Conditions: Tuberculosis Tuberculosis, Pulmonary Bacterial Infections Respiratory Lung Diseases Mycobacterium Infections
Keywords: Anti-Bacterial Agents
Allocation: NA | Intervention model: SINGLE_GROUP
Enrollment: 4
Healthy volunteers: yes

Brief summary:
This study is a Phase 1, single-center, open-label study to investigate the absorption, metabolism, and excretion of BTZ-043 after a single oral administration of 500 mg BTZ-043 containing 3.7 MBq of \[14C\]BTZ-043 in 4 healthy adult male subjects

Detailed description:
A total of 4 evaluable subjects completing all procedures are required. Six (6) subjects will be enrolled in the cohort in order to have 4 evaluable subjects.

The study will consist of a screening period (Day -21 to -2), a baseline period (Day -1), a single dose treatment on Day 1 with a minimum of 96 hours (=4 days) post dose in-house observation period (Days -1 up to afternoon Day 5), and a follow-up visit 30 days (±2 days) after the \[14C\]BTZ-043 dose.

Subjects will be administered a single 500 mg \[14C\]BTZ-043 dose as drinking suspension. Subjects will be confined to the clinical site for at least 96 hours following drug administration (ie, afternoon of Day 5). During this time, blood, feces, and urine samples for measurement of \[14C\]BTZ-043 and metabolites will be collected.

The subjects will be released from the clinic approximately 96 hours to 168 hours after dose administration and upon satisfactory recovery of radioactivity (at least 90%) approved by the Sponsor's scientific advisor after consulation of the Sponsor.

Arms:
arm 1: 4 subjects to receive a single oral administration of 14C-labeled radioactive 500mg BTZ-043

Interventions: intervention 1: 500mg BTZ-043 containing 3.7 MBq of [14C]BTZ-043
Intervention descriptions: intervention 1: Single oral administration of 14C-labeled radioactive 500mg BTZ-043

Protocol document:
Prot_000.pdf: 
  
CLINICAL STUDY PROTOCOL 
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Page 1 of 57 
CLINICAL STUDY PROTOCOL 
A SINGLE-CENTER, OPEN-LABEL STUDY TO INVESTIGATE THE MASS BALANCE, 
EXCRETION PATHWAYS AND METABOLITES AFTER A SINGLE ORAL DOSE OF 
500 MG, 3.7 MBq, [14C]BTZ-043 IN HEALTHY MALE VOLUNTEERS 
 
CONFIDENTIAL
 
Sponsor code: LMU-IMPH-BTZ-043-03 
PRA code: NVS543EC-185431 
EudraCT number: 2021-000449-42 
 
 
Investigational product: 
BTZ-043 
Clinical phase: 
Phase 1 study 
Indication to be studied: 
Not applicable 
 
Sponsor: 
LMU Klinikum  
 
Marchioninistr. 15 
 
81377 Munich 
 
Germany 
 
 
 
Contract Research 
PRA Health Sciences – Early Development Services 
Organization and 
Van Swietenlaan 6 
Clinical Site: 
9728 NZ Groningen 
 
The Netherlands 
 
Principal Investigator: 
Jan Jaap van Lier, MD  
 
 
Phone: +31 50 402 2325  
 
Fax: 
+31 50 402 2223  
 
E-mail: lierjanjaapvan@prahs.com  
 
 
 
Version 1.0, 18 Mar 2021 
This study will be performed in compliance with the principles of Good Clinical Practice. 
 
 


  
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Page 1 of 57 
CLINICAL STUDY PROTOCOL 
A SINGLE-CENTER, OPEN-LABEL STUDY TO INVESTIGATE THE MASS BALANCE, 
EXCRETION PATHWAYS AND METABOLITES AFTER A SINGLE ORAL DOSE OF 
500 MG, 3.7 MBq, [14C]BTZ-043 IN HEALTHY MALE VOLUNTEERS 
 
CONFIDENTIAL
 
Sponsor code: LMU-IMPH-BTZ-043-03 
PRA code: NVS543EC-185431 
EudraCT number: 2021-000449-42 
 
 
Investigational product: 
BTZ-043 
Clinical phase: 
Phase 1 study 
Indication to be studied: 
Not applicable 
 
Sponsor: 
LMU Klinikum  
 
Marchioninistr. 15 
 
81377 Munich 
 
Germany 
 
 
 
Contract Research 
PRA Health Sciences – Early Development Services 
Organization and 
Van Swietenlaan 6 
Clinical Site: 
9728 NZ Groningen 
 
The Netherlands 
 
Principal Investigator: 
Jan Jaap van Lier, MD  
 
 
Phone: +31 50 402 2325  
 
Fax: 
+31 50 402 2223  
 
E-mail: lierjanjaapvan@prahs.com  
 
 
 
Version 1.0, 18 Mar 2021
This study will be performed in compliance with the principles of Good Clinical Practice. 
 
 






  
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SERIOUS ADVERSE EVENT CONTACT INFORMATION 
In case of a serious adverse event (see Appendix 8.2), the Principal Investigator will send a 
report within 24 hours of notification to: 
Contract Research Organization 
PRA Health Sciences Drug Safety Center EAPA
E-mail: 
mhgsafety@prahs.com  
Fax: 
+44 1792 525 720 
Phone: 
+49 621 8782 154 
 
Sponsor’s Medical Expert 
Leoni Matt, MD 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
E-mail: 
matt@lrz.uni-muenchen.de 
Fax: 
+49 89336038 
Phone: 
+49 89 4400 59847 
Mobile:  
+49 176 74717577 
 
 
 


  
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CONTACT INFORMATION 
Sponsor
LMU Klinikum  
Marchioninistr. 15 
81377 Munich 
Germany 
Sponsor’s Delegated Person 
Michael Hoelscher, MD, FRCP (Lond) 
Director 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
Phone: 
+49 89 4400 59800   
E-mail: 
hoelscher@lrz.uni-muenchen.de 
 
Sponsor’s Project Managers
Julia Dreisbach, VMD 
Clinical Project Manager 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
Phone: 
+49 89 4400 59825    
Mobile: 
+49 179 7036391 
Fax: 
+49 89 336038  
E-mail: 
dreisbach@lrz.uni-muenchen.de 
 
Petra Groß-Demel, PhD 
Clinical Project Manager 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
Phone: 
+49 89 4400 59825      
Mobile: 
+49 152 26309142 
Fax: 
+49 89 336038  
E-mail: 
gross-demel@lrz.uni-muenchen.de 
 


  
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Sponsor’s Medical Experts
Norbert Heinrich, MD 
Sponsor’s Medical Expert and Sponsor’s Delegated Person Deputy 
Senior Scientist, Tuberculosis and Emerging Diseases 
Division of Infectious Diseases and Tropical Medicine        
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich                   
Phone: 
+49 89 4400 59805 
Fax: 
+49 89 336038 
E-mail: 
heinrich@lrz.uni-muenchen.de  
 
Leoni Matt, MD  
Sponsor’s Medical Expert 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
Phone: 
+49 89 4400 59847 
Mobile:  
+49 176 74717577 
Fax: 
+49 89 336038 
E-mail: 
matt@lrz.uni-muenchen.de 
 
Client 
NUVISAN GmbH 
Wegenerstrasse 13 
89231 Neu-Ulm 
Germany 
 
Sponsor’s Scientific Advisor  
Piet Swart, PhD, Dip PharMed, Pharmacologist 
Chief Scientific Officer, Nuvisan Pharma Holding GmbH 
NUVISAN Pharma Holding GmbH 
Wegenerstrasse 13 
89231 Neu-Ulm 
Germany 
Mobile: 
+49 1525 6637 802 
E-mail: 
piet.swart@nuvisan.com 
 
Client’s Clinical Project Manager  
Flavia Koch, PhD, MSc 
Astrid Patzlaff, PhD 
Senior Project Manager/Clinical Operations 
NUVISAN GmbH Site Gauting


  
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Robert-Koch-Allee 29
82131 Gauting 
Germany 
Phone: 
+49 89 89356 9536925 
Fax: 
+49 89 89356922 
E-mail:
flavia.kochastrid.patzlaff@nuvisan.com
Client’s Nonclinical Project Manager 
Dieter Gallemann, PhD 
NUVISAN GmbH Site Grafing  
Am Feld 32 
85567 Grafing  
Germany 
Phone: 
+49 80 9270 0813+49 8092 7008 13 
Home office:+49 89 8946 0419
Fax: 
+49 80 92700823+49 8092 7008 23 
E-mail: 
dieter.gallemann@nuvisan.com 
 
Promotor 
Leibniz Institute for Natural Product Research and Infection Biology 
Hans Knöll Institut (HKI) 
Adolf-Reichwein-Straße 23 
07745 Jena 
Germany 
 
Promotor’s Contact 
Florian Kloss, PhD 
Head of Transfer Group Anti-infectives 
Phone: 
+49 3641 532 1356                                 
E-mail: 
florian.kloss@hki-jena.de/florian.kloss@leibniz-hki.de                                                      
 
 
Contract Research Organization, Contract Manufacturing Organization, and Clinical Site 
PRA Health Sciences (PRA) – Early Development Services (EDS) 
Van Swietenlaan 6 
9728 NZ Groningen 
The Netherlands 
Phone: 
+31 50 402 2222 
Fax: 
+31 50 402 2223 
 
Principal Investigator
Jan Jaap van Lier, MD  
Phone: 
+31 50 402 2325  
Fax: 
+31 50 402 2223  
E-mail: 
lierjanjaapvan@prahs.com 
 


  
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Project Manager
Marloes ter Veer, PhD 
Phone: 
+31 50 402 2292 
E-mail: 
veermarloester@prahs.com 
 
GMP-QA Responsible Person
Roos Imbos, PhD 
Phone: 
+31 59 230 3430 
Mobile: 
+31 6 227 889 05 
E-mail: 
imbosroos@prahs.com 
Laboratories 
Clinical Laboratory 
PRA-EDS
Van Swietenlaan 6 
9728 NZ Groningen 
The Netherlands 
Phone: 
+31 50 402 2555 
Fax: 
+31 50 402 2223 
Laboratory for Analysis of Plasma and Urine for BTZ-043 and Main Metabolites 
Bioanalytical Department of Nuvisan Grafing 
Melanie Tausch 
Am Feld 32 
85567 Grafing 
Germany 
Phone: 
+49 8092 7008 79  
Mobile: 
+49 152 06657818 
Fax: 
+49 8092 7008 23 
E-mail: 
melanie.tausch@nuvisan.com 
 
Laboratory for Analysis of Total Radioactivity in Whole blood, Plasma, Urine, and Feces 
PRA Bioanalytical Laboratory
Erik Vermeulen 
Amerikaweg 18 
9407 TK Assen 
The Netherlands 
Phone: 
+31 592 303 222 
Fax: 
+31 592 303 223 
E-mail: 
vermeulenerik@prahs.com 
 
Laboratory for Metabolite Profiling and Metabolite Identification in Plasma, Urine, and Feces 
PK Sciences and Biotransformation Department of Nuvisan Pharma Services GmbH
Matthias Bader, PhD 
Am Feld 32 


  
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85567 Grafing
Germany 
Phone: 
+49 8092 7008 48 
Fax: 
+49 8092 7008 23 
E-mail: 
matthias.bader@nuvisan.com 
Medical Screening Centers  
PRA-EDS 
Van Swietenlaan 6 
9728 NZ Groningen 
The Netherlands 
Phone: 
+31 50 850 5798 
Fax: 
+31 50 850 5782 
PRA-EDS 
Yin Yang Building 
Euclideslaan 1 
3584 BL Utrecht 
The Netherlands 
Phone: 
+31 50 851 5830 
Study Monitor 
Oliver Simonides 
Monitoring/LCRA 
NUVISAN GmbH Wegenerstrasse 13  
89231 Neu-Ulm 
Germany 
Phone: 
+49 731 9840 138 
Fax: 
+49 731 9840 571 
E-mail:  
oliver.simonides@nuvisan.com 
 


  
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SUMMARY OF CHANGES 
The following changes have been introduced in this Version 2.0 of the protocol (dated 01 Jul 2021) 
and are given as a combination of double underlined and italic text. Deleted text is given as double 
strikethrough. 
The rationale for these changes is the change in blood sample volumes and timepoints, and changes 
in the reporting of metabolite profiling/identification and PK. In addition, contact information of the 
Client’s Clinical Project Manager has been altered.  
The changes can be found in: 
 
Contact Information 
 
Synopsis  
 
Table 1 - Schedule of Assessments  
 
Section 3.5.1.3 Total Blood Volume: Table 3 - Number and Volume of Blood Samples and 
Total Blood Volume Collected per Subject 
 
Section 3.5.3.3 Exploratory Variables 
 
Section 3.5.4 Drug Concentration Measurements 
 
Section 3.6.2.1 Pharmacokinetic Evaluation 
 
Section 3.6.2.3 Evaluation of Exploratory Variables  
 
 


  
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SYNOPSIS 
Study Title 
A SINGLE-CENTER, OPEN-LABEL STUDY TO INVESTIGATE MASS BALANCE, EXCRETION 
PATHWAYS AND METABOLITES AFTER A SINGLE ORAL DOSE OF 500 MG, 3.7 MBq, [14C]BTZ-043 IN 
HEALTHY MALE VOLUNTEERS 
 
Study Codes 
Sponsor code 
: LMU-IMPH-BTZ-043-03  
PRA code 
: NVS543EC-185431 
EudraCT number 
: 2021-000449-42 
 
Sponsor 
LMU Klinikum, Marchioninistr. 15, 81377 Munich, Germany 
Sponsor’s Delegated Person 
 
: Michael Hoelscher, MD, FRCP (Lond) 
Sponsor’s Clinical Project Managers 
: Julia Dreisbach, VMD and Petra Gross-Demel, PhD 
 
Promotor 
Leibniz Institute for Natural Product Research and Infection Biology, Hans Knöll Institut (HKI),  
Adolf-Reichwein-Strasse 23, 07745 Jena, Germany 
Promotor’s contact 
  
 
: Florian Kloss, PhD  
 
Client 
Nuvisan GmbH, Wegenerstrasse 13, 89231, Neu-Ulm, Germany 
Client’s Clinical Project Manager 
: Flavia Koch, PhD, MSc Astrid Patzlaff, PhD 
 
Contract Research Organization and Clinical Site 
PRA Health Sciences – Early Development Services, Van Swietenlaan 6, 9728 NZ Groningen, 
The Netherlands 
 
Principal Investigator 
Jan Jaap van Lier, MD 
 
Objectives 
Primary 
: To determine the rates and routes of excretion of [14C]BTZ-043-related 
radioactivity, including mass balance of total drug-related radioactivity in urine 
and feces (and vomit, if applicable), following the oral administration of a single 
500 mg dose of [14C]BTZ-043 in healthy male volunteers. 
 
: To determine the pharmacokinetics (PK) of total radioactivity in blood and in 
plasma. 
 
: To characterize the plasma PK of BTZ-043 and main metabolites by liquid 
chromatography-mass spectrometry (LC-MS), if applicable. 
 
: To characterize the urine concentrations of BTZ-043 and main metabolites by 
LC-MS, if applicable. 
Secondary 
: To assess the safety and tolerability of a single 500 mg oral dose of BTZ-043 
administered to healthy volunteers. 
Exploratory 
: To identify BTZ-043’s metabolites in plasma, and to quantify metabolites not 
measured in the primary objectives by radiometry in order to elucidate key 
biotransformation pathways and clearance mechanisms of BTZ-043 in humans. 


  
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: To identify BTZ-043’s metabolites in excreta (urine and feces) and to quantify 
metabolites not measured in the primary objectives by radiometry in order to 
elucidate key biotransformation pathways and clearance mechanisms of BTZ-
043 in humans. 
 
Design and Treatments 
This study is a Phase 1, single-center, open-label study to investigate the absorption, metabolism, and 
excretion of BTZ-043 after a single oral administration of 500 mg BTZ-043 containing 3.7 MBq of 
[14C]BTZ-043 in 4 healthy adult male subjects. 
 
A total of 4 evaluable subjects completing all procedures are required. Six (6) subjects will be enrolled in the 
cohort in order to have 4 evaluable subjects. If the cohort of 4 subjects administered with 500 mg BTZ-043 
containing 3.7 MBq of [14C]BTZ-043 leads to less than 4 evaluable subjects, the 2 back-up subjects can be 
dosed subsequently to ensure that 4 subjects have completed all study procedures and are evaluable. If any 
of the 4 subjects vomits within 4 hours after study drug administration (approximately 2 times the median 
Tmax of BTZ-043), the subject will be replaced. The 2 back-up subjects will stay in the clinical research center 
until the morning of Day 2. 
 
The study will consist of a screening period (Day -21 to -2), a baseline period (Day -1), a single dose 
treatment on Day 1 with a minimum of 96 hours (=4 days) post dose in-house observation period (Days -1 
up to afternoon Day 5), and a follow-up visit 30 days (±2 days) after the [14C]BTZ-043 dose. The subjects 
will be admitted to the clinical site on Day -1, at least 17 hours before the administration of [14C]BTZ-043 for 
baseline evaluation. Subjects will be administered a single 500 mg [14C]BTZ-043 dose as drinking 
suspension. Subjects will be confined to the clinical site for at least 96 hours following drug administration 
(ie, afternoon of Day 5). During this time, blood, feces, and urine samples for measurement of [14C]BTZ-043 
and metabolites will be collected. The radioactivity excreted in urine and feces as well as radioactivity in 
blood and plasma should be measured and the data should be communicated daily to Matthias Bader or his 
designee. The subjects will be released from the clinic approximately 96 hours to 168 hours after dose 
administration and upon satisfactory recovery of radioactivity (at least 90%) approved by the Sponsor’s 
scientific advisor after consulation of the Sponsor. This will depend on the radioactivity excretion balance as 
assessed by a normal counting method or real time rapid analysis on site (‘quick counting’ procedure, 
utilizing liquid scintillation counter, or sample combustion apparatus) of radioactivity in plasma, urine, and 
feces accumulated until 96 hours, as feasible. The data has to be available before subjects will be released 
from the clinic.  
 
The in-house period should be prolonged up to the morning of Day 8 if at least 1 of the following applies: 
 
Total radioactivity in plasma is higher than 5% of total radioactivity-Cmax and/or PK parameters for 
determination of the total radioactivity-elimination phase cannot be determined reliably. 
 
The mass balance in urine and feces is not complete by Day 5 (ie, <90%).  
 
The combined urinary and fecal excretion is not 1% of the administered dose per 24 hours for 
2 consecutive days based on 14C-radioactivity “quick counts” or normal counts, as feasible. 
 
In the event of an incomplete mass balance on the morning of Day 8, subjects may be discharged from the 
clinical site upon discretion of the Investigator. Subjects with an incomplete mass balance (<90% of dose 
administered and/or combined urinary and fecal excretion >1% of the administered dose per 24 hours for 
2 consecutive days based on 14C-radioactivity “quick counts” or normal counts, as feasible) on Day 8 will be 
asked to return to the clinical site for one or two 24-hour visits (Day 11-12 and Day 15-16). 
 


  
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In the event of subjects not defecating for 2 days, measures will be taken to ensure defecation (eg, additional 
fibers, plum juice, dried prunes, and/or sufficient hydration). 
 
Study Schedule 
Screening 
: Between Day -21 and Day -2.  
Treatment period 
: From Day -1 (admission) to approximately 96 hours after study drug 
administration (Day -1 up to afternoon Day 5) with possible extension (Day 6 to 
Day 8) as well as possible 24-hour visits (Day 11-12, Day 15-16).  
Follow-up 
: 30 days (±2 days) after the [14C]BTZ-043 dose. 
 
Subjects 
4 healthy male subjects 
 
Main Criteria for Inclusion 
Age 
: 18 years to 55 years, inclusive, at screening 
Weight 
: 55 to 90 kg, inclusive, at screening 
Body mass index 
: 18.0 to 29.0 kg/m2, inclusive, at screening  
 
Study Drug 
Active Medication 
Active substance 
: BTZ-043 
Activity 
: Inhibition of Mycobacterium tuberculosis DprE1 
In development for 
: Tuberculosis 
Dose 
:  1 x 100 mL (equivalent to 500 mg BTZ-043) 
Strength 
: 5 mg/mL BTZ-043 containing 0.037 MBq/mL of [14C]-BTZ-043 
 Dosage form 
: Oral suspension 
Manufacturer 
: Prepared by pharmacy at PRA 
 
Variables 
Safety 
: Adverse events, clinical laboratory, vital signs, 12-lead electrocardiogram, 
physical examination  
Pharmacokinetics 
: Total radioactivity concentrations in plasma, whole blood, urine, and feces 
 
 Total radioactivity in plasma and whole blood PK parameters estimated using 
noncompartmental analysis, as appropriate : Cmax, tmax, t1/2, AUCinf, AUC0-t, and 
whole blood to plasma ratios  
 
 Concentrations of BTZ-043 and main metabolites in plasma and urine 
 
 Plasma and urine BTZ-043 PK parameters estimated using noncompartmental 
analysis, as appropriate: Cmax, tmax, t1/2, kel, AUCinf, AUC0-t, CL/F, Vz/F, Aeurine, 
Feurine, and CLr 
 
 Plasma 
and 
urine 
metabolites PK 
parameters estimated using 
noncompartmental analysis, as appropriate: Cmax, tmax, t1/2, kel, AUCinf, AUC0-t, 
Aeurine, and Feurine 
 
 Urine and feces (and vomit, if available) radioactivity PK parameters estimated 
using noncompartmental analysis, as appropriate: Aeurine, Feurine, Aefeces, Fefeces, 
and, if available, Aevomit, and Fevomit
Exploratory 
: Metabolite profiles, metabolite identification, as feasible 
 
 %AUCtotal radioactivity of BTZ-043 and metabolites 
 
 
 


  
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Statistical Methods 
Safety parameters 
: Descriptive statistics 
PK parameters 
: Descriptive statistics for all relevant PK parameters: n, number of missing 
values to be reported (m), geometric mean, geometric coefficient of variation, 
mean, coefficient of variation, SD, median, minimum, and maximum  
Exploratory parameters : Descriptive statistics 
 




AE=adverse event; BMI=body mass index; ECG=electrocardiogram; HBsAg=hepatitis B surface antigen; HCV=hepatitis C virus;PCR=polymerase chain reaction; PK=pharmacokinetic(s) 
a 
 
b 
c
 
d 
 
e 
f 
g 
 
h 
i 
 
 
j 
 
 
k 
 
 
l 
m 
Subjects will be in the clinic from Day -1 (predose) until Day 5 with possible extension (Day 6 to Day 8) as well as possible 24-hour visits (Day 11-12, Day 15-16). The 2 back-up subjects will stay 
in the clinic until the morning of Day 2. 
Subjects will be released from the clinic 96 hours after dose administration and upon satisfactory recovery of radioactivity. 
Complete physical examinations will be conducted at screening, Day -1, and follow-up/early termination. Symptom driven physical examinations may be conducted at any time, per the Investigator’s 
discretion.  
Sampling of nasal and throat mucosal cells. If deemed necessary, additional tests may be conducted during the study. The nonclinical project manager will be informed immediately upon availability  
of Day 11 results, as this will qualify the feces samples for metabolite profiling with regard to potential COVID-19 infectivity. 
Clinical laboratory tests (including clinical chemistry, hematology, and urinalysis): at screening; on Day -1 (admission), Day 2, and Day 4; and at follow-up.
12-lead ECG: at screening; Day -1, and 1 hour, 4 hours, and 24 hours postdose; at discharge; and at follow-up.  
Vital signs (supine systolic and diastolic blood pressure, pulse, body temperature, and respiratory rate): at screening; at admission; at predose and at 1 hour, 4 hours, and 24 hours postdose; at 
discharge; and at follow-up. 
Plasma sampling for PK of BTZ-043 and main metabolites in plasma: at predose and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, and 48 hours postdose. 
Total radioactivity in whole blood and plasma, and metabolite profiling in plasma: at predose and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, 48, 72, and 96 hours postdose (in case total radioactivity 
recovered is still <90%: in addition 120, 144, and 168 hours postdose and at further 24-hour visits [264 and 360 hours postdose] until study completion). Metabolite profiling in plasma: at predose 
and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, 48, 72, and 96 hours postdose (in case total radioactivity recovered is still <90%: in addition 120, 144, and 168 hours postdose. 
Urine collection and pooling for BTZ-043 and main metabolites, total radioactivity, and metabolite profiling: at predose (within 12 hours prior to dosing) and over 0-24, 24-48, 48-72, and 72-96 hours 
postdose collection intervals (in case total radioactivity recovered is still <90%, in addition 96-120, 120-144, and 144-168 hours postdose and at further 24-hour intervals [Day 11-12 and Day 15-16] 
until study completion). 
Feces collection for total radioactivity and metabolite profiling; predose feces collection to be collected at any time within 48 hours prior to study drug administration (at home or in the clinic). After 
administration of BTZ-043, feces will be collected in 24 hour collection intervals up to 96 hours postdose (in case total radioactivity recovered is still <90%: in addition 120, 144, and 168 hours 
postdose and at further 24-hour intervals [Day 11-12 and Day 15-16] until study completion). 
If produced, vomit will be collected up to 8 hours after the administration of the study drug for total radioactivity determination.
Day 6-8 procedures will only be conducted in case that the recovery of radioactivity was not satisfactory until Day 5. 24-hour visits procedures will only be conducted in case that the recovery of 
radioactivity was not satisfactory until Day 8. For the 24-hour visits (if applicable): feces collection to be collected at home within 48 hours prior to admission.


  
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TABLE OF CONTENTS 
TITLE PAGE ..............................................................................................................................1 
SPONSOR AUTHORIZATION OF CLINICAL STUDY PROTOCOL ................................... 2 
AUTHORIZATION OF CLINICAL STUDY PROTOCOL BY CONTRACT RESEARCH 
ORGANIZATION ............................................................................................................... 3 
SERIOUS ADVERSE EVENT CONTACT INFORMATION................................................... 4 
CONTACT INFORMATION ..................................................................................................... 5 
SUMMARY OF CHANGES.................................................................................................... 10
SYNOPSIS .............................................................................................................................. 11 
TABLE OF CONTENTS......................................................................................................... 17 
TABLE OF TABLES .............................................................................................................. 20 
LIST OF ABBREVIATIONS................................................................................................... 21 
1. INTRODUCTION.............................................................................................................. 22 
1.1 
Background.............................................................................................................. 22 
1.1.1 Nonclinical Summary ...................................................................................... 22 
1.1.2 Clinical Summary ............................................................................................ 24 
1.2
Risk-benefit Assessment......................................................................................... 26
1.3 
Study Rationale........................................................................................................ 27 
2.
OBJECTIVES...................................................................................................................28
2.1 
Primary ..................................................................................................................... 28 
2.2 
Secondary ................................................................................................................ 28 
2.3 
Exploratory............................................................................................................... 28 
3. INVESTIGATIONAL PLAN ............................................................................................. 29 
3.1 
Overall Study Design and Plan ................................................................................ 29 
3.1.1 Type of Study ................................................................................................. 29 
3.1.2 Screening Period ............................................................................................ 29 
3.1.3 Treatment Period ............................................................................................ 29 
3.1.4 Follow-up........................................................................................................ 30 


  
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3.2
Discussion of Study Design.................................................................................... 30
3.2.1 COVID-19 Risk Mitigation ............................................................................... 30 
3.3
Selection of Study Population................................................................................. 31
3.3.1 Inclusion Criteria ............................................................................................. 31 
3.3.2 Exclusion Criteria ............................................................................................ 32 
3.3.3 Removal of Subjects from Assessment............................................................ 33 
3.4 
Treatments ............................................................................................................... 34 
3.4.1 Treatments Administered ................................................................................ 34 
3.4.2 Identity of Investigational Product .................................................................... 34 
3.4.3 Method of Assigning Subjects to Treatment Groups......................................... 34 
3.4.4 Selection of Doses and Meals in the Study ...................................................... 35 
3.4.5 Radiation Safety of Clinical and Laboratory Staff ............................................. 35 
3.4.6 Timing of Doses in the Study........................................................................... 36 
3.4.7 Meals During the Study ................................................................................... 36 
3.4.8 Blinding .......................................................................................................... 37 
3.4.9 Concomitant Medication and Other Restrictions During the Study .................... 37 
3.4.10 Treatment Compliance .................................................................................... 38 
3.5 
Pharmacokinetic, Safety, and Exploratory Measurements and Variables ............. 38 
3.5.1 Pharmacokinetic and Safety Measurements Assessed and Schedule of 
Assessments .................................................................................................. 38 
3.5.1.1 
Pharmacokinetic Measurements ...................................................... 38 
3.5.1.1.1 
Blood Sampling ............................................................. 38 
3.5.1.1.2 
Urine Collection ............................................................. 39 
3.5.1.1.3 
Feces Collection............................................................ 39 
3.5.1.2 
Safety and Tolerability Measurements ............................................. 39 
3.5.1.2.1 
Adverse Events ............................................................. 40 
3.5.1.2.2 
Clinical Laboratory......................................................... 40 
3.5.1.2.3 
Vital Signs ..................................................................... 41 
3.5.1.2.4 
Electrocardiogram ......................................................... 41 
3.5.1.2.5 
Physical Examination .................................................... 41 
3.5.1.3 
Total of Blood Volume ..................................................................... 41
3.5.2 Appropriateness of Measurements .................................................................. 42 
3.5.2.1 
Timing of Assessments.................................................................... 42 
3.5.3 Pharmacokinetic, Safety, and Exploratory Variables ........................................ 42 
3.5.3.1 
Pharmacokinetic Variables .............................................................. 42 
3.5.3.2 
Safety Variables .............................................................................. 44 
3.5.3.3 
Exploratory Variables ...................................................................... 44 
3.5.4 Drug Concentration Measurements ................................................................. 44 
3.5.5 Retention of Blood, Plasma, Urine, and Feces Samples................................... 44 
3.6 
Statistical Procedures and Determination of Sample Size ..................................... 45 
3.6.1 Analysis Sets .................................................................................................. 45 
3.6.1.1 
Safety Set ....................................................................................... 45 
3.6.1.2 
Pharmacokinetic Set........................................................................ 45 
3.6.2 Statistical and Analytical Plan for Pharmacokinetic, Safety, and Exploratory 
Evaluation....................................................................................................... 45 
3.6.2.1 
Pharmacokinetic Evaluation............................................................. 45 


  
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3.6.2.2
Evaluation of Safety and Tolerability ................................................ 45
3.6.2.2.1 
Adverse Events ............................................................. 45 
3.6.2.2.2 
Clinical Laboratory......................................................... 46 
3.6.2.2.3 
Vital Signs and Electrocardiograms................................ 46 
3.6.2.2.4 
Physical Examination .................................................... 46 
3.6.2.3 
Evaluation of Exploratory Variables.................................................. 46 
3.6.3 Determination of Sample Size ......................................................................... 46 
3.7 
Data Quality Assurance ........................................................................................... 46 
4. ETHICS............................................................................................................................. 48 
4.1 
Independent Ethics Committee ............................................................................... 48 
4.2 
Ethical Conduct of the Study................................................................................... 48 
4.3 
Subject Information and Consent............................................................................ 49 
4.4 
Privacy ..................................................................................................................... 49 
5. STUDY ADMINISTRATIVE STRUCTURE..................................................................... 50 
5.1 
Distribution of Activities .......................................................................................... 50 
5.1.1 Preparation of Study Drug ............................................................................... 50 
5.1.2 Laboratory Assessments ................................................................................. 50 
5.1.3 Electronic Case Report Form Design............................................................... 50 
5.1.4 Data Management .......................................................................................... 50 
5.1.5 Statistics ......................................................................................................... 50 
5.1.6 Clinical Study Report Writing ........................................................................... 50 
5.2 
Documentation......................................................................................................... 51 
5.2.1 Archiving ........................................................................................................ 51 
5.2.2 Recording of Data in Source Documents and Electronic Case Report Forms ... 51 
6. CONFIDENTIALITY AND PUBLICATION POLICY ...................................................... 52 
7. REFERENCES................................................................................................................. 53 
8. APPENDICES .................................................................................................................. 54 
8.1 
Drug Accountability ................................................................................................. 54 
8.2 
Adverse Events and Serious Adverse Events Evaluation and Reporting .............. 54 
8.2.1 Adverse Events............................................................................................... 54 
8.2.2 Serious Adverse Events .................................................................................. 55 
8.2.3 Suspected Unexpected Serious Adverse Reactions......................................... 56 
8.2.4 Follow-up of Adverse Events ........................................................................... 57 
 


  
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TABLE OF TABLES  
Table 1
Schedule of Assessments.......................................................................................... 15
Table 2 
Geometric mean and range of AUC0-24h for BTZ-043 in Phase 1a and Phase 1b studies
.................................................................................................................................. 26 
Table 3 
Number and Volume of Blood Samples and Total Blood Volume Collected per Subject42 
Table 4 
Pharmacokinetic Parameters ..................................................................................... 43 
 


  
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LIST OF ABBREVIATIONS 
AME
absorption, metabolism, and excretion
AE
adverse event 
ALT
alanine aminotransferase
AST
aspartate aminotransferase
AUC
area under the concentration-time curve
BMI
body mass index
CA
Competent Authority
CCMO
Centrale Commissie Mensgebonden Onderzoek (Central Committee on 
Research Involving Human Subjects) 
CHMP
Committee for Medicinal Products for Human Use 
GMP
Good Manufacturing Practice
CSP
clinical study protocol
CSR
clinical study report
CTCAE
Common Terminology Criteria for Adverse Events
CTD
Clinical Trial Directive
DprE1
decaprenylphosphoryl- - -ribose-2 -epimerase 
ECG
electrocardiogram
eCRF
electronic case report form
EDS
Early Development Services
EMA
European Medicines Agency
FDA
Food and Drug Administration
GCP
Good Clinical Practice
GDPR
General Data Protection Regulation
HBsAg
hepatitis B surface antigen
HCV
hepatitis C virus
HED
human equivalent dose
HKI
Hans Knöll Institut
ICF
informed consent form
ICH
International Council for Harmonisation
IEC
Independent Ethics Committee
MEB 
Medicine Evaluation Board
MedDRA
Medical Dictionary for Regulatory Activities
NOAEL
no observed adverse effect level
PCR
polymerase chain reaction
PK
pharmacokinetic(s)
PRA
PRA Health Sciences
SAE
serious adverse event
SAP
statistical analysis plan
SOP
standard operating procedure
SUSAR
suspected unexpected serious adverse reaction
TEAE
treatment-emergent adverse event
WMA
World Medical Association
WMO
Wet medisch-wetenschappelijk onderzoek met mensen (medical research 
involving human subjects act) 
Note: Definitions of pharmacokinetic (PK) parameters are provided in Section 3.5.3. 
 


