x Doseiv)/ (AUC(0-∞)_iv x Doseinh).  
The parameter AUC(0-t) or other common partial area may be used to determine Finh, metabolite 
and total radioactivity ratios if AUC(0-∞) cannot be reliably calculated for the majority of 
subjects. 
9.2 Criteria for the Calculation of Apparent Terminal Elimination Rate 
Constant and Half-life 
The start of the terminal elimination phase for each subject will be defined by visual inspection 
and generally will be the first point at which there is no systematic deviation from the log-
linear decline in concentrations. 
The apparent terminal elimination rate constant (λz) will only be calculated when a reliable 
estimate can be obtained using at least 3 data points, preferably not including Cmax, and the 
adjusted coefficient for determination of exponential fit (R2-adj) of the regression line is 
≥0.7.Parameters requiring λz for their calculation (e.g., AUC(0-∞), t1/2, CL, Vz and Vdss) will only 
be calculated if the R2-adj value of the regression line is ≥0.7. If this rule of r²adj ≥0.7 is not 
respected, this should be well described in the Data Review Report (DRR). 
The following regression-related diagnostic PK parameters will be determined, when possible: 


 
 
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Parameter 
Units 
Definition 
λz 
1/h 
apparent terminal elimination rate constant 
λz Upper 
h 
end of exponential fit 
λz Lower 
h 
start of exponential fit 
λz N 
 
number of data points included in the log-linear regression 
λz Span Ratio 
 
time period over which λz was determined as a ratio of t1/2 
R2-adj 
 
adjusted coefficient for determination of exponential fit 
Where possible, the span of time used in the determination of λz (i.e., the difference between 
λz Upper and λz Lower) should be ≥2 half-lives. If the λz Span Ratio is <2, the robustness of 
the t1/2 values will be discussed in the Data Review Report (DRR). 
9.3 Criteria for Calculation and Reporting of Area Under the 
Concentration-time Curve 
The minimum requirement for the calculation of area under the concentration-time curve 
(AUC) will be the inclusion of at least 3 consecutive concentrations above the lower limit of 
quantification. If there are only 3 consecutive concentrations, at least 1 should follow Cmax. An 
exception may be made for metabolites, where Cmax may be the last timepoint.  
If the extrapolated area is >20%, AUC(0-∞) (and derived parameter i.e. Plasma [14C] - 
CHF6001) / Total Radioactivity Ratio) may be excluded from the summary statistics at the 
discretion of the sponsor or pharmacokineticist and should be well described in the Data 
Review Report (DRR). 
9.4 Calculation of [14C]-CHF6001 and [14C]-total radioactivity Urine 
Parameters  
The amount of the dose administered recovered (Aeu) in urine as [14C]-CHF6001 for each urine 
collection interval (t1-t2) will be calculated as the product of urine concentration and urine 
volume.  Where only urine sample weight is supplied, a specific gravity of 1 g/mL will be 
assumed, and it will be considered equivalent to urine volume.  A total cumulative Aeu 0-x h will 
be calculated by summing across collection intervals over the 0-x h interval, where x = end of 
the last collection time interval. 
The percentage of the dose administered recovered in urine over the time interval t1 to t2 (feu t1-
t2) as [14C]-CHF6001 will be calculated for each urine collection interval as follows: 
feu t1-t2 = (Aeu t1-t2 / doseIV) × 100 
Cumulative feu (Feu) will be calculated by summing the feu t1-t2 values across collection intervals 
in the same manner as cumulative Aeu. 
Renal plasma clearance (CLR) will be calculated according to the following formula: 
CLR = cumulative Aeu / AUC(0-∞)_iv 


 
 
