Assessing the Safety, Tolerability and Pharmacokinetics of Benfo-Oxythiamine (B-OT) in Healthy Volunteers - An Open Label, Phase I Study
Phase: PHASE1
Conditions: Healthy Volunteers
Sponsor: Benfovir AG

Official Study Title: Assessing the safety, tolerability and pharmacokinetics of benfo-
oxythiamine (B-OT) in healthy volunteers – an open-label, phase I study. 
NCT Number: NCT07450833 
Document Date: 13 July 2022 
Document Type: Study Protocol 
 

Study Protocol 
 
 
         Study number: BV-01-101 
 
  
   Version No. : 3.0 
 
 
 
Date: 13 Jul 2022 
CONFIDENTIAL  
 
 
 
 
 
 
 
Page 1 of 101 
 
Protocol title 
 
Assessing the safety, tolerability and pharmacokinetics of 
Benfo-Oxythiamine (B-OT) in healthy volunteers  An Open 
label, phase I Study 
Study number (Protocol ID)
BV-01-101 
Author 
Lilyana Mircheva 
Document type 
Clinical Study Protocol  
Study Phase 
I (First in Human) 
 
Clinical Research 
Organization 
Convex Ltd  
6, Tri Ushi str., 1000 Sofia, Bulgaria 
Sponsor 
benfovir AG 
Gräfenhäuserstr. 26, 64293 Darmstadt, Germany 
Document status/version 
Version 3.0 
Date: 
13-Jul-2022 
EudraCT 
2021-005616-60 
Confidentiality Statement 
 
The information contained in this document, especially unpublished data, is the property of the 
sponsor of this study, benfovir AG. It is therefore provided to you in confidence as an 
investigator, potential investigator, or consultant, for review by you, your staff, and an Ethics 
Committee or Institutional Review Board. It is understood that this information will not be 
disclosed to others without written authorization from benfovir AG, except to the extent 
necessary to obtain informed consent from those persons to whom the study drug may be 
administered. 

Study Protocol 
 
 
         Study number: BV-01-101 
 
  
   Version No. : 3.0 
 
 
 
Date: 13 Jul 2022 
CONFIDENTIAL  
 
 
 
 
 
 
 
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3 
TABLE OF CONTENTS 
1 
Signature Page for Benfovir AG ........................................................................................ 4 
2 
Signature Page for Investigator .......................................................................................... 5 
3 
Table of Contents ............................................................................................................... 6 
4 
List of Abbreviations ....................................................................................................... 11 
5 
protocol synopsis ............................................................................................................. 14 
Table 1: Visit and Assessment Schedule of the Single Ascending Dose PArt ........................ 22 
Table 2: Visit and Assessment Schedule of the Multiple Ascending Dose PArt .................... 24 
6 
Introduction and Rationale ............................................................................................... 26 
6.1 
Introduction ............................................................................................................. 26 
6.2 
Risk-Benefit and Ethical Assessment .................................................................... 29 
7 
Study Objectives .............................................................................................................. 30 
8 
STUDY Design ................................................................................................................ 31 
8.1 
Overall Study Design and plan .............................................................................. 31 
8.1.1 
Screening (SAD and MAD) ............................................................................... 33 
8.1.2 
Treatment and observation period (SAD and MAD) ......................................... 33 
8.1.3 
Follow-up (SAD and MAD) .............................................................................. 34 
8.2 
Justification for population and design ................................................................. 34 
8.3 
Proposed Safe Starting Dose .................................................................................. 35 
8.4 
Dose Escalation Strategy ........................................................................................ 36 
8.5 
Stopping Criteria for Dose Escalation ................................................................... 37 
8.5.1 
Stopping Rules for an Individual Subject, at any Time in the Study ................. 37 
8.5.2 
Stopping Rules for a Whole Cohort, Dose Escalation, Progression to Next Study 
Part and Termination of Study ......................................................................................... 37 
8.6 
Safety Review ........................................................................................................... 37 
8.6.1 
Data Reviewed by Safety Review Committee for Dose Escalation Decision ... 38 
8.6.2 
Safety Review Committee Decision on Next Dose ........................................... 38 
8.6.3 
Blinding at Safety Review Committee Meeting ................................................ 39 
8.6.4 
Assessments Adaptation .................................................................................... 39 
9 
Study Population .............................................................................................................. 40 
9.1 
Subject population .................................................................................................. 40 
9.2 
Inclusion criteria ..................................................................................................... 40 
9.3 
Exclusion Criteria ................................................................................................... 40 
9.4 
Prior and concomitant medication ........................................................................ 41 
9.5 
Dietary and other restrictions prior and during the study.................................. 42 

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9.5.1 
Contraception requirements ............................................................................... 43 
9.6 
Study drug discontinuation and subject withdrawal ........................................... 44 
9.6.1 
Study drug interruption or discontinuation ........................................................ 44 
9.6.2 
Subject withdrawal............................................................................................. 45 
9.6.3 
Replacement policy ............................................................................................ 46 
10 
Investigational product .................................................................................................... 47 
10.1 Investigational drug and comparative drug ......................................................... 47 
10.1.1 Investigational test drug ..................................................................................... 47 
10.1.2 Investigational comparative drug ....................................................................... 47 
10.2 Study drug up- and down-titration ....................................................................... 47 
10.3 Packaging and labelling .......................................................................................... 47 
10.4 Drug accountability ................................................................................................. 48 
10.5 Treatment assignment and blinding ...................................................................... 48 
10.5.1 Randomization and treatment assignment ......................................................... 48 
10.5.2 Blinding.............................................................................................................. 48 
10.6 Dosing Procedure .................................................................................................... 48 
10.6.1 Benfo-oxythiamine (B-OT) ............................................................................... 49 
10.7 Treatment Compliance ........................................................................................... 49 
11 
Study Endpoints ............................................................................................................... 50 
11.1 Primary endpoints ................................................................................................... 50 
11.2 Secondary endpoints ............................................................................................... 50 
11.3 Exploratory endpoints ............................................................................................ 50 
12 
Study assessmEnts (SAD and MAD) .............................................................................. 51 
12.1 Visit Procedures (SAD ) .......................................................................................... 51 
12.1.1 Procedures during the Screening Visit (Day -28 to -2)...................................... 51 
12.1.2 Procedures on Day -1 ......................................................................................... 51 
12.1.3 Procedures on Day 1 .......................................................................................... 52 
12.1.4 Procedures on Day 2 .......................................................................................... 52 
12.1.5 Procedures on Day 3 .......................................................................................... 53 
12.1.6 Procedures on Day 4 .......................................................................................... 53 
12.1.7 Procedures on Follow-up visit (Day 8) .............................................................. 53 
12.2 Visit Procedures (MAD ) ........................................................................................ 54 
12.2.1 Procedures during the Screening Visit (Day -28 to -2)...................................... 54 
12.2.2 Procedures on Day -1 ......................................................................................... 55 
12.2.3 Procedures on Day 1 .......................................................................................... 55 

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12.2.4 Procedures on Day 2 .......................................................................................... 55 
12.2.5 Procedures on Day 3 .......................................................................................... 56 
12.2.6 Procedures on Day 4 .......................................................................................... 56 
12.2.7 Procedures on Day 5 .......................................................................................... 57 
12.2.8 Procedures on Day 6 .......................................................................................... 57 
12.2.9 Procedures on Day 7 .......................................................................................... 58 
12.2.10 Procedures on Day 8 .......................................................................................... 58 
12.2.11 Procedures on follow-up Day 11 ....................................................................... 58 
12.2.12 Procedures on End of study visit (Day 14) ........................................................ 59 
12.3 Safety and tolerability assessments ........................................................................ 59 
12.3.1 Vital Signs .......................................................................................................... 60 
12.3.2 Weight and height .............................................................................................. 60 
12.3.3 Physical examination ......................................................................................... 60 
12.3.4 Electrocardiography ........................................................................................... 60 
12.3.5 Laboratory assessments, drug screen and breath alcohol test ............................ 60 
12.3.6 Concomitant medications................................................................................... 62 
12.4 Pharmacokinetic and pharmacodynamics assessments ...................................... 63 
12.4.1 Blood samples collection ................................................................................... 63 
12.4.2 Labelling ............................................................................................................ 67 
12.4.3 Shipping procedures........................................................................................... 67 
12.4.4 Determination of plasma drug concentrations and measurement of the 
transketolase activity ........................................................................................................ 68 
13 
Safety reporting ................................................................................................................ 70 
13.1 Definitions of adverse events .................................................................................. 70 
13.1.1 Severity of adverse events ................................................................................. 71 
13.1.2 Relationship to study drug ................................................................................. 71 
13.1.3 Chronicity of adverse events .............................................................................. 72 
13.1.4 Action ................................................................................................................. 72 
13.1.5 Outcome of adverse event .................................................................................. 72 
13.1.6 Reporting of adverse events and serious adverse events ................................... 72 
13.1.7 Follow-up of adverse events .............................................................................. 74 
13.2 Temporary halt for reasons of subject safety ....................................................... 74 
14 
STATISTICAL METHODOLOGY AND ANALYSES ................................................ 75 
14.1 Statistical analysis plan ........................................................................................... 75 
14.2 Protocol violations/deviations ................................................................................ 76 

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14.3 Power calculation .................................................................................................... 76 
14.4 Missing, unused and spurious data ........................................................................ 76 
14.5 Analysis sets ............................................................................................................. 77 
14.5.1 Safety set ............................................................................................................ 77 
14.5.2 Pharmacokinetic and pharmacodynamic analysis set ........................................ 77 
14.6 Subject disposition .................................................................................................. 78 
14.7 Baseline parameters ................................................................................................ 78 
14.7.1 Demographics and baseline variables ................................................................ 78 
14.7.2 Medical history .................................................................................................. 78 
14.8 Safety and tolerability endpoints ........................................................................... 78 
14.8.1 Adverse events ................................................................................................... 78 
14.8.2 Vital signs .......................................................................................................... 79 
14.8.3 ECG.................................................................................................................... 79 
14.8.4 Clinical laboratory tests ..................................................................................... 79 
14.9 Pharmacokinetic and pharmacodynamic endpoints analysis ............................. 79 
14.9.1 Dose proportionality .......................................................................................... 81 
14.9.2 Inferential methods ............................................................................................ 81 
14.10 
Interim analyses ................................................................................................... 81 
15 
GOOD CLINICAL PRACTICE, ETHICS AND ADMINISTRATIVE PROCEDURES
 
82 
15.1 Good clinical practice.............................................................................................. 82 
15.1.1 Ethics and good clinical practice ....................................................................... 82 
15.1.2 Ethics committee / institutional review board ................................................... 82 
15.1.3 Informed consent ............................................................................................... 82 
15.1.4 Insurance ............................................................................................................ 82 
15.2 Study funding .......................................................................................................... 83 
15.3 Data handling and record keeping ........................................................................ 83 
15.3.1 Data collection and confidentiality .................................................................... 83 
15.3.2 Database management and quality control ........................................................ 84 
15.4 Access to source data and documents .................................................................... 85 
15.5 Quality control and quality assurance .................................................................. 85 
15.5.1 Monitoring ......................................................................................................... 86 
15.6 Protocol amendments .............................................................................................. 87 
15.6.1 Substantial amendment ...................................................................................... 87 
15.6.2 Non-substantial amendment............................................................................... 87 

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15.6.3 Urgent amendment ............................................................................................. 87 
15.7 Clinical study report ............................................................................................... 88 
15.8 End of study report ................................................................................................. 88 
15.9 Public disclosure and publication policy ............................................................... 88 
16 
REFERENCES ................................................................................................................ 89 
17 
Annexes ........................................................................................................................... 92 
17.1 Annex No 1 ............................................................................................................... 92 
17.2 Annex No 2 ............................................................................................................... 97 
 
 