  
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1. 
INTRODUCTION 
1.1 
Background 
BTZ-043 is a promising new antibiotic for the treatment of tuberculosis. Its mechanism 
of action is based on a reduction of BTZ-043 to a transient nitroso moiety, which then 
reacts with an essential cysteine residue in the enzyme decaprenylphosphoryl- - -
ribose-2 -epimerase (DprE1), forming a semimercaptal as covalent adduct. DprE1 itself 
is required for the biosynthesis of decaprenylphosphoryl- - -arabinose, an essential 
component for the cell wall assembly of mycobacteria. Formation of the covalent adduct 
between BTZ-043 and DprE1 results in inhibition of cell wall biosynthesis and loss of 
viability of M. tuberculosis. Due to this mechanism BTZ-043 is highly selective for 
mycobacteria species (and some other members of Actinomycetales), but does not 
affect the gut flora.1 
 
In 2018 a Phase 1a study entitled “A Randomized, Double blind, Placebo-controlled, 
Single 
Ascending 
Dose 
Study 
to 
Evaluate 
Safety, 
Tolerability, 
and 
Pharmacokinetics (PK) of Single Doses of BTZ-043 in Healthy Adult Volunteers” 
(LMU-IMPH-BTZ-043-01) was conducted in humans and currently a Phase 1a/2b study 
entitled “A Prospective Phase 1b/2a, Active-controlled, Randomized, Open-label Study 
to Evaluate the Safety, Tolerability, Extended Early Bactericidal Activity and PK of 
Multiple Oral Doses of BTZ-043 Tablets in Subjects With Newly Diagnosed, 
Uncomplicated, 
Smear-positive, 
Drug-susceptible 
Pulmonary 
Tuberculosis” 
(PanACEA-BTZ-043-02) is ongoing. This human absorption, metabolism, and excretion 
study is conducted during Phase 2 proof of concept studies and before a Phase 3 study 
is initiated. 
1.1.1 
Nonclinical Summary 
Safety 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 


  
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Pharmacokinetics
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


  
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1.1.2 
Clinical Summary 
Safety 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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1.2
Risk-benefit Assessment
There is no expected clinical benefit for the healthy subjects who will participate in this 
study. The information obtained in this study can be used for the further clinical 
development of BTZ-043. As depicted in Table 2, BTZ-043 has been well tolerated in 
much higher exposures as proposed in this study.  
 
Overall, on the basis of the available nonclinical and clinical data, the risk-benefit profile 
of BTZ-043 is judged acceptable for the proposed clinical study. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


  
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The risk-benefit assessment for the subjects receiving BTZ-043 remains unchanged in 
relation to the corona virus disease-19 (COVID-19) pandemic as available clinical and 
nonclinical results do not suggest that administration of BTZ-043 will lead to suppression 
or modulation of the immune system. In addition, the mode of action does not appear to 
have a substantial effect on the respiratory or cardiovascular system critically affected 
by a SARS-CoV-2 infection. As the subjects to be included in this study are in general 
young to middle aged without major comorbidities, the study population is not considered 
to be a high-risk population for serious COVID-19 disease. Only persons with a negative 
SARS-CoV-2 test at admission to the clinical research center will be allowed to 
participate in the study. In addition, all appropriate measures to prevent SARS-CoV-2 
infection during the study will be taken as detailed in Section 3.2.1. 
1.3
Study Rationale
This study will determine the disposition and metabolism of BTZ-043 in humans after 
oral administration. The use of radiolabeled molecules (usually [14C]) is a common 
method to ascertain information on the elimination routes and metabolic fate of a 
compound at an early stage of development. This evaluation will provide an estimate of 
PK parameters, the routes and rates of elimination of radioactivity, and identification of 
metabolites and metabolic pathways. These data will be compared with similar data 
obtained from nonclinical studies, both from PK and metabolic points of view. 
The rationale for the proposed dose can be found in Section 3.4.4. 
 
 


  
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2. 
OBJECTIVES 
2.1 
Primary  
To determine the rates and routes of excretion of [14C]BTZ-043-related radioactivity, 
including mass balance of total drug-related radioactivity in urine and feces (and 
vomit, if applicable), following the oral administration of a single 500 mg dose of 
[14C]BTZ-043 in healthy male volunteers. 
 
To determine the PK of total radioactivity in whole blood and in plasma. 
 
To characterize the plasma PK of BTZ-043 and main metabolites by liquid 
chromatography-mass spectrometry (LC-MS), if applicable. 
 
To characterize the urine concentrations of BTZ-043 and main metabolites by 
LC-MS, if applicable. 
2.2
Secondary
 
To assess the safety and tolerability of a single 500 mg oral dose of BTZ-043 
administered to healthy male volunteers. 
2.3 
Exploratory 
 
To identify BTZ-043’s metabolites in plasma, and to quantify metabolites not 
measured in the primary objectives by radiometry in order to elucidate key 
biotransformation pathways and clearance mechanisms of BTZ-043 in humans.  
 
To identify BTZ-043’s metabolites in excreta (urine and feces) and to quantify 
metabolites not measured in the primary objectives by radiometry in order to 
elucidate key biotransformation pathways and clearance mechanisms of BTZ-043 in 
humans. 
 
 
 


  
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3. 
INVESTIGATIONAL PLAN 
3.1 
Overall Study Design and Plan 
3.1.1
Type of Study
This will be a Phase 1, single-center, open-label, absorption, metabolism, and excretion 
(AME) study in 4 healthy adult male subjects. 
3.1.2 
Screening Period 
Subjects will report to the medical screening facility/clinical site for the eligibility 
screening (see Section 3.3 for inclusion and exclusion criteria) within 3 weeks prior to 
drug administration.  
 
Subjects will sign the study-specific informed consent form (ICF) prior to any 
study-specific screening procedures being performed. The written informed consent will 
be obtained for all subjects, regardless of their eligibility for the study; the signed ICFs 
will be retained and archived at PRA Health Sciences (PRA) and a copy will be provided 
to the subject.  
 
Eligibility screening will consist of the assessments as presented in the schedule of 
assessments (Table 1). 
3.1.3 
Treatment Period 
Subjects will be admitted to the clinical research center on Day -1, which is the day prior 
to Day 1, the day of drug administration.  
 
Subjects will receive a single oral dose of 500 mg BTZ-043 containing 3.7 MBq of 
[14C]BTZ-043 on Day 1. Subjects will be discharged from the clinical research center on 
Day 5 (approximately 96 hours after drug administration) upon satisfactory recovery of 
radioactivity. If the criteria for discharge are not met on Day 5, subjects will be required 
to remain confined for a maximum of 3 additional days (Days 6 to 8) until the criteria are 
met (daily check by quick counts or normal counts, as feasible). The in-house period 
should be prolonged up to the morning of Day 8 if at least 1 of the following applies:  
 
Total radioactivity in plasma is higher than 5% of radioactivity-Cmax and/or 
PK parameters for determination of the total radioactivity-elimination phase 
cannot be determined reliably.  
 
The mass balance in urine and feces is not complete by Day 5 (ie, >90%). 
 
The combined urinary and fecal excretion is not 1% of the administered dose 
per 24 hours for 2 consecutive days based on 14C-radioactivity “quick counts” or 
normal counts, as feasible. 
If the criteria for discharge are still not met on Day 8, subjects may be discharged from 
the clinical site upon discretion of the Investigator. To ensure that adequate 
consideration is given to the protection of the subject’s interests, subjects with an 
incomplete mass balance on Day 8 will be asked to return to the clinical site for one or 
two 24-hour visits (Day 11-12 and Day 15-16). 
 


  
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On the day of study drug administation (Day 1), an FDA-recommended high-fat (or a 
standardized breakfast, if applicable, as outlined in Section 3.4.4 and Section 3.4.7) with 
calculated caloric content will be provided. 
 
Assessments during the treatment period will be performed as presented in the schedule 
of assessments (Table 1). 
3.1.4 
Follow-up 
The follow-up assessments will be performed 30 days (±2 days) after the [14C]BTZ-043 
dose.  
 
Assessments during follow-up will be performed as presented in the schedule of 
assessments (Table 1). 
3.2 
Discussion of Study Design 
The current design is commonly used for AME studies and in accordance with the current 
EMA “Guideline on the investigation of drug interactions”. 2 Since the present study is a 
descriptive study aimed at assessing excretion routes of BTZ-043 and evaluating mass 
balance, a formal statistical sample size calculation was not performed. A sample size 
of 4 subjects is a minimum, accepted number of subjects for AME studies and is 
considered sufficient to achieve the study objectives. Early-termination subjects may be 
replaced if the total number of completers drops below 4, after mutual agreement 
between the Sponsor and PRA.  
3.2.1 
COVID-19 Risk Mitigation 
This study will be conducted in accordance with guidance from the Central Committee 
on Research Involving Human Subjects (CCMO [Centrale Commissie Mensgebonden 
Onderzoek], ie, the Dutch competent authority [CA]) on conducting Phase 1 trials in 
clinical research centers in The Netherlands during the COVID-19 pandemic. 
 
During the entire study, the clinical research center will implement all recommendations 
issued by the Dutch government, including specific guidelines related to clinical research 
executed in clinical research centers with respect to minimizing the risk of disease 
spreading (eg, social distancing, disinfection, hygiene, and wearing of personal 
protection equipment by study staff). Details on specific procedures are described in the 
site specific manual.  
 
In cases where subjects are not able to attend study visits due to an infection with 
SARS-CoV-2, the Investigator will discuss with the Sponsor potential mitigation 
approaches (including, but not limited to, extending the visit window, conducting 
evaluations via video link or phone call, allowing for safety procedures to be conducted 
at a local facility). The rationale (eg, the specific limitation imposed by the SARS-CoV-2 
infection that led to the inability to perform the protocol-specified assessment) and 
outcome of the discussion will be documented in the electronic case report form (eCRF). 
 
In addition, the following containment measures will be taken during the study: 


  
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Polymerase chain reaction (PCR) testing for SARS-CoV-2 will be performed at the 
time points indicated in Table 1.
 
A subject should not be admitted if there was any contact with a person who tested 
positive for SARS-CoV-2 or a COVID-19 patient within the last 2 weeks prior to 
admission to the clinical research center. 
 
If a subject is tested to be SARS-CoV-2 positive on Day -1, the subject will be 
excluded from participation with reference to Exclusion Criterion 13, and referred 
for treatment. 
 
Physical examinations will be limited. 
 
If a subject becomes ill and/or is tested to be SARS-CoV-2 positive after the 
administration of study treatment, study assessments will be stopped (see also 
Section 3.3.3.1). A safety follow-up can be done if deemed necessary by the 
Investigator. The subject will be kept in-house in quarantine with appropriate 
medical intervention until complete elimination of the study compound or will be 
asked to quarantine at home according to guidelines of the Dutch government. 
3.3 
Selection of Study Population 
3.3.1 
Inclusion Criteria 
Note: Restrictions that apply to the period after admission are described in Section 3.4.9, 
except when they concern a statement of willingness. 
 
The following inclusion criteria must be met for a subject to be eligible for inclusion in the 
study: 
1. 
Sex 
: male 
2. 
Age 
: 18 years to 55 years, inclusive, at screening. 
3. 
Body mass index (BMI) : 18.0 to 29.0 kg/m2, inclusive, at screening.  
4. 
Weight 
: 55 to 90 kg, inclusive, at screening. 
5. 
Status 
: healthy subjects. 
6. 
Male subjects, if not surgically sterilized, must agree to use adequate contraception 
and not donate sperm from admission to the clinical research center until 90 days 
after the follow-up visit. Adequate contraception for the male subject (and his 
female partner, if she is of childbearing potential) is defined as using hormonal 
contraceptives or an intrauterine device combined with at least 1 of the following 
forms of contraception: a diaphragm, a cervical cap, or a condom. Total abstinence, 
in accordance with the lifestyle of the subject, is also acceptable.  
7. 
All prescribed medication must have been stopped at least 30 days prior to 
admission to the clinical research center.  
8. 
All over-the-counter medications, vitamin preparations (especially vitamin C), other 
food supplements, and herbal medications (eg, St. John’s wort) must have been 
stopped at least 14 days prior to admission to the clinical research center. An 
exception is made for paracetamol, which is allowed up to 48 hours prior to study 
drug administration. 
9. 
No vaccination within 14 days prior to study drug administration. 


  
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10.
Ability and willingness to abstain from alcohol from 48 hours (2 days) prior to 
screening and admission to the clinical research center. 
11. Ability and willingness to abstain from methylxanthine-containing beverages or food 
(coffee, tea, cola, chocolate, and energy drinks), grapefruit (juice), corn (whole corn 
kernels and popcorn), cruciferous vegetables, and bitter oranges from 48 hours 
(2 days) prior to admission to the clinical research center.  
12. Good physical and mental health on the basis of medical history, physical 
examination, clinical laboratory, ECG, and vital signs, as judged by the 
Investigator.  
13. Willing and able to sign the ICF. 
3.3.2 
Exclusion Criteria 
Note: Restrictions that apply to the period after admission are described in Section 3.4.9, 
except when they concern a statement of willingness. 
 
A subject who meets any of the following exclusion criteria will not be eligible for inclusion 
in the study: 
1. 
Participation in another study with a radiation burden of >0.1 mSv and 1 mSv in 
the period of 1 year prior to screening; a radiation burden of >1.1 mSv and 2 mSv 
in the period of 2 years prior to screening; a radiation burden of >2.1 mSv and 
3 mSv in the period of 3 years prior to screening, etc. 
2. 
Exposure to radiation for diagnostic reasons (except dental X-rays and plain X-rays 
of thorax and bony skeleton [excluding spinal column]), or during work within 1 year 
prior to drug administration. 
3. 
Irregular defecation pattern (less than once per day on average). 
4. 
Employee of PRA, Nuvisan, or the Sponsor. 
5. 
History of relevant drug and/or food allergies. 
6. 
Using tobacco products within 60 days prior to drug administration.  
7. 
History of alcohol abuse or drug addiction (including soft drugs like cannabis 
products). 
8. 
Positive drug and alcohol screen (opiates, methadone, cocaine, amphetamines 
[including 
ecstasy], 
cannabinoids, 
barbiturates, 
benzodiazepines, 
gamma-hydroxybutyric acid, tricyclic antidepressants, and alcohol) at screening or 
admission to the clinical research center. 
9. 
Average intake of more than 24 grams of alcohol per day. 
10. Positive screen for hepatitis B surface antigen (HBsAg), hepatitis C virus (HCV) 
antibodies, or HIV 1 and 2 antibodies. 
11. Participation in a drug study within 30 days prior to drug administration in the 
current study. Participation in more than 4 drug studies in the 12 months prior to 
drug administration in the current study. 
12. Donation or loss of more than 450 mL of blood within 60 days prior to drug 
administration. Donation or loss of more than 1.5 liters of blood in the 10 months 
prior to drug administration in the current study. 
13. Significant and/or acute illness within 5 days prior to drug administration that may 
impact safety assessments, in the opinion of the Investigator. 


  
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14.
Unwillingness to consume the Food and Drug Administration (FDA)-recommended
high-fat breakfast. 
15. Unsuitable veins for infusion or blood sampling. 
16. Positive nasopharyngeal PCR test for SARS-CoV-2 on Day -1. 
 
Please note that subjects should refrain from consumption of any foods containing poppy 
seeds, corn (whole corn kernels and popcorn), cruciferous vegetables, and bitter 
oranges within 48 hours (2 days) prior to screening and admission to the clinical 
research center to avoid false positive drug screen results, difficult feces texture, or 
induction of drug metabolizing enzymes. In addition, they should refrain from strenuous 
exercise within 96 hours (4 days) prior to screening and admission as this could result in 
abnormal clinical laboratory values.  
3.3.3 
Removal of Subjects from Assessment 
Participation in the study is strictly voluntary. A subject has the right to withdraw from the 
study at any time for any reason. 
 
The Investigator has the right to terminate participation of a subject for any of the 
following reasons: difficulties in obtaining blood samples, violation of the protocol, 
severe AEs or SAEs, or for any other reason relating to the subject’s safety or integrity 
of the study data. 
 
If a subject is withdrawn from the study, Nuvisan and LMU Klinikum will be informed 
immediately. If there is a medical reason for withdrawal, the subject will remain under 
the supervision of the Investigator until satisfactory health has returned, or the condition 
has stabilized with no more change being likely. 
 
After signing informed consent, subjects who drop out or withdraw for any reason without 
completing all screening evaluations successfully as well as subjects who drop out or 
withdraw prior to the first dose of study drug will be considered screening failures. 
 
A subject who is withdrawn or voluntarily withdraws from the study for any reason, 
whether related to the study drug or not, after having received the study drug, will be 
considered an early-termination subject. If a subject is withdrawn from the study, the 
early-termination subject will be replaced after mutual agreement between the Sponsor 
and PRA if the number of evaluable subjects drops below 4 (with a maximum of 
2 replacements).  
 
The decision regarding the replacement of subjects will be documented. 
 
PRA will make every effort to ensure that early-termination subjects who have received 
the study drug complete the early-termination assessments/safety follow-up 
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Early-termination subjects may be replaced if the total number of completers drops 
below 4, after mutual agreement between Nuvisan, LMU Klinikum, and PRA.   
3.3.3.1 
Stopping Rules for Individual Subjects 
In case of a positive SARS-CoV-2 virus PCR test or evidence for COVID-19 during the 
study, the subject will be withdrawn from the study. The subject will be isolated from 
other study participants and referred for treatment. The subject will be followed up in 
quarantine in the clinical research center until complete elimination of the study 
compound or will be asked to quarantine at home according to guidelines of the Dutch 
government. 
3.4
Treatments
3.4.1
Treatments Administered
Subjects will receive a single oral dose of 500 mg BTZ-043 containing 3.7 MBq of 
[14C]BTZ-043 on Day 1. 
3.4.2 
Identity of Investigational Product 
Active Medication 
Active substance 
: BTZ-043 
Activity 
: Inhibition of M. tuberculosis DprE1 
In development for 
: Tuberculosis 
Dose 
:  1 x 100 mL (equivalent to 500 mg BTZ-043) 
Strength 
: 5 mg/mL BTZ-043 containing 0.037 MBq of [14C]BTZ-043 
Dosage form 
: Oral suspension 
Manufacturer 
: Sourced by pharmacy at PRA 
 
The active pharmaceutical ingredient will be provided by the Promotor.  
 
[14C]BTZ-043 is a drug under development and not approved yet. It will be manufactured 
and analysed according to Good Manufacturing Practice (GMP) requirements. A GMP 
certificate of the manufacturer of the [14C]BTZ-043 suspension and an Investigator’s 
Brochure that summarizes preclinical and clinical data with respect to BTZ-043 are in 
place. A certificate of analysis and a certificate of conformity will be provided by PRA 
prior to clinical part. 
 
For details concerning drug storage and drug accountability see Appendix 8.1. 
3.4.3 
Method of Assigning Subjects to Treatment Groups 
After obtaining informed consent, subjects will receive a screening number and will be 
screened according to the inclusion and exclusion criteria. Subjects who have met all 
eligibility criteria will receive a subject number upon inclusion in the study 
(Subject Numbers 001-004). They will receive the subject number just prior to dosing. 
The subject number will ensure identification throughout the study after study drug 
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replaced, increased by 100 (eg, 101 replacement number for Subject Number 001), and 
will be administered the same treatment. 
 
After signing informed consent, subjects who drop out or withdraw for any reason without 
completing all screening evaluations successfully as well as subjects who drop out or 
withdraw prior to the first dose of study drug will be considered screening failures. Such 
subjects, and also subjects who are eligible for inclusion in the study but do not receive 
the study drug, will not receive a subject number, and only applicable data will be entered 
in the eCRFs.  
3.4.4 
Selection of Doses and Meals in the Study 
The targeted dose of 500 mg is considered a clinically relevant dose for the human AME 
study (Table 2) since this exposure achieves a maximal bactericidal effect and has 
shown to be tolerable in the 14-day Phase 1b study. It is proposed that the active 
pharmaceutical ingredient is taken after a high-fat breakfast (see Section 3.4.7), as 
absorption is then much improved. As shown in Table 2, the expected exposure is well 
below the maximum and well tolerated exposure achieved in the Phase 1b study. In the 
unexpected case that the currently conducted Phase 2a study with 250 mg, 500 mg, and 
1000 mg BTZ-043 administered after a high-fat breakfast shows any dose-related safety 
alert, the study team and the Sponsor will take measures to reduce the exposure. 
Reduction of exposure will be achieved by adapting the food intake (up to fasting) before 
the administration of [14C]BTZ-043. This protocol describes the drug administration of 
500 mg [14C]BTZ-043 after a high-fat breakfast as default. However, the study team and 
the Sponsor will decide whether a reduction in food intake is necessary to reduce the 
exposure to [14C]BTZ-043. 
 
In case it is indeed decided to administer 500 mg [14C]BTZ-043 after a 
standardized breakfast or in the fasted state rather than after a high-fat breakfast 
as planned, this decision and the associated rationale will be laid down in a Note to File, 
to be signed by the Sponsor and the PI, and to be submitted to the IEC for information 
only. 
 
The dose of 14C (approximately 3.7 MBq) to be administered was chosen to ensure 
successful profiling and identification of metabolites. 
 
In the present study, the estimated effective radiation burden after a single oral 
radioactivity dose of 3.7 MBq is approximately 0.16 mSv. 3 For biomedical investigations 
in small groups of human volunteers, an effective dose of 0.1 to 1.0 mSv is considered 
acceptable. 
3.4.5 
Radiation Safety of Clinical and Laboratory Staff 
It is expected that the clinical and laboratory staff handling the radioactive material and 
samples will not be exposed to any relevant radiation and associated health risk. The 
[14C] isotope emits low energy beta particles, which are not able to penetrate the 
dose containers or sample 
container walls. 
Therefore, there will 
be 
no 
“external” exposure from sample handling. The samples from the participants after 
y (Table 2) since this exposure achieves a maximal bactericidal effect and has 
shown to be tolerable in the 14-day Phase 1b study. It is proposed that the active 
pharmaceutical ingredient is taken after a high-fat breakfast (see Section 3.4.7), as 
absorption is then much improved. As shown in Table 2, t


  
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radiolabeled dose administration will contain low levels of radioactivity. The radioactive 
dose of 3.7 MBq [14C] per subject is below the legal handling limit, above which a special 
laboratory and special procedures for handling radioactivity would be required. 
Therefore, 
the 
staff 
can 
handle 
the 
study 
samples 
like 
any 
other 
non-radioactive biological material. Common practice of safe laboratory technique and 
hygiene will be exercised in handling all biological materials from this study.  
3.4.6 
Timing of Doses in the Study 
On Day 1, the study drug will be administered with the subject in the upright position. 
The study drug will be administered to subjects between 08:00 and 11:00 in the morning. 
After an overnight fast of at least 10 hours, subjects will receive an FDA-recommended 
high-fat breakfast (or a standardized breakfast, if applicable as outlined in Section 3.4.4) 
that will have to be finished within 20 minutes. The entire breakfast must be consumed 
by the subjects. The actual consumed food will be documented.  
 
Dosing will occur at 30 minutes after the start of the high-fat breakfast (or the 
standardized breakfast, if applicable, or dosing will occur in a fasted state, if applicable 
as outlined in Section 3.4.4).  
 
The study drug will be given as an oral suspension of 100 mL, which will be provided in 
an amber glass bottle from which the subjects can drink. Thereafter the bottle will be 
rinsed twice with 70 mL of water, and the subjects will be required to drink the bottle 
contents after rinsing. Thereafter, the subjects are required to drink additional 
non-carbonated water until they have ingested a total volume of 380 mL. 
 
Following drug administration, subjects will fast for a period of 4 hours until lunch. During 
fasting, no fluids are allowed except water; however, water is not allowed from 1 hour 
predose until 1 hour postdose (apart from the water taken with the dose).  
 
Subjects will not lie down for 4 hours after drug administration, except when required for 
assessments that need to be performed. 
3.4.7 
Meals During the Study 
A fasting period of at least 4 hours is required before obtaining clinical laboratory 
blood samples at the time points indicated in the schedule of assessments (Table 1). 
 
An FDA-recommended high-fat (or a standardized breakfast, if applicable, as outlined in 
Section 3.4.4) with calculated caloric content will be provided on Day 1.  
 
The FDA-recommended high-fat breakfast of 918 kcal consists of: 
 
2 fried eggs (in 15 g butter/margarine) (approximately 100 g) 
 
1 portion of bacon (40 g) (or brie 60+ for vegetarians) 
 
1 portion of fried potatoes (115 g) 
 
2 slices of (toasted) (wheat) bread with 15 g margarine 
 
1 glass of whole milk (240 mL) 


  
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The total of 918 kcal (vegetarian version 915 kcal) can be broken down as follows: 
 
39 g protein 
 
= 156 kcal 
 
59 g fat 
 
= 527 kcal 
 
59 g carbohydrates  
= 235 kcal 
 
The standardized breakfast of 572-755 kcal (depending on choices made) consists of: 
 
3 slices of whole-wheat bread  
 
1 Dutch rusk  
 
20 g halvarine/margarine  
 
1 portion of cheese/old cheese/cheese spread  
 
1 portion of ham/chicken breast/smoked meat/spreadable liver sausage/baloney/filet 
American/salami (or cheese, cheese spread, or peanut butter for vegetarians) 
 
2 portions of jam or other sweet spread  
 
2 mugs of decaffeinated coffee/tea (cream and sugar optional, max 2 of both) 
 
The total of 572-755 kcal (vegetarian version 589-805 kcal) can be broken down as 
follows: 
 
18-19 g protein 
 
= 71-75 kcal 
 
26-32 g fat 
 
= 233-292  kcal 
 
67-97 g carbohydrates 
= 268-388 kcal 
 
Other meals and snacks (such as decaffeinated coffee, herbal tea, fruit, and biscuits) 
will be provided according to PRA standard operating procedures (SOPs), taking into 
account the restrictions as described in Section 3.4.9 and what has been described in 
Section 3.4.6.   
3.4.8 
Blinding 
This is an open-label study. 
3.4.9 
Concomitant Medication and Other Restrictions During the Study 
Note: Restrictions that apply to the period before admission are described in 
Section 3.3.1 and Section 3.3.2. 
 
The use of all prescribed medication is not allowed from admission to the clinical research 
center until follow-up. The use of all over-the-counter medications, vaccines, 
vitamin preparations (especially vitamin C), other food supplements, and herbal 
medications (eg, St. John’s wort) is not allowed from admission to the clinical research 
center until follow-up. An exception is made for paracetamol: from admission onwards, 
the Investigator may permit a limited amount of paracetamol for the treatment of 
headache or any other pain. Other medication to treat AEs may only be prescribed if 
deemed necessary by the Investigator. If medication is used, the name of the drug, the 
dose, and dosage regimen will be recorded in the eCRF.  
 


  
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The use of methylxanthine-containing beverages or food (coffee, tea, cola, chocolate, and 
energy drinks), grapefruit (juice), kiwi, and tobacco products is not allowed during the stay 
in the clinical research center. 
 
The use of alcohol is not allowed during the stay in the clinical research center and within 
48 hours (2 days) prior to the 24-hour return visits (Days 11-12 and 15-16, as needed) and 
prior to follow-up. 
 
Strenuous exercise is not allowed within 96 hours (4 days) prior to admission and 
follow-up. Strenuous exercise is also not allowed during the stay in the clinical research 
center.  
 
Subjects should not consume any foods containing poppy seeds, corn (whole corn 
kernels and popcorn), cruciferous vegetables, and bitter oranges within 48 hours 
(2 days) prior to admission to the clinical research center as this could cause a false 
positive drug screen result. 
 
Male subjects, if not surgically sterilized, are required to use adequate contraception 
(see Inclusion Criterion 6) and not donate sperm from admission to the clinical research 
center until 90 days after the follow-up visit. 
 
Subjects must not donate blood during the study until the follow-up visit (other than the 
blood sampling planned for this study). 
3.4.10 
Treatment Compliance 
Study drug will be administered in the clinical research center. To ensure treatment 
compliance, administration of the study drug will be supervised by the Investigator or an 
authorized designee. Compliance will be further confirmed by bioanalytical assessment 
of BTZ-043 and total radioactivity in plasma and urine samples (see Section 3.5.4). 
 
The exact times of study drug administration and the number of units administered will 
be recorded in the eCRF.  
3.5
Pharmacokinetic, Safety, and Exploratory Measurements and Variables
3.5.1 
Pharmacokinetic and Safety Measurements Assessed and Schedule of 
Assessments  
A schedule of assessments is presented in Table 1. 
3.5.1.1 
Pharmacokinetic Measurements  
3.5.1.1.1 Blood Sampling  
At the time points defined in the schedule of assessments (Table 1), blood samples will 
be taken for the analysis of BTZ-043 and metabolites in plasma samples, total 
radioactivity in whole blood and plasma samples, and metabolite profiling and 
identification in plasma samples. 


  
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The blood samples will be taken via an indwelling iv catheter or by direct venipuncture. 
The exact times of blood sampling will be recorded in the eCRF. 
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA and Nuvisan. 
3.5.1.1.2 Urine Collection  
During the intervals defined in the schedule of assessments, urine will be collected for 
the analysis of BTZ-043 and metabolites, for total radioactivity, for quick counts or normal 
counts of total radioactivity (as feasible), and for metabolite profiling and identification. 
The subjects will be instructed to empty their bladders completely before drug 
administration and at the end of each collection interval. A baseline urine sample will be 
collected within 12 hours prior to drug administration. The exact times of urine collection 
and the urine weight of the entire interval will be recorded in the eCRF. 
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA and Nuvisan. 
3.5.1.1.3 Feces Collection  
During the intervals defined in the schedule of assessments, all fecal excretions will be 
collected for the analysis of total radioactivity, for quick or normal counts of total 
radioactivity (as feasible), and for metabolite profiling and identification. A blank fecal 
sample will be collected within 48 hours prior to study drug administration. The exact 
times of feces collection and the fecal weight will be recorded in the eCRF.  
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA and Nuvisan. 
3.5.1.1.4 
Vomit Collection  
If a subject vomits within the time period from drug administration up to 8 hours after the 
administration of the study drug, the vomit should, if possible, be collected. Any collected 
vomit will be included in the total radioactivity measurements and in the determination of 
the mass balance. The exact times of vomit production will be recorded in the eCRF.  
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA and Nuvisan. 
3.5.1.2 
Safety and Tolerability Measurements 
Safety and tolerability assessments will consist of AEs, clinical laboratory, vital signs, 
12-lead ECG, and physical examination. Assessments will be performed in accordance 
with the schedule of assessments. 


  
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3.5.1.2.1
Adverse Events
AEs will be recorded from signing the ICF until completion of the follow-up visit. Any 
clinically significant observations, as determined by the Investigator, in results of clinical 
laboratory, 12-lead ECGs, vital signs, or physical examinations will be recorded as AEs. 
 
A TEAE is defined as any event not present prior to administration of the study drug or 
any event already present that worsens in either severity or frequency following 
exposure to the study drug.  
 
An AE that occurs prior to administration of the study drug will be considered a 
pretreatment AE.  
 
At several time points before and after drug administration, subjects will be asked 
nonleading questions to determine the occurrence of AEs. Subjects will be asked in 
general terms about any AEs at regular intervals during the study. In addition, all AEs 
reported spontaneously during the course of the study will be recorded. All answers will 
be interpreted by the Investigator using the Medical Dictionary for Regulatory Activities 
(MedDRA; most recent version) for AEs and will be recorded in the eCRF as reported 
terms.  
 