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Alternatively, AUC(0-t) may be used if AUC(0-∞) cannot be reliably calculated following the 
formula cumulative Ae(0 t) / AUC(0-t), where ‘t’ is the longest common interval with quantifiable 
concentrations in plasma and urine. 
The fraction of the relevant metabolites recovered in urine calculated as amount recovered 
(Fr_Urine) will be calculated as follows: 
Fr_Urine = (Aru / DoseIV) x 100 
9.5 Calculation of [14C]-CHF6001 and [14C]-total radioactivity Feces 
Parameters  
The amount of the dose administered recovered (Aef) in feces as [14C]-CHF6001 for each faecal 
collection interval (t1-t2) will be calculated as concentration in faecal collected sample and 
faecal collected sample weight.  A total cumulative Aef 0-x h will be calculated by summing 
across collection intervals over the 0-x h interval, where x = end of the last collection time 
interval. 
The percentage of the dose administered recovered in feces over the time interval t1 to t2 (fef t1-
t2) as [14C]-CHF6001 will be calculated for each faecal collection interval as follows: 
fef t1-t2 = (Aef t1-t2 / DoseIV) × 100 
Cumulative fef (Fef) will be calculated by summing the fef t1-t2 values across collection intervals 
in the same manner as cumulative Aef. 
The fraction of the relevant metabolites recovered in feces calculated as amount recovered 
(Fr_feces) will be calculated as follows: 
Fr_feces = (Arf / DoseIV) x 100 
9.6 Criteria for Handling Below the Limit of Quantification or Missing 
Concentrations for Pharmacokinetic Analysis 
Plasma and whole blood concentrations below the limit of quantification will be reported as 
BLQ in the concentration and parameters tables. All BLQ values will be considered as 0.00 for 
pharmacokinetic and descriptive statistical analyses. 
All concentration data-points with time deviations outside the permitted ranges (see Table in 
section 7.2.1.1 and 7.2.1.2 of study protocol) will be excluded from the descriptive statistics 
on plasma, whole blood and urine concentrations, explained by a footnote in the appropriate 
tables and figures, but kept in PK parameters estimation unless very large deviations not 
allowing for a proper calculation of PK parameters are found: these cases will be discussed 
during the Data Review Meeting and decision will be fully documented in the DRR. 
General rules to handle missing PK data are detailed below, but a case by case evaluation will 
be performed during the DRM in order to properly handle every specific situation (e.g., missing 
values at time points close to expected tmax). Decisions will be fully documented in the DRR. 
Rules on how to deal with missing values when deriving PK AUC: 


 
 
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● If intermediate values (not the pre-dose, and not more than 3 consecutive) are missing, 
they will be considered as missing for the PK analysis and automatically imputed by 
linear interpolation by Phoenix WNL software. 
● If multiple (>1) adjacent values are missing or more than 3 values are missing, then the 
AUCs will be reported, flagged in the result tables and excluded from the statistical 
analyses. However, such situations will be discussed case by case at the discretion of the 
clinical pharmacologist (CP) during the review of the data and fully documented in the 
DRR. 
● If the predose value is missing, then the predose value should be set to “0” for the 
estimation of the PK parameters. In case of missing predose sample, the Phoenix WNL 
software will automatically use the value of 0 for the estimation of the PK parameters;  
● For an estimation of the urine and stool PK parameters, missing predose value do not 
affect the final parameter estimates because they are not used in the calculation of Ae. 
The following rules apply with special situations defined below: 
● If an entire concentration-time profile is BLQ, it will be excluded from PK analysis. 
● Where 2 or more consecutive concentrations are BLQ at the end of a profile, the profile 
will be deemed to have terminated and any further quantifiable concentrations will be set 
to missing for the calculation of the PK parameters, unless they are considered to be a 
true characteristic of the profile of the drug. 
● If a predose plasma and whole blood concentration is missing, it will be set to 0 by default 
within Phoenix WNL. 
Urine and feces concentrations that are missing or BLQ will be set to zero for the calculation 
of Aeu t1-t2 and Aef t1-t2. 
9.7 Treatment of Outliers in Pharmacokinetic Analysis 
If a value is considered to be anomalous due to being inconsistent with the expected PK profile, 
it may be appropriate to exclude the value from the PK analysis. However, the exclusion of 
any data be fully documented in the DRR and discussed in the CSR. 
Any quantifiable predose concentration value will be considered anomalous and set to missing 
for the PK analysis. This will be set to 0 by default within Phoenix WNL. 
9.8 Presentation of Pharmacokinetic Data 
For plasma whole blood and urine concentration data, the following rules will apply: 
● Values that are BLQ will be set to 0 for the calculation of summary statistics. In listings 
BLQ values will be reported as “<LLQ” where LLQ will be replaced with the value for 
the lower limit of quantification 
● Arithmetic mean or median values that are BLQ will be presented as 0. 