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4 
LIST OF ABBREVIATIONS 
ADR 
Adverse Drug Reaction 
AE 
Adverse Event 
ALT 
alanine aminotransferase/serum glutamic pyruvic transaminase (SGPT)
ANOVA 
Analysis of variance 
AST/SGOT 
Aspartate aminotransferase 
ATD 
Anticipated Therapeutic Dose Range 
AUC 
Area Under the Curve  
BASO 
Basophil 
B-OT 
Benfo-oxythiamine 
BMI 
Body Mass Index 
BP 
Blood Pressure 
Bpm 
beats per minute 
BUN 
Blood Urea Nitrogen 
CA 
Competent authority  
Cl/F 
Clearance 
Cmax 
Maximum concentration 
EC 
Ethics Committee 
eCRF 
electronic Case Report Form 
EOS 
Eosinophil 
CSP 
Clinical Study Protocol 
CSR 
Clinical Study Report 
CV 
Coefficient of Variation 
DBP 
Diastolic Blood Pressure 
DCC 
Diagnostics and Consultation Centre 
DMP 
Data Management Plan 
EC 
Ethics Committee  
ECG 
Electrocardiogram 
EDTA 
Ethylene Diamine Tetra-acetic Acid 
EOS 
End of study 
EU 
European Union 
FIH 
First in human 
GCP 
Good Clinical Practice 
GLP 
Good Laboratory Practices 
Gamma-GT 
Gamma-glutamyltransferase 
HCT 
Hematocrit 
HGB 
Hemoglobin 
Hem 
Hematology 
Hep B 
Hepatitis B 
Hep C 
Hepatitis C 
HIV 
Human Immunodeficiency Virus 

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HR 
Heart Rate 
hr. 
Hour 
IB 
Investigator  Brochure 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonization 
IMP 
Investigational Medicinal Product 
kg 
Kilogram 
LLOQ 
Lower Limit of Quantification 
LYMP 
Lymphocyte 
MAD 
Multiple Ascending Dose 
MedDRA 
Medical Dictionary for Regulatory Activities 
MCV 
Mean Corpuscular volume 
MCH 
Mean Corpuscular Haemoglobin 
MCHC 
Mean corpuscular haemoglobin concentration 
MONO 
Monocyte 
NEUT 
Neutrophil 
NOAEL 
No Observed Adverse Effect Level 
OTC 
Over-the-counter 
OT 
Oxythiamine 
PAD 
Pharmacologically Active Dose 
PD 
Pharmacodynamic 
PE 
Physical examination 
PI 
Principal Investigator 
PK 
Pharmacokinetic 
PLT 
Platelet 
PPP 
Pentose phosphate pathway 
PR 
Pulse Rate 
PT 
Preferred Term 
RBC 
Red Cell Count 
R5P 
Ribose-5-Phosphate 
SAD 
Single Ascending Dose 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SAR 
Serious Adverse Reaction 
SAS 
Statistical Analysis Software 
SOC 
System Organ Class 
SOP 
Standard Operating Procedure 
SRC 
Safety Review Committee 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
TEAE 
Treatment Emergent Adverse Event 
TK 
Toxicokinetic 

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TKT 
Transketolase 
TKTL1 
Transketolase-like 1 
t1/2 
Terminal half life 
tmax 
Time of the maximum concentration 
UADR 
Unexpected Adverse Drug Reaction 
UAE 
Unexpected Adverse Event 
ULOQ 
Upper limit of quantification 
WBC 
White Cell Count 
WHO 
World Health Organization 
 
 

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5 
PROTOCOL SYNOPSIS 
 
Study Title 
Assessing the safety, tolerability and pharmacokinetics of Benfo-
Oxythiamine (B-OT) in healthy volunteers  An Open label, phase 
I Study 
Study Type 
Monocentric, open label, first-in-man phase I study  
Protocol No. / Short 
Title 
BV-01-101 / BOOST 
EUDRA-CT No 
2021-005616-60 
Targeted Indication 
SARS-CoV-2 infection / Covid-19 
Investigational Drug 
Benfo-oxythiamine (B-OT) 
B-OT is a novel thiamine analogue, which functions as a prodrug 
and releases oxythiamine in the human body. The introduction of a 
benfo group into a molecule improves its cell permeability and may 
ensure better bioavailability. The thiamine antagonist oxythiamine 
is an irreversible inhibitor of thiamine-dependent enzymes, such as 
transketolases, and efficiently inhibits the sugar metabolism and the 
accumulation of Ribose-5-Phosphate (R5P) building blocks.  
Sponsor 
benfovir AG 
Gräfenhäuserstr. 26,  
64293 Darmstadt 
Germany 
 
 

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Objectives and 
end point 
Primary Objectives: 
To assess the safety and tolerability of single and multiple ascending 
doses of B-OT 
 Endpoints: 
o safety evaluations including safety laboratory 
assessments, physical examination (PE), 
electrocardiograms (ECGs), vital signs, adverse events 
(AEs) and serious adverse events (SAEs). 
Secondary Objectives 
 To evaluate the pharmacokinetics (PK) of benfo-oxythiamine
(B-OT) and its active metabolite oxythiamine (OT) in plasma 
following single doses administered to healthy volunteers  
 To evaluate the pharmacokinetics (PK) of benfo-oxythiamine 
(B-OT) active metabolite oxythiamine (OT) in plasma following 
multiple doses administered to healthy volunteers. 
 Endpoints: 
 
o SAD: Plasma PK parameters, including Cmax, tmax, 
Vz/F, AUC0-inf and AUC last as 
appropriate 
 
o MAD: The following PK endpoints will be derived 
from plasma oxythiamine (OT) concentration versus 
time data following administration according to the 
7: 
Vz/F, Ctrough, Cav, Cmin,  t1/2, AUCinf and AUClast 
as appropriate 
 
 
Exploratory Objectives 
 To evaluate the pharmacodynamics (PD) by measuring the 
transketolase activity in  
 To analyse PK parameters of other B-OT derivative 
metabolite(s), if any. 
 
 

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Study Design,  
Mode 
of 
administration, 
starting dose and 
dose level, 
Stopping rule 
This is a monocentric, open-label, non-randomized, uncontrolled, first-
in-human study to assess the safety, tolerability, and pharmacokinetics 
/pharmacodynamics of B-OT in healthy subjects.  
This study has two parts: a single ascending dose (SAD) part and a 
multiple ascending dose (MAD) part. 
Administration and dose levels: 
First Part of the study: SAD 
In the SAD part, a single dose of B-OT will be administered per oral 
(p.o.) once daily on day 1 followed by 7 days without dosing.  
Up to 5 cohorts will be enrolled and dosed. Dosing is based on 
NOAEL x HED x 1/10 (No Observed Adverse Effect Level x Human 
Equivalent Dose [dog/human] x Safety Factor). As NOAEL was 
established in dog after a 7-day treatment and B-OT has already been 
used in compassionate use at a minimum daily dose of 3 mg for 7 days 
in 5 Covid-19 patients and 5 late-stage prostate cancer patients without 
any safety issue, only 3 subjects will receive the two first lowest doses.
The first dose of B-OT will be 0.5 mg (0.0083 mg/kg body weight, 
average 60 kg adult). After the first cohort, enrolment will be halted. If 
no safety issue is observed, the second dose level will be 1 mg (0.016 
mg/kg body weight). The first and second dose level cohort will have 3 
subjects. The human PK/PD model will be refined before enrolling to 
the next dose level. As this is the first study testing B-OT in humans, 
study design adjustments may be made based on emerging data from 
each dose cohort based on review of preliminary safety, tolerability, and 
PK/PD results. If these data allow for, the dose escalation scheme of 2 
mg, 3 mg and 5 mg will follow in 3 additional cohorts of 6 subjects.   
Each cohort will start with a sentinel subject (subject #1).  
Subject #2 - #3 or #2 - #6 of the same cohort will start treatment after a 
minimum of 48 hours if safety data indicate that dosing of subsequent 
subjects of this cohort is justified. 
 

Study Protocol
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The following figure represents the study design of the SAD part: 
The filled box represents a dosing event, a white box a represents a 
dosing free day. Single dose treatment will be administered on Day 1.
Dose escalation to the next dose level will occur if three (first and 
second cohort) or six subjects (3 next cohorts) have completed the 
single dose treatment and if PK as well as safety data allow an escalation 
to the next dose level.
Second Part of the study: MAD
In the MAD part, a single dose of B-OT will be administered orally 
(p.o.) once daily for 7 days (D1-D7) followed by 7 days without dosing. 
The first dose will be 1 mg, as the PK and safety data in the SAD part 
of the study have shown an acceptable safety margin at this dose (see 
IB).
Each cohort will start with a sentinel subject (subject #1). 
Subjects #2 - #6 of the same cohort will start treatment after a minimum 
of 72 hours if no adverse event is observed in the sentinel subject.
The following figure represents the study design of the MAD part:

Study Protocol
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Definite Stopping rules
Dose escalation will not continue to the next dose level if (a) maximal 
OT plasma levels are achieved, i.e. further increase is not justified from 
a scientific point of view, or if (b) toxicities are observed that do not 
justify continuation from an ethical point of view.
The study will be halted if one or more subjects experience a serious 
adverse event (SAE) that is considered as related to B-OT or 2 or more 
subjects in the same group experience severe AEs that are considered 
to be related to B-OT.
Dose-escalation will stop if clinically relevant signs or symptoms of 
similar nature occur in 2 or more subjects in a group that, in the opinion 
of the PI, warrant stopping of dose-escalation.
Inclusion criteria
Subjects will be enrolled if they meet all the following inclusion 
criteria:
- Signed and dated informed consent form
- Subjects capable to understand the purposes and risks of the study, and 
who are willing and able to participate in the study 
- Male volunteers who are willing to comply with contraception 
requirements
- Aged 18-60 years, inclusive
- Healthy participants, as determined by screening assessments and 
Principal Investigator's judgment
- Healthy status is defined by the absence of evidence of any active or 
chronic disease following a detailed medical and surgical history, a 
complete physical examination including vital signs, 12-lead ECG, 
hematology, blood chemistry, serology, and urinalysis
- Body Mass Index (BMI) of 18-30 kg/m2 inclusive
Contraception Requirements

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- Male participants with female partners of childbearing potential are 
eligible to participate if they agree to one of the following during the 
protocol defined time frame:  
- Be abstinent from penile-vaginal intercourse as their usual and 
preferred lifestyle (abstinent on a long term and persistent basis) and 
agree to remain abstinent 
- Use a male condom plus partner use of a contraceptive method with a 
failure rate of < 1% per year when having penile-vaginal intercourse 
with a woman of childbearing potential who is not currently pregnant. 
Note: Men with a pregnant or breastfeeding partner must agree to 
remain abstinent from penile-vaginal intercourse or use a male condom 
during each episode of penile penetration. 
Exclusion 
criteria 
- Subjects who meet any of the following exclusion criteria at 
Screening (unless otherwise stated) will not be eligible for participation: 
- Subjects who have a clinically relevant history as determined by the 
Investigator, or presence of respiratory, gastrointestinal, renal, hepatic, 
hematological, lymphatic, neurological, cardiovascular, psychiatric, 
musculoskeletal, 
genitourinary, 
immunological, 
dermatological, 
endocrine, connective tissue diseases, or disorders. 
- Fridericia's correction factor for QT (QTcF) > 450 ms or history of QT 
interval prolongation. 
- Have acute gastrointestinal symptoms (e.g., nausea, vomiting, 
diarrhea, heartburn) at the time of screening or admission. 
- Has any abnormal laboratory value of Grade 2 or higher, which is 
considered as clinically significant by the PI (or designee). 
- Has values of 10 % above the upper limit of normal for the following 
laboratory analytes: 
- 
alanine aminotransferase (ALT/SGPT), alkaline phosphatase 
(serum), aspartate aminotransferase (AST/SGOT), at 
Screening and Day-1  
- 
creatinine and urea concentration in blood at Screening and 
Day-1 
- Have a clinically relevant surgical history, as determined by the 
Investigator. 
- Have a history of relevant drug hypersensitivity. 
- Have a history of alcoholism or drug abuse. 
- Have a significant infection or known inflammatory process at 
screening or admission. 