The severity of the AEs will be rated as mild, moderate, or severe; the relationship 
between the AEs and the study drug will be indicated as related or not related. Details 
on rating the severity of AEs and relationship to study treatment are given in 
Appendix 8.2. 
 
Pregnancy of female partners of male subjects will be monitored along with follow-up, if 
warranted (see Appendix 8.3). 
3.5.1.2.2 Clinical Laboratory  
Blood and urine samples for clinical laboratory assessments will be collected according 
to PRA SOPs. 
The following parameters will be measured: 
 
Clinical chemistry (serum quantitatively): 
 
total bilirubin, alkaline phosphatase, gamma glutamyl transferase, AST, ALT, lactate 
dehydrogenase, creatinine, urea, total protein, glucose, inorganic phosphate, 
sodium, potassium, calcium, and chloride.  
 
Hematology (blood quantitatively): 
 
leukocytes, erythrocytes, hemoglobin, hematocrit, thrombocytes, absolute partial 
automated differentiation (lymphocytes, monocytes, eosinophils, basophils, and 
neutrophils), mean corpuscular volume, mean corpuscular hemoglobin, and mean 
corpuscular hemoglobin concentration. 
 
Urinalysis (urine qualitatively): 
 
hemoglobin, urobilinogen, ketones, glucose, protein. 
 
Serology: 
 
HBsAg, HCV antibodies, and HIV 1 and 2 antibodies. 


  
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Drug and alcohol screen: 
 
opiates, methadone, cocaine, gamma hydroxybutyric acid, amphetamines (including 
ecstasy), cannabinoids, barbiturates, benzodiazepines, tricyclic antidepressants, 
nicotine metabolites, and alcohol. 
 
Nasal and throat mucosal cell samples will be collected according to PRA work 
instructions detailed in the site specific manual. The samples will be tested for 
SARS-CoV-2 virus using PCR tests. The nonclinical project manager will be informed 
immediately upon availability of Day 11 results, as this will qualify the feces samples for 
metabolite profiling with regard to potential COVID-19 infectivity. 
 
In case of unexplained or unexpected clinical laboratory test values, the tests will be 
repeated as soon as possible and followed up until the results have returned to the 
normal range and/or an adequate explanation for the abnormality is found. The clinical 
laboratory will clearly mark all laboratory test values that are outside the normal range, 
and the Investigator will indicate which of these deviations are clinically significant. 
Clinically significant laboratory result deviations will be recorded as AEs, and the 
relationship to the treatment will be indicated (see also Appendix 8.2). 
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA. 
3.5.1.2.3 Vital Signs 
Systolic and diastolic blood pressure and pulse will be recorded after the subject has 
been resting for at least 5 minutes in the supine position. These assessments will be 
made using an automated device whenever possible. Body temperature and respiratory 
rate will also be measured. 
3.5.1.2.4 Electrocardiogram 
A standard 12-lead ECG will be recorded after the subject has been resting for at least 
5 minutes in the supine position. The ECG will be recorded using an ECG machine 
equipped with computer-based interval measurements. The following ECG parameters 
will be recorded: heart rate, PR-interval, QRS-duration, QT-interval, QTc-interval 
(Fridericia’s), and the interpretation of the ECG profile by the Investigator. 
3.5.1.2.5 Physical Examination 
Physical examination will be performed according to PRA SOPs. In addition, body weight 
and height will be measured according to PRA SOPs. 
3.5.1.3 
Total of Blood Volume 
Table 3 presents the number and volume of blood samples and the total volume of blood 
that will be collected throughout the study per subject.  
 
If deemed necessary by the Investigator or the Sponsor, the number and/or volume of 
blood samples per assessment may be increased, as long as the total volume of blood 


  
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drawn for a subject does not surpass 500 mL (except when extra blood samples need 
to be taken for safety reasons). 
 
Table 3 
Number and Volume of Blood Samples and Total Blood Volume 
Collected per Subject 
Assessment
Maximum # 
Samples 
Volume of Blood per 
Sample (mL) 
Total Volume of 
Blood (mL) 
Pharmacokinetics 
 - BTZ-043 and metabolites in 
Plasma 
- Total Radioactivity in Plasma 
 - Total Radioactivity in Whole 
Blood 
 - Metabolite Profiling in Plasma 
 
14 
 
21 
21 
 
2119 
42 
24
1 
 
1215 
 
5628 
4284
21 
 
252285 
Clinical Chemistry 
4
3.5  
14
Hematology  
4 
3  
12 
Serology 
1 
5  
5 
Total Volume of Blood Drawn 
 
 
402
 
3.5.2 
Appropriateness of Measurements 
The assessments that will be made in this study are standard and generally recognized 
as reliable, accurate, and relevant.  
3.5.2.1 
Timing of Assessments  
For PK, predose samples will be obtained between waking up and dosing. Postdose 
samples up to 20 minutes postdose will be obtained with a time window of ±1 minute. 
Thereafter, postdose samples will be obtained with time margins of ±5% of the time that 
has passed since dosing. The ±5% time window also applies to the start and end times 
of urine collection intervals and to the total duration of each collection interval. 
 
For safety assessments, predose assessments will be performed between waking up 
and dosing. For safety assessments up to 2.5 hours postdose, a time window of 
±15 minutes is allowed. Thereafter, serial postdose assessments (eg, multiple 
assessments within any given day) will be performed with time margins of ±10% of the 
time that has passed since dosing. 
 
When assessments are planned for the same scheme time, the order of the 
assessments should be arranged in such a way that PK blood sampling will be done 
after the ECG and vital signs recordings have been conducted, with PK blood sampling 
exactly on time. 
3.5.3 
Pharmacokinetic, Safety, and Exploratory Variables  
3.5.3.1 
Pharmacokinetic Variables  
PK variables will be the plasma and urine concentrations of BTZ-043 and 
main metabolites, the total radioactivity concentrations in plasma, whole blood, urine, 
and feces (and in vomit, if produced) and their PK parameters. The PK parameters to be 


  
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determined or calculated using noncompartmental analysis include, but are not limited 
to, the parameters as given in Table 4. A complete list of PK parameters will be provided 
in the PK statistical analysis plan (SAP). 
 
Table 4
Pharmacokinetic Parameters 
Parameter 
Description 
Parent 
Metabolites
TRA 
Cmax
Maximum 
observed 
plasma 
concentration 
X
X
X
tmax
Time to attain maximum observed 
plasma concentration 
X 
X 
X
AUC0-t 
Area under the plasma concentration-
time curve up to time t, where t is the last 
point with concentrations above the 
LLOQ  
X 
X 
X
AUCinf 
Area under the plasma concentration-
time curve from time 0 to infinity 
calculated as: AUC0-inf=AUC0-t+Clast/kel, 
where Clast is the last measurable plasma 
concentration  
X 
X 
X
Kel 
Terminal elimination rate constant 
X 
X 
X
t1/2
Terminal elimination half-life, calculated 
as 0.693/kel 
X 
X 
X
CL/F 
Apparent oral clearance, calculated as 
dose/AUC0-inf 
X 
 
 
Vz/F 
Apparent volume of distribution at 
terminal phase  
X 
 
 
CLR 
Renal clearance 
X 
 
 
Aeurine 
Cumulative amount of drug excreted in 
urine  
X 
X 
X
Aefeces
Cumulative amount of drug excreted in 
feces 
 
 
X
Aevomit 
Cumulative amount of drug excreted in 
vomit, if produced 
 
 
X
Aetotal 
Total 
amount 
of drug 
excreted, 
calculated 
as 
Aetotal=Aeurine+Aefeces(+Aevomit)
 
 
X
feurine
Fraction of the dose administered 
excreted in urine (%) 
X
X
X
fefeces 
Fraction of the dose administered 
excreted in feces (%) 
 
 
X
fevomit 
Fraction of the dose administered 
excreted in vomit (%) 
 
 
X
fetotal 
Fraction of the dose administered 
excreted in urine, feces, and vomit (if 
produced) (%) 
 
 
X
TRA=total radioactivity; LLOQ=lower limit of quantification 
 


  
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3.5.3.2
Safety Variables
The safety variables to be measured include, but are not limited to, the variables as given 
below. A complete list of safety variables will be provided in the SAP.
 
 
AEs 
 
Clinical laboratory 
 
Vital signs 
 
12-lead ECG 
 
Physical examination  
3.5.3.3 
Exploratory Variables 
Metabolite profiling and metabolite identification will be conducted under a separate 
protocol/bioanalytical study plan, by Nuvisan Grafing, and the results will be reported 
separately. The metabolite profiling/identification report will be included in the clinical 
study 
report 
(CSR) 
as 
an 
appendix, 
and 
results 
from 
the 
metabolite profiling/identification report will be briefly summarized in the CSR. 
3.5.4 
Drug Concentration Measurements 
The analysis of BTZ-043 and main metabolites in plasma and urine samples will be 
performed at the Bioanalytical Department of Nuvisan Grafing using validated/qualified 
LC-MS. The analysis of whole blood, plasma, urine, and feces (and vomit, if produced) 
for total radioactivity will be conducted at the Bioanalytical Laboratory of PRA using a 
validated liquid scintillation counting method. The bioanalytical reports for the 
determinations will be included as appendices in the CSR.  
 
The analysis of quick counts will be conducted at the Bioanalytical Laboratory of PRA 
using a validated method. 
 
Metabolite profiling in plasma, urine, and feces samples by radiometry will be conducted 
at the PK Sciences & Biotransformation Department of Nuvisan Grafing. The 
bioanalytical report for the metabolite profiling and identification will be included as 
appendix in the CSR. 
3.5.5 
Retention of Blood, Plasma, Urine, and Feces Samples 
Blood and urine samples remaining after clinical laboratory assessments have been 
performed will not be stored for future use and will be destroyed after analysis as per 
laboratory procedure. 
 
Blood, plasma, urine, and feces (and vomit, if applicable) samples remaining after total 
radioactivity measurements and after sample aliquotation for bioanalytical and 
metabolite identification have been performed, will be stored at the site of the Promotor  
and destroyed no later than 10 years after the date of the final CSR.  


  
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3.6 
Statistical Procedures and Determination of Sample Size 
3.6.1
Analysis Sets
3.6.1.1 
Safety Set 
All subjects who have received the single oral dose of 14C-BTZ-043. 
3.6.1.2 
Pharmacokinetic Set 
All subjects who have received the single oral dose of 14C-BTZ-043 and provided 
sufficient bioanalytical assessment results to calculate reliable estimates of the PK 
parameters. 
3.6.2 
Statistical and Analytical Plan for Pharmacokinetic, Safety, and Exploratory 
Evaluation  
A safety SAP will be generated by the Biostatistics Department of PRA and a PK SAP 
will be generated by Nuvisan; the SAPs will be finalized prior to database lock. Full 
details of the analysis to be performed will be included in the SAPs. 
 
Any deviation from the SAPs will be reported in the section “Changes in Planned 
Analysis” in the CSR. 
3.6.2.1 
Pharmacokinetic Evaluation 
The PK parameters and their statistical evaluation of BTZ-043 and main metabolites will 
be reported as an integrated PK report, which will be included as an appendix and briefly 
summarized in the CSR. part of the bioanalytical report (included as appendix in the 
CSR). The PK parameters and their statistical evaluation of total radioactivity will be part 
of the mass balance report (included as an appendix in the CSR). A separate integrated 
PK report will incorporate the BTZ-043 and main metabolites’ PK, the total radioactivity 
PK, and the metabolite profiling PK data for an integrated evaluation. 
 
All data will be summarized using descriptive statistics and will be listed and summarized 
in tabular and/or graphical form. 
3.6.2.2 
Evaluation of Safety and Tolerability 
Safety and tolerability will be assessed through AEs, clinical laboratory, vital signs, 
ECGs, physical examination findings, and any other parameter that is relevant for safety 
assessment. 
3.6.2.2.1 Adverse Events 
A listing of all individual AEs will be provided. Summary tables of TEAEs will be 
presented by system organ class based on the MedDRA terminology list (preferred 
terms): 1 containing the number of TEAEs (frequency of occurrence, number of subjects 
experiencing the event) by treatment and 1 containing the number of drug-related TEAEs 
(frequency of occurrence, number of subjects experiencing the event) per treatment. 
Additional tables of total counts by treatment and relationship and by treatment and 
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3.6.2.2.2
Clinical Laboratory
Clinical laboratory data will be listed accompanied by an indication if the parameter is 
outside the reference range. A summary of all data outside the reference range of the 
clinical laboratory will be provided. Clinical laboratory data will be presented 
descriptively, where applicable. 
3.6.2.2.3 Vital Signs and Electrocardiograms 
Vital signs and ECG parameters will be listed and they will be presented descriptively, 
where applicable. 
3.6.2.2.4 Physical Examination 
Changes from baseline for physical examination will be described and listed. 
3.6.2.3 
Evaluation of Exploratory Variables  
The PK parameters and their statistical evaluation of metabolites not measured for the 
primary objectives will be reported in the a separate integrated PK report., which will be 
included as an appendix and briefly summarized in the CSR. 
 
All data will be summarized using descriptive statistics and will be listed and summarized 
in tabular and/or graphical form. 
 
The metabolite profiling and identification results will be reported separately 
(see Section 3.5.4). 
3.6.3 
Determination of Sample Size 
Since the present study is a descriptive study aimed at assessing excretion routes of 
BTZ-043 by evaluating mass balance, a formal statistical sample size calculation was 
not performed. A sample size of 4 subjects is a minimum accepted number of subjects 
for AME studies and is considered sufficient to achieve the study objectives. 
Early-termination subjects may be replaced if the total number of completers drops 
below 4, after mutual agreement between the Sponsor and PRA. 
3.7 
Data Quality Assurance 
The study may be audited by the Quality Assurance Department at PRA to assess 
adherence to the clinical study protocol (CSP) and Quality System. During the conduct 
of the study, process-related audits may be performed. An audit certificate will be 
provided in the appendices of the final CSR outlining any audits and other related 
activities performed.  
 
The clinical research site will be monitored by the study monitor to ensure correct 
performance of the study procedures and assure that the study is conducted according 
to the relevant regulatory requirements. The eCRF entries will be verified with the source 
documentation, if applicable (in some cases data are captured directly on the eCRF, 
therefore source data verification is not necessary). 
 


  
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Regulatory authorities, the Independent Ethics Committee (IEC), and/or the Sponsor’s 
clinical quality assurance group may request access to all source documents, eCRFs, 
and other study documentation for on-site audit or inspection. Direct access to these 
documents must be guaranteed by the Investigator, who must provide support at all 
times for these activities. 
 
Quality control principles will be applied throughout the performance of this study. 
Review procedures will be followed at PRA for all documents that are generated in 
relation with the study.  
 
An explanation will be given for all missing, unused, and spurious data in the relevant 
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4. 
ETHICS 
4.1 
Independent Ethics Committee 
The submission package including the CSP and the ICFs will be submitted for review 
and approval by the IEC of the foundation “Beoordeling Ethiek Biomedisch Onderzoek” 
(English translation: “Evaluation of Ethics in Biomedical Research”) (Dr. Nassaulaan 10, 
9401 HK Assen, The Netherlands) prior to the eligibility screening. The composition of 
the IEC is in accordance with the recommendations of the World Health Organization, 
the International Council for Harmonisation (ICH) E6(R2) Guideline for Good Clinical 
Practice (GCP) (European Medicines Agency [EMA]/Committee for Medicinal Products 
for Human Use [CHMP]/ICH/135/1995)4, and the EU Clinical Trial Directive (CTD) 
(Directive 2001/20/EC)5 (see below). The submission package will also be shared with 
the CA in the Netherlands for a statement of no objection. 
 
PRA will keep the IEC informed about the progress of the study. All changes in research 
activities and all unanticipated problems involving risks to human subjects will be 
immediately reported to the IEC. In accordance with Section 10, Subsection 1, of the 
Dutch law on Medical Research in Human Subjects (WMO, revised Dec 2015)6, PRA 
will inform the subjects and the IEC if anything occurs on the basis of which it appears 
that the disadvantages of participation may be significantly greater than was foreseen in 
the research proposal, or if further recruitment of subjects in the study has been put on 
hold for that reason, whichever occurs first. The study may be suspended pending 
further review by the IEC, except insofar as suspension would jeopardize the subjects’ 
health. PRA will take care that all subjects are kept informed.  
 
No changes will be made to the study without IEC approval, except when required to 
eliminate apparent immediate hazards to human subjects. 
 
Notification of the end of the study will be sent by PRA to the CA in The Netherlands and 
to the IEC within 90 days after completion of follow-up for the last subject. In case a 
study is temporarily halted, PRA will notify the IEC immediately, including the reason for 
this. In case a study is ended prematurely, PRA will notify the IEC and the CA in The 
Netherlands within 15 days, including the reasons for the premature termination. A 
summary of the results of the study will be sent by PRA to the CA and the IEC within 
1 year after the end of the study.  
 
The written documentation of approval of the CSP and ICFs will be provided to the 
Sponsor before the start of the study. Any communication with the IEC and/or CA before, 
during, and/or after the study will be provided to the Sponsor.    
4.2 
Ethical Conduct of the Study 
This study will be conducted in accordance with the ethical principles that have their 
origin in the World Medical Association (WMA) Declaration of Helsinki, adopted by the 
18th
WMA General Assembly,
Helsinki, Finland, Jun 1964, and subsequent 
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This study is also designed to comply with ICH E6(R2) Guideline for GCP 
(EMA/CHMP/ICH/135/1995)4, and the EU CTD Directive 2001/20/EC5, as incorporated 
into Dutch Law.6 
 
Guidelines adopted by the ICH and other relevant international guidelines, 
recommendations, and requirements will be taken into account as comprehensively as 
possible, as long as they do not violate local law. 
 
Whenever the term “Investigator” is noted in the CSP text, it may refer to either the 
Investigator at the site or an appropriately qualified, trained, and delegated individual of 
the investigational site. 
4.3 
Subject Information and Consent 
All subjects will be informed verbally and in writing regarding the objectives, procedures, 
and risks of study participation. The subjects will be given the time to ask questions. 
The subjects will have to sign the Dutch or English version of the ICF before any 
study-related procedures are started. The signed ICFs will be retained and archived at 
PRA and a copy will be provided to the subject. The ICF contains information about the 
objectives of the study, the procedures followed during the study, and the risks and 
restrictions of the study, with special reference to possible side effects of the study drug 
and potential interactions. In addition, insurance coverage provided during the study is 
explained. The elements addressed in the ICF are according to the ICH E6(R2) 
Guideline for GCP (EMA/CHMP/ICH/135/1995).4
4.4 
Privacy 
All personal details will be treated as confidential by the Investigator and staff at PRA, 
the Sponsor, and any sub-contractors involved, and handling of personal data will be in 
compliance with the EU General Data Protection Regulation.8
 


  
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5. 
STUDY ADMINISTRATIVE STRUCTURE 
5.1 
Distribution of Activities 
5.1.1
Preparation of Study Drug
The study drug will be prepared at the PRA Pharmacy. 
5.1.2 
Laboratory Assessments 
The analysis of BTZ-043 and main metabolites in plasma and urine samples will be 
performed at the Bioanalytical Department of Nuvisan Grafing (Grafing, Germany) by 
using the Nuvisan study protocol G-A-LAB-21-004.  
 
The analysis of total radioactivity in whole blood, plasma, urine, and feces (and vomit, if 
produced) will be performed at the Bioanalytical Laboratory of PRA (Assen, 
The Netherlands).  
 
The analysis of clinical laboratory samples will be performed at the PRA Clinical 
Laboratory.  
 
Metabolite profiling and identification will be performed at the PK Sciences and 
Biotransformation Department of Nuvisan Pharma Services GmbH (Grafing, Germany) 
by using the Nuvisan study protocol G-A-MET-21-002. 
5.1.3 
Electronic Case Report Form Design 
The eCRF design will be performed with the computer program Oracle Clinical (Oracle, 
Redwood Shores, Redwood City, CA, US) by the Database Programming Department 
of PRA. 
5.1.4 
Data Management 
Data management will be performed with the computer programs Oracle Clinical 
(Oracle, Redwood Shores, Redwood City, CA, US), SAS® (SAS Institute Inc, Cary, NC, 
US), and EXACT (Kinship EXACT™, Kinship Technologies, a technology subsidiary of 
PRA) by the Data Management Department of PRA. 
5.1.5 
Statistics 
A safety SAP will be provided by the Biostatistics Department of PRA. The safety 
analysis will be conducted by the Biostatistics Department of PRA. Statistical analysis 
will be performed with the computer program SAS® (SAS Institute Inc, Cary, NC, US). A 
PK SAP will be provided by Nuvisan Grafing. Statistical evaluation of PK parameters will 
be conducted by Nuvisan Grafing. PK parameters will be calculated using Phoenix 
WinNonlin (Pharsight, Mountain View, CA, US). All individual results will be provided to 
the Sponsor after completion of the study. 
5.1.6 
Clinical Study Report Writing 
The CSR, structured in accordance with the guideline “Structure and Content of Clinical 
Study Reports - ICH E3”9, will be written by PRA and a draft version prepared for 


  
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approval by the Sponsor. The final version will be provided as paper version to the 
Sponsor. 
5.2 
Documentation 
5.2.1 
Archiving 
All documents concerning the study will be kept on file in the Central Archives of PRA 
for at least 25 years after conduct of the study. The Sponsor will receive the completed 
eCRFs as PDF file. 
5.2.2 
Recording of Data in Source Documents and Electronic Case Report Forms 
Wherever possible, all data will be entered directly into the eCRFs. Source documents 
will be used in some cases. 
 
A data management plan will be written by the Data Management Department of PRA, 
which will be finalized prior to performing any data validation. An appendix to the data 
management plan (origin of source data list for data entry) will identify any data to be 
recorded directly in the eCRF (ie, no prior written or electronic record of data) and which 
data should be considered source data.  
 


  
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6. 
CONFIDENTIALITY AND PUBLICATION POLICY 
All information generated in this study is considered highly confidential and must not be 
disclosed to any person or entity not directly involved with the study unless prior written 
consent is gained from the Sponsor and Promotor. However, authorized regulatory 
officials, the Sponsor and its authorized representatives are allowed full access to the 
records. 
 
All study subjects must be informed that their personal study-related data will be used 
by the Sponsor in accordance with local data protection law. The level of disclosure must 
also be explained to the subject, who will be required to give consent for their data to be 
used as described in the ICF. The subjects must be informed that their medical records 
may be examined by auditors or other authorized personnel appointed by the Sponsor, 
by appropriate IEC members, and by inspectors from regulatory authorities. 
 
Identification of subjects and eCRFs shall be by unique subject numbers only.  
 
All personal details will be treated as confidential by the Investigator and staff at PRA. 
 
All relevant aspects regarding publication will be part of the contract (or similar 
document) between the Sponsor and PRA.  
 


  
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7. 
REFERENCES 
1.
Investigator’s Brochure BTZ-043. Edition 04. 10 Nov 2020.
2. 
CPMP/EWP/560/95/Rev. 1 Corr. 2** Committee for Human Medicinal Products 
(CHMP). Guideline on the investigation of drug interactions. 21 Jun 2012. 
3. 
Radiation Burder Calculation Report EDS-NL. Dosimetry BTZ-043. 27 Nov 2018. 
4. 
International Council for Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, E6(R2): Integrated Addendum to ICH E6(R1): Guideline for Good 
Clinical Practice. Adopted by the European Medicines Agency (EMA), 
Committee 
for 
Human 
Medicinal 
Products, 
Document 
Reference 
EMA/CHMP/ICH/135/1995), 14 Jun 2017. 
5. 
Directive 2001/20/EC of the European Parliament and of the Council of 
4 Apr 2001 on the approximation of the laws, regulations and administrative 
provisions of the Member States relating to the implementation of good clinical 
practice in the conduct of clinical trials on medicinal products for human use. 
6. 
Medical 
Research 
Involving 
Human 
Subjects 
Act 
(WMO, 
Wet 
Medisch-Wetenschappelijk Onderzoek met Mensen), revision Dec 2015. 
7. 
World Medical Association (WMA) Declaration of Helsinki – Ethical Principles for 
Medical Research Involving Human Subjects (18th WMA General Assembly 
1964), revised at 64th WMA General Assembly, Fortaleza, Brazil, Oct 2013. 
8. 
The General Data Protection Regulation (GDPR). Regulation (EU) 2016/679 of 
the European Parliament and the Council of the European Union, 27 Apr 2016, 
applicable as of 25 May 2018. 
9. 
International Council for Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, E3: Structure and Content of Clinical Study Reports. Note for 
Guidance on Structure and Content of Clinical Study Reports, Adopted by the  
Committee for Human Medicinal Products, European Medicines Agency (EMA), 
Document Reference CPMP/ICH/137/95, Jul 1996. 
10. 
International Council for Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, E2A: Clinical Safety Data Management: Definitions and Standards for 
Expedited Reporting. Note for Guidance on Clinical Safety Data Management, 
Adopted by the Committee for Human Medicinal Products, European Medicines 
Agency (EMA), Document Reference CPMP/ICH/377/95, Jun 1995.


  
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8. 
APPENDICES 
8.1
Drug Accountability 
Upon receipt of the study drug, it will be inspected and counted by the responsible 
pharmacist. If necessary, all study drug will be repacked per dosing occasion and labeled 
according to PRA SOPs. 
 
The study drug will be kept in the PRA Pharmacy or in a locked and secured storage 
facility accessible to the pharmacist and the pharmacy assistant only. Temperature is 
monitored 24/7 with an automatic environmental monitoring system. In case of an alarm, 
the pharmacist or pharmacist on duty is informed immediately by email and phone. 
 
The responsible pharmacist will keep an inventory. This will include a description of the 
formulation and the quantity of study drug received for the study and a record of what is 
dispensed, to whom, and when. 
 
On termination of the study, the responsible pharmacist will conduct a final inventory of 
the study drug supply and will record the results of this inventory in the 
Drug Accountability Form. Unused study drug will be returned to the Promotor upon 
notice is given to Promotor and Sponsor or will be locally destroyed according to PRA 
standard procedures. Final accountability needs to be verified by monitor before return 
and/or destruction of medication. 
8.2
Adverse Events and Serious Adverse Events Evaluation and Reporting 
8.2.1 
Adverse Events 
An AE is any unfavorable and unintended sign (including an abnormal laboratory finding), 
symptom, or disease temporally associated with the use of a medical treatment or 
procedure that may or may not be considered related to the medical treatment or 
procedure. AE definitions will be followed as stated in the “Note for Guidance on Clinical 
Safety Data Management: Definitions and Standards for Expedited Reporting” (ICH topic 
E2A).10
 
All AEs reported by the subjects or apparent from their physical appearance during the 
clinical study will be reported on the AE eCRF page. 
 
The severity of AEs will be graded using the most current version of the MedDRA: 
 
Mild: An AE that is easily tolerated by the subject, causes minimal discomfort, does 
not interfere with everyday activities, and does not require intervention. 
 
Moderate: An AE that is sufficiently discomforting to interfere with normal everyday 
activities; intervention may be needed. 
 
Severe: An AE that prevents normal everyday activities; treatment or other 
intervention usually needed. 
 
If an AE has multiple aspects, the aspect with the highest severity will be graded. 


  
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It is emphasized that the term severe is a measure of severity; thus, a severe AE is not 
necessarily serious. For example, itching for several days may be rated as severe, but 
may not be clinically serious. 
 
In addition, clinically significant changes in physical examination findings and abnormal 
objective test findings (eg, laboratory, x-ray, ECG) should also be recorded as AEs. Test 
findings and physical examination findings can result in AEs if they:  
 
Are associated with accompanying symptoms, and/or 
 
Require additional diagnostic testing or medical/surgical intervention, and/or 
 
Lead to a change in study dosing or discontinuation from the study; result in the 
addition of significant additional concomitant drug treatment or other therapy, and/or 
 
Lead to any of the outcomes included in the definition of an SAE, and/or 
 
Are considered to be an AE by the Investigator or Sponsor. 
 
Reporting as an AE should not be triggered by: 
 
Merely repeating an abnormal test, or  
 
Any abnormal test result that is determined to be an error. 
 
The relationship of any AE to the study drug will be assessed and graded as related or 
not related. 
 
Related:   
 
The AE follows a reasonable temporal sequence to study drug administration, and 
cannot be reasonably explained by the subject’s clinical state or other factors (eg, 
disease under study, concurrent diseases, or concomitant medications). 
 
The AE follows a reasonable temporal sequence to study drug administration, and is 
a known reaction to the drug under study or a related chemical group, or is predicted 
by known pharmacology. 
 
Not Related: 
The AE does not follow a reasonable sequence from study drug administration, or can 
be reasonably explained by the subject’s clinical state or other factors (eg, disease under 
study, concurrent diseases, and concomitant medications).  
8.2.2 
Serious Adverse Events 
An SAE is any untoward medical occurrence that, on the basis of medical and scientific 
judgment: 
 
Results in death, or 
 
Is life-threatening (this refers to an event in which the patient was at risk of death at 
the time of the event; it does not refer to an event which hypothetically might have 
caused death if it were more severe), or  
 
Requires inpatient hospitalization for a medical reason or prolongation of existing 
hospitalization (this refers to hospital admission required for treatment of the AE). 
(Note: this does not include confinement in, for example, a respite unit; a skilled 


  
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nursing unit; rehabilitation facility; the clinical research center; or confinement due to 
planned or unplanned reason unrelated to study), or 
 
Results in persistent or significant disability/incapacity, or 
 
Is a congenital anomaly/birth defect. 
 
Medical and scientific judgment should be exercised in deciding whether expedited 
reporting is appropriate in other situations, such as important medical events that 
may not be immediately life-threatening or result in death or hospitalization, but may 
jeopardize the patient or may require intervention to prevent one of the other 
outcomes listed in the definition above. These should also usually be considered 
serious. 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 hospitalization. 
 
SAEs will be collected from admission until the final study visit. SAEs that are related to 
the investigational drug and continue beyond the normal collection period (ie, are 
ongoing at the time a subject exits the study) will be followed until resolution or until 
stabilized with sequelae. SAEs that begin after the subject’s participation in the study is 
complete, but that the Investigator considers to be related to study drug, may be reported 
at any time.  
 
The Investigator or clinical site personnel must notify the Sponsor’s Medical Monitor and 
PRA Drug Safety Center of all SAEs, regardless of relationship to the investigational 
drug, within 24 hours of clinical site personnel becoming aware of the event. The 
Investigator will provide the initial notification by sending a completed “SAE Notification 
Form,” which must include the Investigator’s assessment of the relationship of the event 
to investigational drug and must be signed by the Investigator. 
 
In addition, notification is sent by PRA to the IEC and the subject’s General Practitioner. 
 
Follow-up information, or new information regarding an ongoing SAE, must be provided 
promptly to the Sponsor’s Medical Monitor and PRA Drug Safety Center. 
All SAE reports should be sent to the contacts provided on Page 4: SAE Contact 
Information. 
8.2.3 
Suspected Unexpected Serious Adverse Reactions 
An SAE that is also an unexpected adverse drug reaction is called a suspected 
unexpected serious adverse reaction (SUSAR). Unexpected adverse reactions are 
adverse reactions of which the nature or severity is not consistent with the applicable 
product information (eg, investigator’s brochure for an unapproved investigational 
medicinal product or the Summary of Product Characteristics for an authorized medicinal 
product). 
 
The Sponsor or its representative (eg, PRA if agreed to before start of the study) will 
promptly report (expedited reporting) the following SUSARs to the IEC: 
 
SUSARs that have arisen in the current clinical study that was assessed by the IEC 


  
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SUSARs that have arisen in other clinical studies of the same Sponsor and with the 
same medicinal product, and that could have consequences for the safety of the 
subjects involved in the current clinical study that was assessed by the IEC 
 
The Sponsor will promptly report (expedited reporting) all SUSARs to the CA and the 
Medicine Evaluation Board (MEB) of the country where this study is conducted and to 
the CAs in other Member States, as applicable. 
 
SUSARs that have already been reported to the EMA Eudravigilance database do not 
have to be reported again to the CA and the MEB because they have direct access to 
the Eudravigilance database. 
 
Expedited reporting will occur no later than 15 calendar days after the Sponsor or its 
representative has first knowledge of the adverse reactions. For fatal or life-threatening 
cases, the term will be maximally 7 calendar days for a preliminary report with another 
8 days for completion of the report.  
8.2.4 
Follow-up of Adverse Events 
Follow-up of AEs will continue until resolution, stabilization, or death. In case of ongoing 
AEs at database closure, the data obtained at database closure will be used in the 
statistical analysis. The follow-up of the AE will be documented in the source documents 
and will be described in the final CSR only if considered relevant by the Investigator. 
8.3
Pregnancy 
If the Investigator becomes aware of a pregnancy occurring in the partner of a male 
subject participating in the study up to 90 days after the last dose of study drug of the 
male subject, the pregnancy should be reported to the Sponsor within 1 working day of 
obtaining written consent from the pregnant partner. The Investigator may make 
arrangements for the partner to be counseled by a specialist, to discuss the risks of 
continuing with the pregnancy and the possible effects on the fetus. Monitoring of the 
partner should continue untile the outcome of the pregnancy is known.  