 
 
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● If any BLQ results (treated as 0) are in a series of summarized data, geometric mean and 
CV% of geometric mean will be reported as not calculated (NC). 
For PK parameters the following rule will apply: 
● Geometric mean and coefficient of variation will not be calculated for tlast and tmax. 
Concentration (mass equivalent in mL or mass in mL)/time curves for plasma total [14C], [14C]-
CHF6001 and non-radiolabelled CHF6001 will be presented in linear/linear and log/linear 
scale. Plots will be presented based on arithmetic means and by subject i.e. all individual 
plasma concentration/time curves will be presented in one graph. 
Arithmetic means (+ SD) showing [14C]-total recovery (Fe_f, Fe_u, Fe_tot in the same graph) 
and 14C-CHF6001 recovery (Fe_f, Fe_u, Fe_tot in the same graph) over the time will be 
presented using line charts. 
All individual concentration data (mass equivalent per mL or mass per mL) and PK parameters 
will be listed.  
In addition, a listing of the actual sampling times relative to drug inhalation will be provided. 
Concentrations will be summarized by scheduled sampling time by using n, arithmetic mean, 
SD, CV, median, minimum and maximum. 
For each applicable listing, appropriate flag will be added in case of a concentration reported 
as ND (i.e., not done) or NS (i.e., no sample) or of value BLQ or for PK parameters not 
estimated / Not calculated. 
 
10 Safety Analyses 
10.1 Extent of Exposure 
Not applicable since this is a single dose study. 
 
10.2 Adverse Events 
An AE will be classified as pre-treatment AE if it starts after the informed consent signature 
and before the time of first inhalation of the study drug (date of informed consent < AE onset 
date and AE onset date-time < date-time of first study drug inhalation). 
All adverse events starting on or after the time of first study drug inhalation will be classified 
as treatment emergent adverse event (TEAE) (date-time of first study drug inhalation ≤ AE 
onset date-time ≤ date of discharge). 
An AE will be classified as a post-study AE if it starts after the date of discharge (AE onset 
date > date of discharge). 
An adverse drug reaction (ADR) is an AE judged as related to the study medication. 
A serious ADR is a serious AE (SAE) judged as related to the study medication. 
A severe AE is an AE with severe intensity. 
An AE leading to discontinuation will be identified based on the information collected in study 
termination form (i.e. reason for discontinuation =Adverse event). 
An AE leading to death is an AE with outcome equal to “Fatal”. 


 
 
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Pre-treatment AEs, TEAEs and post-study AEs will be presented separately. Pre-treatment AEs 
and post-study AEs will be presented in the listings only. 
The number of TEAEs, drug-related TEAE, serious TEAE and TEAE leading to study 
discontinuation and the number and the percentage of subjects experiencing TEAEs, drug-
related TEAEs, serious TEAEs and TEAEs leading to study discontinuation will be 
summarised.  
The MedDRA SOC and PT will also be used for tabulation. The number and percentage of 
subjects with at least one AE and the number of AEs will be presented by SOC and PT for the 
above categories of AEs. 
Derived variables presented in the listings: 
The relative day of AE onset will be calculated as follows: 
 for pre-treatment AEs: 
o AE onset date - date of first inhalation of study drug (if AE onset date is 
completely known); 
o missing (if AE onset date is incomplete or unknown). 
 For TEAEs and post-Study AEs: 
o AE onset date – date-time of first inhalation of study drug +1 (if AE onset date 
is completely known); 
o missing (if AE onset date is incomplete or unknown). 
 
The duration of an AE (hours) will be calculated as follows: 
 (AE end date-time – AE onset date-time) /3600 (when both date-time are completely 
known) 
The duration of an AE (days) will be calculated as follows: 
If duration in hours ≥ 24 hours:  
 AE end date – AE onset date +1  (when both dates are completely known); 
 date of completion/discontinuation – AE onset date + 1 (when the AE onset date is fully 
known but the AE is not resolved at the end of the trial): in this case the duration will 
be presented as “>x days” in the listing rather than “x days”; 
 missing (when the AE onset date is incomplete or unknown, or when the AE has 
resolved but with an incomplete or unknown end date, or when the AE onset date is > 
date of completion/discontinuation and the AE is not resolved). 
If duration in hours <24 hours: 
 duration (hours)/24  
Duration will be displayed in days with values <1 days in case of short term AE. 
 