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- Have used any prescription or non-prescription medicines within 2 
weeks of admission, unless in the investigator's opinion it will not affect 
the determination of safety or other study assessments. 
- Have received any investigational drug within 30 days prior to 
screening. 
- Have used tobacco or nicotine products within 3 months of screening 
- Has a positive alcohol or drug screen at Screening or the Day1 visit  
- Have donated or received any blood or blood products within the 3 
months prior to screening. 
- Cannot communicate reliably with the investigator. 
- Have been vaccinated with a Covid-19 vaccine within 2 weeks prior 
to screening and/or during participation in the clinical trial 
- Are unlikely to co-operate with the requirements of the study.  
Statistical 
considerations 
and sample size 
The study is exploratory and is not powered to address any pre-defined 
hypotheses. The assessment of safety and tolerability will be performed 
on the safety analysis set, which includes all subjects who received at 
least one dose of B-OT. Complete description of all statistical analyses 
and methods will be presented in the Statistical Analysis Plan (SAP). 
The SAP will be reviewed and approved by the Sponsor and will be 
finalized prior to database lock. Plans for PK analyses will be included 
in the SAP. 
Determination of Sample Size:  
The sample sizes for the SAD and MAD cohorts are typical for a Phase 
1 first in human (FIH) study. 
The total number of subjects accrued in the study will depend on the 
number of dose levels, but is currently set to a maximum of 48 subjects 
(in the SAD: 2 cohorts with 3 subjects and 3 cohorts with 6 subjects 
each and in the MAD: 4 cohorts of 6 subjects each). The safety and 
tolerability of B-OT will be analyzed by appropriate descriptive 
statistics. All other secondary endpoints will be summarized 
descriptively. 
Study Populations: 
Safety Population: All subjects who receive at least one dose of study 
drug. 
Pharmacokinetic Population: All subjects receiving study drug who 
comply sufficiently with protocol requirements (i.e., no major protocol 
deviations) and for whom enough analyzable PK samples have been 
obtained to allow the determination of the PK profile for B-OT/OT and 
the statistical analysis of the results. 

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Safety Analyses: Statistical methods for the safety analyses will be
descriptive in nature. Safety data, including AEs, clinical laboratory 
data, vital signs, ECG parameters, and PEs. All appropriate AEs will be 
graded using the DMID toxicity scale (March 2014). Change from 
baseline will be included in summary tables for laboratory parameters. 
All laboratory data will be included in the data listings and all test values 
outside the normal range will be flagged. 
Pharmacokinetic Analyses: Plasma PK parameters for each dose level 
in the SAD part of the study will be calculated from the concentrations 
of B-OT and OT measured in pre-dose and post-dose plasma samples. 
For each dose level, descriptive statistics will be presented. Figures will 
be created to display mean and individual subject B-OT and OT 
concentration versus time. Plasma PK parameters for each dose level in 
the MAD part of the study will be calculated from the concentrations of 
OT and possible derivate metabolites. Figures will be created to display 
mean and individual subject OT concentration versus time. 
Interim Analyses: No formal interim analyses are planned for this 
study. Data from SAD cohorts were frozen and analyzed prior to 
conducting the second MAD part of the study. 

Study Protocol 
 
 
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TABLE 1: VISIT AND ASSESSMENT SCHEDULE OF THE SINGLE ASCENDING DOSE PART 
Visit 
Screening  
(Days -28 to -
2) 
Day -
1 
 
 
 
Day 1 
 
(T=0) 
  
Day 2 
 
(24 hr)
+/-
30min 
Day 3
 
(48 
hr) 
+/-1hr
Day 4
 
(72 
hr) 
+/-1hr
Day 8 
(EOS) 
 
(168 hr)
+/-2hr 
  
 
Informed consent 
x 
  
  
  
  
  
  
  
Demography 
x 
  
  
  
  
  
  
  
I/E; Medical history 
x 
x 
  
  
  
  
  
  
Physical 
examination-PE1 
x 
x 
x2 
x 
  
x 
x 
1: a complete PE will be performed at Screening 
and/or Check-in and EOS; on all other days, a 
limited PE will target any change from the 
previous status, or any body system deemed 
pertinent to that subject by the PI (or designee) 
 
2: at 4 hr post-dose 
Vital signs3 
x 
x 
x 
x 
x 
x 
x 
3: vital signs include pulse rate, respiratory rate, 
blood pressure and body temperature; on Day1, 
VS should be taken pre-dose and at 2,4, 8 and 12 
hr post-dose 
Laboratory 
assessment  
(hematology, 
biochemistry) 
x 
x4 
  
x 
  
x 
x 
4: Results for safety blood samples must be 
available and reviewed before drug is 
administered  
Urine analysis 
x 
x 
  
x 
x 
x 
x 
  
ECG 
x 
  
x5 
x 
  
  
x 
5: the baseline ECG should be conducted prior to 
dosing on day 1 
SARS CoV antigen 
x 
  
  
  
  
 x 
  
SARS CoV PCR 
 
x 
 
 
 
x 
 
 

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Visit 
Screening  
(Days -28 to -
2) 
Day -
1 
 
 
 
Day 1 
 
(T=0) 
  
Day 2 
 
(24 hr)
+/-
30min 
Day 3
 
(48 
hr) 
+/-1hr
Day 4
 
(72 
hr) 
+/-1hr
Day 8 
(EOS) 
 
(168 hr)
+/-2hr 
  
 
Viral screening 
(HIV, Hepatitis) 
x 
  
  
  
  
  
  
  
Drug screening  
x 
x 
  
  
  
  
x 
  
Breath alcohol test 
x 
x 
 
 
 
 
x 
 
Weight/height (BMI)
x 
x6 
  
  
  
  
x6 
 6: only weight on Day -1 and 8  
Confinement 
  
x 
x 
x 
x 
x 
  
  
Ambulatory visit 
x 
  
  
  
  
  
x 
  
Administer study 
drug 
  
  
x7 
  
  
  
  
7: T=0 is time of drug administration 
Plasma PK sample 
collection 
  
  
x8 
x9 
x9 
x9 
x9 
8: PK samples should be collected pre-dose and at 
0.25, 0.5,0.75,1,1.5,2,4,8,12 hr. depending on 
ongoing PK data review, timepoints may be 
modified or removed. 
 
9: PK samples should be collected at 24,36,48, 72 
and 168 hr post-dose on Day 2,3, 4 and 8 
PD (TKT actvity) 
  
  
x10 
x 
x 
x 
x 
10: on Day 1, TKT blood sample should be taken 
pre-dose and at 1,2,4,8, and 12 hr and then at 24, 
48, 72 and 168 hr post-dose. 
Adverse Events 
x11 
x11 
x11 
x 
x 
x 
x 
11: Adverse events occurring prior to study drug 
administration should be recorded as medical 
history unless the event is directly related to 
study specific procedure 
Prior and/or 
Concomitant 
medications 
x 
x 
x 
x 
x 
x 
x 
  

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TABLE 2: VISIT AND ASSESSMENT SCHEDULE OF THE MULTIPLE ASCENDING DOSE PART  
 
Visit 
Screening 
Day 
-1 
Day 1
Day 2
Day 3
Day 4
Day 5
Day 6
Day 7
Day 8
Day 
11
Day 14 
(EOS)
  
(Days -28 to -2)
  
Informed consent 
x 
  
  
  
  
  
  
  
  
  
  
  
  
Demography 
x 
  
  
  
  
  
  
  
  
  
  
  
  
I/E; Medical history 
x 
x 
  
  
  
  
  
  
  
  
  
  
  
Ambulatory visit 
x 
  
  
  
  
  
  
  
  
  
x 
x 
  
Confinement 
  
x 
x 
x 
x 
x 
x 
x 
x 
x 
  
  
  
Administer study drug 
  
  
x 
x 
x 
x 
x 
x 
x 
  
  
  
  
Physical examination-PE1
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
1: a complete PE will be performed at Screening and/or 
Check-in and EOS; on all other days, a limited PE will 
target any change from the previous status, or any body 
system deemed pertinent to that subject by the PI (or 
designee) 
Vital signs2 
x 
x 
x 
x3 
x 
x 
x3 
x 
x3 
x 
x 
x 
2: vital signs include pulse rate, respiratory rate, blood 
pressure and body temperature; 
if not otherwise indicated, VS should be taken either at 
pre-dose on dosing days or at time of visit on ambulatory 
visit days 
3: on Day 2, 5 and 7, VS should be taken pre-dose and at 4 
hr post-dose 
Laboratory assessment 
x 
x4 
  
x4 
  
  
x4 
  
  
x 
x 
x 
4: Results for safety blood samples must be available and 
reviewed before drug is administered on Day 1, 3 and 6 
(hematology, 
biochemistry, coagulation)
Urine analysis 
x 
x 
  
x 
  
  
x 
  
  
x 
x 
x 
  
ECG 
x 
  
x5 
x 
  
x 
  
  
x 
  
  
x 
5: the baseline ECG should be conducted prior to dosing 
on Day 1, 
and subsequent ECGs should be taken at pre-dose on the 
rest of the dosing days and at time of visit on ambulatory 
visit days 
SARS CoV antigen 
x 
x 
  
  
  
  
  
  
  
  
  
x 
  

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Visit 
Screening 
Day 
-1 
Day 1
Day 2
Day 3
Day 4
Day 5
Day 6
Day 7
Day 8
Day 
11
Day 14 
(EOS)
  
(Days -28 to -2)
  
Viral screening 
x 
  
  
  
  
  
  
  
  
  
  
  
  
(HIV, Hepatitis) 
Drug screening  
x 
x 
  
  
  
  
  
  
  
  
  
x 
  
Breath alcohol test 
x 
x 
 
 
 
 
 
 
 
 
 
x 
 
Weight/height (BMI) 
x 
x6 
  
  
  
x6 
  
  
  
x6 
x6
x6 
 6: only weight  
Plasma PK sample 
collection 
  
  
x7 
x8 
x8 
x8 
x8 
x8 
x7 
x8 
x8
x8 
7: PK samples should be collected pre-dose and at 0.25, 
0.5,0.75,1,1.5,2,4,8,12 hr, on Day 1 and Day 7. Depending 
on ongoing PK data review, timepoints may be modified 
or removed. 
8: PK samples should be collected pre-dose on Day 2, 3, 
4,5 and 6 and 24hr +/- 30 min (Day 8), 96hr +/- 4hr (Day 
11) and 168hr +/- 4hr (Day 14) after last dosing of Day 7. 
PD9 
  
  
x9 
x 
  
x9 
x 
  
x9 
x 
  
  
9: PD blood sample should be taken pre-dose and at 
1,2,4,8, and 12 hr after dosing on Day 1, Day 4 and Day 7. 
 