  
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SERIOUS ADVERSE EVENT CONTACT INFORMATION 
In case of a serious adverse event (see Appendix 8.2), the Principal Investigator will send a 
report within 24 hours of notification to: 
Contract Research Organization 
PRA Health Sciences Drug Safety Center EAPA 
E-mail: 
mhgsafety@prahs.com  
Fax: 
+44 1792 525 720 
Phone: 
+49 621 8782 154 
 
Sponsor’s Medical Expert 
Leoni Matt, MD 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
E-mail: 
matt@lrz.uni-muenchen.de 
Fax: 
+49 89336038 
Phone: 
+49 89 4400 59847 
Mobile:  
+49 176 74717577 
 
 
 


  
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CONTACT INFORMATION 
Sponsor 
LMU Klinikum  
Marchioninistr. 15 
81377 Munich 
Germany 
Sponsor’s Delegated Person 
Michael Hoelscher, MD, FRCP (Lond) 
Director 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
Phone: 
+49 89 4400 59800   
E-mail: 
hoelscher@lrz.uni-muenchen.de 
 
Sponsor’s Project Managers
Julia Dreisbach, VMD 
Clinical Project Manager 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
Phone: 
+49 89 4400 59825    
Mobile: 
+49 179 7036391 
Fax: 
+49 89 336038  
E-mail: 
dreisbach@lrz.uni-muenchen.de 
 
Petra Groß-Demel, PhD 
Clinical Project Manager 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
Phone: 
+49 89 4400 59825      
Mobile: 
+49 152 26309142 
Fax: 
+49 89 336038  
E-mail: 
gross-demel@lrz.uni-muenchen.de 
 


  
CLINICAL STUDY PROTOCOL 
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Sponsor’s Medical Experts  
Norbert Heinrich, MD 
Sponsor’s Medical Expert and Sponsor’s Delegated Person Deputy 
Senior Scientist, Tuberculosis and Emerging Diseases 
Division of Infectious Diseases and Tropical Medicine        
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich                   
Phone: 
+49 89 4400 59805 
Fax: 
+49 89 336038 
E-mail: 
heinrich@lrz.uni-muenchen.de  
 
Leoni Matt, MD  
Sponsor’s Medical Expert 
Division of Infectious Diseases and Tropical Medicine                           
LMU Klinikum 
Leopoldstrasse 5 
80802 Munich 
Germany 
Phone: 
+49 89 4400 59847 
Mobile:  
+49 176 74717577 
Fax: 
+49 89 336038 
E-mail: 
matt@lrz.uni-muenchen.de 
 
Client 
NUVISAN GmbH 
Wegenerstrasse 13 
89231 Neu-Ulm 
Germany 
 
Sponsor’s Scientific Advisor  
Piet Swart, PhD, Dip PharMed, Pharmacologist 
Chief Scientific Officer, Nuvisan Pharma Holding GmbH 
NUVISAN Pharma Holding GmbH 
Wegenerstrasse 13 
89231 Neu-Ulm 
Germany 
Mobile: 
+49 1525 6637 802 
E-mail: 
piet.swart@nuvisan.com 
 
Client’s Clinical Project Manager  
Flavia Koch, PhD, MSc 
Astrid Patzlaff, PhD
Senior Project Manager/Clinical Operations 
NUVISAN GmbH Site Gauting 


  
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Robert-Koch-Allee 29 
82131 Gauting 
Germany 
Phone:
+49 89 89356 9536925
Fax: 
+49 89 89356922 
E-mail: 
flavia.kochastrid.patzlaff@nuvisan.com 
Client’s Nonclinical Project Manager 
Dieter Gallemann, PhD
NUVISAN GmbH Site Grafing  
Am Feld 32 
85567 Grafing  
Germany 
Phone: 
+49 80 9270 0813+49 8092 7008 13 
Home office:+49 89 8946 0419 
Fax: 
+49 80 92700823+49 8092 7008 23 
E-mail: 
dieter.gallemann@nuvisan.com 
 
Promotor 
Leibniz Institute for Natural Product Research and Infection Biology 
Hans Knöll Institut (HKI) 
Adolf-Reichwein-Straße 23 
07745 Jena 
Germany 
 
Promotor’s Contact 
Florian Kloss, PhD 
Head of Transfer Group Anti-infectives 
Phone: 
+49 3641 532 1356                                 
E-mail: 
florian.kloss@hki-jena.de/florian.kloss@leibniz-hki.de                                                      
 
 
Contract Research Organization, Contract Manufacturing Organization, and Clinical Site 
PRA Health Sciences (PRA) – Early Development Services (EDS) 
Van Swietenlaan 6 
9728 NZ Groningen 
The Netherlands 
Phone: 
+31 50 402 2222 
Fax: 
+31 50 402 2223 
 
Principal Investigator 
Jan Jaap van Lier, MD  
Phone: 
+31 50 402 2325  
Fax: 
+31 50 402 2223  
E-mail: 
lierjanjaapvan@prahs.com 
 


  
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Project Manager 
Marloes ter Veer, PhD 
Phone: 
+31 50 402 2292 
E-mail: 
veermarloester@prahs.com 
 
GMP-QA Responsible Person 
Roos Imbos, PhD 
Phone: 
+31 59 230 3430 
Mobile: 
+31 6 227 889 05 
E-mail: 
imbosroos@prahs.com 
Laboratories
Clinical Laboratory 
PRA-EDS 
Van Swietenlaan 6 
9728 NZ Groningen 
The Netherlands 
Phone: 
+31 50 402 2555 
Fax: 
+31 50 402 2223 
Laboratory for Analysis of Plasma and Urine for BTZ-043 and Main Metabolites 
Bioanalytical Department of Nuvisan Grafing
Melanie Tausch 
Am Feld 32 
85567 Grafing 
Germany 
Phone: 
+49 8092 7008 79  
Mobile: 
+49 152 06657818 
Fax: 
+49 8092 7008 23 
E-mail: 
melanie.tausch@nuvisan.com 
 
Laboratory for Analysis of Total Radioactivity in Whole blood, Plasma, Urine, and Feces 
PRA Bioanalytical Laboratory 
Erik Vermeulen 
Amerikaweg 18 
9407 TK Assen 
The Netherlands 
Phone: 
+31 592 303 222 
Fax: 
+31 592 303 223 
E-mail: 
vermeulenerik@prahs.com 
 
Laboratory for Metabolite Profiling and Metabolite Identification in Plasma, Urine, and Feces 
PK Sciences and Biotransformation Department of Nuvisan Pharma Services GmbH 
Matthias Bader, PhD 
Am Feld 32 


  
CLINICAL STUDY PROTOCOL 
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85567 Grafing 
Germany 
Phone: 
+49 8092 7008 48 
Fax: 
+49 8092 7008 23 
E-mail: 
matthias.bader@nuvisan.com 
Medical Screening Centers  
PRA-EDS 
Van Swietenlaan 6 
9728 NZ Groningen 
The Netherlands 
Phone: 
+31 50 850 5798 
Fax: 
+31 50 850 5782 
PRA-EDS 
Yin Yang Building 
Euclideslaan 1 
3584 BL Utrecht 
The Netherlands 
Phone: 
+31 50 851 5830 
Study Monitor 
Oliver Simonides 
Monitoring/LCRA 
NUVISAN GmbH Wegenerstrasse 13  
89231 Neu-Ulm 
Germany 
Phone: 
+49 731 9840 138 
Fax: 
+49 731 9840 571 
E-mail:  
oliver.simonides@nuvisan.com
 


  
CLINICAL STUDY PROTOCOL 
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VERSION 1.02.0 – 18-Mar-01-Jul-2021 
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SUMMARY OF CHANGES 
The following changes have been introduced in this Version 2.0 of the protocol (dated 01 Jul 2021) 
and are given as a combination of double underlined and italic text. Deleted text is given as double 
strikethrough. 
The rationale for these changes is the change in blood sample volumes and timepoints, and changes 
in the reporting of metabolite profiling/identification and PK. In addition, contact information of the 
Client’s Clinical Project Manager has been altered.  
The changes can be found in:
Contact Information
Synopsis 
Table 1 - Schedule of Assessments 
 
Section 3.5.1.3 Total Blood Volume: Table 3 - Number and Volume of Blood Samples and 
Total Blood Volume Collected per Subject 
 
Section 3.5.3.3 Exploratory Variables 
 
Section 3.5.4 Drug Concentration Measurements 
 
Section 3.6.2.1 Pharmacokinetic Evaluation 
 
Section 3.6.2.3 Evaluation of Exploratory Variables  
 
 


  
CLINICAL STUDY PROTOCOL 
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SYNOPSIS 
Study Title 
A SINGLE-CENTER, OPEN-LABEL STUDY TO INVESTIGATE MASS BALANCE, EXCRETION 
PATHWAYS AND METABOLITES AFTER A SINGLE ORAL DOSE OF 500 MG, 3.7 MBq, [14C]BTZ-043 IN 
HEALTHY MALE VOLUNTEERS 
 
Study Codes 
Sponsor code 
: LMU-IMPH-BTZ-043-03  
PRA code 
: NVS543EC-185431 
EudraCT number 
: 2021-000449-42 
 
Sponsor 
LMU Klinikum, Marchioninistr. 15, 81377 Munich, Germany 
Sponsor’s Delegated Person 
 
: Michael Hoelscher, MD, FRCP (Lond) 
Sponsor’s Clinical Project Managers 
: Julia Dreisbach, VMD and Petra Gross-Demel, PhD 
 
Promotor 
Leibniz Institute for Natural Product Research and Infection Biology, Hans Knöll Institut (HKI),  
Adolf-Reichwein-Strasse 23, 07745 Jena, Germany 
Promotor’s contact 
  
 
: Florian Kloss, PhD  
 
Client 
Nuvisan GmbH, Wegenerstrasse 13, 89231, Neu-Ulm, Germany 
Client’s Clinical Project Manager 
: Flavia Koch, PhD, MSc Astrid Patzlaff, PhD 
 
Contract Research Organization and Clinical Site 
PRA Health Sciences – Early Development Services, Van Swietenlaan 6, 9728 NZ Groningen, 
The Netherlands 
 
Principal Investigator 
Jan Jaap van Lier, MD 
 
Objectives 
Primary 
: To determine the rates and routes of excretion of [14C]BTZ-043-related 
radioactivity, including mass balance of total drug-related radioactivity in urine 
and feces (and vomit, if applicable), following the oral administration of a single 
500 mg dose of [14C]BTZ-043 in healthy male volunteers. 
 
: To determine the pharmacokinetics (PK) of total radioactivity in blood and in 
plasma. 
 
: To characterize the plasma PK of BTZ-043 and main metabolites by liquid 
chromatography-mass spectrometry (LC-MS), if applicable. 
 
: To characterize the urine concentrations of BTZ-043 and main metabolites by 
LC-MS, if applicable. 
Secondary 
: To assess the safety and tolerability of a single 500 mg oral dose of BTZ-043 
administered to healthy volunteers. 
Exploratory 
: To identify BTZ-043’s metabolites in plasma, and to quantify metabolites not 
measured in the primary objectives by radiometry in order to elucidate key 
biotransformation pathways and clearance mechanisms of BTZ-043 in humans. 


  
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: To identify BTZ-043’s metabolites in excreta (urine and feces) and to quantify 
metabolites not measured in the primary objectives by radiometry in order to 
elucidate key biotransformation pathways and clearance mechanisms of BTZ-
043 in humans. 
 
Design and Treatments 
This study is a Phase 1, single-center, open-label study to investigate the absorption, metabolism, and 
excretion of BTZ-043 after a single oral administration of 500 mg BTZ-043 containing 3.7 MBq of 
[14C]BTZ-043 in 4 healthy adult male subjects.
 
A total of 4 evaluable subjects completing all procedures are required. Six (6) subjects will be enrolled in the 
cohort in order to have 4 evaluable subjects. If the cohort of 4 subjects administered with 500 mg BTZ-043 
containing 3.7 MBq of [14C]BTZ-043 leads to less than 4 evaluable subjects, the 2 back-up subjects can be 
dosed subsequently to ensure that 4 subjects have completed all study procedures and are evaluable. If any 
of the 4 subjects vomits within 4 hours after study drug administration (approximately 2 times the median 
Tmax of BTZ-043), the subject will be replaced. The 2 back-up subjects will stay in the clinical research center 
until the morning of Day 2. 
 
The study will consist of a screening period (Day -21 to -2), a baseline period (Day -1), a single dose 
treatment on Day 1 with a minimum of 96 hours (=4 days) post dose in-house observation period (Days -1 
up to afternoon Day 5), and a follow-up visit 30 days (±2 days) after the [14C]BTZ-043 dose. The subjects 
will be admitted to the clinical site on Day -1, at least 17 hours before the administration of [14C]BTZ-043 for 
baseline evaluation. Subjects will be administered a single 500 mg [14C]BTZ-043 dose as drinking 
suspension. Subjects will be confined to the clinical site for at least 96 hours following drug administration 
(ie, afternoon of Day 5). During this time, blood, feces, and urine samples for measurement of [14C]BTZ-043 
and metabolites will be collected. The radioactivity excreted in urine and feces as well as radioactivity in 
blood and plasma should be measured and the data should be communicated daily to Matthias Bader or his 
designee. The subjects will be released from the clinic approximately 96 hours to 168 hours after dose 
administration and upon satisfactory recovery of radioactivity (at least 90%) approved by the Sponsor’s 
scientific advisor after consulation of the Sponsor. This will depend on the radioactivity excretion balance as 
assessed by a normal counting method or real time rapid analysis on site (‘quick counting’ procedure, 
utilizing liquid scintillation counter, or sample combustion apparatus) of radioactivity in plasma, urine, and 
feces accumulated until 96 hours, as feasible. The data has to be available before subjects will be released 
from the clinic.  
 
The in-house period should be prolonged up to the morning of Day 8 if at least 1 of the following applies: 
 
Total radioactivity in plasma is higher than 5% of total radioactivity-Cmax and/or PK parameters for 
determination of the total radioactivity-elimination phase cannot be determined reliably. 
 
The mass balance in urine and feces is not complete by Day 5 (ie, <90%).  
 
The combined urinary and fecal excretion is not 1% of the administered dose per 24 hours for 
2 consecutive days based on 14C-radioactivity “quick counts” or normal counts, as feasible. 
 
In the event of an incomplete mass balance on the morning of Day 8, subjects may be discharged from the 
clinical site upon discretion of the Investigator. Subjects with an incomplete mass balance (<90% of dose 
administered and/or combined urinary and fecal excretion >1% of the administered dose per 24 hours for 
2 consecutive days based on 14C-radioactivity “quick counts” or normal counts, as feasible) on Day 8 will be 
asked to return to the clinical site for one or two 24-hour visits (Day 11-12 and Day 15-16). 
 


  
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In the event of subjects not defecating for 2 days, measures will be taken to ensure defecation (eg, additional 
fibers, plum juice, dried prunes, and/or sufficient hydration). 
 
Study Schedule
Screening 
: Between Day -21 and Day -2.  
Treatment period 
: From Day -1 (admission) to approximately 96 hours after study drug 
administration (Day -1 up to afternoon Day 5) with possible extension (Day 6 to 
Day 8) as well as possible 24-hour visits (Day 11-12, Day 15-16).  
Follow-up 
: 30 days (±2 days) after the [14C]BTZ-043 dose. 
 
Subjects 
4 healthy male subjects 
 
Main Criteria for Inclusion 
Age 
: 18 years to 55 years, inclusive, at screening 
Weight 
: 55 to 90 kg, inclusive, at screening 
Body mass index 
: 18.0 to 29.0 kg/m2, inclusive, at screening  
 
Study Drug 
Active Medication 
Active substance 
: BTZ-043 
Activity 
: Inhibition of Mycobacterium tuberculosis DprE1 
In development for 
: Tuberculosis 
Dose 
:  1 x 100 mL (equivalent to 500 mg BTZ-043) 
Strength 
: 5 mg/mL BTZ-043 containing 0.037 MBq/mL of [14C]-BTZ-043 
 Dosage form 
: Oral suspension 
Manufacturer 
: Prepared by pharmacy at PRA 
 
Variables 
Safety 
: Adverse events, clinical laboratory, vital signs, 12-lead electrocardiogram, 
physical examination  
Pharmacokinetics 
: Total radioactivity concentrations in plasma, whole blood, urine, and feces 
 
 Total radioactivity in plasma and whole blood PK parameters estimated using 
noncompartmental analysis, as appropriate : Cmax, tmax, t1/2, AUCinf, AUC0-t, and 
whole blood to plasma ratios  
 
 Concentrations of BTZ-043 and main metabolites in plasma and urine 
 
 Plasma and urine BTZ-043 PK parameters estimated using noncompartmental 
analysis, as appropriate: Cmax, tmax, t1/2, kel, AUCinf, AUC0-t, CL/F, Vz/F, Aeurine, 
Feurine, and CLr
 
 Plasma 
and 
urine 
metabolites PK 
parameters estimated using 
noncompartmental analysis, as appropriate: Cmax, tmax, t1/2, kel, AUCinf, AUC0-t, 
Aeurine, and Feurine 
 
 Urine and feces (and vomit, if available) radioactivity PK parameters estimated 
using noncompartmental analysis, as appropriate: Aeurine, Feurine, Aefeces, Fefeces, 
and, if available, Aevomit, and Fevomit
Exploratory 
: Metabolite profiles, metabolite identification, as feasible 
 
 %AUCtotal radioactivity of BTZ-043 and metabolites 
 
 
 


  
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Statistical Methods 
Safety parameters 
: Descriptive statistics 
PK parameters 
: Descriptive statistics for all relevant PK parameters: n, number of missing 
values to be reported (m), geometric mean, geometric coefficient of variation, 
mean, coefficient of variation, SD, median, minimum, and maximum  
Exploratory parameters : Descriptive statistics 
 
 


  
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Table 1 
Schedule of Assessments  
Visit 
Screening 
Treatment Perioda 
24-hour Visitsa 
Follow-up  
Study Day
Days 
-21 to -2 
Day -1 
 
Day 1
 
Day 2
Day 3
Day 4
Day 5
Day 6m Day 7m
 
Day 8m
 
Day 11m 
 
Day 12m Day 15m Day 16m 
Day 31 
(±2 days)  
Confinementa
 
X 
X 
X 
X 
X 
X 
(X) 
(X) 
(X) 
(X)
(X) 
(X) 
(X) 
 
Ambulatory 
X
 
 
 
 
X 
Admission
 
X 
 
(X)
 
(X) 
 
Dischargeb
 
 
 
X 
(X) 
(X) 
(X) 
 
(X) 
(X) 
 
Informed Consent
X
 
 
Medical History
X
Demographics
X
Physical Examinationc 
X
X
X
Body Weight  
X
X 
 
 
Height and BMI 
Calculation 
X 
 
 
 
 
 
 
 
Serology (HBsAg, anti-
HCV, anti-HIV 1 and 2)
X 
 
 
 
 
 
 
 
 
 
SARS-CoV-2 PCRd
 
X 
 
X 
(X)
 
(X) 
 
Drug and Alcohol Screen 
X
X 
 
 
 
 
Clinical Laboratorye
X
X
X
X
 
 
X
Urinalysis
X
X
X
X
 
 
X
12-lead ECGf
X
X
X
X
X
(X) 
(X) 
(X) 
 
(X) 
(X) 
X
Vital Signsg
X
X
X
X
X
(X) 
(X) 
(X) 
 
(X) 
(X) 
X
Eligibility Check
X
X
X
 
 
Study Drug 
Administration 
 
 
X 
 
 
 
 
 
Plasma Sampling for 
BTZ-043 and Main 
Metabolitesh 
 
 
X 
X 
X 
 
 
 
 
 
 
 
Whole Blood Sampling 
for Radioactivityi
 
 
X 
X 
X 
X 
X 
(X) 
(X) 
(X) 
 
(X) 
 
 
(X) 
 
Plasma Sampling for 
Total Radioactivity and 
Metabolite Profilingi
 
 
X 
X 
X 
X 
X 
(X) 
(X) 
(X) 
 
(X) 
 
 
(X) 
 


  
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Visit
Screening
Treatment Perioda
24-hour Visitsa 
Follow-up
Study Day 
Days 
-21 to -2 
Day -1 
 
Day 1 
 
Day 2 
Day 3 
Day 4 
Day 5 Day 6m Day 7m 
 
Day 8m 
 
Day 11m
 
Day 12m  Day 15m Day 16m
Day 31 
(±2 days)  
Urine Collection and 
Pooling for BTZ-043 and 
Main Metabolites, Total 
Radioactivity, and 
Metabolite Profilingj
 
X 
X 
X 
X 
X 
X 
 (X) 
 
(X) 
 
(X) 
 
 
(X) 
 
 
(X) 
 
(X) 
 
(X) 
 
 
Feces Collection for 
Total Radioactivity and 
Metabolite Profilingk 
X
X
X
X
X
X
X
(X)
(X)
(X)
 
(X)
 
(X)
(X)
(X)
Vomit Collection for Total 
Radioactivity, if availablel 
 
 
X 
 
 
 
 
 
 
 
Previous and 
Concomitant Medication 
X
X 
X 
X 
X 
X 
X 
(X) 
(X) 
(X) 
 
(X) 
 
(X) 
(X) 
 
(X) 
X 
AE Monitoring
X
X
X
X
X
X
(X) 
(X) 
(X) 
(X)
(X) 
(X) 
(X) 
X
AE=adverse event; BMI=body mass index; ECG=electrocardiogram; HBsAg=hepatitis B surface antigen; HCV=hepatitis C virus;PCR=polymerase chain reaction; PK=pharmacokinetic(s) 
a 
 
b
c 
 
d 
 
e 
f 
g 
 
h 
i 
 
 
j 
 
 
k 
 
 
l 
m
Subjects will be in the clinic from Day -1 (predose) until Day 5 with possible extension (Day 6 to Day 8) as well as possible 24-hour visits (Day 11-12, Day 15-16). The 2 back-up subjects will stay 
in the clinic until the morning of Day 2. 
Subjects will be released from the clinic 96 hours after dose administration and upon satisfactory recovery of radioactivity.
Complete physical examinations will be conducted at screening, Day -1, and follow-up/early termination. Symptom driven physical examinations may be conducted at any time, per the Investigator’s 
discretion.  
Sampling of nasal and throat mucosal cells. If deemed necessary, additional tests may be conducted during the study. The nonclinical project manager will be informed immediately upon availability  
of Day 11 results, as this will qualify the feces samples for metabolite profiling with regard to potential COVID-19 infectivity. 
Clinical laboratory tests (including clinical chemistry, hematology, and urinalysis): at screening; on Day -1 (admission), Day 2, and Day 4; and at follow-up.
12-lead ECG: at screening; Day -1, and 1 hour, 4 hours, and 24 hours postdose; at discharge; and at follow-up.  
Vital signs (supine systolic and diastolic blood pressure, pulse, body temperature, and respiratory rate): at screening; at admission; at predose and at 1 hour, 4 hours, and 24 hours postdose; at 
discharge; and at follow-up. 
Plasma sampling for PK of BTZ-043 and main metabolites in plasma: at predose and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, and 48 hours postdose. 
Total radioactivity in whole blood and plasma, and metabolite profiling in plasma: at predose and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, 48, 72, and 96 hours postdose (in case total radioactivity 
recovered is still <90%: in addition 120, 144, and 168 hours postdose and at further 24-hour visits [264 and 360 hours postdose] until study completion). Metabolite profiling in plasma: at predose 
and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, 48, 72, and 96 hours postdose (in case total radioactivity recovered is still <90%: in addition 120, 144, and 168 hours postdose. 
Urine collection and pooling for BTZ-043 and main metabolites, total radioactivity, and metabolite profiling: at predose (within 12 hours prior to dosing) and over 0-24, 24-48, 48-72, and 72-96 hours 
postdose collection intervals (in case total radioactivity recovered is still <90%, in addition 96-120, 120-144, and 144-168 hours postdose and at further 24-hour intervals [Day 11-12 and Day 15-16] 
until study completion). 
Feces collection for total radioactivity and metabolite profiling; predose feces collection to be collected at any time within 48 hours prior to study drug administration (at home or in the clinic). After 
administration of BTZ-043, feces will be collected in 24 hour collection intervals up to 96 hours postdose (in case total radioactivity recovered is still <90%: in addition 120, 144, and 168 hours 
postdose and at further 24-hour intervals [Day 11-12 and Day 15-16] until study completion). 
If produced, vomit will be collected up to 8 hours after the administration of the study drug for total radioactivity determination.  
Day 6-8 procedures will only be conducted in case that the recovery of radioactivity was not satisfactory until Day 5. 24-hour visits procedures will only be conducted in case that the recovery of 
radioactivity was not satisfactory until Day 8. For the 24-hour visits (if applicable): feces collection to be collected at home within 48 hours prior to admission. 


  
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TABLE OF CONTENTS 
TITLE PAGE ..............................................................................................................................1 
SPONSOR AUTHORIZATION OF CLINICAL STUDY PROTOCOL ................................... 2 
AUTHORIZATION OF CLINICAL STUDY PROTOCOL BY CONTRACT RESEARCH 
ORGANIZATION ............................................................................................................... 3 
SERIOUS ADVERSE EVENT CONTACT INFORMATION................................................... 4 
CONTACT INFORMATION ..................................................................................................... 5 
SUMMARY OF CHANGES.................................................................................................... 10 
SYNOPSIS .............................................................................................................................. 11 
TABLE OF CONTENTS......................................................................................................... 17 
TABLE OF TABLES .............................................................................................................. 20 
LIST OF ABBREVIATIONS................................................................................................... 21
1. INTRODUCTION.............................................................................................................. 22 
1.1 
Background.............................................................................................................. 22 
1.1.1 Nonclinical Summary ...................................................................................... 22 
1.1.2 Clinical Summary ............................................................................................ 24 
1.2 
Risk-benefit Assessment......................................................................................... 26 
1.3 
Study Rationale........................................................................................................ 27 
2. OBJECTIVES................................................................................................................... 28 
2.1 
Primary ..................................................................................................................... 28 
2.2 
Secondary ................................................................................................................ 28 
2.3 
Exploratory............................................................................................................... 28 
3. INVESTIGATIONAL PLAN ............................................................................................. 29 
3.1 
Overall Study Design and Plan ................................................................................ 29 
3.1.1 Type of Study ................................................................................................. 29 
3.1.2 Screening Period ............................................................................................ 29 
3.1.3 Treatment Period ............................................................................................ 29 
3.1.4 Follow-up........................................................................................................ 30 


  
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3.2 
Discussion of Study Design .................................................................................... 30 
3.2.1 COVID-19 Risk Mitigation ............................................................................... 30 
3.3 
Selection of Study Population ................................................................................. 31 
3.3.1 Inclusion Criteria ............................................................................................. 31 
3.3.2 Exclusion Criteria ............................................................................................ 32 
3.3.3 Removal of Subjects from Assessment............................................................ 33 
3.4 
Treatments ............................................................................................................... 34 
3.4.1 Treatments Administered ................................................................................ 34 
3.4.2 Identity of Investigational Product .................................................................... 34 
3.4.3 Method of Assigning Subjects to Treatment Groups......................................... 34 
3.4.4 Selection of Doses and Meals in the Study ...................................................... 35 
3.4.5 Radiation Safety of Clinical and Laboratory Staff ............................................. 35 
3.4.6 Timing of Doses in the Study........................................................................... 36 
3.4.7 Meals During the Study ................................................................................... 36 
3.4.8 Blinding .......................................................................................................... 37 
3.4.9 Concomitant Medication and Other Restrictions During the Study .................... 37 
3.4.10 Treatment Compliance .................................................................................... 38 
3.5 
Pharmacokinetic, Safety, and Exploratory Measurements and Variables ............. 38 
3.5.1 Pharmacokinetic and Safety Measurements Assessed and Schedule of 
Assessments .................................................................................................. 38 
3.5.1.1 
Pharmacokinetic Measurements ...................................................... 38 
3.5.1.1.1 
Blood Sampling ............................................................. 38 
3.5.1.1.2 
Urine Collection ............................................................. 39 
3.5.1.1.3 
Feces Collection............................................................ 39 
3.5.1.2 
Safety and Tolerability Measurements ............................................. 39 
3.5.1.2.1 
Adverse Events ............................................................. 40 
3.5.1.2.2 
Clinical Laboratory......................................................... 40 
3.5.1.2.3 
Vital Signs ..................................................................... 41 
3.5.1.2.4 
Electrocardiogram ......................................................... 41 
3.5.1.2.5 
Physical Examination .................................................... 41 
3.5.1.3 
Total of Blood Volume ..................................................................... 41 
3.5.2 Appropriateness of Measurements .................................................................. 42 
3.5.2.1 
Timing of Assessments.................................................................... 42 
3.5.3 Pharmacokinetic, Safety, and Exploratory Variables ........................................ 42 
3.5.3.1 
Pharmacokinetic Variables .............................................................. 42 
3.5.3.2 
Safety Variables .............................................................................. 44 
3.5.3.3 
Exploratory Variables ...................................................................... 44 
3.5.4 Drug Concentration Measurements ................................................................. 44 
3.5.5 Retention of Blood, Plasma, Urine, and Feces Samples................................... 44 
3.6 
Statistical Procedures and Determination of Sample Size ..................................... 45 
3.6.1 Analysis Sets .................................................................................................. 45 
3.6.1.1 
Safety Set ....................................................................................... 45 
3.6.1.2 
Pharmacokinetic Set........................................................................ 45 
3.6.2 Statistical and Analytical Plan for Pharmacokinetic, Safety, and Exploratory 
Evaluation....................................................................................................... 45 
3.6.2.1 
Pharmacokinetic Evaluation............................................................. 45 


  
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3.6.2.2 
Evaluation of Safety and Tolerability ................................................ 45 
3.6.2.2.1 
Adverse Events ............................................................. 45 
3.6.2.2.2 
Clinical Laboratory......................................................... 46 
3.6.2.2.3 
Vital Signs and Electrocardiograms................................ 46 
3.6.2.2.4 
Physical Examination .................................................... 46 
3.6.2.3 
Evaluation of Exploratory Variables.................................................. 46 
3.6.3 Determination of Sample Size ......................................................................... 46 
3.7 
Data Quality Assurance ........................................................................................... 46 
4.
ETHICS............................................................................................................................. 48
4.1 
Independent Ethics Committee ............................................................................... 48 
4.2 
Ethical Conduct of the Study................................................................................... 48 
4.3 
Subject Information and Consent............................................................................ 49 
4.4
Privacy ..................................................................................................................... 49
5. STUDY ADMINISTRATIVE STRUCTURE..................................................................... 50 
5.1 
Distribution of Activities .......................................................................................... 50 
5.1.1 Preparation of Study Drug ............................................................................... 50 
5.1.2 Laboratory Assessments ................................................................................. 50 
5.1.3 Electronic Case Report Form Design............................................................... 50 
5.1.4 Data Management .......................................................................................... 50 
5.1.5 Statistics ......................................................................................................... 50 
5.1.6 Clinical Study Report Writing ........................................................................... 50 
5.2 
Documentation......................................................................................................... 51 
5.2.1 Archiving ........................................................................................................ 51 
5.2.2 Recording of Data in Source Documents and Electronic Case Report Forms ... 51 
6. CONFIDENTIALITY AND PUBLICATION POLICY ...................................................... 52 
7. REFERENCES................................................................................................................. 53 
8. APPENDICES .................................................................................................................. 54 
8.1 
Drug Accountability ................................................................................................. 54 
8.2 
Adverse Events and Serious Adverse Events Evaluation and Reporting .............. 54 
8.2.1 Adverse Events............................................................................................... 54
8.2.2 Serious Adverse Events .................................................................................. 55 
8.2.3 Suspected Unexpected Serious Adverse Reactions......................................... 56 
8.2.4 Follow-up of Adverse Events ........................................................................... 57 
 


  
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TABLE OF TABLES  
Table 1 
Schedule of Assessments .......................................................................................... 15 
Table 2 
Geometric mean and range of AUC0-24h for BTZ-043 in Phase 1a and Phase 1b studies
.................................................................................................................................. 26 
Table 3 
Number and Volume of Blood Samples and Total Blood Volume Collected per Subject42 
Table 4 
Pharmacokinetic Parameters ..................................................................................... 43 
 


  
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LIST OF ABBREVIATIONS 
AME
absorption, metabolism, and excretion
AE
adverse event 
ALT
alanine aminotransferase
AST
aspartate aminotransferase
AUC
area under the concentration-time curve
BMI
body mass index
CA
Competent Authority
CCMO
Centrale Commissie Mensgebonden Onderzoek (Central Committee on 
Research Involving Human Subjects) 
CHMP
Committee for Medicinal Products for Human Use 
GMP
Good Manufacturing Practice
CSP
clinical study protocol
CSR
clinical study report
CTCAE
Common Terminology Criteria for Adverse Events
CTD
Clinical Trial Directive
DprE1
decaprenylphosphoryl- - -ribose-2 -epimerase 
ECG
electrocardiogram
eCRF
electronic case report form
EDS
Early Development Services
EMA
European Medicines Agency
FDA
Food and Drug Administration
GCP
Good Clinical Practice
GDPR
General Data Protection Regulation
HBsAg
hepatitis B surface antigen
HCV
hepatitis C virus
HED
human equivalent dose
HKI
Hans Knöll Institut
ICF
informed consent form
ICH
International Council for Harmonisation
IEC
Independent Ethics Committee
MEB 
Medicine Evaluation Board
MedDRA
Medical Dictionary for Regulatory Activities
NOAEL
no observed adverse effect level
PCR
polymerase chain reaction
PK
pharmacokinetic(s)
PRA
PRA Health Sciences
SAE
serious adverse event
SAP
statistical analysis plan
SOP
standard operating procedure
SUSAR
suspected unexpected serious adverse reaction
TEAE
treatment-emergent adverse event
WMA
World Medical Association
WMO
Wet medisch-wetenschappelijk onderzoek met mensen (medical research 
involving human subjects act) 
Note: Definitions of pharmacokinetic (PK) parameters are provided in Section 3.5.3. 
 