 


 
 
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10.3 Vital Signs 
For the purpose of safety analyses, the mean of triplicate measurements of systolic blood 
pressure (SBP) and diastolic blood pressure (DBP) will be used. The mean SBP and DBP 
values at baseline and at 2.5 hours post- dose will be summarized using descriptive statistics. 
Change from baseline to 2.5 hours post-dose in mean SBP and DBP will also be summarized. 
Number and percentage of subjects with clinically relevant changes from baseline to 2.5 
hours post-dose in SBP and DBP will be summarized. Clinically relevant changes are defined 
as: 
 Change from baseline in SBP > 10 mmHg 
 Change from baseline in DBP > 20 mmHg 
 
10.4 12-lead Safety Electrocardiogram 
Triplicate ECG measurements will be averaged on available values and the average value will 
be used to calculate summary statistics for heart rate (HR), PR interval (PR), QRS interval 
(QRS) and QTcF interval (QTcF)..  
Baseline value (i.e. pre dose value) and 2.5 hours post dose will be summarized. 
Number and percentage of subjects with clinically relevant change in HR at 2.5 hours post-
dose defined as change from baseline > 20 bpm, will also be summarized. 
The number and the percentage of subjects with QTcF abnormalities defined as: 
 QTcF >450 ms,  
 QTcF>480 ms  
 QTcF>500 ms; 
 change from baseline to 2.5 Hours post dose in QTcF >30 ms  
 change from baseline to 2.5 Hours post dose in QTcF >60 ms; 
will be summarized. 
 
10.5  Laboratory Data 
Quantitative chemistry and haematology parameters and fasting glucose values will be 
summarized descriptively at screening and discharge. Shift tables from screening with 
reference to normal ranges will be presented.  
Serology and urinalysis results will only be listed.  
Abnormal laboratory data will be flagged in the listings. 
 
11 Other Analyses 
No other analyses planned. 
 
12 Changes in the Planned Analyses from Study 
Protocol 
There are no changes to the protocol defined analysis. 


 
 
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13 Output 
13.1 Software 
SAS version 9.4 will be used to perform all the statistical analyses. PK analyses will be 
carried out using WNL Phoenix 8.1 or higher. 
 
 


 
 
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13.2 Reporting Conventions 
13.2.1 Treatment, Visit and Subgroup Descriptors 
In the tables, listings and figures, the treatments and the visits will be identified as described 
below. 
Treatment group 
Descriptor for treatment 
CHF6001 800 µg/ 20 µg, 4 inhalations + [14C] 
CHF6001 IV Infusion 18.5 µg 
CHF6001 
 
Output 
Descriptor for visits 
Tables 
Screening, Day -1, Day 1, Day 2–10, Day 11, Follow-up 
Listings 
Same as above. 
Figures 
Same as ‘Tables’. 
 
13.2.2 Decimal places 
Quantitative variables except PK variables will be listed with the same number of decimal 
places as in the actual data.  
In case of PK variables, plasma concentrations as well as calculated PK parameters will be 
presented with 3 decimal points except for tmax, tmax_iv, and tmax_iv which will be presented with 
2 decimals. Values ≥1000 will be presented without the decimals and the whole number will 
be reported without roundings (i.e. 1237 is not rounded to 1240 and 12322 is not rounded to 
12300). 
The following rules on decimal places will be considered in the listings for the derived variables 
(in the analyses rounding will not be performed): 
 BMI: whole numbers; 
 average of triplicate measurements (ECG and Blood Pressure): same as the actual 
variable 
 change from baseline: same as the variable considered. 
 AE duration (days): whole number if duration (hours)≥24 and 2 decimals in case of AE 
duration (hours)<24 hours 
The following rules on decimal places will be considered for the results of the analyses (if the 
analyses are performed on derived variables, the level of precision of the actual data is derived 
from the previous list): 
 min, max: same as actual data; 
 mean, SD, median, geometric mean: actual data + 1 decimal place; 
 percentage, geometric CV%,  CV%: 1 decimal place; 
 
13.2.3 Other reporting conventions 
Unless otherwise stated, listings will be sorted by Subject ID. 
In a listing, in the case that a subject’s record has been continued to the next page, an 
appropriate identification (e.g., the subject ID number) must be presented at the beginning of 
that page. 