One sample per day should be taken on Day 2 and Day 5 
pre-dose, corresponding to 24 h after previous day dosing. 
One sample should be taken 24 h after previous day 
dosing on Day 8. 
Adverse Events 
x10 
x10 
x10 
x 
x 
x 
x 
x 
x 
x 
x 
x 
10: Adverse events occurring prior to study drug 
administration should be recorded as medical history 
unless the event is directly related to study specific 
procedure 
Prior and/or Concomitant 
medications 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
  

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6 
INTRODUCTION AND RATIONALE 
6.1 
Introduction 
Benfo-oxythiamine (B-OT) is a novel inhibitory thiamine analogue that acts as a prodrug and 
releases oxythiamine in the human body. B-OT is a previously unknown dosage form of 
oxythiamine which, like the drug benfotiamine (trade name Milgamma), has a lipophilic side 
group, the benfo group. Its introduction into a molecule improves its cell permeability and can 
thus ensure better bioavailability [Bitsch et al.,1991]. 
The active substance in the investigational medicinal product capsules is the small molecule 
benfo-oxythiamine (B-OT). B-OT represents a novel inhibitory thiamine (vitamin B1) analog 
and prodrug which is converted in the human body into oxythiamine. Oxythiamine is a 
thiamine antagonist and inhibits transketolase, the enzyme that controls the nonoxidative 
branch of the pentose phosphate pathway (PPP). Via the intermediate ribose-5-phosphate, the 
PPP supplies ribonucleotides for SARS CoV 2 replication. Therefore, it is hypothesized that 
the inhibition of transketolase by B-OT will reduce the capacity for viral replication. 
SARS-CoV-2 (severe acute respiratory syndrome coronavirus type 2) is a new coronavirus that 
was identified as the cause of COVID-19 in early 2020. In March 2020, the WHO declared the 
outbreak of SARS-CoV-2 infections a pandemic.  
The course of a SARS-CoV-2-infection varies in symptoms and severity, from asymptomatic 
infections to severe pneumonia with lung failure, multiple organ failure and death. The most 
common symptoms include cough, fever, rhinitis, sore throat, gastrointestinal symptoms and 
neurologically associated symptoms such as the loss of the sense of smell or taste [Lechien JR, 
et al. 2019; Yan CH, et al. 2020; Tong Jy, et al. 2020]. Other neurologically associated 
symptoms include headache, dizziness, cognitive impairment, and fatigue, which can also 
occur after mild courses and can manifest long after the acute SARS-CoV-2 infection has 
already resolved [Schenk M 2020; Li YC, et al. 2020; Whittaker A, et al. 2020; Kennedy M, et 
al. 2020; Hampshire A, et al. 2020]. This can significantly compromise the long-term health 
and quality of life of patients. 
Some SARS-CoV-2 patients develop COVID-19 disease, which is associated with lung 
damage, oxygen deficiency and respiratory distress. Pneumonia may develop, usually in the 
second week of illness, and may progress to ARDS (Acute Respiratory Distress Syndrome), 
requiring ventilation and intensive care and possibly oxygenation of the blood outside the body 
(ECMO) [Yang X, et al. 2020; Graselli G, et al. 2020; Rieg S, et al. 2020]. Moreover, COVID-
19 can also manifest in other organ systems. The sites of manifestation depend, among others, 
on the density of ACE-2 receptors (entry portal of the virus into the cell) in the tissues. High 
ACE-2 density exists in the respiratory tract, as well as in the intestine, vascular cells, kidney, 
heart muscle and other organs. 
Severe courses of SARS-CoV-2 infection are often associated with an excessive reaction of the 
macrophages and induce production of viral nucleic acid by use of cellular enzymes. The virus 
infection causes a metabolic switch from mitochondrial energy release (OxPhos) to a glucose-
based energy release (aerobic glycolysis). This metabolic switch stimulates the release of large 
amounts of cytokines, as well as activates macrophages, dendritic cells, and others, leading to 
multisystem organ failure and death. Inhibition of glucose metabolism by 2-deoxy-glucose 
(2DG) inhibits virus replication in infected cells confirming the critical role of glucose 
metabolism. 

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Drugs that inhibit immune system activity, such as dexamethasone, show positive effects on 
severe disease courses. Nevertheless, a high proportion of patients undergoing a severe course 
of SARS-CoV-2 infection die. According to the study of critical illness courses during the first 
COVID-19 epidemic, 47% of cases treated with intensive care died [Schilling J, et al. 2020]. 
Containment of the SARS-CoV-2 pandemic is massively complicated by the fact that, firstly, 
SARS-CoV-2 infections can also proceed asymptomatic, and therefore apparently healthy 
people infect others, thus facilitating the further expansion of the virus.  Secondly, new and 
faster-spreading viral mutations have now emerged and can lead to a sudden increase in the 
number of cases. This poses the risk of overloading the healthcare system and hospitals  as is 
currently the case, for example, in the UK, Portugal and Brazil. 
B-OT is a novel drug candidate in development as a therapy for COVID-19. It is intended that 
patients diagnosed with COVID-19, i.e., people with disease symptoms and confirmed 
diagnosis of SARS-CoV-2 viral infection, will be treated with B-OT capsules. It is anticipated 
that B OT will actively interfere with viral replication and lead to fast recovery and cure of 
COVID 19. In consequence, the risk for disease aggravation and hospitalization will be 
reduced, as well as long-term fatigue and similar lingering symptoms experienced by some 
COVID-19 patients. 
The clinical development program will investigate the potential of B-OT to shorten the time to 
recovery from COVID-19 and to prevent hospitalization. It will further generate data on safety 
and potential adverse reactions. 
In the FIH clinical trial, safety will be assessed through the incidence and severity of adverse 
events at each of the dose levels tested. Furthermore, for clinical and laboratory parameters 
change from baseline will be assessed, to assess the overall safety profile and tolerability of B-
OT. 
In subsequent clinical investigations  to be conducted in patients with SARS-CoV-2 infection 
and COVID-19 
 the above evaluations will be complemented with clinical efficacy 
assessments, such as, but not limited to, overall response, prevention of progression, and 
reduction in the need for oxygen supplementation. 
To support the clinical development of B-OT, several non-clinical safety studies have been 
performed using the oral route of administration, according to the requirements of the M3(R2) 
ICH guideline (Nonclinical safety studies for the conduct of human clinical trials) [M3 (R2) 
ICH Guideline S9, EMA 2009]. All pivotal toxicology and toxicokinetic (TK) studies reported 
in 
Practice (GLP) principles. Pilot, exploratory, and ADME studies were not conducted under 
strict GLP procedures but were conducted using appropriate protocols and documentation to 
ensure data integrity. In the in vivo studies as performed, B-OT was administered as a solution 
in drinking water, by oral (gavage) administration.  
The clinical development of B-OT has started. The single ascending dose (SAD) part of this 
study assessing the safety, tolerability, and pharmacokinetics of B-OT in healthy volunteers 
has been completed and the multiple ascending part (MAD) will start as soon as the 
amendment of this protocol has been approved.  
The SAD part of the study showed good tolerability and there were no side effects detected 
after once-daily administration of B-OT at an escalating dose ranging from 0.5 mg to 5 mg. 
No B-OT was detected at any dose level in the plasma of the healthy volunteers. 

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At the maximum dose of 5 mg, the maximum concentration of OT in human plasma was 15.45± 
3.76 ng/ml. The time to reach the maximum concentration ranged at the different dose between 
1.67±0,29 and 2.50±1,18 hours.  
Following single administrations of B-OT, the exposure to OT in terms of Cmax and AUC0-t 
increased with ascending doses from 0.5 to 5 mg in a slightly over proportional manner. 
Further details can be found in the IB. 
Study Rationale 
This is a FIH clinical study and as such is conducted primarily to obtain information on safety, 
PK and PD properties of the tested compound. The scientific rationale of the study is based on 
the information that RNA viruses such as SARS-CoV-2 bring their own RNA polymerase into 
the cell. However, after infecting the cell and releasing their viral RNA, they depend on 
precursors necessary for replication of their viral RNA. The virus is dependent on ribose-5-
phosphate (R5P) from the host cell as an essential building block for their replication. 
The central switch for the regulation of cell cycle and cell proliferation as well as for the 
sufficient supply of R5P building blocks is provided by transketolase-like 1 (TKTL1) [Li et al., 
2019], which therefore represents a good target for antiviral therapy. 
Transketolases (TKT) are key enzymes of the pentose-phosphate-pathway (PPP). They form 
dimers and are activated by the binding of thiamine diphosphate and a divalent cation. Li et al. 
have shown that during cell cycle TKTL1 is overexpressed in late G1 phase and S phases. This 
results in the formation of stable TKT/TKTL1-heterodimers which exhibit a different 
transketolase activity than TKT-homodimers. The formation of TKT/TKTL1 heterodimers 
shifts glucose metabolism towards R5P formation, resulting in an accumulation of R5P in the 
cell [Li et al. 2019; Coy JF 2017]. In addition, it has been shown in cancer cells that TKTL1 is 
involved in the regulation of cell division via a feed-forward loop mechanism, i.e., that the end 
product of the enzyme reaction further enhances the reaction [Sun W et al. 2010]. In terms of 
metabolism towards R5P formation) appears to be judicious: during the S phase, in which DNA 
duplication occurs in the cell, the DNA is unpackaged and accordingly extremely vulnerable 
to mutations. It is therefore essential to complete this phase as quickly as possible in order to 
minimize the risk of DNA damage. Thus, by means of a binary regulatory mechanism, the cell 
ensures that cell division - once it has begun - can proceed safely and completely and that 
sufficient building blocks (R5P) are available for DNA synthesis and DNA repair during this 
phase. These R5P building blocks are also required by RNA viruses for replication and form 
the basis for the replication of RNA viruses in the cell. 
Since transketolases are essential for the formation of R5P and thus for the replication of 
SARS-CoV-2 in the host cell, they represent an attractive target for drug therapy: Substances 
that inhibit the activity of the transketolases prevent the accumulation of R5P, the essential 
building block for virus replication, in the cell. In this way, the virus can be deprived of its 
essential basis for its replication. Thus, this therapeutic approach, which fundamentally 
prevents viral replication - unlike a vaccine - is also independent of mutations of the virus. 
Thiamine antagonists, such as oxythiamine (OT), are irreversible inhibitors of thiamine-
dependent enzymes such as transketolases and efficiently inhibit sugar metabolism [Tylicki et 
al. 2018].  Since the TKT/TKTL1 heterodimer, which is responsible for enhanced R5P 
synthesis, is being dependent on thiamine, the use of OT or B-OT enables an inhibition of the 
TKT/TKTL1 heterodimer thereby reducing the R5P synthesis in a dose dependent manner. 
Thus, it can be assumed that ribose-5-phosphate can be reduced to a certain critical low level, 
limitation of ribose-5-phosphate concentration in cells as a building block for virus replication. 

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Recent studies in SARS-CoV-2 infected Caco-2 cell lines [Bojkova D et al. 2020 and 2021] 
have shown that a) the pentose phosphate pathway was activated after SARS-CoV-2 infection 
and b) a dose-dependent inhibition of SARS-CoV-2 replication was detectable when non-toxic 
B-OT concentrations were applied. 
6.2 
Risk-Benefit and Ethical Assessment 
B-OT represents a novel inhibitory thiamine (vitamin B1) analog and prodrug which is 
converted in the human body into oxythiamine. Oxythiamine is a thiamine antagonist and 
inhibits transketolase, the enzyme that controls the nonoxidative branch of the pentose 
phosphate pathway (PPP). Via the intermediate ribose-5-phosphate, the PPP supplies 
ribonucleotides for SARS CoV2 replication. Consequently, inhibition of transketolase by B-
OT will reduce the capacity for viral replication. Subjects in this study should not expect to 
receive any benefit from the study intervention provided. If benfo-oxythiamine (B-OT) is 
shown to be safe and efficacious in future studies, there is potential benefit for future patients. 
Based on the effects observed in the nonclinical studies, adverse effects can be detected in 
biochemistry and hematology parameters. Therefore, these parameters will be carefully 
checked in this study.  Safety laboratory assessments, consisting of a complete blood count 
with differential (red and white blood cells), a comprehensive panel of coagulation (Fibrinogen, 
PT and APTT) and biochemistry (with a specific attention to liver function parameters) as well 
as a urinalysis, will be conducted prior to dosing (baseline), during treatment and at the end of 
treatment. Abnormal laboratory parameters will be followed-up until return to normal or 
baseline values.  
Based on the safety results and PK results of single ascending dose part of this study as well as 
on the multiple dose non-clinical data, the overall risks for the subjects are considered to be 
small. The risks are further reduced by the chosen study design and risk mitigations. 
 
For more information, please refer to the IB (
). 