  
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1. 
INTRODUCTION 
1.1 
Background 
BTZ-043 is a promising new antibiotic for the treatment of tuberculosis. Its mechanism 
of action is based on a reduction of BTZ-043 to a transient nitroso moiety, which then 
reacts with an essential cysteine residue in the enzyme decaprenylphosphoryl- - -
ribose-2 -epimerase (DprE1), forming a semimercaptal as covalent adduct. DprE1 itself 
is required for the biosynthesis of decaprenylphosphoryl- - -arabinose, an essential 
component for the cell wall assembly of mycobacteria. Formation of the covalent adduct 
between BTZ-043 and DprE1 results in inhibition of cell wall biosynthesis and loss of 
viability of M. tuberculosis. Due to this mechanism BTZ-043 is highly selective for 
mycobacteria species (and some other members of Actinomycetales), but does not 
affect the gut flora.1 
 
In 2018 a Phase 1a study entitled “A Randomized, Double blind, Placebo-controlled, 
Single 
Ascending 
Dose 
Study 
to 
Evaluate 
Safety, 
Tolerability, 
and 
Pharmacokinetics (PK) of Single Doses of BTZ-043 in Healthy Adult Volunteers” 
(LMU-IMPH-BTZ-043-01) was conducted in humans and currently a Phase 1a/2b study 
entitled “A Prospective Phase 1b/2a, Active-controlled, Randomized, Open-label Study 
to Evaluate the Safety, Tolerability, Extended Early Bactericidal Activity and PK of 
Multiple Oral Doses of BTZ-043 Tablets in Subjects With Newly Diagnosed, 
Uncomplicated, 
Smear-positive, 
Drug-susceptible 
Pulmonary 
Tuberculosis” 
(PanACEA-BTZ-043-02) is ongoing. This human absorption, metabolism, and excretion 
study is conducted during Phase 2 proof of concept studies and before a Phase 3 study 
is initiated. 
1.1.1 
Nonclinical Summary 
Safety 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 


  
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Pharmacokinetics 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


  
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1.1.2 
Clinical Summary 
Safety
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
. 
 
  
 
 


  
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1.2 
Risk-benefit Assessment  
There is no expected clinical benefit for the healthy subjects who will participate in this 
study. The information obtained in this study can be used for the further clinical 
development of BTZ-043. As depicted in Table 2, BTZ-043 has been well tolerated in 
much higher exposures as proposed in this study.  
 
Overall, on the basis of the available nonclinical and clinical data, the risk-benefit profile 
of BTZ-043 is judged acceptable for the proposed clinical study. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


  
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The risk-benefit assessment for the subjects receiving BTZ-043 remains unchanged in 
relation to the corona virus disease-19 (COVID-19) pandemic as available clinical and 
nonclinical results do not suggest that administration of BTZ-043 will lead to suppression 
or modulation of the immune system. In addition, the mode of action does not appear to 
have a substantial effect on the respiratory or cardiovascular system critically affected 
by a SARS-CoV-2 infection. As the subjects to be included in this study are in general 
young to middle aged without major comorbidities, the study population is not considered 
to be a high-risk population for serious COVID-19 disease. Only persons with a negative 
SARS-CoV-2 test at admission to the clinical research center will be allowed to 
participate in the study. In addition, all appropriate measures to prevent SARS-CoV-2 
infection during the study will be taken as detailed in Section 3.2.1. 
1.3 
Study Rationale 
This study will determine the disposition and metabolism of BTZ-043 in humans after 
oral administration. The use of radiolabeled molecules (usually [14C]) is a common 
method to ascertain information on the elimination routes and metabolic fate of a 
compound at an early stage of development. This evaluation will provide an estimate of 
PK parameters, the routes and rates of elimination of radioactivity, and identification of 
metabolites and metabolic pathways. These data will be compared with similar data 
obtained from nonclinical studies, both from PK and metabolic points of view. 
The rationale for the proposed dose can be found in Section 3.4.4. 
 
 


  
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2. 
OBJECTIVES 
2.1 
Primary  
 
To determine the rates and routes of excretion of [14C]BTZ-043-related radioactivity, 
including mass balance of total drug-related radioactivity in urine and feces (and 
vomit, if applicable), following the oral administration of a single 500 mg dose of 
[14C]BTZ-043 in healthy male volunteers.
 
To determine the PK of total radioactivity in whole blood and in plasma. 
 
To characterize the plasma PK of BTZ-043 and main metabolites by liquid 
chromatography-mass spectrometry (LC-MS), if applicable. 
 
To characterize the urine concentrations of BTZ-043 and main metabolites by 
LC-MS, if applicable. 
2.2 
Secondary 
 
To assess the safety and tolerability of a single 500 mg oral dose of BTZ-043 
administered to healthy male volunteers. 
2.3 
Exploratory 
 
To identify BTZ-043’s metabolites in plasma, and to quantify metabolites not 
measured in the primary objectives by radiometry in order to elucidate key 
biotransformation pathways and clearance mechanisms of BTZ-043 in humans.  
 
To identify BTZ-043’s metabolites in excreta (urine and feces) and to quantify 
metabolites not measured in the primary objectives by radiometry in order to 
elucidate key biotransformation pathways and clearance mechanisms of BTZ-043 in 
humans. 
 
 
 


  
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3. 
INVESTIGATIONAL PLAN 
3.1 
Overall Study Design and Plan 
3.1.1 
Type of Study 
This will be a Phase 1, single-center, open-label, absorption, metabolism, and excretion 
(AME) study in 4 healthy adult male subjects. 
3.1.2 
Screening Period 
Subjects will report to the medical screening facility/clinical site for the eligibility 
screening (see Section 3.3 for inclusion and exclusion criteria) within 3 weeks prior to 
drug administration.  
 
Subjects will sign the study-specific informed consent form (ICF) prior to any 
study-specific screening procedures being performed. The written informed consent will 
be obtained for all subjects, regardless of their eligibility for the study; the signed ICFs 
will be retained and archived at PRA Health Sciences (PRA) and a copy will be provided 
to the subject.  
 
Eligibility screening will consist of the assessments as presented in the schedule of 
assessments (Table 1). 
3.1.3 
Treatment Period 
Subjects will be admitted to the clinical research center on Day -1, which is the day prior 
to Day 1, the day of drug administration.  
 
Subjects will receive a single oral dose of 500 mg BTZ-043 containing 3.7 MBq of 
[14C]BTZ-043 on Day 1. Subjects will be discharged from the clinical research center on 
Day 5 (approximately 96 hours after drug administration) upon satisfactory recovery of 
radioactivity. If the criteria for discharge are not met on Day 5, subjects will be required 
to remain confined for a maximum of 3 additional days (Days 6 to 8) until the criteria are 
met (daily check by quick counts or normal counts, as feasible). The in-house period 
should be prolonged up to the morning of Day 8 if at least 1 of the following applies:  
 
Total radioactivity in plasma is higher than 5% of radioactivity-Cmax and/or 
PK parameters for determination of the total radioactivity-elimination phase 
cannot be determined reliably.  
 
The mass balance in urine and feces is not complete by Day 5 (ie, >90%). 
 
The combined urinary and fecal excretion is not 1% of the administered dose 
per 24 hours for 2 consecutive days based on 14C-radioactivity “quick counts” or 
normal counts, as feasible. 
If the criteria for discharge are still not met on Day 8, subjects may be discharged from 
the clinical site upon discretion of the Investigator. To ensure that adequate 
consideration is given to the protection of the subject’s interests, subjects with an 
incomplete mass balance on Day 8 will be asked to return to the clinical site for one or 
two 24-hour visits (Day 11-12 and Day 15-16). 
 


  
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On the day of study drug administation (Day 1), an FDA-recommended high-fat (or a 
standardized breakfast, if applicable, as outlined in Section 3.4.4 and Section 3.4.7) with 
calculated caloric content will be provided. 
 
Assessments during the treatment period will be performed as presented in the schedule 
of assessments (Table 1). 
3.1.4 
Follow-up 
The follow-up assessments will be performed 30 days (±2 days) after the [14C]BTZ-043 
dose.  
 
Assessments during follow-up will be performed as presented in the schedule of 
assessments (Table 1). 
3.2 
Discussion of Study Design 
The current design is commonly used for AME studies and in accordance with the current 
EMA “Guideline on the investigation of drug interactions”. 2 Since the present study is a 
descriptive study aimed at assessing excretion routes of BTZ-043 and evaluating mass 
balance, a formal statistical sample size calculation was not performed. A sample size 
of 4 subjects is a minimum, accepted number of subjects for AME studies and is 
considered sufficient to achieve the study objectives. Early-termination subjects may be 
replaced if the total number of completers drops below 4, after mutual agreement 
between the Sponsor and PRA.  
3.2.1 
COVID-19 Risk Mitigation 
This study will be conducted in accordance with guidance from the Central Committee 
on Research Involving Human Subjects (CCMO [Centrale Commissie Mensgebonden 
Onderzoek], ie, the Dutch competent authority [CA]) on conducting Phase 1 trials in 
clinical research centers in The Netherlands during the COVID-19 pandemic. 
 
During the entire study, the clinical research center will implement all recommendations 
issued by the Dutch government, including specific guidelines related to clinical research 
executed in clinical research centers with respect to minimizing the risk of disease 
spreading (eg, social distancing, disinfection, hygiene, and wearing of personal 
protection equipment by study staff). Details on specific procedures are described in the 
site specific manual.  
 
In cases where subjects are not able to attend study visits due to an infection with 
SARS-CoV-2, the Investigator will discuss with the Sponsor potential mitigation 
approaches (including, but not limited to, extending the visit window, conducting 
evaluations via video link or phone call, allowing for safety procedures to be conducted 
at a local facility). The rationale (eg, the specific limitation imposed by the SARS-CoV-2 
infection that led to the inability to perform the protocol-specified assessment) and 
outcome of the discussion will be documented in the electronic case report form (eCRF). 
 
In addition, the following containment measures will be taken during the study: 


  
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Polymerase chain reaction (PCR) testing for SARS-CoV-2 will be performed at the 
time points indicated in Table 1.
 
A subject should not be admitted if there was any contact with a person who tested 
positive for SARS-CoV-2 or a COVID-19 patient within the last 2 weeks prior to 
admission to the clinical research center. 
 
If a subject is tested to be SARS-CoV-2 positive on Day -1, the subject will be 
excluded from participation with reference to Exclusion Criterion 13, and referred 
for treatment. 
 
Physical examinations will be limited. 
 
If a subject becomes ill and/or is tested to be SARS-CoV-2 positive after the 
administration of study treatment, study assessments will be stopped (see also 
Section 3.3.3.1). A safety follow-up can be done if deemed necessary by the 
Investigator. The subject will be kept in-house in quarantine with appropriate 
medical intervention until complete elimination of the study compound or will be 
asked to quarantine at home according to guidelines of the Dutch government. 
3.3 
Selection of Study Population 
3.3.1 
Inclusion Criteria 
Note: Restrictions that apply to the period after admission are described in Section 3.4.9, 
except when they concern a statement of willingness. 
 
The following inclusion criteria must be met for a subject to be eligible for inclusion in the 
study: 
1. 
Sex 
: male 
2. 
Age 
: 18 years to 55 years, inclusive, at screening. 
3. 
Body mass index (BMI) : 18.0 to 29.0 kg/m2, inclusive, at screening.
4. 
Weight 
: 55 to 90 kg, inclusive, at screening. 
5. 
Status 
: healthy subjects. 
6. 
Male subjects, if not surgically sterilized, must agree to use adequate contraception 
and not donate sperm from admission to the clinical research center until 90 days 
after the follow-up visit. Adequate contraception for the male subject (and his 
female partner, if she is of childbearing potential) is defined as using hormonal 
contraceptives or an intrauterine device combined with at least 1 of the following 
forms of contraception: a diaphragm, a cervical cap, or a condom. Total abstinence, 
in accordance with the lifestyle of the subject, is also acceptable.  
7. 
All prescribed medication must have been stopped at least 30 days prior to 
admission to the clinical research center.  
8. 
All over-the-counter medications, vitamin preparations (especially vitamin C), other 
food supplements, and herbal medications (eg, St. John’s wort) must have been 
stopped at least 14 days prior to admission to the clinical research center. An 
exception is made for paracetamol, which is allowed up to 48 hours prior to study 
drug administration. 
9. 
No vaccination within 14 days prior to study drug administration. 


  
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10. Ability and willingness to abstain from alcohol from 48 hours (2 days) prior to 
screening and admission to the clinical research center. 
11. Ability and willingness to abstain from methylxanthine-containing beverages or food 
(coffee, tea, cola, chocolate, and energy drinks), grapefruit (juice), corn (whole corn 
kernels and popcorn), cruciferous vegetables, and bitter oranges from 48 hours 
(2 days) prior to admission to the clinical research center.  
12. Good physical and mental health on the basis of medical history, physical 
examination, clinical laboratory, ECG, and vital signs, as judged by the 
Investigator.  
13. Willing and able to sign the ICF. 
3.3.2 
Exclusion Criteria 
Note: Restrictions that apply to the period after admission are described in Section 3.4.9, 
except when they concern a statement of willingness. 
 
A subject who meets any of the following exclusion criteria will not be eligible for inclusion 
in the study: 
1. 
Participation in another study with a radiation burden of >0.1 mSv and 1 mSv in 
the period of 1 year prior to screening; a radiation burden of >1.1 mSv and 2 mSv 
in the period of 2 years prior to screening; a radiation burden of >2.1 mSv and 
3 mSv in the period of 3 years prior to screening, etc. 
2. 
Exposure to radiation for diagnostic reasons (except dental X-rays and plain X-rays 
of thorax and bony skeleton [excluding spinal column]), or during work within 1 year 
prior to drug administration. 
3. 
Irregular defecation pattern (less than once per day on average). 
4. 
Employee of PRA, Nuvisan, or the Sponsor. 
5. 
History of relevant drug and/or food allergies. 
6. 
Using tobacco products within 60 days prior to drug administration.  
7. 
History of alcohol abuse or drug addiction (including soft drugs like cannabis 
products). 
8. 
Positive drug and alcohol screen (opiates, methadone, cocaine, amphetamines 
[including 
ecstasy], 
cannabinoids, 
barbiturates, 
benzodiazepines, 
gamma-hydroxybutyric acid, tricyclic antidepressants, and alcohol) at screening or 
admission to the clinical research center. 
9. 
Average intake of more than 24 grams of alcohol per day. 
10. Positive screen for hepatitis B surface antigen (HBsAg), hepatitis C virus (HCV) 
antibodies, or HIV 1 and 2 antibodies. 
11. Participation in a drug study within 30 days prior to drug administration in the 
current study. Participation in more than 4 drug studies in the 12 months prior to 
drug administration in the current study. 
12. Donation or loss of more than 450 mL of blood within 60 days prior to drug 
administration. Donation or loss of more than 1.5 liters of blood in the 10 months 
prior to drug administration in the current study. 
13. Significant and/or acute illness within 5 days prior to drug administration that may 
impact safety assessments, in the opinion of the Investigator. 


  
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14. Unwillingness to consume the Food and Drug Administration (FDA)-recommended 
high-fat breakfast. 
15. Unsuitable veins for infusion or blood sampling. 
16. Positive nasopharyngeal PCR test for SARS-CoV-2 on Day -1. 
 
Please note that subjects should refrain from consumption of any foods containing poppy 
seeds, corn (whole corn kernels and popcorn), cruciferous vegetables, and bitter 
oranges within 48 hours (2 days) prior to screening and admission to the clinical 
research center to avoid false positive drug screen results, difficult feces texture, or 
induction of drug metabolizing enzymes. In addition, they should refrain from strenuous 
exercise within 96 hours (4 days) prior to screening and admission as this could result in 
abnormal clinical laboratory values.  
3.3.3 
Removal of Subjects from Assessment 
Participation in the study is strictly voluntary. A subject has the right to withdraw from the 
study at any time for any reason. 
 
The Investigator has the right to terminate participation of a subject for any of the 
following reasons: difficulties in obtaining blood samples, violation of the protocol, 
severe AEs or SAEs, or for any other reason relating to the subject’s safety or integrity 
of the study data. 
 
If a subject is withdrawn from the study, Nuvisan and LMU Klinikum will be informed 
immediately. If there is a medical reason for withdrawal, the subject will remain under 
the supervision of the Investigator until satisfactory health has returned, or the condition 
has stabilized with no more change being likely. 
 
After signing informed consent, subjects who drop out or withdraw for any reason without 
completing all screening evaluations successfully as well as subjects who drop out or 
withdraw prior to the first dose of study drug will be considered screening failures. 
 
A subject who is withdrawn or voluntarily withdraws from the study for any reason, 
whether related to the study drug or not, after having received the study drug, will be 
considered an early-termination subject. If a subject is withdrawn from the study, the 
early-termination subject will be replaced after mutual agreement between the Sponsor 
and PRA if the number of evaluable subjects drops below 4 (with a maximum of 
2 replacements).  
 
The decision regarding the replacement of subjects will be documented. 
 
PRA will make every effort to ensure that early-termination subjects who have received 
the study drug complete the early-termination assessments/safety follow-up 
assessments. 
 


  
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Early-termination subjects may be replaced if the total number of completers drops 
below 4, after mutual agreement between Nuvisan, LMU Klinikum, and PRA.   
3.3.3.1 
Stopping Rules for Individual Subjects 
In case of a positive SARS-CoV-2 virus PCR test or evidence for COVID-19 during the 
study, the subject will be withdrawn from the study. The subject will be isolated from 
other study participants and referred for treatment. The subject will be followed up in 
quarantine in the clinical research center until complete elimination of the study 
compound or will be asked to quarantine at home according to guidelines of the Dutch 
government. 
3.4 
Treatments 
3.4.1 
Treatments Administered 
Subjects will receive a single oral dose of 500 mg BTZ-043 containing 3.7 MBq of 
[14C]BTZ-043 on Day 1. 
3.4.2 
Identity of Investigational Product 
Active Medication
Active substance 
: BTZ-043 
Activity 
: Inhibition of M. tuberculosis DprE1 
In development for 
: Tuberculosis 
Dose 
:  1 x 100 mL (equivalent to 500 mg BTZ-043) 
Strength 
: 5 mg/mL BTZ-043 containing 0.037 MBq of [14C]BTZ-043 
Dosage form 
: Oral suspension 
Manufacturer 
: Sourced by pharmacy at PRA 
 
The active pharmaceutical ingredient will be provided by the Promotor.  
 
[14C]BTZ-043 is a drug under development and not approved yet. It will be manufactured 
and analysed according to Good Manufacturing Practice (GMP) requirements. A GMP 
certificate of the manufacturer of the [14C]BTZ-043 suspension and an Investigator’s 
Brochure that summarizes preclinical and clinical data with respect to BTZ-043 are in 
place. A certificate of analysis and a certificate of conformity will be provided by PRA 
prior to clinical part. 
 
For details concerning drug storage and drug accountability see Appendix 8.1. 
3.4.3 
Method of Assigning Subjects to Treatment Groups 
After obtaining informed consent, subjects will receive a screening number and will be 
screened according to the inclusion and exclusion criteria. Subjects who have met all 
eligibility criteria will receive a subject number upon inclusion in the study 
(Subject Numbers 001-004). They will receive the subject number just prior to dosing. 
The subject number will ensure identification throughout the study after study drug 
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replaced, increased by 100 (eg, 101 replacement number for Subject Number 001), and 
will be administered the same treatment. 
 
After signing informed consent, subjects who drop out or withdraw for any reason without 
completing all screening evaluations successfully as well as subjects who drop out or 
withdraw prior to the first dose of study drug will be considered screening failures. Such 
subjects, and also subjects who are eligible for inclusion in the study but do not receive 
the study drug, will not receive a subject number, and only applicable data will be entered 
in the eCRFs.  
3.4.4 
Selection of Doses and Meals in the Study 
The targeted dose of 500 mg is considered a clinically relevant dose for the human AME 
study (Table 2) since this exposure achieves a maximal bactericidal effect and has 
shown to be tolerable in the 14-day Phase 1b study. It is proposed that the active 
pharmaceutical ingredient is taken after a high-fat breakfast (see Section 3.4.7), as 
absorption is then much improved. As shown in Table 2, the expected exposure is well 
below the maximum and well tolerated exposure achieved in the Phase 1b study. In the 
unexpected case that the currently conducted Phase 2a study with 250 mg, 500 mg, and 
1000 mg BTZ-043 administered after a high-fat breakfast shows any dose-related safety 
alert, the study team and the Sponsor will take measures to reduce the exposure. 
Reduction of exposure will be achieved by adapting the food intake (up to fasting) before 
the administration of [14C]BTZ-043. This protocol describes the drug administration of 
500 mg [14C]BTZ-043 after a high-fat breakfast as default. However, the study team and 
the Sponsor will decide whether a reduction in food intake is necessary to reduce the 
exposure to [14C]BTZ-043.
 
In case it is indeed decided to administer 500 mg [14C]BTZ-043 after a 
standardized breakfast or in the fasted state rather than after a high-fat breakfast 
as planned, this decision and the associated rationale will be laid down in a Note to File, 
to be signed by the Sponsor and the PI, and to be submitted to the IEC for information 
only. 
 
The dose of 14C (approximately 3.7 MBq) to be administered was chosen to ensure 
successful profiling and identification of metabolites. 
 
In the present study, the estimated effective radiation burden after a single oral 
radioactivity dose of 3.7 MBq is approximately 0.16 mSv. 3 For biomedical investigations 
in small groups of human volunteers, an effective dose of 0.1 to 1.0 mSv is considered 
acceptable. 
3.4.5 
Radiation Safety of Clinical and Laboratory Staff 
It is expected that the clinical and laboratory staff handling the radioactive material and 
samples will not be exposed to any relevant radiation and associated health risk. The 
[14C] isotope emits low energy beta particles, which are not able to penetrate the 
dose containers or sample 
container walls. 
Therefore, there will 
be no 
“external” exposure from sample handling. The samples from the participants after 


  
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radiolabeled dose administration will contain low levels of radioactivity. The radioactive 
dose of 3.7 MBq [14C] per subject is below the legal handling limit, above which a special 
laboratory and special procedures for handling radioactivity would be required. 
Therefore, 
the 
staff 
can 
handle 
the 
study 
samples 
like 
any 
other 
non-radioactive biological material. Common practice of safe laboratory technique and 
hygiene will be exercised in handling all biological materials from this study.  
3.4.6 
Timing of Doses in the Study 
On Day 1, the study drug will be administered with the subject in the upright position. 
The study drug will be administered to subjects between 08:00 and 11:00 in the morning. 
After an overnight fast of at least 10 hours, subjects will receive an FDA-recommended 
high-fat breakfast (or a standardized breakfast, if applicable as outlined in Section 3.4.4) 
that will have to be finished within 20 minutes. The entire breakfast must be consumed 
by the subjects. The actual consumed food will be documented.  
 
Dosing will occur at 30 minutes after the start of the high-fat breakfast (or the 
standardized breakfast, if applicable, or dosing will occur in a fasted state, if applicable 
as outlined in Section 3.4.4).  
 
The study drug will be given as an oral suspension of 100 mL, which will be provided in 
an amber glass bottle from which the subjects can drink. Thereafter the bottle will be 
rinsed twice with 70 mL of water, and the subjects will be required to drink the bottle 
contents after rinsing. Thereafter, the subjects are required to drink additional 
non-carbonated water until they have ingested a total volume of 380 mL. 
 
Following drug administration, subjects will fast for a period of 4 hours until lunch. During 
fasting, no fluids are allowed except water; however, water is not allowed from 1 hour 
predose until 1 hour postdose (apart from the water taken with the dose).  
 
Subjects will not lie down for 4 hours after drug administration, except when required for 
assessments that need to be performed. 
3.4.7 
Meals During the Study 
A fasting period of at least 4 hours is required before obtaining clinical laboratory 
blood samples at the time points indicated in the schedule of assessments (Table 1). 
 
An FDA-recommended high-fat (or a standardized breakfast, if applicable, as outlined in 
Section 3.4.4) with calculated caloric content will be provided on Day 1.  
 
The FDA-recommended high-fat breakfast of 918 kcal consists of: 
 
2 fried eggs (in 15 g butter/margarine) (approximately 100 g) 
 
1 portion of bacon (40 g) (or brie 60+ for vegetarians) 
 
1 portion of fried potatoes (115 g) 
 
2 slices of (toasted) (wheat) bread with 15 g margarine 
 
1 glass of whole milk (240 mL) 


  
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The total of 918 kcal (vegetarian version 915 kcal) can be broken down as follows: 
 
39 g protein 
 
= 156 kcal 
 
59 g fat 
 
= 527 kcal 
 
59 g carbohydrates  
= 235 kcal 
 
The standardized breakfast of 572-755 kcal (depending on choices made) consists of: 
 
3 slices of whole-wheat bread  
 
1 Dutch rusk  
 
20 g halvarine/margarine  
 
1 portion of cheese/old cheese/cheese spread  
 
1 portion of ham/chicken breast/smoked meat/spreadable liver sausage/baloney/filet 
American/salami (or cheese, cheese spread, or peanut butter for vegetarians) 
 
2 portions of jam or other sweet spread  
 
2 mugs of decaffeinated coffee/tea (cream and sugar optional, max 2 of both) 
 
The total of 572-755 kcal (vegetarian version 589-805 kcal) can be broken down as 
follows: 
 
18-19 g protein 
 
= 71-75 kcal 
 
26-32 g fat 
 
= 233-292  kcal 
 
67-97 g carbohydrates 
= 268-388 kcal 
 
Other meals and snacks (such as decaffeinated coffee, herbal tea, fruit, and biscuits) 
will be provided according to PRA standard operating procedures (SOPs), taking into 
account the restrictions as described in Section 3.4.9 and what has been described in 
Section 3.4.6.   
3.4.8 
Blinding 
This is an open-label study. 
3.4.9 
Concomitant Medication and Other Restrictions During the Study 
Note: Restrictions that apply to the period before admission are described in 
Section 3.3.1 and Section 3.3.2. 
 
The use of all prescribed medication is not allowed from admission to the clinical research 
center until follow-up. The use of all over-the-counter medications, vaccines, 
vitamin preparations (especially vitamin C), other food supplements, and herbal 
medications (eg, St. John’s wort) is not allowed from admission to the clinical research 
center until follow-up. An exception is made for paracetamol: from admission onwards, 
the Investigator may permit a limited amount of paracetamol for the treatment of 
headache or any other pain. Other medication to treat AEs may only be prescribed if 
deemed necessary by the Investigator. If medication is used, the name of the drug, the 
dose, and dosage regimen will be recorded in the eCRF.  
 


  
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The use of methylxanthine-containing beverages or food (coffee, tea, cola, chocolate, and 
energy drinks), grapefruit (juice), kiwi, and tobacco products is not allowed during the stay 
in the clinical research center. 
 
The use of alcohol is not allowed during the stay in the clinical research center and within 
48 hours (2 days) prior to the 24-hour return visits (Days 11-12 and 15-16, as needed) and 
prior to follow-up. 
 
Strenuous exercise is not allowed within 96 hours (4 days) prior to admission and 
follow-up. Strenuous exercise is also not allowed during the stay in the clinical research 
center.  
 
Subjects should not consume any foods containing poppy seeds, corn (whole corn 
kernels and popcorn), cruciferous vegetables, and bitter oranges within 48 hours 
(2 days) prior to admission to the clinical research center as this could cause a false 
positive drug screen result. 
 
Male subjects, if not surgically sterilized, are required to use adequate contraception 
(see Inclusion Criterion 6) and not donate sperm from admission to the clinical research 
center until 90 days after the follow-up visit. 
 
Subjects must not donate blood during the study until the follow-up visit (other than the 
blood sampling planned for this study). 
3.4.10 
Treatment Compliance 
Study drug will be administered in the clinical research center. To ensure treatment 
compliance, administration of the study drug will be supervised by the Investigator or an 
authorized designee. Compliance will be further confirmed by bioanalytical assessment 
of BTZ-043 and total radioactivity in plasma and urine samples (see Section 3.5.4). 
 
The exact times of study drug administration and the number of units administered will 
be recorded in the eCRF.  
3.5 
Pharmacokinetic, Safety, and Exploratory Measurements and Variables  
3.5.1 
Pharmacokinetic and Safety Measurements Assessed and Schedule of 
Assessments  
A schedule of assessments is presented in Table 1. 
3.5.1.1 
Pharmacokinetic Measurements  
3.5.1.1.1 Blood Sampling  
At the time points defined in the schedule of assessments (Table 1), blood samples will 
be taken for the analysis of BTZ-043 and metabolites in plasma samples, total 
radioactivity in whole blood and plasma samples, and metabolite profiling and 
identification in plasma samples. 


  
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The blood samples will be taken via an indwelling iv catheter or by direct venipuncture. 
The exact times of blood sampling will be recorded in the eCRF. 
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA and Nuvisan. 
3.5.1.1.2 Urine Collection  
During the intervals defined in the schedule of assessments, urine will be collected for 
the analysis of BTZ-043 and metabolites, for total radioactivity, for quick counts or normal 
counts of total radioactivity (as feasible), and for metabolite profiling and identification. 
The subjects will be instructed to empty their bladders completely before drug 
administration and at the end of each collection interval. A baseline urine sample will be 
collected within 12 hours prior to drug administration. The exact times of urine collection 
and the urine weight of the entire interval will be recorded in the eCRF. 
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA and Nuvisan. 
3.5.1.1.3 Feces Collection  
During the intervals defined in the schedule of assessments, all fecal excretions will be 
collected for the analysis of total radioactivity, for quick or normal counts of total 
radioactivity (as feasible), and for metabolite profiling and identification. A blank fecal 
sample will be collected within 48 hours prior to study drug administration. The exact 
times of feces collection and the fecal weight will be recorded in the eCRF.  
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA and Nuvisan. 
3.5.1.1.4 
Vomit Collection  
If a subject vomits within the time period from drug administration up to 8 hours after the 
administration of the study drug, the vomit should, if possible, be collected. Any collected 
vomit will be included in the total radioactivity measurements and in the determination of 
the mass balance. The exact times of vomit production will be recorded in the eCRF.  
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA and Nuvisan. 
3.5.1.2 
Safety and Tolerability Measurements 
Safety and tolerability assessments will consist of AEs, clinical laboratory, vital signs, 
12-lead ECG, and physical examination. Assessments will be performed in accordance 
with the schedule of assessments. 


  
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3.5.1.2.1 Adverse Events 
AEs will be recorded from signing the ICF until completion of the follow-up visit. Any 
clinically significant observations, as determined by the Investigator, in results of clinical 
laboratory, 12-lead ECGs, vital signs, or physical examinations will be recorded as AEs. 
 
A TEAE is defined as any event not present prior to administration of the study drug or 
any event already present that worsens in either severity or frequency following 
exposure to the study drug.  
 
An AE that occurs prior to administration of the study drug will be considered a 
pretreatment AE.  
 
At several time points before and after drug administration, subjects will be asked 
nonleading questions to determine the occurrence of AEs. Subjects will be asked in 
general terms about any AEs at regular intervals during the study. In addition, all AEs 
reported spontaneously during the course of the study will be recorded. All answers will 
be interpreted by the Investigator using the Medical Dictionary for Regulatory Activities 
(MedDRA; most recent version) for AEs and will be recorded in the eCRF as reported 
terms.  
 
The severity of the AEs will be rated as mild, moderate, or severe; the relationship 
between the AEs and the study drug will be indicated as related or not related. Details 
on rating the severity of AEs and relationship to study treatment are given in 
Appendix 8.2. 
 
Pregnancy of female partners of male subjects will be monitored along with follow-up, if 
warranted (see Appendix 8.3). 
3.5.1.2.2 Clinical Laboratory  
Blood and urine samples for clinical laboratory assessments will be collected according 
to PRA SOPs. 
The following parameters will be measured: 
 
Clinical chemistry (serum quantitatively): 
 
total bilirubin, alkaline phosphatase, gamma glutamyl transferase, AST, ALT, lactate 
dehydrogenase, creatinine, urea, total protein, glucose, inorganic phosphate, 
sodium, potassium, calcium, and chloride.  
 
Hematology (blood quantitatively): 
 
leukocytes, erythrocytes, hemoglobin, hematocrit, thrombocytes, absolute partial 
automated differentiation (lymphocytes, monocytes, eosinophils, basophils, and 
neutrophils), mean corpuscular volume, mean corpuscular hemoglobin, and mean 
corpuscular hemoglobin concentration. 
 