 
 
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In general, dates will be presented on listings in the format ddmmmyyyy (date9.) and time in 
the format hh:mm (time5.). In case of partial dates or times, missing information will be 
replaced by dashes. 
 
13.3 Format 
In the top left portion of each table/listing, a table/listing number followed by the title of the 
table/listing will be presented. After the title line, optional sub-title or analysis set information 
can be presented. Horizontal lines will appear before and after the column heading of the 
table/listing. Footnotes will be put under the main body of text at the bottom of the page.  
The sponsor name, protocol number, programmer’s User ID, status of the table/listing (i.e. draft 
or final) and SAS program name will appear bottom left in a string and the page number will 
appear on the bottom right corner of each table/listing. The date and time of creation of 
table/listing will appear bottom left under the sponsor name. The source listing number will 
appear bottom left.  
Tables and listings will be produced in rich text format (i.e., they will tabular in format). 
A landscape layout will be used for both tables and listing. 
The left and right margins of all tables and listings will be a minimum of 2.1 cm from the left 
and 1.9 cm from the right. The top and bottom margins will be a minimum 2.92 cm. Header 
and footer will be both 1.27 cm. 
A 9-point font size for tables and 7 or 8-point font size for listings will be used using Courier 
New font. A maximum SAS line size=141 and page size=44 for 8-point font size, and line 
size=161 and page size=50 for 7-point will be used so as to fit on both UK and US paper sizes. 
A portrait layout will be used for figures. 
Titles and footnote will not be included in the body of figure. 
The size of the figures (except forest plot) will be: width=16.3 cm height=12.2 cm. The size of 
forest plot figures will be: width=16.3 cm height=20 cm. The resolution will be set using the 
option IMAGE_DPI=400. 
All tables, listings and figures will be collated into three Microsoft Word complete documents. 
If the listings are too large to be included in one file they will be separated into manageable 
sized files. The Microsoft Word documents will be subsequently converted in PDF format. 
Both, Word and PDF documents will include a table of contents with hyperlinks. 
 
13.4 Quality Control 
The following steps will be taken to ensure the quality of the outputs: 
 The author of each table/listing/figure program will review the program and will verify 
that no error message is highlighted in the ‘LOG’ file.  
 Tables and figures will be independently programmed form the raw datasets by a 
second statistician/programmer and outputs will be compared either electronically (by 
comparing the data being tabulated using PROC COMPARE) or by manually 
comparing the data in the two independent outputs. 


 
 
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 Listings will be checked either electronically (by comparing the data being listed using 
PROC COMPARE) or by manually comparing the data in the listings to the raw/derived 
data. 
 All outputs will be compared to the shells. 
 Related outputs will be compared for consistency. 
 
14 SAS Code 
14.1 Tables that need descriptive statistics 
PROC UNIVARIATE DATA=dset NOPRINT; 
    VAR var1 var2 var3 …varn; 
   BY byvar; (optional) 
   OUTPUT OUT=outname 
   N=n MEAN=mean MIN=min MAX=max MEDIAN=median STD=std CV=cv; 
RUN; 
 
Notes:  
– var1, var2 represent the variables for which summary statistics will be calculated; 
– byvar represents by variables to be used if any 
– out=outname specifies the output dataset that should be created 
 
For tables that need also geometric mean and geometric CV, the original variable will be LOG 
transformed and the geometric mean will be calculated using exp (mean_Log_var) and 
Geometric CV will be calculated using sqrt{exp[std_Log_var ^2]-1} 
 
14.2 Tables that need frequency counts 
PROC FREQ DATA=dset NOPRINT; 
  
BY byvar; (optional) 
   TABLES var1*var2;(for two ways table) 
 
TABLES var1 var2; (for one way table) 
  
OUTPUT OUT=outname; 
RUN; 
Notes:  
– byvar represents by variables to be used for frequency tabulations, if any 
– var1, var2 represent the variables to be tabulated; 
 
 
15 References 
NA