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7 
STUDY OBJECTIVES 
Primary objective:  
 To assess the safety and tolerability of single and multiple ascending doses of B-OT. 
Secondary objectives: 
 To evaluate the pharmacokinetics (PK) of benfo-oxythiamine (B-OT) and its active 
metabolite oxythiamine (OT) in plasma following single doses administered to healthy 
volunteers. 
 To evaluate the pharmacokinetics (PK) of benfo-oxythiamine (B-OT) active metabolite 
oxythiamine (OT) in plasma following multiple doses administered to healthy 
volunteers. 
Exploratory Objectives 
 To evaluate the pharmacodynamics (PD) by measuring the transketolase activity 
 To analyse PK parameters of other B-OT derivative metabolite(s), if any.  

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8
STUDY DESIGN
8.1
Overall Study Design and plan
This is a First-in-Human (FIH) single centre, open label study to assess safety, tolerability, 
pharmacokinetic and pharmacodynamic properties of benfo-oxythiamine (B-OT) after a single
and multiple oral dose administration under fasting conditions in healthy male subjects. 
This study has two parts:  a single ascending dose (SAD) part and a multiple ascending dose 
(MAD) part.
The SAD part of the study will consist of an ambulatory screening visit, treatment period 
comprised of four nights (day -1 until day 4) in the clinical site where the study will be 
conducted during which safety parameters will be measured and numerous blood samples will 
be drawn for assessment of benfo-oxythiamine (B-OT) and oxythiamine PK and PD properties 
and a final ambulatory follow-up visit
(day 8). After the 
IMP administration during the treatment visit, safety, pharmacokinetic and pharmacodynamics 
data will be collected and reviewed 72 hours post-dose.
The following figure represents the study design of the SAD study: 
Figure 1 SAD design
The filled box represents a dosing event, a white box a represents a dosing free day. Single 
dose treatment will be administered on Day 1.
The total duration of the SAD study for each subject will be up to 5 weeks divided as follows:
Screening: Up to 28 days before the first day of the treatment visit;
Treatment and study assessments: Admission in the clinical site on day -1 and discharge 
on day 4

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Final follow-up visit (End of study (EOS)): three days after treatment visit completion 
on day 8.
The following description of activities during each study visit concerns each subject, regardless 
of whether it is the sentinel subject or not.
After confirmation of eligibility during the screening visit, each subject will undergo one 
treatment visit. The treatment visit will consist of five days (4 nights): on the first day subject(s) 
will come to the site for admission in the afternoon under the condition that a negative Sars-
CoV2 PCR test has been confirmed after the performance of a test on the same day in the 
morning at the responsible laboratory location. On the second day (day 1) subject(s) will be 
administered the assigned IMP in the morning and the serial PK and PD sampling that has 
started pre-dose will continue post-dose until day 4 (72nd hour). On the fifth day (day 4) after 
the last PK sample (72 h post-dose) subject(s) will be discharged from the unit. During the 
treatment visit each subject will undergo not only serial PK and PD sampling but all
assessments listed in the Schedule of Activities (Table 1).
A follow up visit will be held by each subject 3 days after the last day of the treatment visit
during which they will complete their participation. All safety events that are reported as not 
resolved during the follow-up visit will be observed until resolution, but not longer than 30 
days thereafter or after the end of the study (defined as last subject last visit).
In the MAD part of the study, B-OT will be administered orally (p.o.) once daily for 7 days 
(D1-D7), followed by 7 days without dosing. The first dose will be 1 mg, as the PK and safety 
data have shown a very low concentration of OT at 0.5 mg dose and no safety issues were
observed at any dosage in the SAD part.
Each cohort will start with a sentinel subject (subject #1). 
Subjects #2 - #6 of the same cohort will start treatment after a minimum of 72 hours if no 
adverse event is observed in the sentinel subject.
The following figure represents the study design of the MAD part:
The total duration of the MAD part of the study for each subject will be up to 6 weeks divided 
as follows:
Screening: Up to 28 days before the first day of the treatment visit;

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 confinement visit: Admission in the clinical site on Day -1, seven treatment days 
(Day1,2,3,4,5,6,7) and discharge on Day 8 
 Ambulatory follow-up visit: four days after last treatment day on Day 11 
 Final follow-up visit (End of study (EOS)): seven days after last treatment day visit on 
Day 14 
 
After confirmation of eligibility during the screening visit, one treatment visit will follow. The 
confinement visit will consist of 9 days (8 nights): on the first day the subject will come to the 
site for the confinement period in the afternoon, under the condition that a negative Sars-CoV2 
antigen test has been confirmed on the same day. Over the next seven days (Day 1 until Day 
7) the subject will be administered the assigned IMP in the morning, and the serial PK and PD 
sampling that has started pre-dose will continue post-dose until Day 8. On Day 8, after the last 
PK sample, the subject will be discharged from the unit.  
A follow-up visit will be held 4 days after the last day of treatment and a second follow-up 
during which the subject will complete their participation will be held seven days after the last 
day of the treatment visit. All safety events that are reported as not resolved during the follow-
up visit will be observed until resolution, but not longer than 30 days thereafter. 
A detailed description of all visits and procedures in the MAD part of the study is depicted in 
Table 2 Visit and Assessment Schedule of the MAD Part. 
8.1.1 Screening (SAD and MAD) 
During the screening visit, the study subjects will undergo a complete physical examination as 
well as ECG, vital signs (pulse rate, respiratory rate, blood pressure, body temperature), and 
relevant safety laboratory tests (hematology, biochemistry, urine analysis, viral screening) to 
assess and check if they meet the eligibility criteria as defined in sections 9.2 and 9.3. The 
screening is to take place within maximum 28 days and minimum 2 days prior to the first day 
(day -1) of the treatment visit. 
All test results will have to be within acceptable (normal) ranges as set out in the CSP before a 
subject can be included in the study. Screening results that are outside the normal range will be 
delegated physician will then determine whether or not the respective abnormal test result is 
clinically significant and/or exclusionary before the subject can be enrolled in the study.  
 
8.1.2 Treatment and observation period (SAD and MAD) 
A total number of 24 subjects will be enrolled in the SAD part of the study in 5 cohorts: 
 
Cohort 1 (0.5 mg B-OT): 3 subjects  
Cohort 2 (1 mg B-OT): 3 subjects  
Cohort 3 (2mg B-OT): 6 subjects 
Cohort 4 (3 mg B-OT): 6 subjects  
Cohort 5 (5 mg B-OT): 6 subjects  
 
Each cohort will start with a sentinel subject (subject #1). Subject #2 - #3 or #2 - #6 of the same 
cohort will start treatment after a minimum of 48 hours if safety data indicate that dosing of 
subsequent subjects of this cohort is justified. Dose escalation to the next dose level will occur 
if three (first and second cohort) or six subjects (3 next cohorts) have completed the single dose 
treatment and if PK at Day 2 as well as safety data allow an escalation to the next dose level. 
 

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A total of 24 subjects will be enrolled in the MAD part of the study and assigned to multiple 
ascending dose groups in consecutive order: 
Group 1 (1 mg B-OT): 6 subjects  
Group 2 (2 mg B-OT): 6 subjects  
Group 3 (3 mg B-OT): 6 subjects 
Group 4 (5 mg B-OT): 6 subjects  
 
Each cohort will start with a sentinel subject (subject #1). Subject #2 - #6 of the same cohort 
will start treatment after a minimum of 72 hours if safety data indicate that dosing of subsequent 
subjects of this cohort is justified. Dose escalation to the next dose level will occur if six 
subjects have completed the multiple dose treatment and if PK at Day 5 as well as safety data 
allow an escalation to the next dose level. 
 
The health status of the trial subjects in the SAD and in the MAD part of the study will be 
assessed once again against the inclusion and exclusion criteria, as set by the study protocol, 
prior to study drug administration (Day -1). When considered suitable, the subjects will be 
assigned a unique study identifier and will be invited for treatment visit on day -1. Detailed 
description of all trial activities that need to be carried out are listed in the Visit and Assessment 
Schedule (Table 1 for the SAD part and Table 2 for the MAD part). 
 
8.1.3 Follow-up (SAD and MAD) 
Each subject in the SAD part of the study will complete the study with a final study visit which 
will take place on Day 8. This visit will include all procedures listed in the Visit and Assessment 
Schedule (Table 1). 
Each subject in the MAD part of the study will visit the site for two follow-up visits  one on 
Day 11 and one end of study visit on Day 14. These visits will include all procedures listed in 
the Visit and Assessment Schedule (Table 2). 
8.2 
Justification for population and design 
The study is based on established designs for FIH studies and will comprise of single ascending 
dose (SAD) and multiple ascending dose (MAD) cohorts in sequential order. It will be 
conducted in healthy volunteers to evaluate the safety, tolerability and PK, of oral benfo-
oxythiamine (B-OT) administration. 
The sentinel dosing maximizes the chance that, in the event of a significant unanticipated safety 
risk, dosing will be discontinued after exposure of a single healthy volunteers to a given dose 
level of benfo-oxythiamine (B-OT). 
The sequential nature of the cohorts requires that the necessary data is reviewed by the SRC 
before the next cohort/sub-part/part can begin. 
The study is to be conducted in healthy male subjects. 24 subjects represented in the SAD PK 
Analysis Set and another 24 subjects represented in the MAD PK Analysis Set will be required 
to complete the study. A healthy subject population with carefully considered 
inclusion/exclusion criteria will avoid the potential for interaction of benfo-oxythiamine (B-
OT) with any underlying disease state or concomitant medication that might be necessary to 
administer to patients, while ensuring that subjects are fit and well enough for participation in 
the study.  

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The eligibility criteria are designed to select subjects for whom protocol treatment and 
procedures are considered appropriate. All relevant medical and non-medical conditions should 
be taken into consideration when deciding whether this protocol is suitable for a particular 
subject. 
 
8.3 
Proposed Safe Starting Dose 
According to current CHMP 
Mitigate Risks for First-in-Human and Early Clinical Trials with Investigational Medicinal 
-in-human dose will be 
calculated considering results of the planned toxicology studies. No further considerations on 
the expected biological effective dose from completed pharmacology studies with B-OT were 
included, since efficacy studies have been primarily performed in rodents, which have been 
proven being less sensitive to B-OT compared to other species, including the dog. Therefore, 
considerations in the FIH dose are based on safety data only. 
B-OT rapidly degraded in plasma, and high levels of the active metabolite oxythiamine were 
detected. The in vitro formation rate of the metabolite in rat and dog plasma was approximately 
two-fold higher than in human plasma. Plasma protein binding in human plasma is high (92.8% 
- 94.5%). B-OT is not well transported passively over Caco-2 cells or across the blood-brain 
barrier. The compound does not bind to red blood cells. B-OT as well as oxythiamine do not 
inhibit human CYP1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1 and 3A4 isoenzymes up to 200 
µM (98 µg/mL B-OT or XX µg/mL OT). Two major metabolic pathways were found in in 
vitro metabolism studies with mouse, rat and human hepatocytes. 
In both species tested (rat and dog), no plasma concentrations of B-OT could be measured in 
DRF studies at doses up to 1000 mg/kg in rats and 1 mg/kg in dogs, respectively. Therefore, in 
repeat dose toxicity studies only TK of the (main) metabolite oxythiamine was measured. In 
the repeat-dose study in rats the exposure (Cmax and AUC0-t) to oxythiamine was roughly 
dose-proportional. No major sex differences were observed nor accumulation in Cmax. AUC-
values were similar or slightly under-proportional after repeated dosing. The toxicokinetic data 
from the repeat dose study in dogs showed a slightly under proportional increase of Cmax with 
increasing dose, while for AUC0-t linearly proportional increase was observed in males and 
apparently slightly over-proportional increase in females. No sex differences in exposure were 
observed. No difference in Cmax values were observed after multiple dosing, while for AUC 
no or a slight over-proportional increase was found (less than two folds accumulation after 
repeated doses). 
The safety and tolerability of B-OT were assessed through a series of non-clinical toxicity 
studies, which included safety pharmacology investigations. The safety pharmacology 
investigations in rats (i.e., Irwin study) showed effects in female animals (1000 mg/kg/day) 
which were in a bad condition at the moment of the investigations. In the dog cardiovascular 
investigations, no effects on ECG by conventional ECG measurement. In a jacketed telemetry 
measurement, increases in heart rate were observed, at a supra-NOAEL dose level. In the same 
study no effect on respiratory functions were recorded. 
Two-cycle (i.e., 1-week treatment followed by a 1-week treatment free period per cycle) repeat 
dose toxicity studies with B-OT were performed in rats and dogs. Female rats at the highest 
tested dose level (1000 mg/kg/day) were all prematurely euthanized on Day 7 of the study 
because of moribund condition. Other effects noted in the rats were lower body weight, 
transient hematological effects and some effects on cholesterol and triglyceride levels. In dogs 
one female animal was found dead on Day 5 in the second treatment cycle. Effects in surviving 