Urinalysis (urine qualitatively): 
 
hemoglobin, urobilinogen, ketones, glucose, protein. 
 
Serology: 
 
HBsAg, HCV antibodies, and HIV 1 and 2 antibodies. 


  
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Drug and alcohol screen: 
 
opiates, methadone, cocaine, gamma hydroxybutyric acid, amphetamines (including 
ecstasy), cannabinoids, barbiturates, benzodiazepines, tricyclic antidepressants, 
nicotine metabolites, and alcohol. 
 
Nasal and throat mucosal cell samples will be collected according to PRA work 
instructions detailed in the site specific manual. The samples will be tested for 
SARS-CoV-2 virus using PCR tests. The nonclinical project manager will be informed 
immediately upon availability of Day 11 results, as this will qualify the feces samples for 
metabolite profiling with regard to potential COVID-19 infectivity. 
 
In case of unexplained or unexpected clinical laboratory test values, the tests will be 
repeated as soon as possible and followed up until the results have returned to the 
normal range and/or an adequate explanation for the abnormality is found. The clinical 
laboratory will clearly mark all laboratory test values that are outside the normal range, 
and the Investigator will indicate which of these deviations are clinically significant. 
Clinically significant laboratory result deviations will be recorded as AEs, and the 
relationship to the treatment will be indicated (see also Appendix 8.2). 
 
Details on sample collection, handling, storage, and shipping will be described in the 
laboratory manual prepared by PRA. 
3.5.1.2.3 Vital Signs 
Systolic and diastolic blood pressure and pulse will be recorded after the subject has 
been resting for at least 5 minutes in the supine position. These assessments will be 
made using an automated device whenever possible. Body temperature and respiratory 
rate will also be measured. 
3.5.1.2.4 Electrocardiogram 
A standard 12-lead ECG will be recorded after the subject has been resting for at least 
5 minutes in the supine position. The ECG will be recorded using an ECG machine 
equipped with computer-based interval measurements. The following ECG parameters 
will be recorded: heart rate, PR-interval, QRS-duration, QT-interval, QTc-interval 
(Fridericia’s), and the interpretation of the ECG profile by the Investigator. 
3.5.1.2.5 Physical Examination 
Physical examination will be performed according to PRA SOPs. In addition, body weight 
and height will be measured according to PRA SOPs. 
3.5.1.3 
Total of Blood Volume 
Table 3 presents the number and volume of blood samples and the total volume of blood 
that will be collected throughout the study per subject.  
 
If deemed necessary by the Investigator or the Sponsor, the number and/or volume of 
blood samples per assessment may be increased, as long as the total volume of blood 


  
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drawn for a subject does not surpass 500 mL (except when extra blood samples need 
to be taken for safety reasons). 
 
Table 3 
Number and Volume of Blood Samples and Total Blood Volume 
Collected per Subject 
Assessment
Maximum # 
Samples 
Volume of Blood per 
Sample (mL) 
Total Volume of 
Blood (mL) 
Pharmacokinetics 
 - BTZ-043 and metabolites in 
Plasma 
- Total Radioactivity in Plasma 
 - Total Radioactivity in Whole 
Blood 
 - Metabolite Profiling in Plasma 
 
14 
 
21 
21 
 
2119
42 
24 
1 
 
1215
 
5628 
 
4284 
21 
 
252285
Clinical Chemistry 
4
3.5  
14
Hematology  
4 
3  
12 
Serology 
1 
5  
5 
Total Volume of Blood Drawn 
 
 
402
 
3.5.2 
Appropriateness of Measurements 
The assessments that will be made in this study are standard and generally recognized 
as reliable, accurate, and relevant.  
3.5.2.1 
Timing of Assessments  
For PK, predose samples will be obtained between waking up and dosing. Postdose 
samples up to 20 minutes postdose will be obtained with a time window of ±1 minute. 
Thereafter, postdose samples will be obtained with time margins of ±5% of the time that 
has passed since dosing. The ±5% time window also applies to the start and end times 
of urine collection intervals and to the total duration of each collection interval. 
 
For safety assessments, predose assessments will be performed between waking up 
and dosing. For safety assessments up to 2.5 hours postdose, a time window of 
±15 minutes is allowed. Thereafter, serial postdose assessments (eg, multiple 
assessments within any given day) will be performed with time margins of ±10% of the 
time that has passed since dosing. 
 
When assessments are planned for the same scheme time, the order of the 
assessments should be arranged in such a way that PK blood sampling will be done 
after the ECG and vital signs recordings have been conducted, with PK blood sampling 
exactly on time. 
3.5.3 
Pharmacokinetic, Safety, and Exploratory Variables  
3.5.3.1 
Pharmacokinetic Variables  
PK variables will be the plasma and urine concentrations of BTZ-043 and 
main metabolites, the total radioactivity concentrations in plasma, whole blood, urine, 
and feces (and in vomit, if produced) and their PK parameters. The PK parameters to be 


  
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determined or calculated using noncompartmental analysis include, but are not limited 
to, the parameters as given in Table 4. A complete list of PK parameters will be provided 
in the PK statistical analysis plan (SAP). 
 
Table 4
Pharmacokinetic Parameters 
Parameter
Description
Parent
Metabolites
TRA
Cmax
Maximum 
observed 
plasma 
concentration 
X 
X 
X
tmax
Time to attain maximum observed 
plasma concentration 
X 
X 
X
AUC0-t 
Area under the plasma concentration-
time curve up to time t, where t is the last 
point with concentrations above the 
LLOQ  
X 
X 
X
AUCinf 
Area under the plasma concentration-
time curve from time 0 to infinity 
calculated as: AUC0-inf=AUC0-t+Clast/kel, 
where Clast is the last measurable plasma 
concentration  
X 
X 
X
Kel 
Terminal elimination rate constant 
X 
X 
X
t1/2
Terminal elimination half-life, calculated 
as 0.693/kel 
X
X
X
CL/F
Apparent oral clearance, calculated as 
dose/AUC0-inf 
X
Vz/F
Apparent volume of distribution at 
terminal phase  
X
CLR 
Renal clearance 
X 
 
 
Aeurine 
Cumulative amount of drug excreted in 
urine  
X 
X 
X
Aefeces
Cumulative amount of drug excreted in 
feces 
X
Aevomit
Cumulative amount of drug excreted in 
vomit, if produced 
X
Aetotal
Total 
amount 
of drug 
excreted, 
calculated 
as 
Aetotal=Aeurine+Aefeces(+Aevomit)
X
feurine
Fraction of the dose administered 
excreted in urine (%)
X 
X 
X
fefeces 
Fraction of the dose administered 
excreted in feces (%) 
 
 
X
fevomit 
Fraction of the dose administered 
excreted in vomit (%) 
 
 
X
fetotal 
Fraction of the dose administered 
excreted in urine, feces, and vomit (if 
produced) (%) 
 
 
X
TRA=total radioactivity; LLOQ=lower limit of quantification 
 


  
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3.5.3.2 
Safety Variables 
The safety variables to be measured include, but are not limited to, the variables as given 
below. A complete list of safety variables will be provided in the SAP. 
 
 
AEs 
 
Clinical laboratory 
 
Vital signs 
 
12-lead ECG 
 
Physical examination  
3.5.3.3 
Exploratory Variables 
Metabolite profiling and metabolite identification will be conducted under a separate 
protocol/bioanalytical study plan, by Nuvisan Grafing, and the results will be reported 
separately. The metabolite profiling/identification report will be included in the clinical 
study 
report 
(CSR) 
as 
an 
appendix, 
and 
results 
from 
the 
metabolite profiling/identification report will be briefly summarized in the CSR. 
3.5.4
Drug Concentration Measurements
The analysis of BTZ-043 and main metabolites in plasma and urine samples will be 
performed at the Bioanalytical Department of Nuvisan Grafing using validated/qualified 
LC-MS. The analysis of whole blood, plasma, urine, and feces (and vomit, if produced) 
for total radioactivity will be conducted at the Bioanalytical Laboratory of PRA using a 
validated liquid scintillation counting method. The bioanalytical reports for the 
determinations will be included as appendices in the CSR.  
 
The analysis of quick counts will be conducted at the Bioanalytical Laboratory of PRA 
using a validated method. 
 
Metabolite profiling in plasma, urine, and feces samples by radiometry will be conducted 
at the PK Sciences & Biotransformation Department of Nuvisan Grafing. The 
bioanalytical report for the metabolite profiling and identification will be included as 
appendix in the CSR. 
3.5.5
Retention of Blood, Plasma, Urine, and Feces Samples
Blood and urine samples remaining after clinical laboratory assessments have been 
performed will not be stored for future use and will be destroyed after analysis as per 
laboratory procedure. 
 
Blood, plasma, urine, and feces (and vomit, if applicable) samples remaining after total 
radioactivity measurements and after sample aliquotation for bioanalytical and 
metabolite identification have been performed, will be stored at the site of the Promotor  
and destroyed no later than 10 years after the date of the final CSR.  


  
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3.6 
Statistical Procedures and Determination of Sample Size 
3.6.1 
Analysis Sets 
3.6.1.1 
Safety Set 
All subjects who have received the single oral dose of 14C-BTZ-043. 
3.6.1.2 
Pharmacokinetic Set 
All subjects who have received the single oral dose of 14C-BTZ-043 and provided 
sufficient bioanalytical assessment results to calculate reliable estimates of the PK 
parameters. 
3.6.2 
Statistical and Analytical Plan for Pharmacokinetic, Safety, and Exploratory 
Evaluation  
A safety SAP will be generated by the Biostatistics Department of PRA and a PK SAP 
will be generated by Nuvisan; the SAPs will be finalized prior to database lock. Full 
details of the analysis to be performed will be included in the SAPs. 
 
Any deviation from the SAPs will be reported in the section “Changes in Planned 
Analysis” in the CSR. 
3.6.2.1 
Pharmacokinetic Evaluation 
The PK parameters and their statistical evaluation of BTZ-043 and main metabolites will 
be reported as an integrated PK report, which will be included as an appendix and briefly 
summarized in the CSR. part of the bioanalytical report (included as appendix in the 
CSR). The PK parameters and their statistical evaluation of total radioactivity will be part 
of the mass balance report (included as an appendix in the CSR). A separate integrated 
PK report will incorporate the BTZ-043 and main metabolites’ PK, the total radioactivity 
PK, and the metabolite profiling PK data for an integrated evaluation. 
All data will be summarized using descriptive statistics and will be listed and summarized 
in tabular and/or graphical form. 
3.6.2.2 
Evaluation of Safety and Tolerability 
Safety and tolerability will be assessed through AEs, clinical laboratory, vital signs, 
ECGs, physical examination findings, and any other parameter that is relevant for safety 
assessment. 
3.6.2.2.1 Adverse Events 
A listing of all individual AEs will be provided. Summary tables of TEAEs will be 
presented by system organ class based on the MedDRA terminology list (preferred 
terms): 1 containing the number of TEAEs (frequency of occurrence, number of subjects 
experiencing the event) by treatment and 1 containing the number of drug-related TEAEs 
(frequency of occurrence, number of subjects experiencing the event) per treatment. 
Additional tables of total counts by treatment and relationship and by treatment and 
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3.6.2.2.2 Clinical Laboratory 
Clinical laboratory data will be listed accompanied by an indication if the parameter is 
outside the reference range. A summary of all data outside the reference range of the 
clinical laboratory will be provided. Clinical laboratory data will be presented 
descriptively, where applicable. 
3.6.2.2.3 Vital Signs and Electrocardiograms 
Vital signs and ECG parameters will be listed and they will be presented descriptively, 
where applicable. 
3.6.2.2.4 Physical Examination 
Changes from baseline for physical examination will be described and listed. 
3.6.2.3 
Evaluation of Exploratory Variables  
The PK parameters and their statistical evaluation of metabolites not measured for the 
primary objectives will be reported in the a separate integrated PK report., which will be 
included as an appendix and briefly summarized in the CSR.
 
All data will be summarized using descriptive statistics and will be listed and summarized 
in tabular and/or graphical form. 
 
The metabolite profiling and identification results will be reported separately 
(see Section 3.5.4). 
3.6.3 
Determination of Sample Size 
Since the present study is a descriptive study aimed at assessing excretion routes of 
BTZ-043 by evaluating mass balance, a formal statistical sample size calculation was 
not performed. A sample size of 4 subjects is a minimum accepted number of subjects 
for AME studies and is considered sufficient to achieve the study objectives. 
Early-termination subjects may be replaced if the total number of completers drops 
below 4, after mutual agreement between the Sponsor and PRA. 
3.7 
Data Quality Assurance 
The study may be audited by the Quality Assurance Department at PRA to assess 
adherence to the clinical study protocol (CSP) and Quality System. During the conduct 
of the study, process-related audits may be performed. An audit certificate will be 
provided in the appendices of the final CSR outlining any audits and other related 
activities performed.  
 
The clinical research site will be monitored by the study monitor to ensure correct 
performance of the study procedures and assure that the study is conducted according 
to the relevant regulatory requirements. The eCRF entries will be verified with the source 
documentation, if applicable (in some cases data are captured directly on the eCRF, 
therefore source data verification is not necessary). 
 


  
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Regulatory authorities, the Independent Ethics Committee (IEC), and/or the Sponsor’s 
clinical quality assurance group may request access to all source documents, eCRFs, 
and other study documentation for on-site audit or inspection. Direct access to these 
documents must be guaranteed by the Investigator, who must provide support at all 
times for these activities. 
 
Quality control principles will be applied throughout the performance of this study. 
Review procedures will be followed at PRA for all documents that are generated in 
relation with the study.  
 
An explanation will be given for all missing, unused, and spurious data in the relevant 
sections of the CSR. 


  
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4. 
ETHICS 
4.1 
Independent Ethics Committee 
The submission package including the CSP and the ICFs will be submitted for review 
and approval by the IEC of the foundation “Beoordeling Ethiek Biomedisch Onderzoek” 
(English translation: “Evaluation of Ethics in Biomedical Research”) (Dr. Nassaulaan 10, 
9401 HK Assen, The Netherlands) prior to the eligibility screening. The composition of 
the IEC is in accordance with the recommendations of the World Health Organization, 
the International Council for Harmonisation (ICH) E6(R2) Guideline for Good Clinical 
Practice (GCP) (European Medicines Agency [EMA]/Committee for Medicinal Products 
for Human Use [CHMP]/ICH/135/1995)4, and the EU Clinical Trial Directive (CTD) 
(Directive 2001/20/EC)5 (see below). The submission package will also be shared with 
the CA in the Netherlands for a statement of no objection. 
 
PRA will keep the IEC informed about the progress of the study. All changes in research 
activities and all unanticipated problems involving risks to human subjects will be 
immediately reported to the IEC. In accordance with Section 10, Subsection 1, of the 
Dutch law on Medical Research in Human Subjects (WMO, revised Dec 2015)6, PRA 
will inform the subjects and the IEC if anything occurs on the basis of which it appears 
that the disadvantages of participation may be significantly greater than was foreseen in 
the research proposal, or if further recruitment of subjects in the study has been put on 
hold for that reason, whichever occurs first. The study may be suspended pending 
further review by the IEC, except insofar as suspension would jeopardize the subjects’ 
health. PRA will take care that all subjects are kept informed.  
 
No changes will be made to the study without IEC approval, except when required to 
eliminate apparent immediate hazards to human subjects. 
 
Notification of the end of the study will be sent by PRA to the CA in The Netherlands and 
to the IEC within 90 days after completion of follow-up for the last subject. In case a 
study is temporarily halted, PRA will notify the IEC immediately, including the reason for 
this. In case a study is ended prematurely, PRA will notify the IEC and the CA in The 
Netherlands within 15 days, including the reasons for the premature termination. A 
summary of the results of the study will be sent by PRA to the CA and the IEC within 
1 year after the end of the study.  
 
The written documentation of approval of the CSP and ICFs will be provided to the 
Sponsor before the start of the study. Any communication with the IEC and/or CA before, 
during, and/or after the study will be provided to the Sponsor.    
4.2 
Ethical Conduct of the Study 
This study will be conducted in accordance with the ethical principles that have their 
origin in the World Medical Association (WMA) Declaration of Helsinki, adopted by the 
18th
WMA General Assembly, Helsinki, Finland, Jun 1964, and subsequent 
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This study is also designed to comply with ICH E6(R2) Guideline for GCP 
(EMA/CHMP/ICH/135/1995)4, and the EU CTD Directive 2001/20/EC5, as incorporated 
into Dutch Law.6
 
Guidelines adopted by the ICH and other relevant international guidelines, 
recommendations, and requirements will be taken into account as comprehensively as 
possible, as long as they do not violate local law. 
 
Whenever the term “Investigator” is noted in the CSP text, it may refer to either the 
Investigator at the site or an appropriately qualified, trained, and delegated individual of 
the investigational site. 
4.3 
Subject Information and Consent 
All subjects will be informed verbally and in writing regarding the objectives, procedures,
and risks of study participation. The subjects will be given the time to ask questions. 
The subjects will have to sign the Dutch or English version of the ICF before any 
study-related procedures are started. The signed ICFs will be retained and archived at 
PRA and a copy will be provided to the subject. The ICF contains information about the 
objectives of the study, the procedures followed during the study, and the risks and 
restrictions of the study, with special reference to possible side effects of the study drug 
and potential interactions. In addition, insurance coverage provided during the study is 
explained. The elements addressed in the ICF are according to the ICH E6(R2) 
Guideline for GCP (EMA/CHMP/ICH/135/1995).4
4.4 
Privacy 
All personal details will be treated as confidential by the Investigator and staff at PRA, 
the Sponsor, and any sub-contractors involved, and handling of personal data will be in 
compliance with the EU General Data Protection Regulation.8 
 


  
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5. 
STUDY ADMINISTRATIVE STRUCTURE 
5.1 
Distribution of Activities 
5.1.1 
Preparation of Study Drug 
The study drug will be prepared at the PRA Pharmacy. 
5.1.2 
Laboratory Assessments 
The analysis of BTZ-043 and main metabolites in plasma and urine samples will be 
performed at the Bioanalytical Department of Nuvisan Grafing (Grafing, Germany) by 
using the Nuvisan study protocol G-A-LAB-21-004.  
 
The analysis of total radioactivity in whole blood, plasma, urine, and feces (and vomit, if 
produced) will be performed at the Bioanalytical Laboratory of PRA (Assen, 
The Netherlands).  
 
The analysis of clinical laboratory samples will be performed at the PRA Clinical 
Laboratory.  
 
Metabolite profiling and identification will be performed at the PK Sciences and 
Biotransformation Department of Nuvisan Pharma Services GmbH (Grafing, Germany) 
by using the Nuvisan study protocol G-A-MET-21-002. 
5.1.3 
Electronic Case Report Form Design 
The eCRF design will be performed with the computer program Oracle Clinical (Oracle, 
Redwood Shores, Redwood City, CA, US) by the Database Programming Department 
of PRA. 
5.1.4 
Data Management 
Data management will be performed with the computer programs Oracle Clinical 
(Oracle, Redwood Shores, Redwood City, CA, US), SAS® (SAS Institute Inc, Cary, NC, 
US), and EXACT (Kinship EXACT™, Kinship Technologies, a technology subsidiary of 
PRA) by the Data Management Department of PRA. 
5.1.5 
Statistics 
A safety SAP will be provided by the Biostatistics Department of PRA. The safety 
analysis will be conducted by the Biostatistics Department of PRA. Statistical analysis 
will be performed with the computer program SAS® (SAS Institute Inc, Cary, NC, US). A 
PK SAP will be provided by Nuvisan Grafing. Statistical evaluation of PK parameters will 
be conducted by Nuvisan Grafing. PK parameters will be calculated using Phoenix 
WinNonlin (Pharsight, Mountain View, CA, US). All individual results will be provided to 
the Sponsor after completion of the study. 
5.1.6 
Clinical Study Report Writing 
The CSR, structured in accordance with the guideline “Structure and Content of Clinical 
Study Reports - ICH E3”9, will be written by PRA and a draft version prepared for 


  
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approval by the Sponsor. The final version will be provided as paper version to the 
Sponsor. 
5.2 
Documentation 
5.2.1 
Archiving 
All documents concerning the study will be kept on file in the Central Archives of PRA 
for at least 25 years after conduct of the study. The Sponsor will receive the completed 
eCRFs as PDF file. 
5.2.2 
Recording of Data in Source Documents and Electronic Case Report Forms 
Wherever possible, all data will be entered directly into the eCRFs. Source documents 
will be used in some cases. 
 
A data management plan will be written by the Data Management Department of PRA, 
which will be finalized prior to performing any data validation. An appendix to the data 
management plan (origin of source data list for data entry) will identify any data to be 
recorded directly in the eCRF (ie, no prior written or electronic record of data) and which 
data should be considered source data.  
 


  
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6. 
CONFIDENTIALITY AND PUBLICATION POLICY 
All information generated in this study is considered highly confidential and must not be 
disclosed to any person or entity not directly involved with the study unless prior written 
consent is gained from the Sponsor and Promotor. However, authorized regulatory 
officials, the Sponsor and its authorized representatives are allowed full access to the 
records. 
 
All study subjects must be informed that their personal study-related data will be used 
by the Sponsor in accordance with local data protection law. The level of disclosure must 
also be explained to the subject, who will be required to give consent for their data to be 
used as described in the ICF. The subjects must be informed that their medical records 
may be examined by auditors or other authorized personnel appointed by the Sponsor, 
by appropriate IEC members, and by inspectors from regulatory authorities. 
 
Identification of subjects and eCRFs shall be by unique subject numbers only.  
 
All personal details will be treated as confidential by the Investigator and staff at PRA. 
 
All relevant aspects regarding publication will be part of the contract (or similar 
document) between the Sponsor and PRA.  
 


  
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7. 
REFERENCES 
1. 
Investigator’s Brochure BTZ-043. Edition 04. 10 Nov 2020. 
2. 
CPMP/EWP/560/95/Rev. 1 Corr. 2** Committee for Human Medicinal Products 
(CHMP). Guideline on the investigation of drug interactions. 21 Jun 2012. 
3. 
Radiation Burder Calculation Report EDS-NL. Dosimetry BTZ-043. 27 Nov 2018. 
4. 
International Council for Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, E6(R2): Integrated Addendum to ICH E6(R1): Guideline for Good 
Clinical Practice. Adopted by the European Medicines Agency (EMA), 
Committee 
for 
Human 
Medicinal 
Products, 
Document 
Reference 
EMA/CHMP/ICH/135/1995), 14 Jun 2017. 
5. 
Directive 2001/20/EC of the European Parliament and of the Council of 
4 Apr 2001 on the approximation of the laws, regulations and administrative 
provisions of the Member States relating to the implementation of good clinical 
practice in the conduct of clinical trials on medicinal products for human use. 
6. 
Medical 
Research 
Involving 
Human 
Subjects 
Act 
(WMO, 
Wet 
Medisch-Wetenschappelijk Onderzoek met Mensen), revision Dec 2015. 
7. 
World Medical Association (WMA) Declaration of Helsinki – Ethical Principles for 
Medical Research Involving Human Subjects (18th WMA General Assembly 
1964), revised at 64th WMA General Assembly, Fortaleza, Brazil, Oct 2013. 
8. 
The General Data Protection Regulation (GDPR). Regulation (EU) 2016/679 of 
the European Parliament and the Council of the European Union, 27 Apr 2016, 
applicable as of 25 May 2018. 
9. 
International Council for Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, E3: Structure and Content of Clinical Study Reports. Note for 
Guidance on Structure and Content of Clinical Study Reports, Adopted by the  
Committee for Human Medicinal Products, European Medicines Agency (EMA), 
Document Reference CPMP/ICH/137/95, Jul 1996. 
10. 
International Council for Harmonisation of Technical Requirements for 
Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite 
Guideline, E2A: Clinical Safety Data Management: Definitions and Standards for 
Expedited Reporting. Note for Guidance on Clinical Safety Data Management, 
Adopted by the Committee for Human Medicinal Products, European Medicines 
Agency (EMA), Document Reference CPMP/ICH/377/95, Jun 1995.


  
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8. 
APPENDICES 
8.1
Drug Accountability 
Upon receipt of the study drug, it will be inspected and counted by the responsible 
pharmacist. If necessary, all study drug will be repacked per dosing occasion and labeled 
according to PRA SOPs. 
 
The study drug will be kept in the PRA Pharmacy or in a locked and secured storage 
facility accessible to the pharmacist and the pharmacy assistant only. Temperature is 
monitored 24/7 with an automatic environmental monitoring system. In case of an alarm, 
the pharmacist or pharmacist on duty is informed immediately by email and phone. 
 
The responsible pharmacist will keep an inventory. This will include a description of the 
formulation and the quantity of study drug received for the study and a record of what is 
dispensed, to whom, and when. 
 
On termination of the study, the responsible pharmacist will conduct a final inventory of 
the study drug supply and will record the results of this inventory in the 
Drug Accountability Form. Unused study drug will be returned to the Promotor upon 
notice is given to Promotor and Sponsor or will be locally destroyed according to PRA 
standard procedures. Final accountability needs to be verified by monitor before return 
and/or destruction of medication. 
8.2
Adverse Events and Serious Adverse Events Evaluation and Reporting 
8.2.1 
Adverse Events 
An AE is any unfavorable and unintended sign (including an abnormal laboratory finding), 
symptom, or disease temporally associated with the use of a medical treatment or 
procedure that may or may not be considered related to the medical treatment or 
procedure. AE definitions will be followed as stated in the “Note for Guidance on Clinical 
Safety Data Management: Definitions and Standards for Expedited Reporting” (ICH topic 
E2A).10
 
All AEs reported by the subjects or apparent from their physical appearance during the 
clinical study will be reported on the AE eCRF page. 
 
The severity of AEs will be graded using the most current version of the MedDRA: 
 
Mild: An AE that is easily tolerated by the subject, causes minimal discomfort, does 
not interfere with everyday activities, and does not require intervention. 
 
Moderate: An AE that is sufficiently discomforting to interfere with normal everyday 
activities; intervention may be needed. 
 
Severe: An AE that prevents normal everyday activities; treatment or other 
intervention usually needed. 
 
If an AE has multiple aspects, the aspect with the highest severity will be graded. 


  
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It is emphasized that the term severe is a measure of severity; thus, a severe AE is not 
necessarily serious. For example, itching for several days may be rated as severe, but 
may not be clinically serious. 
 
In addition, clinically significant changes in physical examination findings and abnormal 
objective test findings (eg, laboratory, x-ray, ECG) should also be recorded as AEs. Test 
findings and physical examination findings can result in AEs if they:  
 
Are associated with accompanying symptoms, and/or 
 
Require additional diagnostic testing or medical/surgical intervention, and/or 
 
Lead to a change in study dosing or discontinuation from the study; result in the 
addition of significant additional concomitant drug treatment or other therapy, and/or 
 
Lead to any of the outcomes included in the definition of an SAE, and/or 
 
Are considered to be an AE by the Investigator or Sponsor. 
 
Reporting as an AE should not be triggered by: 
 
Merely repeating an abnormal test, or  
 
Any abnormal test result that is determined to be an error. 
 
The relationship of any AE to the study drug will be assessed and graded as related or 
not related. 
 
Related:   
 
The AE follows a reasonable temporal sequence to study drug administration, and 
cannot be reasonably explained by the subject’s clinical state or other factors (eg, 
disease under study, concurrent diseases, or concomitant medications). 
 
The AE follows a reasonable temporal sequence to study drug administration, and is 
a known reaction to the drug under study or a related chemical group, or is predicted 
by known pharmacology. 
 
Not Related: 
The AE does not follow a reasonable sequence from study drug administration, or can 
be reasonably explained by the subject’s clinical state or other factors (eg, disease under 
study, concurrent diseases, and concomitant medications).  
8.2.2 
Serious Adverse Events 
An SAE is any untoward medical occurrence that, on the basis of medical and scientific 
judgment: 
 
Results in death, or 
 
Is life-threatening (this refers to an event in which the patient was at risk of death at 
the time of the event; it does not refer to an event which hypothetically might have 
caused death if it were more severe), or  
 
Requires inpatient hospitalization for a medical reason or prolongation of existing 
hospitalization (this refers to hospital admission required for treatment of the AE). 
(Note: this does not include confinement in, for example, a respite unit; a skilled 


  
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nursing unit; rehabilitation facility; the clinical research center; or confinement due to 
planned or unplanned reason unrelated to study), or 
 
Results in persistent or significant disability/incapacity, or 
 
Is a congenital anomaly/birth defect. 
 
Medical and scientific judgment should be exercised in deciding whether expedited 
reporting is appropriate in other situations, such as important medical events that 
may not be immediately life-threatening or result in death or hospitalization, but may 
jeopardize the patient or may require intervention to prevent one of the other 
outcomes listed in the definition above. These should also usually be considered 
serious. 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 hospitalization. 
 
SAEs will be collected from admission until the final study visit. SAEs that are related to 
the investigational drug and continue beyond the normal collection period (ie, are 
ongoing at the time a subject exits the study) will be followed until resolution or until 
stabilized with sequelae. SAEs that begin after the subject’s participation in the study is 
complete, but that the Investigator considers to be related to study drug, may be reported 
at any time.  
 
The Investigator or clinical site personnel must notify the Sponsor’s Medical Monitor and 
PRA Drug Safety Center of all SAEs, regardless of relationship to the investigational 
drug, within 24 hours of clinical site personnel becoming aware of the event. The 
Investigator will provide the initial notification by sending a completed “SAE Notification 
Form,” which must include the Investigator’s assessment of the relationship of the event 
to investigational drug and must be signed by the Investigator. 
 
In addition, notification is sent by PRA to the IEC and the subject’s General Practitioner. 
 
Follow-up information, or new information regarding an ongoing SAE, must be provided 
promptly to the Sponsor’s Medical Monitor and PRA Drug Safety Center. 
All SAE reports should be sent to the contacts provided on Page 4: SAE Contact 
Information. 
8.2.3 
Suspected Unexpected Serious Adverse Reactions 
An SAE that is also an unexpected adverse drug reaction is called a suspected 
unexpected serious adverse reaction (SUSAR). Unexpected adverse reactions are 
adverse reactions of which the nature or severity is not consistent with the applicable 
product information (eg, investigator’s brochure for an unapproved investigational 
medicinal product or the Summary of Product Characteristics for an authorized medicinal 
product). 
 
The Sponsor or its representative (eg, PRA if agreed to before start of the study) will 
promptly report (expedited reporting) the following SUSARs to the IEC: 
 
SUSARs that have arisen in the current clinical study that was assessed by the IEC 


  
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SUSARs that have arisen in other clinical studies of the same Sponsor and with the 
same medicinal product, and that could have consequences for the safety of the 
subjects involved in the current clinical study that was assessed by the IEC 
 
The Sponsor will promptly report (expedited reporting) all SUSARs to the CA and the 
Medicine Evaluation Board (MEB) of the country where this study is conducted and to 
the CAs in other Member States, as applicable. 
 
SUSARs that have already been reported to the EMA Eudravigilance database do not 
have to be reported again to the CA and the MEB because they have direct access to 
the Eudravigilance database. 
 
Expedited reporting will occur no later than 15 calendar days after the Sponsor or its 
representative has first knowledge of the adverse reactions. For fatal or life-threatening 
cases, the term will be maximally 7 calendar days for a preliminary report with another 
8 days for completion of the report.  
8.2.4 
Follow-up of Adverse Events 
Follow-up of AEs will continue until resolution, stabilization, or death. In case of ongoing 
AEs at database closure, the data obtained at database closure will be used in the 
statistical analysis. The follow-up of the AE will be documented in the source documents 
and will be described in the final CSR only if considered relevant by the Investigator. 
8.3
Pregnancy 
If the Investigator becomes aware of a pregnancy occurring in the partner of a male 
subject participating in the study up to 90 days after the last dose of study drug of the 
male subject, the pregnancy should be reported to the Sponsor within 1 working day of 
obtaining written consent from the pregnant partner. The Investigator may make 
arrangements for the partner to be counseled by a specialist, to discuss the risks of 
continuing with the pregnancy and the possible effects on the fetus. Monitoring of the 
partner should continue untile the outcome of the pregnancy is known.  