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animals were lower body weight and food consumption, which were partially reversed after a 
1-week treatment-free period, as well as some transient changes in clinical pathology 
parameters. Furthermore, some non-adverse histopathological effects in thymus tissue were 
found. The No Observed Adverse Effect Level (NOAEL) for the rat study was set at 1000 
mg/kg/day (males) and 300 mg/kg/day (females), and the NOAEL for the dog study was set at 
0.15 mg/kg/day for both sexes. 
Dosing is based on NOAEL x HED x 1/10 (No Observed Adverse Effect Level x Human 
Equivalent Dose [dog/human] x Safety Factor). As NOAEL was established in dog after a 7-
day treatment and B-OT has already been used in compassionate use at a minimum daily dose 
of 3 mg for 7 days in 5 Covid-19 patients and 5 late-stage prostate cancer patients without any 
safety issue, only 3 subjects in the SAD part will receive the two first lowest doses. 
In the SAD part, the first dose of B-OT will be 0.5 mg (0.0083 mg/kg body weight). After the 
first cohort, enrolment will be halted. If no safety issue is observed, the second dose level will 
be 1 mg (0.016 mg/kg body weight). The first and second dose level cohort will have 3 subjects. 
The human PK/PD model will be refined before enrolling to the next dose level may resume. 
As this is the first study testing B-OT in humans, study design adjustments may be made based 
on emerging data from each dose cohort based on review of preliminary safety, tolerability, 
and PK/PD results. If these data allow for, the dose escalation scheme of 2 mg, 3 mg and 5 mg 
will follow in 3 additional cohorts of 6 subjects.  
Based on the SAD PK results, a starting dose of 1 mg in the MAD part provides a margin of 
exposure of 7.6 to steady-state exposure values observed at the NOAEL in dogs and is 
considered acceptable. If these data allow for, the dose escalation scheme of 2 mg, 3 mg and 5 
mg will follow in 3 additional cohorts of 6 subjects.   
For more information, please refer to the IB. 
 
8.4 
Dose Escalation Strategy 
In general, a safety review committee (SRC) approval of commencement of a cohort is 
contingent on acceptable safety, tolerability, and PK results. 
Each cohort will start with a sentinel subject (subject #1). The rest of the subjects of the same 
cohort will start treatment after a minimum of 48 hours in the SAD part or 72 hours in the 
MAD part if safety data indicate that dosing of subsequent subjects of this cohort is justified. 
In the SAD part of the study, dose escalation to the next dose level will occur if three (first and 
second cohort) or six subjects (3 next cohorts) have completed the single dose treatment and if 
PK as well as safety data allow an escalation to the next dose level. 
In the MAD part of the study, dose escalation to the next dose level will occur if six subjects 
(each of the 4 cohorts) have completed the multiple dose treatment and if PK as well as safety 
data allow an escalation to the next dose level. 
The general dose increase is planned to follow a modified Fibonacci scheme. It may be adapted 
to the PK and safety profile analysis. However, a maximum multiplication factor of 3 will not 
be used. 
Each cohort, after the first, may only commence after SRC review of results from the preceding 
cohort.  

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8.5 
Stopping Criteria for Dose Escalation  
Dose escalation will not continue to the next dose level if (a) maximal OT plasma levels are 
achieved, i.e. further increase is not justified from a scientific point of view, or if (b) toxicities 
are observed that do not justify continuation from an ethical point of view. 
The study will be halted if one or more subjects experience a serious adverse event (SAE) that 
is considered as related to B-OT or 2 or more subjects in the same group experience severe 
AEs that are considered to be related to B-OT. 
Dose-escalation will stop if clinically relevant signs or symptoms of similar nature occur in 2 
or more subjects in a group that, in the opinion of the PI, warrant stopping of dose-escalation. 
8.5.1 Stopping Rules for an Individual Subject, at any Time in the Study  
Dosing for any individual subject will be stopped if the subject experiences a possible drug-
related SAE or a possibly drug-related significant non-serious AE, which in the opinion of the 
SRC or investigator warrants discontinuation of the subject from the active protocol for his or 
her well-being. 
8.5.2 Stopping Rules for a Whole Cohort, Dose Escalation, Progression to Next 
Study Part and Termination of Study 
Stopping rules as detailed below are applicable for each of the following situation: 
- 
Stopping dosing for a whole cohort. 
o When subjects in a cohort are dosed staggered. 
- 
Stopping dose escalation. 
- 
Stopping progression to the next part of the study. 
- 
Final stop of study. 
The study will be halted (i.e., no further dosing will occur) if one or more subjects experience 
an SAE that is considered to be related to B-OT or 2 or more subjects in the same group 
experience severe AEs that are considered to be related to B-OT. If, following an internal safety 
review, the Sponsor deems it appropriate to restart the study, this can be done following 
approval of a substantial protocol amendment. 
Dose-escalation will stop if clinically relevant signs or symptoms of similar nature occur in 2 
or more subjects in a group that, in the opinion of the PI, warrant stopping of dose-escalation. 
The risk to all participating subjects will be evaluated thoroughly prior to a decision to terminate 
the study prematurely or continue dosing in agreement with the regulatory authorities. 
The SRC will carefully review the totality of available data, taking into account moderate non-
serious AEs at least possibly related to IMP administration in unblinded fashion, the number 
of subjects in which they occur, concurrency of more than one within the same subject and 
potential safety signals identified for other IMPs in the same class (mechanistic and/or 
chemical). Although changes outside normal ranges that apply for healthy volunteers are most 
relevant, changes from baseline measurements will also be considered observing any trends. 
8.6 
Safety Review  
The SRC will consist of the following core members: 
Principal investigator, sponsor safety representative, sponsor head of clinical department, 
sponsor PM. 

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The SRC may also request to have attendance of or off-line support and input from the 
following functions as required: CRO PM, sponsor preclinical department representative, CRO 
statistician and PK analyst. 
Written statements and conclusions by the SRC will be in place before allowing study 
progression at the noted times as per CSP. This includes documentation of appropriate quality 
control checks on the data reviewed. 
 
8.6.1 Data Reviewed by Safety Review Committee for Dose Escalation Decision  
The SRC will decide on the next dose levels throughout the study, considering at each decision 
point the degree of uncertainty. The choice of the dose levels will take into consideration an 
estimate of exposure levels to be achieved and potential adverse effects. Emerging Cmax and 
AUC data will always be benchmarked to exposures expected at key human predicted doses 
(PAD/ATD). Late emerging safety issues that may have occurred after the time point for the 
dose escalation decision will always be considered. 
 
In the SAD part of the study, at every SRC meeting, cumulative data for all cohorts will be 
reviewed. The following data will be reviewed for each individual cohort: 
 
 
Safety and tolerability data for 4 days for at least 3 healthy volunteers in the 2 first 
cohorts and 5 healthy volunteers in the next cohorts. 
 
PK data for 48 hours for at least 3 or 5 healthy volunteers in the first or next 
cohorts respectively. 
 
Appropriate data collection up to 7 days in a given dosing cohort should be complete to proceed 
to the next dose cohort. 
 
In the MAD part of the study, at every SRC meeting, cumulative data for all cohorts will be 
reviewed. The following data will be reviewed for each individual cohort: 
 
 
Safety and tolerability data for 8 days for at least 5 healthy volunteers in each cohort. 
 
PK data for 96 hours for at least 5 healthy volunteers in each cohort. 
 
Appropriate data collection up to 14 days in a given dosing cohort should be complete to 
proceed to the next dose cohort. 
 
 
8.6.2 Safety Review Committee Decision on Next Dose 
The SRC will take the decision for the next dose level or whether to stop the study after 
reviewing all the pertinent safety and any other relevant data. 
If consensus among the voting SRC members cannot be reached then the PI, who has the 
ultimate responsibility for the safety of the healthy volunteers, will make the final decision on 
the next dose level or whether to stop the study. In any event, dose escalation can only occur if 
agreed by the PI. 
The decisions of the SRC on the next dose level will be documented and provided to all the 
appropriate parties involved with the study, including the Pharmacist to enable IMP preparation 
for the next scheduled dosing day. 
 

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Where stopping criteria have not been met, the decision of the SRC may be to give 
the next higher dose according to the predefined dose increment, a smaller dose 
than previously given, a repeated dose or to stop dosing. 
 
A dose in which the safety stopping criteria have been met will not be repeated and further 
dose escalation must not occur. 
 
In this case, the SRC will review the totality of data and restart of dosing is 
possible without a substantial amendment (e.g., in the case of a laboratory 
error) if SRC review concludes that the relevant stopping criterion was not
fulfilled. 
 
A lower dose level expected to be tolerable and not to meet the stopping 
criteria, would be acceptable in this case. 
 
If the safety stopping criteria are met and the SRC decides that there are 
reasons that the dose level should be repeated or further dose escalation is 
warranted a summary of the data and justification (CSP amendment) will be 
submitted to the ESB, the Regulatory Authority and the IEC/IRB for their 
approval before further dosing. 
 
In some circumstances, consideration can be given to test fractionated or split doses. 
 
8.6.3 Blinding at Safety Review Committee Meeting 
Data will be reviewed unblinded. 
 
8.6.4 Assessments Adaptation 
Following review of data from a cohort of healthy volunteers, the timing of assessments and/or 
blood samples may be adjusted for subsequent cohorts. 
Additional assessment or sampling times may be added if indicated by the data; however, the 
maximum blood volume taken from each healthy volunteer will not exceed 500 mL. 

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9 
STUDY POPULATION 
9.1 
Subject population 
Subjects will be recruited from the subject database of the investigational team in the Clinical 
site as well as through referrals.  
Subjects who fail to meet the inclusion criteria or meet any exclusion criterion should not, 
under any circumstances, be included into the study. There can be no exceptions to this rule. If 
subjects fail out of the screen window but did not fail on any inclusion or exclusion criteria, 
they can be rescreened. One repeat assessment may be requested per the discretion of 
investigator at the screening visit and on admission to the Clinical site for the treatment visit. 
9.2 
Inclusion criteria 
The following criteria must be met by all subjects considered for study participation: 
 
1. Signed and dated informed consent form 
2. Subjects capable to understand the purposes and risks of the study, and who are willing 
and able to participate in the study  
3. Male volunteers who are willing to comply with contraception requirements 
4. Aged 18-60 years, inclusive 
5. Healthy participants, as determined by screening assessments and Principal 
Investigator's judgment. Healthy status is defined by the absence of evidence of any 
active or chronic disease following a detailed medical and surgical history, a complete 
physical examination including vital signs, 12-lead ECG, hematology, blood chemistry, 
serology and urinalysis 
6. Body Mass Index (BMI) of 18-30 kg/m2 inclusive 
7. Male participants with female partners of childbearing potential are eligible to 
participate if they agree to one of the following during the protocol defined time frame:  
 Be abstinent from penile-vaginal intercourse as their usual and preferred 
lifestyle (abstinent on a long term and persistent basis) and agree to remain 
abstinent 
 Use a male condom plus partner use of a contraceptive method with a failure 
rate of < 1% per year when having penile-vaginal intercourse with a woman of 
childbearing potential who is not currently pregnant. 
Note: Men with a pregnant or breastfeeding partner must agree to remain abstinent from penile-
vaginal intercourse or use a male condom during each episode of penile penetration. 
9.3 
Exclusion Criteria 
Subjects will be excluded if they meet any of the following criteria: 
 
1. Subjects who have a clinically relevant history as determined by the Investigator, or 
presence of respiratory, gastrointestinal, renal, hepatic, hematological, lymphatic, 
neurological, 
cardiovascular, 
psychiatric, 
musculoskeletal, 
genitourinary, 
immunological, dermatological, endocrine, connective tissue diseases or disorders. 
2. Fridericia's correction factor for QT (QTcF) > 450 ms or history of QT interval 
prolongation. 
3. Have acute gastrointestinal symptoms (e.g., nausea, vomiting, diarrhea, heartburn) at 
the time of screening or admission. 
4. Has any abnormal laboratory value of Grade 2 or higher, which is considered as 
clinically significant by the PI (or designee). 

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5. Has values of 
10 % above the upper limit of normal for the following laboratory 
analytes: 
 alanine aminotransferase (ALT/SGPT), alkaline phosphatase (serum), aspartate 
aminotransferase (AST/SGOT), at Screening and Day-1 
 creatinine and urea concentration in blood at Screening and Day-1 
6. Have a clinically relevant surgical history, as determined by the Investigator. 
7. Have a history of relevant drug hypersensitivity. 
8. Have a history of alcoholism or drug abuse. 
9. Have a significant infection or known inflammatory process at screening or admission. 
10. Have used any prescription or non-prescription medicines within 2 weeks of admission, 
unless in the investigator's opinion it will not affect the determination of safety or other 
study assessments. 
11. Have received any investigational drug within 30 days prior to screening. 
12. Have used tobacco or nicotine products within 3 months of screening 
13. Has a positive alcohol or drug screen at Screening or the Day 1 visit  
14. Have donated or received any blood or blood products within the 3 months prior to 
screening. 
15. Cannot communicate reliably with the investigator. 
16. Are unlikely to co-operate with the requirements of the study. 
17. Have been vaccinated with a Covid-19 vaccine within 2 weeks prior to screening and/or 
during participation in the clinical trial 
 
Abnormal vital signs after 5 minutes supine rest, are defined below. Abnormal values may be 
repeated once at the discretion of the investigator: 
 Systolic BP < 90 mmHg or > 140 mmHg 
 Diastolic BP < 50 mmHg or > 90 mmHg 
 Pulse < 45 or > 85 bpm 
 Body temperature of > 37.7°C (on admission day) 
 
Clinically important abnormalities in rhythm, conduction or morphology of the resting ECG 
and any clinically important abnormalities in the 12-lead ECG are defined below: 
 
 Prolonged QTcF > 450 ms. 
 Family history of long QT syndrome. 
 PR (PQ) interval shortening < 120 ms (PR > 110 ms but < 120 ms is acceptable if there 
is no evidence of ventricular pre-excitation). 
 PR (PQ) interval prolongation (> 240 ms) intermittent second (Wenckebach block 
while asleep is not exclusive) or third-degree AV block, or AV dissociation. 
 Persistent or intermittent complete BBB, IBBB, or IVCD with QRS > 110 ms. 
Volunteers with QRS > 110 ms but < 115 ms are acceptable if there is no evidence of 
eg, ventricular hypertrophy or pre-excitation. 
9.4 
Prior and concomitant medication 
Prior Medication: All prior medications (prescription and over-the-counter [OTC]) taken 
within 90 days of study screening will be recorded. Prescription or non-prescription drugs, 
including vitamins, herbal and dietary supplements should not be taken within 14 days (or 5 
half-lives, whichever is longer) prior to the first dose of Benfo-oxythiamine (B-OT), unless in 
the opinion of the Investigator and Sponsor the medication will not interfere with the study 
procedures or would compromise subject safety. Prescription or non-prescription drugs, 

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including vitamins, herbal and dietary supplements taken during the 14 days before the first 
dose of benfo-oxythiamine (B-OT), and the reason for taking them, will be noted in the 
 
Inclusion of subjects who have taken prior medication will be reviewed on a case-by-case basis 
in relation to the safety aspects and objectives of this study. 
Concomitant Medication: Prescription or non-prescription drugs, including vitamins, herbal 
and dietary supplements should not be taken throughout the duration of the study, with the 
exception of paracetamol (which may be taken as an analgesic to a maximum of 2 g in 24 h 
(500 mg 4 times a day) and ibuprofen (which may be taken as an analgesic to a maximum of 
1.2 g in 24 h (400 mg 3 times a day). 
-being, may be 
given at the discretion of the investigator. 
When any medication is required, it should be prescribed by the investigator. Following 
consultation with benfovir AG, the investigator should determine whether or not the subject 
should continue in the study. Administration of concomitant medications that may influence 
the measurement of the PK and PD endpoints may be documented as a protocol deviation after 
consultation of the investigator with benfovir AG. 
If intake of ANY prior or concomitant medication is necessary during the study, the daily 
 
9.5 
Dietary and other restrictions prior and during the study 
To avoid possible negative effects on the measurement of plasma drug concentrations and/or 
and in the following points. 
1. Subjects should abstain from alcohol for 72 hours prior to check-in at the Clinical site 
and before their final Follow-up Visit. Subjects should consume no more than 2 units 
of alcohol per day during the ambulatory days of participation in the study. 
2. The consumption of xanthine-containing products including coffee, tea, cola, energy 
drinks or chocolates is not allowed 24 hours prior to check-in at the Clinical site until 
discharge. 
3. For consistency, all subjects will receive a light meal immediately prior to commencing 
a 10 hour fast before dosing. During the fast, water will be freely available, except 
during the period 1 hours before dosing to 1 hours after dosing. During this 2-hour 
period only the 240 mL of water required for dosing will be available. Decaffeinated 
tea and coffee are allowed from 4 h post-dose. The subjects will be served standardized 
meals throughout the confinement in the site as follows: 
 Subjects will fast overnight for at least 10 hours before dosing. 
 Lunch will be served: approximately 4 h post-dose. 
 Snack will be served: approximately 8 h post-dose. 
 Dinner will be served: approximately 12 h post-dose. 
4. Poppy seed or foods containing poppy seeds are not permitted from 3 days before drug 
of abuse test, as consumption can lead to a positive opiate result in the drugs of abuse 
test. 

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5. No food or drink containing grapefruit, cranberry, or Seville oranges (including 
marmalade and fruit juices), and/or food or drink, sweets, candies or other 
confectionary containing liquorice will be allowed from 7 days prior to check-in at the 
Clinical site until the final study visit. 
6. Smoking is not allowed during the participation in the clinical study. 
7. Strenuous physical activity (e.g., heavy lifting, weight or fitness training) is not allowed 
within 72 hours prior to check-in day and until the final follow up visit. 
8. Subjects will remain seated or ambulatory for the first 4 hours following study treatment 
administration. However, should adverse events occur at any time, subjects may be 
placed in an appropriate position.  
9. Subjects will be advised that they should not donate blood for at least 3 months after 
the final study visit. 
10. Subjects must not donate sperm for at least 3 months after dosing with benfo-
oxythiamine (B-OT). 
11. During 
periods, they should abstain from consuming high energy 
drinks (e.g., Red Bull®), and food containing poppy seeds and any OTC medication or 
herbal preparations until after their final Follow--up Visit has been completed. Subjects 
should also limit their caffeine intake to equivalent of 3 servings of coffee per day (in 
the US: 1 serving = 12 oz soda, 6 oz coffee, or 8 oz tea; in the UK: 1 serving = 330 mL 
cola, 180 mL coffee, or 240 mL tea).  
During each period of the study, bathrooms will be locked for at least 4 hours after each drug 
administration. If necessary, subjects will be permitted to use the washroom facilities under 
supervision only during this interval. Each subject will be questioned on the specific points at 
admission to the clinical site. If a subject admits a non-compliance with these restrictions, the 
Principal Investigator will decide whether or not the subject will be permitted to remain in the 
study. Non-
 
9.5.1 Contraception requirements  
There is no information about effects that B-OT could have on the development of the foetus 
in humans. Therefore, it is important that WOCBP, who are the partners of male subjects, do 
not become pregnant during the study and for a total period of 3 months after the male subject 
has attended the study Follow-up Visit. 
Male subjects who have been sterilised are required to use one barrier method of contraception 
(condom) from the time of IMP administration until after the follow-up visit. The subject must 
have received medical assessment of the surgical success. 
As a precaution, all non-sterilised male subjects should avoid fathering a child by either true 
abstinence* or use a condom and their female partner/spouse has to be either of non-
childbearing potential or has to use a highly effective contraception form of birth control, 
starting from the time of IMP administration until 3 months after the study Follow-up Visit. 
The female partner/spouse should be stable on their chosen method of birth control for at least 
3 months prior to first dosing with the IMP. 
* Sexual abstinence is defined as refraining from heterosexual intercourse during the entire 
period of risk associated with the study intervention. It is only acceptable if preferred and usual 
lifestyle of the subject. 
Highly effective contraception, i.e., a form of birth control with a failure rate of less than 1% 
per year when used consistently and correctly, which are allowed in this clinical study, are: 
 Combined (oestrogen and progestogen containing) hormonal contraception associated 
with inhibition of ovulation: 

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 Oral 
 Intravaginal 
 Transdermal 
 Progestogen-only hormonal contraception associated with inhibition of ovulation: 
 Oral 
 Injectable 
 Implantable 
 Intrauterine device 
 Intrauterine hormone-releasing system 
 Bilateral tubal occlusion 
In addition, a barrier method must also be used i.e., condom (without spermicidal 
foam/gel/film/cream/suppository or fat- or oil-containing lubricants) or occlusive cap 
(diaphragm or cervical/vault caps) with spermicidal foam/gel/film/cream/suppository. 
Male subjects will be instructed that if their partner becomes pregnant during the study this 
should be reported to the investigator. The investigator should also be notified of pregnancy 
occurring during the study but confirmed after completion of the study. In the event that a 
bsequently found to be pregnant after the subject is included in the study, 
then consent will be sought from the partner and if granted any pregnancy will be followed and 
the status of mother and/or child will be reported to the sponsor after delivery. 
A pregnancy notification form and follow-up will be completed. 
9.6 
Study drug discontinuation and subject withdrawal  
9.6.1 Study drug interruption or discontinuation 
Before IPM is administered, changes in the subject health status including laboratory results if 
applicable, since the previous visit has to be checked. The investigator must temporally 
interrupt or permanently discontinue study participation if continued study participation is 
believed to be contrary to the best interests of the subject. The interruption or premature 
discontinuation of study participation might be triggered by: 
 Lost to follow-up. 
 Any SAE that is considered related to study intervention by the investigator. 
 Any other AE that, in the opinion of the investigator or the sponsor, warrants 
discontinuation of further dosing of study intervention. 
 Pregnancy. 
 Diagnostic or therapeutic procedure 
 Clinically significant abnormal assessment (e.g., ECG, vital signs or laboratory 
abnormalities) 
 Administrative reasons in particular withdrawal of the s
 and/or severe 
non-compliance to the CSP.  
If the study is permanently discontinued, the subject will remain in the study to be evaluated 
for assessments at the follow-up visit, including follow-up of any AEs, unless consent is 