SAP_001.pdf: 
Statistical Analysis Plan 
NVS543EC-185431 
Protocol: LMU-IMPH-BTZ-043-03 
Version Date: 02-Dec-2021  
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Statistical Analysis Plan 
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1.0 Table of Contents 
1.0 Approvals .................................................................................. Error! Bookmark not defined. 
2.0 Table of Contents ................................................................................................................. 2 
3.0 Introduction.......................................................................................................................... 4 
4.0 Changes from Previous Version of Approved SAP ..................................................................... 4 
5.0 Study Objectives .................................................................................................................. 4 
5.1 Primary............................................................................................................................ 4 
5.2 Secondary ....................................................................................................................... 4 
5.3 Exploratory....................................................................................................................... 4 
6.0 Study Design ....................................................................................................................... 5 
6.1 Sample Size Considerations ............................................................................................... 5 
6.2 Randomization.................................................................................................................. 5 
6.3 Method of Assigning Subjects to Treatment Groups................................................................ 6 
7.0 Overview of Planned Analysis................................................................................................. 6 
7.1 Changes from Protocol ...................................................................................................... 6 
7.2 Interim Analysis and Key .................................................................................................... 6 
7.3 Final Analysis ................................................................................................................... 6 
8.0 Data Review ........................................................................................................................ 6 
8.1 Data Management............................................................................................................. 6 
8.2 Acceptance of Data for Summarization ................................................................................. 6 
9.0 Definitions and General Analysis Methods ................................................................................ 7 
9.1 Analysis Data Presentation ................................................................................................. 7 
9.1.1 Rounding ................................................................................................................... 7 
9.1.2 Imputation.................................................................................................................. 7 
9.1.3 Descriptive Statistics ................................................................................................... 7 
9.1.4 Pooling ...................................................................................................................... 7 
9.1.5 Unscheduled Measurements......................................................................................... 8 
9.2 Analysis Data Definitions .................................................................................................... 8 
9.2.1 Baseline Definition ...................................................................................................... 8 
9.2.2 Treatment/Subject Grouping ......................................................................................... 8 
9.2.3 Common Variable Derivations ....................................................................................... 8 
9.2.4 QC............................................................................................................................ 8 
9.2.5 ADaM Datasets and Metadata ...................................................................................... 9 
9.3 Software .......................................................................................................................... 9 
9.4 Statistical Methods ............................................................................................................ 9 
9.4.1 Statistical Outlier Determination..................................................................................... 9 
9.4.2 Predetermined Covariates and Prognostic Factors ........................................................... 9 
9.4.3 Hypothesis Testing...................................................................................................... 9 
9.5 TFL Layout....................................................................................................................... 9 
10.0 Analysis Sets ....................................................................................................................10 
10.1 Safety Set .....................................................................................................................10 
10.2 Pharmacokinetic Set .......................................................................................................10 
11.0 Subject Disposition.............................................................................................................10 
12.0 Protocol Deviations ............................................................................................................10 
13.0 Demographic and Baseline Characteristics ............................................................................11 
13.1 Demographics................................................................................................................11 
13.2 Medical History ..............................................................................................................11 
13.3 Other Baseline Characteristics .........................................................................................11 
14.0 Concomitant Medications ....................................................................................................11 
15.0 Treatment Compliance and Exposure ...................................................................................11 
16.0 Pharmacokinetic Analyses...................................................................................................11 
16.1 Pharmacokinetic Variables...............................................................................................11 
16.1.1 Plasma Variables .....................................................................................................12 
Table 1: Plasma Parameters.......................................................................................................13 


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16.1.2 Blood Variables........................................................................................................14 
Table 2:  Blood Parameters ........................................................................................................14 
16.1.3 Urine Variables ........................................................................................................16 
Table 3: Urine Parameters .........................................................................................................16 
16.1.4 Feces Variables .......................................................................................................17 
Table 4: Feces Parameters ........................................................................................................17 
16.1.5 Vomitus Variables ....................................................................................................18 
Table 5: Vomitus Parameters......................................................................................................18 
16.1.6 Total Excretion Variables ...........................................................................................18 
Table 6: Total Excretion Parameters ............................................................................................19 
16.2 Pharmacokinetic Summaries ............................................................................................20 
16.2.1 Pharmacokinetic Concentrations.................................................................................20 
16.2.2 Pharmacokinetic Parameters .....................................................................................20 
16.2.3 Pharmacokinetic Amounts Excreted ............................................................................21 
17.0 Safety Analyses .................................................................................................................23 
17.1 Safety Variables .............................................................................................................23 
17.1.1 Adverse Events........................................................................................................23 
17.1.2 Deaths and Serious Adverse Events ...........................................................................24 
17.1.3 Laboratory Data .......................................................................................................24 
17.1.4 Vital Signs ...............................................................................................................24 
All vital signs measurements will be listed. .............................................................................24 
17.1.5 Electrocardiograms...................................................................................................24 
17.1.6 Other Observations Related to Safety..........................................................................24 
18.0 References .......................................................................................................................24 
Appendix 1: Glossary of Abbreviations .........................................................................................25 
Appendix 2: Schedule of Assessments .........................................................................................27 
Appendix 3: List of In-Text Outputs ..............................................................................................30 
Appendix 4: List of End of Text Outputs ........................................................................................30 
19.0 Document History ..............................................................................................................33 
 


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2.0 Introduction 
This Statistical Analysis Plan (SAP) describes the statistical methods that will be used during the analysis 
and reporting of data collected under LMU Klinikum Protocol LMU-IMPH-BTZ-043-03. 
This SAP should be read in conjunction with the study protocol and electronic case report form (eCRF). 
This version of the plan has been developed using the protocol dated 01-Jul-2021 (including all 
amendments and notes-to-files up to this protocol date) and the final eCRF(s) dated 15-Sep-21 (Screening) 
and 11-Oct-2021 (during Study). 
An approved and signed SAP is a requirement for database lock.  
This SAP covers the results that will be processed by the ICON/PRA Early Development Services (EDS) 
Biostatistics Department, and the results that will be processed by Nuvisan GmbH. ICON/PRA EDS will 
perform and is responsible for the safety and tolerability evaluation as well as for the total radioactivity in 
all matrices, including mass balance evaluation. Nuvisan will perform and is responsible for the 
pharmacokinetic (PK) evaluation of parent drug and main metabolite data in plasma and urine. 
ICON/PRA EDS will prepare the Tables, Figures and Listings (TFLs) for Total Radioactivity (TR), BTZ-043 
(M0), and main metabolites M1, M2, M4tot, and M10. 
Metabolic profiling (and Fe(%) of metabolites) is not part of this SAP and will be provided in a separate 
document. 
This SAP supersedes the statistical considerations identified in the protocol; where considerations are 
substantially different, they will be so identified. Any post-hoc or unplanned analyses, or significant changes 
from the planned analysis in this SAP performed to provide results for inclusion in the clinical study report 
(CSR) but not included in this SAP, will be clearly identified in Section 9.8.2 of the CSR. Changes to planned 
analyses do not require an updated SAP but should be included in the CSR if significant. 
3.0 Changes from Previous Version of Approved SAP 
This is the first version of the SAP. 
4.0 Study Objectives 
4.1 Primary 
• 
To determine the rates and routes of excretion of [14C]BTZ-043-related radioactivity, including mass 
balance of total drug-related radioactivity in urine and feces (and vomit, if applicable), following the oral 
administration of a single 500 mg dose of [14C]BTZ-043 in healthy male volunteers. 
• 
To determine the pharmacokinetics (PK) of total radioactivity in blood and in plasma. 
• 
To characterize the plasma PK of BTZ-043 and main metabolites by liquid chromatography-mass 
spectrometry (LC-MS), if applicable. 
• 
To characterize the urine concentrations of BTZ-043 and main metabolites by LC-MS, if applicable. 
4.2 Secondary 
• 
To assess the safety and tolerability of a single 500 mg oral dose of BTZ-043 administered to healthy 
male volunteers. 
4.3 Exploratory  
• 
To identify BTZ-043’s metabolites in plasma, and to quantify metabolites not measured in the primary 
objectives by radiometry in order to elucidate key biotransformation pathways and clearance 
mechanisms of BTZ-043 in humans.  


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• 
To identify BTZ-043’s metabolites in excreta (urine and feces) and to quantify metabolites not measured 
in the primary objectives by radiometry in order to elucidate key biotransformation pathways and 
clearance mechanisms of BTZ-043 in humans. 
 
5.0 Study Design 
This study is a Phase 1, single-center, open-label study to investigate the absorption, metabolism, and 
excretion of BTZ-043 after a single oral administration of 500 mg BTZ-043 containing 3.7 MBq of 
[14C]BTZ-043 in 4 healthy adult male subjects. 
A total of 4 evaluable subjects completing all procedures are required. Six (6) subjects will be enrolled in 
the cohort in order to have 4 evaluable subjects. If the cohort of 4 subjects administered with 500 mg 
BTZ-043 containing 3.7 MBq of [14C]BTZ-043 leads to less than 4 evaluable subjects, the 2 back-up 
subjects can be dosed subsequently to ensure that 4 subjects have completed all study procedures and 
are evaluable. If any of the 4 subjects vomits within 4 hours after study drug administration (approximately 
2 times the median Tmax of BTZ-043), the subject will be replaced. 
Subjects will be admitted to the clinical research center on Day -1, which is the day prior to Day 1, the day 
of drug administration.  
Subjects will receive a single oral dose of 500 mg BTZ-043 containing 3.7 MBq of [14C]BTZ-043 on Day 1. 
Subjects will be discharged from the clinical research center on Day 5 (approximately 96 hours after drug 
administration) upon satisfactory recovery of radioactivity. If the criteria for discharge are not met on Day 
5, subjects will be required to remain confined for a maximum of 3 additional days (Days 6 to 8) until the 
criteria are met (daily check by quick counts or normal counts, as feasible). The in-house period should be 
prolonged up to the morning of Day 8 if at least 1 of the following applies:  
• 
Total radioactivity in plasma is higher than 5% of radioactivity-Cmax and/or PK parameters for 
determination of the total radioactivity-elimination phase cannot be determined reliably.  
• 
The mass balance in urine and feces is not complete by Day 5 (i.e., <90%). 
• 
The combined urinary and fecal excretion is not ≤1% of the administered dose per 24 hours for 2 
consecutive days based on 14C-radioactivity “quick counts” or normal counts, as feasible. 
If the criteria for discharge are still not met on Day 8, subjects may be discharged from the clinical site upon 
discretion of the Investigator. To ensure that adequate consideration is given to the protection of the 
subject’s interests, subjects with an incomplete mass balance on Day 8 will be asked to return to the clinical 
site for one or two 24-hour visits (Day 11-12 and Day 15-16). 
The follow up assessments will be performed 30 days (±2 days) after the [14C]BTZ-043 dose. 
The schedule of assessments is shown in Appendix 2: Schedule of Assessments 
5.1 Sample Size Considerations 
Since the present study is a descriptive study aimed at assessing excretion routes of BTZ-043 by evaluating 
mass balance, a formal statistical sample size calculation was not performed. A sample size of 3 subjects 
is a minimum accepted number of subjects for AME studies and thus 4 subjects are considered sufficient 
to achieve the study objectives, even in case of one drop-out. Early termination subjects may be replaced 
if the total number of completers drops below 4, after mutual agreement between the Sponsor and 
ICON/PRA. 
5.2 Randomization 
This is an open label, non-randomized study. 


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5.3 Method of Assigning Subjects to Treatment Groups 
After obtaining informed consent, subjects will be screened according to the inclusion and exclusion criteria. 
Subjects who meet all eligibility criteria receive a subject number upon inclusion in the study (Subject 
numbers 001-004). They receive the subject number just prior to dosing. The subject number ensures 
identification throughout the study. Replacement subjects will receive the number of the subject to be 
replaced, increased by 100 (eg, 101 replacement number for Subject Number 001), and will be 
administered the same treatment. 
Subjects who drop out or withdraw for any reason without completing all screening evaluations successfully, 
are considered “screening failures“. Such subjects do not receive a subject number, and data will not be 
entered in the eCRFs. 
6.0 Overview of Planned Analysis 
6.1 Changes from Protocol 
The protocol mentions a separate PK SAP. It has been decided to provide one integrated SAP, covering 
PK, safety and tolerability. 
The protocol states in section 3.2 that four subjects are the minimum number of subject for AME studies. 
Although 4-6 subjects are preferred, AME data of three subjects are considered to be sufficient to achieve 
the study objectives.  
6.2 Interim Analysis and Key  
Interim analysis or TFLs before Database freeze (softlock) are not planned. 
6.3 Final Analysis  
Draft TFLs will be provided after database lock. After Sponsor comments have been incorporated, the TFLs 
will be finalized and incorporated in the first draft CSR. 
7.0 Data Review 
7.1 Data Management 
Data handling and transfer will take place under the ICON/PRA Data Management Plan for the study. 
7.2 Acceptance of Data for Summarization 
Programming of analysis datasets and TFLs may be ongoing during the data management of the study. 
However, programming of analysis datasets and TFLs will be completed and quality controlled (QC’d) after 
database lock. Only quality assured (QA’d) results released by the Safety Laboratory, Bioanalytical 
Laboratory, or other external data source will be used for the programming of analysis datasets and TFLs 
for the final report. Any data values requiring investigation or corrections that are identified while 
programming the analysis datasets and TFLs will be sent to the project Data Manager. If the issue affects 
the TFLs the Programmer, Statistician (ICON/PRA EDS) or PK Analyst (Nuvisan) who identified the issue 
will follow it to resolution. 


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8.0 Definitions and General Analysis Methods 
8.1 Analysis Data Presentation 
8.1.1 Rounding 
In listings, data will be presented with the same precision as the original data. Derived data will be rounded 
for presentation purposes.  
Baseline, safety and tolerability data: 
For all summaries, all descriptive statistics will be presented with the same precision (number of decimals) 
as the data they are calculated from. Percentages will be presented with one decimal.  
The above rule can be applied directly to collected data. For derived data rounding will occur prior to 
summarization so a specific number of decimal places will have to be assumed to apply the above rounding 
rules for summary statistics. 
PK concentration, parameters and total radioactivity data: 
Derived PK parameters, except tmax, will be reported with a precision of 3 significant digits or as integers 
when the value is ≥1000. The tmax will be reported with 2 decimals. 
Any ratio will be rounded to one place beyond the decimal, no matter the value of the ratio. 
Additional derived data will be rounded in the derived dataset as determined by the statistician. 
8.1.2 Imputation 
The following imputation will be performed: 
• PK concentrations below the lower limit of quantification (LLOQ) (see 16.2.1) 
• If the actual time or dose time is missing, the scheduled time may be substituted in order to calculate the 
PK parameters (see 16.2.2) 
• Last observation carried forward (LOCF) for mass balance/ urine will be applied (see 16.2.3)  
• missing period start or end date/times in Subject-level analysis dataset (ADSL), as appropriate. 
• missing start or end date/times of adverse events (AEs) for the calculation of onset and duration (see 
18.1.1)  
• missing AE severity, relationship and seriousness of AEs (see 17.1.1) 
• Laboratory data that are <x or >x (e.g. “<1.03”, “>1000”) will be imputed to x (respectively to 1.02 and 
1001 in the example, see section 17.1.2).  
Imputations / substitutions will be used in calculations and summaries only. Original data will be listed. If 
imputations / substitutions are listed, they will be listed along with the original data. 
8.1.3 Descriptive Statistics 
Unless otherwise indicated, continuous variables will be summarized with the following descriptive 
statistics: n (number of observations), (arithmetic) mean, standard deviation (SD), coefficient of variation 
(%CV, only for PK data), minimum (min) value, median, and maximum (max) value. 
Categorical data will be summarized with frequencies and percentages. Percentages by categories will be 
based on the number of subjects exposed within a treatment. 
For categorical data the categories will be presented in the tables exactly as they appear in the database.  
8.1.4 Pooling 
Not applicable for this SAP. 


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8.1.5 Unscheduled Measurements 
Unscheduled measurements will be included in the listings. With the exception of unscheduled 
measurements used for baseline, unscheduled measurements will be excluded from the descriptive 
statistics and statistical analysis. In case an unscheduled measurement was performed immediately after 
the scheduled measurement because of a previous measurement error (e.g. equipment failure), this 
repeated measurement will be used, and the original erroneous measurement will be excluded from the 
analysis. 
8.2 Analysis Data Definitions 
8.2.1 Baseline Definition 
Unless otherwise stated, baseline for post-dose evaluations is defined as the last observation recorded 
before the first study drug administration. The last observation can be an unscheduled / repeated 
measurement. If a pre-treatment observation is missing in a given period, then the screening value may be 
used. 
8.2.2 Treatment/Subject Grouping  
 
Label 
Grouping 
Study Drug 
[14C]-labeled BTZ-043 
Treatment 
500 mg BTZ-043 containing 3.7 MBq of [14C]BTZ-043 
Dose Level 
500 mg 
 
8.2.3 Common Variable Derivations 
 
Variable 
Data Type 
Definition/Calculation 
Change from Baseline 
All 
Post-dose Observation minus Baseline 
Observation 
Analysis Study Day 
(Prior to Dose) 
All 
Date of Measurement minus Dose Date 
Analysis Study Day 
(Post Dose) 
All 
Date of Measurement minus Dose Date +1 
Relative Actual time 
from  
 
Dosing 
Actual time calculated as the sampling  date/time 
minus IMP start  of dosing date/time 
8.2.4 QC 
The analysis datasets and the TFLs will be QC’d according to the general ICON/PRA EDS QC plan. 
8.2.4.1 Critical Data 
The QC plan requires datasets be classified as critical or non-critical. Subject level, pharmacokinetic, and 
adverse events (ADSL, ADPC, ADPP, and ADAE) are considered as critical datasets. These datasets will 
be double programmed per the QC Plan. 


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8.2.5 ADaM Datasets and Metadata 
The analysis datasets will be generated in accordance with Clinical Data Interchange Standard Consortium 
(CDISC) Analysis Data Model (ADaM) Version 2.1, and CDISC ADaM Implementation Guide 1.2. The 
following ADaM datasets will be prepared: 
Table 1 ADaM Datasets 
ADaM Dataset 
Name 
Description 
Double Programmed? 
ADSL 
Subject-Level Analysis Dataset 
Yes 
ADAE 
Adverse Events Analysis Dataset 
No 
ADEG 
ECG Analysis Dataset 
No 
ADLB 
Laboratory Test Results Analysis Dataset 
No 
ADPC 
Pharmacokinetic Concentrations Analysis Dataset 
Yes 
ADPP 
Pharmacokinetic Parameters Analysis Dataset 
Yes 
ADVS 
Vital Signs Analysis Dataset 
No 
 
A define.xml file version 2 with the corresponding metadata and Analysis Data reviewers guide (ADRG) will 
be included. Analysis results (analysis displays or program details) metadata are excluded from the 
define.xml. 
8.3 Software 
The statistical analysis and reporting will be done using SAS® for WindowsTM Version 9.4 or higher (SAS 
Institute, Inc.). 
PK parameter calculations will primarily be done using Phoenix® WinNonlin® (WNL) Version 7.0 or higher  
(Certara USA Inc.). For TR in plasma and blood, version 8.1 will be used. Additional PK computations may 
be performed in SAS®. 
8.4 Statistical Methods 
8.4.1 Statistical Outlier Determination 
No statistical outlier analysis is planned. 
8.4.2 Predetermined Covariates and Prognostic Factors 
There are no predetermined covariates or prognostic factors. 
8.4.3 Hypothesis Testing 
No formal hypothesis testing will be done. 
8.5 TFL Layout 
Report layout will be according to the ICON/PRA EDS – ICH E3 compliant – CSR Template. The layout of 
Tables, Figures and Listings (TFLs) will be according to the ICON/PRA EDS standards.  
No table shells will be provided. The TFLs will be provided in Adobe PDF format. 
Format: 
• 
Page size: A4  
• 
Data in listings will be sorted by subject number and time point. 


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• 
Data in tables will be sorted by treatment and time point (where applicable). 
• 
Column titles will be in title case letters. 
• 
All tables and listings will be in landscape format. 
• 
The following treatment label will be used in the TFLs: [14C]BTZ-043  
 PK and mass balance TFLs will be presented with a subtitle for the analyte and specimen type, 
using BTZ-043, M1, M2, M4tot, M10 and Σ(BTZ-043, M1, M2, M4tot, M10) in plasma and urine, 
total radioactivity in plasma and blood, and total radioactivity excretion in urine/feces/vomitus/sum 
of excreta. 
 Σ(BTZ-043, M1, M2, M4tot, M10) will be reported as Sum (BTZ-043, M1, M2, M4tot, M10). 
9.0 Analysis Sets 
 
Analyses 
Safety Set 
Pharmacokinetic Set 
Disposition 
Summaries 
 
 
Safety Assessments 
 
 
Baseline 
Characteristics 
 
 
Primary Analysis 
 
 
PK Concentrations 
 
 
PK Parameters 
 
 
9.1 Safety Set 
The safety set will consist of subjects who receive one dose of [14C]BTZ-043. This set will be used for the 
safety data summaries, baseline characteristic summaries, and disposition summaries. 
9.2 Pharmacokinetic Set 
The PK set will consist of all subjects who receive the complete dose of [14C]BTZ-043 and have sufficient 
concentration-time data to calculate a valid Cmax and AUClast. Summaries of PK concentrations and  
summaries of PK parameters will be based on the PK set. 
10.0 Subject Disposition 
The number and percentage of subjects randomized, dosed, and members of each analysis set will be 
presented. The number and percentage of subjects who withdrew from the study prematurely and a 
breakdown of the corresponding reasons for withdrawal will also be presented. 
11.0 Protocol Deviations 
Protocol deviations will be listed by subject. 


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12.0 Demographic and Baseline Characteristics 
12.1 Demographics 
All demographic data as collected during the screenings visit will be listed by subject. 
Subject demographics will be summarized descriptively for all subjects by treatment. The summary will 
include the subjects’ age (in years), gender, race, ethnicity, weight at screening (in kg), height (in cm), and 
BMI at screening (in kg/m2). Demographics will be summarized for the safety set (and PK set if different 
from safety set). 
12.2 Medical History 
Medical history will be listed. Medical history will be coded according to Medical Dictionary for Regulatory 
Activities (MedDRA), version 24.1. 
12.3 Other Baseline Characteristics 
The results of drug and alcohol screen at screening will be listed. 
The results of serology at screening will be listed. 
The results of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) polymerase chain reaction 
(PCR) tests will be listed. 
Non-compliance to in- or exclusion criteria (if any) will be listed. 
13.0 Concomitant Medications 
Concomitant medications will be listed uncoded by subject. Medications with an end date prior to the first 
dose of study drug will be considered prior medications and will be noted in the listing. If a partial date 
allows a medication to be considered concomitant it will be categorized as such. 
14.0 Treatment Compliance and Exposure 
Exposure data will be listed by subject. 
15.0 Pharmacokinetic Analyses  
Due to the known instability of some BTZ-043 metabolites in native matrices, not every metabolite will be 
quantified by radio-profiling during this hAME study (stabilization measures are not compatible with radio-
analysis sample processing). Instead, BTZ-043 and its main metabolites will be measured in plasma and 
urine samples by validated / qualified LC-MS methods applying appropriate stabilization measures, and all 
‘other metabolites’ will be quantified by radio-profiling. For feces, such stabilization measures are not 
feasible, but labile metabolites are also expected to be of low priority/amounts, as anticipated from rat data. 
Therefore, quantitative PK feces data of BTZ-043 and its metabolites will come from radio-profiling solely. 
Total radioactivity data and its PK evaluation will be generated as a separate report by ICON/PRA. An 
integrated PK evaluation will combine the metabolite quantification data from LC-MS measurements and 
radio-profiling for an overall evaluation of the drug’s metabolic fate and excretion pathways, and compare 
these data with the total radioactivity data.  
Note:  The term ‘other metabolites’ in the following means all BTZ-043 metabolites, except the main 
metabolites M1, M2, M4total and M10. 
15.1 Pharmacokinetic Variables 
PK concentrations will be collected in plasma, urine, and feces. If applicable - TR concentrations will be 
measured in vomitus. 


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15.1.1 Plasma Variables 
15.1.1.1 Concentrations 
• 
Individual plasma concentrations per time point for TR (reported in ICON/PRA TFLs + CSR).  
• 
Individual plasma concentrations per time point for BTZ-043 (as measured by LC-MS, reported in 
ICON/PRA TFLs + CSR) 
• 
Individual plasma concentrations per time point for metabolites M1, M2, M4tot and M10 (as 
measured by LC-MS, reported in ICON/PRA TFLs + CSR) 
• 
Sum of plasma concentrations per time point for BTZ-043 and its metabolites M1, M2, M4tot and 
M10 (reported in ICON/PRA TFLs + CSR) 
• 
Individual plasma concentrations per Hamilton AUC-pool for individual ‘other metabolites’ (as 
measured by radio-profiling– not a part of this SAP (reported in integrated PK report) 
 
 
15.1.1.2 Parameters 
• 
PK parameters for TR, BTZ-043 (M0), M1, M2, M4total, M10 (reported in ICON/PRA TFLs +CSR), 
defined in table 1 
• 
PK parameters Σ(M0, M1, M2, M4total, M10) [reported in ICON/PRA TFLs +CSR] 
• 
PK parameters for ‘other metabolites’, Σ(parent and all metabolites) as applicable, and defined in 
table 1 (reported in integrated PK report) 
      


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Table 1: Plasma Parameters 
 
Parameter 
Description 
Analyte 
SAS Programming 
Notes 
Cmax 
Maximum plasma concentration. Observed peak 
analyte concentration obtained directly from the 
experimental data without interpolation, 
expressed in concentration units 
TR, M0, M1, M2, M4total, 
and M10 
Cmax from WNL 
Tmax 
Time to maximum plasma concentration. First 
observed time to reach peak analyte 
concentration obtained directly from the 
experimental data without interpolation, 
expressed in time units. 
TR, M0, M1, M2, M4total, 
and M10 
Tmax from WNL 
AUClast 
Area under the concentration-time curve (time 0 
to time of last quantifiable concentration). 
TR, M0, M1, M2, M4total, 
and M10, Σ(M0, M1, 
M2, M4total, M10) 
AUClast from WNL 
AUCall 
Area under the curve from the time of dosing to 
the time of the last observation. If the last 
concentration is positive AUC0-tlast = AUCall. 
Otherwise, AUCall will not be equal to AUC0-tlast as 
it includes the additional area from the last 
measurable concentration down to zero or 
negative observations 
TR, M0, M1, M2, M4total, 
and M10, Σ(M0, M1, 
M2, M4total, M10) 
AUCall from WNL 
AUCinf 
Area under the blood concentration-time curve 
(time 0 to infinity), calculated as: 
 AUCinf=AUC0-t+Clast/kel, 
where Clast is the last measurable blood 
concentration.  
TR, M0, M1, M2, M4total, 
and M10, Σ(M0, M1, 
M2, M4total, M10) 
AUCINF_obs from WNL 
 
Λz (Kel) 
Terminal phase rate constant calculated by linear 
regression of the terminal log-linear portion of the 
concentration vs. time curve.  
TR, M0, M1, M2, M4total, 
and M10 
Lambda_z from WNL 
 


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Parameter 
Description 
Analyte 
SAS Programming 
Notes 
t1/2 
Terminal phase half-life expressed in time units.  
TR, M0, M1, M2, M4total, 
and M10  
HL_Lambda_z from 
WNL 
 
AUC0-t 
Area under the blood concentration-time curve 
from time 0 to t hours post-dose. 
 
TR, M0, M1, M2, M4total, 
M10 and individual 
‘other metabolites’, 
Σ(M0, M1, M2, M4total, 
M10), Σ(M0 and all 
metabolites) 
n.a. Part of the 
integrated PK report. 
CL/F 
Apparent oral clearance, calculated as 
dose/AUCinf 
M0 
CL/F from WNL 
Vz/F 
Apparent volume of distribution at terminal phase M0 
Vz/F from WNL 
Note: AUCs will be calculated using linear up / log down, expressed in units of concentration (ng eq/mL) x time (h). 
 
15.1.2 Blood Variables 
15.1.2.1 Concentrations 
• 
Individual blood concentrations per time point for TR.  Reported in ICON/PRA TFLs + CSR.  
15.1.2.2 Parameters 
• 
Individual PK parameters for TR, as defined in table 2 
Table 2:  Blood Parameters 
Parameter 
Description 
Analyte 
SAS Programming 
Notes 
Cmax 
Maximum blood concentration of 
total radioactivity. Observed peak 
analyte concentration obtained 
directly from the experimental data 
TR 
Cmax from WNL 


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Parameter 
Description 
Analyte 
SAS Programming 
Notes 
without interpolation, expressed in 
concentration units 
Tmax 
Time to maximum blood total 
radioactivity concentration. First 
observed time to reach peak 
analyte concentration obtained 
directly from the experimental data 
without interpolation, expressed in 
time units. 
TR 
Tmax from WNL 
AUClast 
Area under the concentration-time 
curve (time 0 to time of last 
quantifiable concentration). 
TR 
AUClast from WNL 
AUCinf 
Area under the blood 
concentration-time curve (time 0 to 
infinity), calculated as: 
 AUCinf=AUC0-t+Clast/kel, 
where Clast is the last measurable 
blood concentration  
TR 
AUCINF_obs from 
WNL 
 
Λz (Kel) 
Terminal phase rate constant 
calculated by linear regression of 
the terminal log-linear portion of the 
concentration vs. time curve 
TR 
Lambda_z from WNL 
 
t1/2 
Terminal phase half-life expressed 
in time units.  
TR 
HL_Lambda_z from 
WNL 
 
AUC0-t 
Area under the blood 
concentration-time curve from time 
0 to t hours post-dose. 
 
TR 
n.a. Part of the 
integrated PK report.  


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15.1.3 Urine Variables 
15.1.3.1 Amounts Excreted 
• 
Individual cumulative amount of TR excreted in urine (Ae,urine). Reported in ICON/PRA TFLs + CSR. 
o Calculated as the urine volume times the urine concentration for each day pool and added up per subject. 
• 
Individual cumulative amount of BTZ-043 (M0) excreted in urine (Ae,urine), as measured by LC-MS. Reported in ICON/PRA TFLs + CSR. 
o Calculated as the urine volume times the urine concentration for each day pool and added up per subject. 
• 
Individual cumulative amount of M1, M2, M4tot and M10, excreted in urine (Ae,urine), as measured by LC-MS. Reported in ICON/PRA TFLs 
+ CSR. 
o Calculated as the urine volume times the urine concentration for each day pool and added up per subject. 
• 
Individual cumulative amount of Σ(M0, M1, M2, M4total, M10), excreted in urine (Ae,urine). Reported in ICON/PRA TFLs + CSR. 
o Calculated as the urine volume times the urine concentration for each day pool and added up per subject. 
• 
Inter-subject average cumulative amounts of individual ‘other metabolites’ in urine (Ae,urine), as measured by radio-profiling (see study 
protocol G-A-MET-21-002). Reported in integrated PK report. 
15.1.3.2 Parameters 
• 
PK parameters for TR, BTZ-043 (M0), M1, M2, M4total, M10, and Σ(M0, M1, M2, M4total, M10) as applicable, as defined in table 3. Reported 
in ICON/PRA TFLs + CSR 
• 
PK parameters for ‘other metabolites’ and Σ(M0 and all metabolites), as applicable, as defined in table 3. Reported in integrated PK report. 
Table 3: Urine Parameters 
Parameter 
Description 
<Analyte 
SAS Programming 
Notes 
Aet (urine) 
Total amount of drug/metabolites excreted into urine to 
time t, obtained by adding the amounts excreted over 
each collection interval per subject. 
TR, M0, M1, M2, M4total, 
M10 and Σ(M0, M1, M2, 
M4total, M10)  
Summation t1-
tn(Concentration (ng 
eq/mL)ti-
t2*volume(mL)t1-t2) 
Aet (urine) 
Total amount of drug/metabolites excreted to time t, as 
obtained from one pool sample across all subjects 
covering ≥ 90% of TR in urine. 
Individual ‘other 
metabolites’, Σ(M0 and 
all metabolites) 
n.a. 
Fe (urine) 
Fraction (%) of the administered dose excreted into 
urine Fe = (Aet (urine) / Dose) * 100 per subject. 
TR, M0, M1, M2, M4total, 
M10, Σ(M0, M1, M2, 
M4total, M10 
Aet/500 mg*100 


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Parameter 
Description 
<Analyte 
SAS Programming 
Notes 
Fe (urine) 
Fraction (%) of the administered dose excreted into 
urine Fe = (Aet (urine) / Dose) * 100, across all subjects. 
Individual ‘other 
metabolites’, Σ(M0 and 
all metabolites) 
n.a. 
CLR 
Renal clearance of parent drug calculated as Ae/AUCinf 
M0 
Ae/AUCinf 
15.1.4 Feces Variables 
15.1.4.1 Amounts Excreted 
• 
Individual cumulative amount of TR excreted in feces (Ae,feces). Reported in ICON/PRA TFLs + CSR. 
o Calculated as the feces homogenate weight times the feces homogenate concentration for each day pool and added up per subject. 
• 
Inter-subject average cumulative amount of BTZ-043 (M0) in feces (Ae,feces), as measured by radio-profiling (see study protocol G-A-MET-
21-002). Reported in integrated PK report. 
• 
Inter-subject average cumulative amounts of M1, M2, M4total, M10 and individual ‘other metabolites’ in feces (Ae,feces, as applicable), as 
measured by radio-profiling (see study protocol G-A-MET-21-002). Reported in integrated PK report. 
15.1.4.2 Parameters 
• 
PK parameters for TR, as applicable, as defined in table 4 
• 
PK parameters for BTZ-043 (M0), and individual metabolites, as applicable, as defined in table 4 
Table 4: Feces Parameters 
Parameter 
Description 
<Analyte> 
SAS Programming 
Notes 
Aet (feces) 
Total amount of radioactivity excreted into feces to time 
t, obtained by adding the amounts excreted over each 
collection interval per subject. 
TR 
Summation t1-
tn(Concentration (ng 
eq/mg)ti-
t2*weight(mg)t1-t2) 
Aet (feces) 
Total amount of drug/metabolites excreted to time t, as 
obtained from one pool sample across all subjects 
covering ≥ 90% of TR in feces. 
M0, and individual 
metabolites, Σ(M0 and 
all metabolites) 
n.a. 
Fe (feces) 
Fraction (%) of the administered dose excreted into 
feces Fe = (Aet (feces) / Dose) * 100. 
TR 
Aet/500mg*100 


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Parameter 
Description 
<Analyte> 
SAS Programming 
Notes 
Fe (feces) 
Fraction (%) of the administered dose excreted 
unchanged / as individual metabolites into feces Fe = 
(Aet (feces) / Dose) * 100 across all subjects and days. 
M0 and individual 
metabolites, Σ(M0 and 
all metabolites) 
n.a. 
15.1.5 Vomitus Variables 
15.1.5.1 Amounts Recovered 
• 
Individual cumulative amount of TR recovered from vomitus (Ae,vomitus), if applicable. Reported in ICON/PRA TFls + CSR. 
o Calculated as the vomitus homogenate volume times the vomitus homogenate concentration for day 1 of each subject. 
15.1.5.2 Parameters 
• 
PK parameters for TR as defined in table 5, if applicable. Reported in ICON/PRA TFls + CSR. 
Table 5: Vomitus Parameters 
Parameter 
Description 
<Analyte> 
SAS Programming 
Notes 
Aet (vomitus) 
Total amount of radioactivity recovered in vomitus to 
time t on Day 1. 
TR 
Summation t1-
tn(Concentration (ng 
eq/mg)ti-
t2*weight(mg)t1-t2) on 
Day 1 only 
Fe (vomitus) 
Fraction (%) of the administered dose recovered from 
vomitus Fe = (Aet (vomitus) / Dose) * 100, on Day 1. 
TR 
Aet/500mg*100 
 
 
15.1.6 Total Excretion Variables 
15.1.6.1 Amounts Recovered 
• 
Individual cumulative amount of TR recovered from urine, feces and vomitus (Ae,total). Reported in ICON/PRA TFLs + CSR. 
o Calculated as the sum of Aeurine, Aefeces and Aevomitus. 


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15.1.6.2 Parameters 
• 
PK parameters for TR as defined in table 6, if applicable. Reported in ICON/PRA TFLs + CSR. 
Table 6: Total Excretion Parameters 
Parameter 
Description 
<Analyte> 
SAS Programming 
Notes 
Aet (total) 
Total amount of radioactivity excreted in / recovered 
from urine, feces and vomitus 
TR 
Aet (total) = Aet (urine) 
+ Aet (feces) + Aet 
(vomitus) 
Fe (total) 
Fraction (%) of the administered dose excreted in / 
recovered from urine, feces and vomitus Fe = (Aet 
(total) / Dose) * 100. 
TR 
Aet (total)/500mg*100 
 


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15.2 Pharmacokinetic Summaries 
15.2.1 Pharmacokinetic Concentrations 
Measured concentrations will be listed and summarized for each sampling time point, analyte and matrix 
with calculation of arithmetic means, standard deviation and coefficients of variation, as applicable. For 
inter-subject pooled samples, descriptive statistics will not be provided. Any results below the limit of 
quantification will be reported as “BLOQ” and samples where no results were obtained will be reported as 
“NR”.  
When calculating means for purpose of descriptive statistics and plots, the following rules apply: 
• 
Numeric values > sum of BLOQ & NR  mean will be reported 
• 
Numeric values ≤ sum of BLOQ & NR and NR ≥ BLOQ  NR will be reported 
• 
Numeric values ≤ sum of BLOQ & NR and BLOQ > NR  BLOQ will be reported 
Descriptive statistics (number of subjects, arithmetic mean, standard deviation, coefficient of variation) will 
be used to summarize the plasma concentrations across the 4 subjects at each scheduled time point.  
Linear and semi-logarithmic plots of the individual plasma concentration by actual sampling time will be 
provided per subject for: 
• 
M0, M1, M2, M4total, and M10 (combined in one plot) 
• 
TR in plasma and whole blood (combined in one plot) 
• 
TR, and Σ(M0, M1, M2, M4total, M10) (combined in one plot) 
In addition, across subject data (arithmetic mean) will be provided for: 
• 
M0, M1, M2, M4total, M10 (combined in one plot) 
• 
TR in plasma and whole blood (combined in one plot) 
• 
TR, and Σ(M0, M1, M2, M4total, M10) (combined in one plot) 
These plots will show time in hours. Individual plots will use the BLOQ handling procedure described above. 
Individual plasma concentration data will be presented together with descriptive statistics. 
A separate listing with time deviations and comments will be provided. 
For MetID and the integrated PK report: 
Concentrations of M0 and metabolites derived from metabolite profiling in Hamilton pool samples of 
individual subjects will be listed in the MetID and in the integrated PK report for the pooling interval. Plasma 
exposure of M0 and all metabolites (AUC0-t) and %AUC0-t of TR, M0 and metabolites will be calculated. 
Descriptive statistics (number of subjects, arithmetic mean, standard deviation) may be used to summarize 
the plasma concentrations and exposures across the 4 subjects for the pooling interval (if appropriate). The 
Σ(M0, M1, M2, M3, M4, M10) as well as Σ(individual ‘other metabolites’) may be listed (if appropriate). 
15.2.2 Pharmacokinetic Parameters 
Pharmacokinetic parameters will be calculated using the software Phoenix WinNonlin 7.0 or higher. TR in 
plasma and blood will be calculated in version 8.1.  
Actual times will be used for the calculation of the parameters. If the actual time or dose time is missing, 
the scheduled time may be substituted in order to calculate the PK parameters. 
The AUC calculation method will be Linear Up Log Down. Pharmacokinetic plasma parameters will be 
calculated on individual data with group means and relative standard deviation, on pool concentration data 
or on sum concentration data, as applicable. All parameters will be rounded to three significant figures 


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before presentation. Plasma concentrations below the limit of quantification (BLOQ) will be handled 
according to the following rules: 
• 
BLOQ values before the first numeric value will be set to zero. 
• 
BLOQ values after the first numeric value and before tmax will be set to missing (i.e. value will be 
ignored by Phoenix and a linear interpolation will be performed) 
• 
BLOQ values after tmax and before the last numeric value will be set to zero (Phoenix will switch 
from log interpolation to linear interpolation in this case). 
• 
BLOQ values after the last numeric value will be set to zero. 
 
Descriptive statistics (number of subjects, arithmetic mean, standard deviation, coefficient of variation) will 
be used to summarize the calculated PK parameters. For tmax (and tlast if applicable), only median will be 
presented. 
According to the Nuvisan SOP BAS_PKM_03, the elimination half-life (t1/2) will be calculated using the 
automatic detection method for profiles with at least 3 numeric data points and a coefficient of determination 
r2≥0.9025. Where these criteria are not fulfilled, the results will be reported as NC in the result tables. 
Parameters based on %AUCextra above 20% will be flagged but not excluded from descriptive statistics. 
15.2.3 Pharmacokinetic Amounts Excreted 
Urinary and fecal TR PK parameters will be calculated using SAS. Excretions in mg equivalent (mg eq) and 
% of dose for each interval (Aet1-t2) will be derived from the urinary and fecal concentrations and the sample 
collection interval volumes or weights respectively, by multiplying the concentration in the interval (C) and 
the volume or weight (V): 
 
Aet1-t2 = C x V 
If the concentration is <LLOQ the excretion will be assumed to be zero. For intervals in which no sample 
was collected (e.g., no defecation), the excretion will be assumed to be zero. If needed, for cumulative 
excretions, the last observation carried forward (LOCF) method will be used up to the last time point.  
Cumulative excretions for each continuously sampled interval will be calculated by summarizing the 
excretions in the current and all previous collection intervals: 
 
 
For subjects with an incomplete mass balance on Day 8, the cumulative excretion up to each interval (Day 
12 and Day 16) will be supplemented by estimating the area under the excretion rate versus time curve 
(AURC) for intervals between the in house period and ambulant visits.  
The excretion rate (∆Ae/∆t) will be calculated by dividing the excretion in each interval by the time difference 
in hours between the end and the start of the collection interval: 
 
 
 
Aeurine and Feurine:  Descriptive statistics (number of subjects, arithmetic mean, standard deviation, 
coefficient of variation) will be used to summarize the urine amounts excreted as TR, BTZ-043 (M0), M1, 
M2, M4total, M10, and Σ(M0, M1, M2, M4total, M10) by the 4 subjects for each day pool, as well as for the 
total collection periods. No statistics will be available for the urine amounts excreted as individual ‘other 
metabolites’ due to the pooled analysis of the radio-profile. The same holds true for the overall sum of 
excreted amounts of metabolites in urine. 
Aefeces and Fefeces:  Descriptive statistics (number of subjects, arithmetic mean, standard deviation, 
coefficient of variation) will be used to summarize the feces amounts excreted as TR by the 4 subjects for 


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each day pool, as well as for the total collection periods. No statistics will be available for the feces amounts 
excreted as BTZ-043 (M0), M1, M2, M4total, M10 and individual ‘other metabolites’ due to the pooled 
analysis of the radio-profile. The same holds true for the sum of excreted amounts of metabolites. 
Aetotal and Fetotal:   Descriptive statistics (number of subjects, arithmetic mean, standard deviation, 
coefficient of variation) will be used to summarize the overall excreted amounts of TR by urine, feces (and 
vomitus, if applicable) of the 4 subjects over the total collection period. No statistics will be available for the 
overall excreted amounts of BTZ-043 (M0), M1, M2, M4total, M10, individual ‘other metabolites’ and overall 
sum of metabolites. 
Linear plots of the cumulative TR (Fe) in urine, feces and total amounts excreted per day pool will be 
provided by subject and as mean over 4 subjects. These plots will show time in hours. 
Individual total radioactivity excretion data (concentrations, volumes, weights, amount excreted and 
excretion rate [if applicable]) will be listed per collection interval. 
 
 


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16.0 Safety Analyses  
16.1 Safety Variables 
The following safety variables will be summarized: 
• 
Adverse Events (AEs) 
• 
Vital Signs 
o Supine Blood Pressure  
 
Systolic Blood Pressure (SBP) 
 
Diastolic Blood Pressure (DBP) 
o Pulse rate 
o Temporal temperature 
o Respiratory rate 
• 
Electrocardiograms (ECG) 
o Heart Rate 
o PR Interval 
o QRS-Duration 
o QT Interval 
o QTc (Fridericia’s) Interval 
• 
Clinical Laboratory Evaluations 
o Clinical Chemistry  
o Hematology 
o Urinalysis 
• 
Physical Examination  
16.1.1 Adverse Events 
All AE summaries will include only treatment emergent adverse events. Treatment-emergent adverse 
events (TEAE) are those which occur after the first dose of study drug. 
A summary of the number of events, and the number and percentage of subjects reporting each TEAE, 
categorized by system organ class (SOC) and preferred term coded according to the MedDRA (Version 
24.1) will be presented by treatment. Subjects will only be counted once within each SOC or preferred term. 
This summary will be provided for all TEAEs and for related TEAEs. 
Summary tables will present the counts in descending order by SOC and preferred term (within a SOC) 
based on the number of subjects experiencing the event.  
A summary of the number of events, and number and percentage of subjects reporting each TEAE, 
categorized by relationship to study drug as recorded on the eCRF, will be provided.  
A summary of the number of events, and number and percentage of subjects reporting each TEAE, 
categorized by severity as recorded on eCRF, will also be provided.  
A listing of adverse events leading to study discontinuation will be provided. 
All adverse events (including non-treatment-emergent events) recorded on the eCRF will be listed. 
The following missing data will be imputed as defined (for calculations only / will not be presented): 
• 
Missing AE start and / or end times for the calculation of onset and duration will be assumed to be 
at 00:01 for a start time and 23:59 for end times. If the missing AE start time occurs at day of dosing, 
it will be set to one minute after dosing  
• 
Missing AE start times for the determination of treatment emergence will be assumed to occur after 
treatment unless partial date documents the AE as happening prior to treatment 
• 
Missing AE start date will be assumed to be after treatment for the determination of TEAE  
• 
Missing AE severity or relationship will be assumed to be severe or related, respectively 
 


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16.1.2 Deaths and Serious Adverse Events 
A listing of deaths and other serious adverse events (SAE) will be provided by subject. 
16.1.3 Laboratory Data 
Clinical laboratory data will be presented using units from the study data tabulation model (SDTM) 
Controlled Terminology. 
All laboratory data and comments will be listed, including laboratory variables not listed in the protocol (ad 
hoc measurements). A separate listing, including out-of-range values will also be provided. Normal ranges 
will be used directly from the clinical laboratory. 
Descriptive statistics summarizing continuous laboratory results of clinical chemistry, hematology and 
coagulation (observed and derived changes from baseline) by treatment and scheduled time will be 
provided. 
Laboratory data that are <x or >x (e.g. “<1.03”, “>1000”): the analysis value or normal limit value will be the 
value of the detection limit itself plus or minus one precision unit for the parameter concerned (respectively 
1.02 and 1001 in the example). The values before imputation (“<1.03”, “>1000”) will only be shown in 
listings. 
16.1.4 Vital Signs 
Descriptive statistics will be provided to summarize vital signs (observed values and changes from 
baseline) by treatment and scheduled time.  
All vital signs measurements will be listed.  
16.1.5 Electrocardiograms 
All ECG parameters and the corresponding abnormalities and physician’s conclusions will be listed by 
subject. 
Descriptive statistics will be provided to summarize ECG parameters (observed values and changes from 
baseline) by treatment and scheduled time. 
16.1.6 Other Observations Related to Safety 
Findings at screening and changes from screening for the physical examination will be listed. 
17.0 References 
SAS Institute, Inc., SAS® Version 9.4 software, Cary, NC. 
Clinical Study Protocol. A SINGLE-CENTER, OPEN-LABEL STUDY TO INVESTIGATE THE MASS 
BALANCE, EXCRETION PATHWAYS AND METABOLITES AFTER A SINGLE ORAL DOSE OF 500 MG, 
3.7 MBq, [14C]BTZ-043 IN HEALTHY MALE VOLUNTEERS. Version 2.0, Final, 01-Jul-2021. 


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Appendix 1: Glossary of Abbreviations 
 
Glossary of Abbreviations: 
AE 
Adverse event 
ADaM 
Analysis data model 
ADRG 
Analysis data reviewer’s guide 
AME 
Absorption, metabolism, and excretion 
BMI 
Body mass index 
BQL 
Below the quantifiable limit 
CDISC 
Clinical Data Interchange Standard Consortium 
CI 
Confidence interval 
CSR 
Clinical study report 
CV 
Coefficient of variation 
DBP 
Diastolic blood pressure 
ECG 
Electrocardiogram 
eCRF 
Electronic case report form 
EDS 
Early Development Services 
ICH 
The International Conference on Harmonization of Technical Requirements 
for Registration of Pharmaceuticals for Human Use 
LLOQ 
Lower limit of quantification 
LOCF 
Last observation carried forward 
max 
Maximum 
MedDRA 
Medical Dictionary for Regulatory Activities 
min 
Minimum 
n 
Number of observations 
PCR 
Polymerase chain reaction 
PK 
Pharmacokinetic 
QA’d 
Quality assured 
QC’d 
Quality controlled 
SAE 
Serious adverse event 
SAP 
Statistical analysis plan 
SARS-CoV-2 
Severe acute respiratory syndrome coronavirus 
SBP 
Systolic blood pressure 
SD 
Standard deviation 
SDTM 
Study data tabulation model 


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SOC 
System Organ Class 
TEAE 
Treatment-emergent adverse event 
TFL(s) 
Tables, figures and listings 
WHO 
World Health Organization 
WNL 
WinNonlin 
For PK parameters please refer to Section 16 


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Appendix 2: Schedule of Assessments 
Table 2 
Schedule of Assessments  
Visit 
Screenin
g 
Treatment Perioda 
 
24-hour Visitsa 
Follow-up  
Study Day 
Days 
-21 to -2 
Day -
1 
 
Day 1 
 
Day 2 
Day 3 Day 4 Day 5 
 
Day 
6m 
 
Day 
7m 
 
Day 
8m 
 
Day 
11m 
 
Day 
12m  
 
Day 
15m 
 
Day 
16m 
Day 31 
(±2 days)  
Confinementa 
 
X 
X 
X 
X 
X 
X 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
 
Ambulatory 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
Admission 
 
X 
 
 
 
 
 
 
 
 
(X) 
 
(X) 
 
 
Dischargeb 
 
 
 
 
 
 
X 
(X) 
(X) 
(X) 
 
(X) 
 
(X) 
 
Informed Consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical History 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Physical Examinationc 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
X 
Body Weight  
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
Height and BMI 
Calculation 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Serology (HBsAg, anti-
HCV, anti-HIV 1 and 2) 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
SARS-CoV-2 PCRd 
 
X 
 
X 
 
 
 
 
 
 
(X) 
 
(X) 
 
 
Drug and Alcohol 
Screen 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical Laboratorye 
X 
X 
 
X 
 
X 
 
 
 
 
 
 
 
 
X 
Urinalysis 
X 
X 
 
X 
 
X 
 
 
 
 
 
 
 
 
X 
12-lead ECGf 
X 
X 
X 
X 
 
 
X 
(X) 
(X) 
(X) 
 
(X) 
 
(X) 
X 
Vital Signsg 
X 
X 
X 
X 
 
 
X 
(X) 
(X) 
(X) 
 
(X) 
 
(X) 
X 
Eligibility Check 
X 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Study Drug 
Administration 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Plasma Sampling for 
BTZ-043 and Main 
Metabolitesh 
 
 
X 
X 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Whole Blood Sampling 
for Radioactivityi 
 
 
X 
X 
X 
X 
X 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
 
 
 
(X) 
 
 
 


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Visit 
Screenin
g 
Treatment Perioda 
 
24-hour Visitsa 
Follow-up  
Study Day 
Days 
-21 to -2 
Day -
1 
 
Day 1 
 
Day 2 
Day 3 Day 4 Day 5 
 
Day 
6m 
 
Day 
7m 
 
Day 
8m 
 
Day 
11m 
 
Day 
12m  
 
Day 
15m 
 
Day 
16m 
Day 31 
(±2 days)  
Plasma Sampling for 
Total Radioactivity and 
Metabolite Profilingi 
 
 
X 
X 
X 
X 
X 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
 
 
 
(X) 
 
 
 
Urine Collection and 
Pooling for BTZ-043 
and Main Metabolites, 
Total Radioactivity, 
and Metabolite 
Profilingj 
 
X 
X 
X 
X 
X 
X 
 
 (X) 
 
 
(X) 
 
 
(X) 
 
 
(X) 
 
 
(X) 
 
 
(X) 
 
 
(X) 
 
 
Feces Collection for 
Total Radioactivity and 
Metabolite Profilingk 
X 
X 
X 
X 
X 
X 
X 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
 
Vomit Collection for 
Total Radioactivity, if 
availablel 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Previous and 
Concomitant 
Medication 
X 
X 
X 
X 
X 
X 
X 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
 
(X) 
X 
AE Monitoring 
 
X 
X 
X 
X 
X 
X 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
(X) 
X 
 
AE=adverse event; BMI=body mass index; ECG=electrocardiogram; HBsAg=hepatitis B surface antigen; HCV=hepatitis C virus;PCR=polymerase chain reaction; 
PK=pharmacokinetic(s) 


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a 
 
b 
c 
 
d 
 
 
e 
f 
g 
 
h 
i 
 
 
 
 
j 
 
 
k 
 
 
 
l 
m
Subjects w ill be in the clinic from Day -1 (predose) until Day 5 w ith possible extension (Day 6 to Day 8) as w ell as possible 24-hour visits (Day 11-12, Day 15-
16). The 2 back-up subjects w ill stay in the clinic until the morning of Day 2. 
Subjects w ill be released from the clinic 96 hours after dose administration and upon satisfactory recovery of radioactivity. 
Complete physical examinations w ill be conducted at screening, Day -1, and follow -up/early termination. Symptom driven physical examinations may be 
conducted at any time, per the Investigator’s discretion.  
Sampling of nasal and throat mucosal cells. If deemed necessary, additional tests may be conducted during the study. The nonclinical project manager w ill be 
informed immediately upon availability  
of Day 11 results, as this w ill qualify the feces samples for metabolite profiling w ith regard to potential COVID-19 infectivity. 
Clinical laboratory tests (including clinical chemistry, hematology, and urinalysis): at screening; on Day -1 (admission), Day 2, and Day 4; and at follow -up. 
12-lead ECG: at screening; Day -1, and 1 hour, 4 hours, and 24 hours postdose; at discharge; and at follow -up.  
Vital signs (supine systolic and diastolic blood pressure, pulse, body temperature, and respiratory rate): at screening; at admission; at predose and at 1 hour, 
4 hours, and 24 hours postdose; at discharge; and at follow -up. 
Plasma sampling for PK of BTZ-043 and main metabolites in plasma: at predose and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, and 48 hours postdose. 
Total radioactivity in w hole blood and plasma: at predose and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, 48, 72, and 96 hours postdose (in case total radioactivity 
recovered is still <90%: in addition 120, 144, and 168 hours postdose and at further 24-hour visits [264 and 360 hours postdose] until study completion). 
Metabolite profiling in plasma: at predose and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, 48, 72, and 96 hours postdose (in case total radioactivity recovered is 
still <90%: in addition 120, 144, and 168 hours postdose. Metabolite profiling in plasma: at predose and 0.25, 0.5, 1, 1.5, 2, 3, 5, 8, 12, 16, 24, 36, 48, 72, and 
96 hours postdose (in case total radioactivity recovered is still <90%: in addition 120, 144, and 168 hours postdose. 
Urine collection and pooling for BTZ-043 and main metabolites, total radioactivity, and metabolite profiling: at predose (w ithin 12 hours prior to dosing) and 
over 0-24, 24-48, 48-72, and 72-96 hours postdose collection intervals (in case total radioactivity recovered is still <90%, in addition 96-120, 120-144, and 144-
168 hours postdose and at further 24-hour intervals [Day 11-12 and Day 15-16] until study completion). 
Feces collection for total radioactivity and metabolite profiling; predose feces collection to be collected at any time w ithin 48 hours prior to study drug 
administration (at home or in the clinic). After administration of BTZ-043, feces w ill be collected in 24 hour collection intervals up to 96 hours postdose (in case 
total radioactivity recovered is still <90%: in addition 120, 144, and 168 hours postdose and at further 24-hour intervals [Day 11-12 and Day 15-16] until study 
completion). 
If produced, vomit w ill be collected up to 8 hours after the administration of the study drug for total radioactivity determination.  
Day 6-8 procedures w ill only be conducted in case that the recovery of radioactivity w as not satisfactory until Day 5. 24-hour visits procedures w ill only be 
conducted in case that the recovery of radioactivity w as not satisfactory until Day 8. For the 24-hour visits (if applicable): feces collection to be collected at 
home w ithin 48 hours prior to admission. 
 
 


Statistical Analysis Plan 
NVS543EC-185431 
Protocol: LMU-IMPH-BTZ-043-03 
Version Date: 02-Dec-2021  
EDSREP 009 T 01 G 
Page 30 of 33 
Appendix 3: List of In-Text Outputs 
 
For PK, TFLs and CSR are restricted to TR, BTZ-043 (M0), M1, M2, M4tot, M10, and Σ(M0, M1, M2, M4total, 
M10).  
List of In Text Tables and Figures: 
Output 
Title  
Population Set 
Table  
Subject Disposition 
Safety  
Table 
Demographic Parameters 
Safety, PK 
Table 
PK parameters of BTZ-043 and Main Metabolites in Plasma 
PK 
Figure 
Arithmetic Mean Plasma Concentrations Versus Time Profiles 
of BTZ-043 and Main Metabolites (Linear and Semi-
Logarithmic Scale) 
PK 
Table 
PK parameters and Total Radioactivity in Plasma and Whole 
Blood 
PK 
Figure 
Arithmetic Mean Plasma and Whole Blood Concentrations 
Versus Time Profiles of Total Radioactivity (Linear and Semi-
Logarithmic Scale) 
PK 
Table 
Excretion Parameters of Total Radioactivity in Urine, Feces 
and Sum of Excreta 
PK 
Figure 
Mean Cumulative Excretion vs Time Profiles of Total 
Radioactivity in Urine, Feces and Sum of Excreta 
PK 
Table  
Urine PK parameters 
PK 
Table 
Treatment Emergent Adverse Events by SOC and Preferred 
Term 
Safety 
Table 
Treatment Emergent Adverse Events by Relationship and 
Severity 
Safety 
Appendix 4: List of End of Text Outputs 
 
For PK, TFLs and CSR are restricted to TR, BTZ-043 (M0), M1, M2, M4tot, M10, and Σ(M0, M1, M2, M4total, 
M10).   
List of End of Text Tables and Figures: 
Output 
Title  
Population Set 
Section 15.1 – Disposition and Demographic Data 
Table 15.1.1 
Summary of Subject Disposition 
Safety  
Table 15.1.2.1 
Summary of Demographics 
Safety 
Table 15.1.2.2 
Summary of Demographics 
PK 
Section 15.2 – Pharmacokinetic Data 
Section 15.2.1 Plasma and Whole Blood 


Statistical Analysis Plan 
NVS543EC-185431 
Protocol: LMU-IMPH-BTZ-043-03 
Version Date: 02-Dec-2021  
EDSREP 009 T 01 G 
Page 31 of 33 
Table 15.2.1.1 
Individual Values and Descriptive Statistics of BTZ-043, Main 
Metabolites and Sum of Analytes Plasma Concentrations 
PK 
Table 15.2.1.2 
Individual Values and Descriptive Statistics of Total 
Radioactivity Plasma and Whole Blood Concentrations 
PK 
Figure 15.2.1.3 
Arithmetic Mean Plasma Concentrations Versus Time Profiles of 
BTZ-043 and Main Metabolites (Linear and Semi-Logarithmic 
Scale) 
PK 
Figure 15.2.1.4 
Arithmetic Mean Plasma and Whole Blood Concentrations 
Versus Time Profiles of Total Radioactivity (Linear and Semi-
Logarithmic Scale) 
PK 
Figure 15.2.1.5 
Arithmetic Mean Plasma Concentrations Versus Time Profiles of 
Total Radioactivity, BTZ-043 (M0) and Sum of Analytes (Linear 
and Semi-Logarithmic Scale) 
PK 
Figure 15.2.1.6 
Individual Plasma Concentrations Versus Time Profiles of 
BTZ-043 and Main Metabolites (Linear and Semi-Logarithmic 
Scale)  
PK 
Figure 15.2.1.7 
Individual Total Radioactivity Plasma and Whole Blood 
Concentrations Versus Time Profiles (Linear and Semi-
Logarithmic Scale) 
PK 
Figure 15.2.1.8 
Individual Plasma Concentrations Versus Time Profiles of Total 
Radioactivity, BTZ-043 (M0) and Sum of Analytes (Linear and 
Semi-Logarithmic Scale) 
PK 
Table 15.2.1.9 
Individual Values and Descriptive Statistics of BTZ-043, 
Metabolites and Sum of Analytes PK Plasma Parameters 
PK 
Table 15.2.1.10 
Individual Values and Descriptive Statistics of Total 
Radioactivity Plasma and Whole Blood PK Parameters 
PK  
Section 15.2.2 Mass balance and Amounts Excreted in Urine 
Table 15.2.2.1 
Individual Values and Descriptive Statistics of BTZ-043 and 
Metabolite Excretion in Urine by Collection Interval 
PK 
Table 15.2.2.2 
Individual Values and Descriptive Statistics of Total 
Radioactivity Excretion in Urine, Feces and Sum of Excreta by 
Collection Interval 
PK 
Table 15.2.2.3 
Individual Values and Descriptive Statistics of Urine PK 
Parameters 
PK 
Table 15.2.2.4 
Individual Values and Descriptive Statistics of Total 
Radioactivity Excretion Parameters in Urine, Feces and Sum of 
Excreta 
PK 
Figure 15.2.2.5 
Arithmetic Mean Profiles of Cumulative Radioactivity Excretion 
in Urine, Feces and Sum of Excreta (Linear Scale) 
PK 
Figure 15.2.2.6 
Individual Profiles of Cumulative Radioactivity Excretion in 
Urine, Feces and Sum of Excreta (Linear Scale) 
PK 
Section 15.3 – Safety Data 
15.3.1 Adverse Events 
Table 15.3.1.1   
Summary of Treatment Emergent Adverse Events by System 
Organ Class and Preferred Term 
Safety 
Table 15.3.1.2 
Summary of Treatment Emergent Adverse Events by SOC and 
Preferred Term 
Safety 
Table 15.3.1.3 
Summary of Treatment Emergent Adverse Events by 
Relationship to Study Drug 
Safety 
Table 15.3.1.4 
Summary of Treatment Emergent Adverse Events by Severity 
Safety 


Statistical Analysis Plan 
NVS543EC-185431 
Protocol: LMU-IMPH-BTZ-043-03 
Version Date: 02-Dec-2021  
EDSREP 009 T 01 G 
Page 32 of 33 
15.3.2 Lists of Deaths, Other Serious and Significant Adverse Events 
Table 15.3.2.1 
Listing of Deaths and Other Serious Adverse Events  
All Subjects 
Table 15.3.2.2 
Not part of TFL – Reserved for Narratives in CSR 
 
Section 15.3.3 –Laboratory Data 
Table 15.3.3.1 
Listing of Abnormal Laboratory Values 
Safety 
Table 15.3.3.2 
Descriptive Statistics of Laboratory Results - Clinical 
Chemistry 
Safety 
Table 15.3.3.3 
Descriptive Statistics of Laboratory Results - Hematology 
Safety 
Table 15.3.3.4 
Descriptive Statistics of Laboratory Results - Urinalysis 
Safety 
Section 15.3.4 Other Safety Parameters 
Table 15.3.4.1 
Descriptive Statistics of Vital Signs 
Safety 
Table 15.3.4.2 
Descriptive Statistics of 12-Lead Electrocardiogram 
Safety 
 
List of End of Text Listings: 
Output 
Title  
Section 16.2.1 – Disposition 
Listing 16.2.1 
Subject Disposition 
Listing 16.2.2 
Protocol deviations 
Section 16.2.3 – Excluded Subjects/ Analysis Sets 
Listing 16.2.3 
Analysis Sets 
Section 16.2.4 – Demographics and Baseline Characteristics 
Listing 16.2.4.1 
Subject Demographics 
Listing 16.2.4.2 
Medical History 
Listing 16.2.4.3 
Prior and Concomitant Medications 
Listing 16.2.4.4 
Drug and Alcohol Screen 
Listing 16.2.4.5 
Serology 
Listing 16.2.4.6 
SARS-CoV-2 PCR Test Results 
Section 16.2.5 - Compliance 
Listing 16.2.5 
Study Drug Administration 
Section 16.2.6 – Response Data 
Listing 16.2.6.1 
Individual PK Plasma and Total Radioactivity Plasma and Whole 
Blood Sampling Time Deviations and Comments 
Listing 16.2.6.2 
Individual Total Radioactivity Excretion Data in Urine and Feces, 
Including Volumes, Weights and Amount Excreted by Collection 
Interval 
Listing 16.2.6.3 
Individual BTZ-043 and Metabolites Excretion Data in Urine  
Including Volumes, and Amount Excreted by Collection Interval 


Statistical Analysis Plan 
NVS543EC-185431 
Protocol: LMU-IMPH-BTZ-043-03 
Version Date: 02-Dec-2021  
EDSREP 009 T 01 G 
Page 33 of 33 
Section 16.2.7 – Adverse Events Data 
Listing 16.2.7.1 
Adverse Events 
Listing 16.2.7.2 
Adverse Events Leading to Withdrawal 
Section 16.2.8 – Laboratory Data 
Listing 16.2.8.1 
Clinical Laboratory Results – Hematology 
Listing 16.2.8.2 
Clinical Laboratory Results – Chemistry 
Listing 16.2.8.3 
Clinical Laboratory Results – Urinalysis 
Listing 16.2.8.4 
Clinical Laboratory Results – Comments 
Section 16.2.9 – Other Safety Data 
Listing 16.2.9.1 
Vital Signs 
Listing 16.2.9.2 
12-Lead Electrocardiogram Results 
Listing 16.2.9.3 
Physical Examination – Findings and Changes 
 
Other Appendix Outputs: 
Output 
Title  
Appendix 16.1.9.2 
Statistical Appendices (if applicable) 
18.0 Document History 
Version Date 
Modified/Reviewed 
By 
Brief Summary of Changes (if created from a 
template, include template code) 
08-Apr-2021 
JWi 
Set up from template, up to section 10. Section 16 will be 
for Nuvisan 
13-Jul-2021 
Nuvisan input  
PK sections provided by Nuvisan 
28-Sep-2021 
JWi 
Version for internal review 
30-Sep-2021 
JWi 
First draft 
28-Oct-2021 
JWi 
2nd draft 
23-Nov-2021 
JWi 
Final