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withdrawn from further study participation, the subject is lost to follow-up, or the subject is 
enrolled in another clinical study. 
The reason for interruption or premature discontinuation of study participation must be 
documented in the source documentation and on the Case Report Form and reported to the 
monitor. 
9.6.2 Subject withdrawal 
Subjects have the right to withdraw from the study at any time for any reason. Should a subject 
decide to withdraw from the study, all efforts should be made to complete and report the 
observations, particularly the follow-up examinations, as thoroughly as possible. 
The clinical investigator may withdraw the subject from the study for one of the following 
reasons: 
 Positive answer to the re-assessment of any exclusion criteria, during the admission at 
the treatment visit or in a subsequent occasion; 
 Non-adherence to the protocol requirements; 
 Adverse events, symptoms or signs of possible toxicity; 
 Any disease requiring medication; 
 Any other condition that, according to the judgment of the investigator, is important for 
maintaining the health of the subjects; 
 Vomit after administration and/or within 2 hours after study product administration; 
 Diarrhoea within the dosing interval (2 hours prior, during dosing, 2 hours post dosing). 
A subject who considers withdrawing from the study must be informed by the investigator 
about modified follow-up options (e.g., telephone contact, a contact with a relative or treating 
physician, or information from medical records). 
If the subject withdraws consent for disclosure of future information, the sponsor may retain 
and continue to use any data collected before such a withdrawal of consent. 
If a subject withdraws from the study, it should be confirmed if he/she still agrees for existing 
samples to be used in line with the original consent. If he/she requests withdrawal of consent 
for use of samples, destruction of any samples taken and not tested should be carried out in line 
with what was stated in the ICF and local regulation. The investigator must document the 
decision on use of existing samples in the Clinical site study records and inform the Sponsor. 
The details and reasons of the withdrawal from the study will be registered in the source 
documentation and on the Case Report Form and reported to the monitor. 
A subject will be considered lost to follow-up if he/she repeatedly fails to return for scheduled 
visit and is unable to be contacted by the Clinical site. The following actions must be taken if 
a subject fails to return to the Clinical site for a required study visit: 
 The Clinical Unit site must attempt to contact the subject and reschedule the missed 
visit as soon as possible (if applicable) and counsel the subject on the importance of 
maintaining the assigned visit schedule and ascertain whether or not the subject wishes 
to and/or should continue in the study. 
 Before a subject is deemed lost to follow-up, the investigator or designee must make 
every effort to regain contact with the subject (where possible, 3 telephone calls and, if 

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equivalent methods). These contact attempts should be documented in the 
medical record. 
9.6.3 Replacement policy 
Subjects who have given informed consent may be replaced only prior to the beginning of the 
first drug administration. No replacement will be allowed thereafter.  

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10 INVESTIGATIONAL PRODUCT 
10.1 Investigational drug and comparative drug 
10.1.1 Investigational test drug 
The investigational test product in this study is called benfo-oxythiamine (B-OT).  
The active substance is benfo-oxythiamine monosodium and is formulated as powder-in-
capsule. 
For the present first-in-human clinical trial of B-OT, a hard gelatine capsule was selected as 
dosage form. Due to the low strength of the IMP of 0.1 mg, 0.5 mg, 3 mg, and 10 mg, excipients 
were added to facilitate filling of the capsule with equal amounts of active substance. The 
standard compendial excipient microcrystalline cellulose was added to the formulation as filler 
as well as some colloidal anhydrous silica as glidant and magnesium stearate as lubricant  
B-OT is hygroscopic and capsule and container closure system have been selected to provide 
protection from humidity. B-OT capsules should be stored at room temperature inside the 
original primary packaging. 
The manufacturer responsible for production, the primary and secondary packaging, batch 
control and release of test investigational medicinal product and for labelling of the IMP is 
benfovir AG, Germany. 
10.1.2 Investigational comparative drug 
Not applicable. 
10.2 Study drug up- and down-titration  
Please refer to section 6.2.8 Dose Escalation Strategy. 
10.3 Packaging and labelling 
Test investigational medicinal products packed for each subject will be prepared, labelled and 
supplied by the Sponsor. The Test IMPs will be packed and labelled by responsible persons 
delegated by the Sponsor according to Annex 13 of GMP.  
The release of the IMP after packaging and labelling will take place in benfovir AG. The IMP 
can be released by the Sponsor once scanned copies of both an Ethics Committee (EC) and the 
Competent Authority (CA) approvals are received by the Sponsor. 
The Test product will be provided as one bottle for each subject labelled with details of the 
product. 
The containers will be labelled in accordance to the applicable regulatory requirements.  
The Sponsor will provide sufficient quantity of the IMPs for administration and reserve 
purposes (to be used i.e. in case of damage or fall of tablets).  
Labelling of these reserved products will be identical to the original one except for Subject 
Number and Period Number which is not written and should be completed by Investigator in 
case these reserved products will be used. The Certificates of Analysis for the Test products 
will accompany the IMPs and will be presented in the final study report. 
Investigational Medicinal Products will be sent to clinical site under temperature monitoring 
conditions. Temperature monitoring data recorded during IMPs shipment will be evaluated by 
Sponsor. IMPs will be used in the study only after written Sponsor approval. 

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Upon arrival at the clinic, the investigational products should be checked for damage and 
proper identity, quantity, integrity of seals and temperature conditions should be verified, any 
deviations or product complaints upon discovery should be reported. All study drugs will be 
dispensed by the investigator or a person under his supervision. The subjects will not be given 
study medication to take home with them. Study drug packaging will be overseen by the 
 and bearing a label with the identification according to 
the requirements of local law and legislation. All drug supplies must be stored in a secure, 
temperature-controlled area with limited access. For batch-specific storage instructions, see the 
packaging. 
The test IMPs will be stored and administered by a medical professional, who is a member of 
the investigational team and in accordance with section 10.6.   
10.4 Drug accountability 
The study drug provided for this study will be used only as directed in the study protocol.  
The study personnel will account for all study drugs dispensed to and returned from the healthy 
volunteer.  
At the end of the study, study site personnel will account for all study drugs received at the site, 
unused study drugs and for appropriate destruction. Destruction must not take place unless the 
Sponsor has approved it. Certificates of delivery and destruction must be signed. The 
investigator is responsible for ensuring that dosing is administered in compliance with the 
protocol. Delegation of this task must be clearly documented and approved by the investigator. 
All study drug administration will occur under medical supervision. 
10.5 Treatment assignment and blinding 
10.5.1 Randomization and treatment assignment  
This is a non-randomized study. 
If a subject, who does not meet the selection criteria, has been dosed before the error is 
identified, the subject should be advised to continue safety assessments to ensure their safety. 
The investigator will inform the Sponsor of the error and a joint decision made as to whether 
the subject should be replaced. 
10.5.2 Blinding  
This is an open label study 
10.6 Dosing Procedure 
After an overnight fasting of at least 10 hours, the IMP will be administered to the subjects 
according to the dosing schedule for the cohort they are part of. 
Dosing of the subjects will be conducted in the following manner: subjects will be dosed one 
by one in a designated room. Prior to each subject being invited to the dosing room, the PI and 
a designated study nurse will check the prepared IMP in accordance with the relevant dosing 
scheme
with the IMP that is prepared and with the dosing scheme again. Upon confirmation by the PI 
and the study nurse that the IMP is the right one and can be administered, the PI will perform 
the administration according to the available instructions. The Investigator will also perform a 
visual IMP identity and intactness check of the IMP prior to the administration. In case of an 
incorrect identity or damaged tablet, the IMP will be replaced with a correct identity or 
undamaged tablet. IMP will be administered orally to each subject in an upright position. IMP 

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administration of the tablet will be done by a dispensation spoon followed by intake of 240 mL 
water as described in sections 10.6.1  10.6.3. 
After 
the tongue, under the lips, in the corners of the mouth and between gums and cheeks, using a 
tongue depressor or a spatula. In case of the Test IMP the PI will confirm that the subject had 
swallowed the administered dose. 
Drug accountability and dispensing records will be maintained at all times. Each activity 
including date and time of each dose will be documented at the time it is performed in the 
source documentation. Copy of labels will be stuck in the source documentation at the time of 
dosing. All empty IMP containers/packaging will be stored in a secure area. 
For each subject, all scheduled post dose activities and assessments will be performed relative 
to the time of study treatment administration. 
For Test Product only the taste assessment will be done by the trained study personnel after 
dosing in each period and will be recorded in the source data form. 
 
10.6.1   Benfo-oxythiamine (B-OT) 
 Depending on the assigned dose, one or more than one tablet of the test product (benfo-
oxythiamine (B-OT)) will be administered with 240 mL of water. 
The tablet(s) must be swallowed whole. The tablet(s) must not be sucked, chewed or broken.  
No water will be allowed one hour before and one hour after administration, except 240 mL of 
water administered together with benfo-oxythiamine (B-OT) administration. 
10.7 Treatment Compliance  
Administration of investigational products will only be performed by authorized clinical 
research staff and will be followed by a hand and oral cavity check in order to confirm the 
swallowing of the product and fluid intake, if applicable.  
Upon dispensing medication, the clinical research staff will record the related information on 
the applicable source documents, to allow investigational product accountability and evaluation 
of subject compliance. 

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11 STUDY ENDPOINTS 
11.1 Primary endpoints 
The outcome measures for the safety endpoints of the study will be safety laboratory 
assessments, physical examination (PE), electrocardiograms (ECGs), vital signs, adverse 
events (AEs) and serious adverse events (SAEs). 
The safety variables will mainly be addressed by the adverse events (AEs) data as determined 
by clinical or biochemical/ hematologic investigations. 
The safety variables will be tabulated for all study participants. AE data will be processed in 
the statistical analysis after coding according to MedDRA dictionary. AEs will be presented in 
summary tables. These tables will show the number of subjects per group presenting an AE 
and the incidence of its occurrence. AEs will be grouped by system organ class and stratified 
by severity and by relation to study treatment. 
Adverse events which will be summarized and will be categorized in subsets of all treatment-
emergent AEs, and of all treatment-related AEs. 
Clinical laboratory and vital signs will be summarized by change from baseline. 
11.2 Secondary endpoints  
In the SAD part of the study, the following PK endpoints will be derived from plasma benfo-
oxythiamine (B-OT) and oxythiamine (OT) concentration versus time data following 
administration according to the dosing schedule: 
-inf 
and AUC last as appropriate. 
In the MAD part of the study the following PK endpoints will be derived from plasma 
oxythiamine (OT) concentration  versus time data following administration according to the 
dosing schedule on Day 1: AUC , Cmax, tmax, on Day 7: AUC , Cmax,ss, tmax,ss, Swing, PTF%, 
CLss/F, Vz/F, Ctrough, Cav, Cmin,  t1/2, AUCinf and AUClast as appropriate and Day7/Day1: 
Ra(AUC ), Ra(Cmax).  
11.3 Exploratory endpoints 
 To evaluate the pharmacodynamics (PD) by measuring the transketolase activity.  
 To analyse PK parameters of other B-OT derivative metabolite(s), if any. 

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12 STUDY ASSESSMENTS (SAD AND MAD) 
12.1 Visit Procedures (SAD) 
The procedures per visit to be performed in the SAD part of the study are listed below. 
12.1.1 Procedures during the Screening Visit (Day -28 to -2) 
 Subjects will come to the Clinical site for an ambulatory screening visit  
 Subjects will be fully informed verbally and in writing about the study procedures and 
their responsibilities/rights while participating in the study  
 If the subject agrees to participate in the study, he or she will sign a written informed 
consent  
 Subjects will provide nasopharyngeal and oropharyngeal swab which will be tested for 
the presence of SARS-COV-2 (COVID  19) infection with a rapid antigen test 
 Subjects will be screened for inclusion and exclusion criteria  
 The Investigator will record any prior and concomitant medical conditions/medications  
 Subjects will provide a detailed medical history and demographic data 
 Subjects will undergo a complete physical examination including documenting weight, 
height, BMI  
 Subjects will have their vital signs (heart rate, blood pressure, respiratory rate, body 
temperature) recorded  
 Subjects will undergo a breath alcohol test 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, serology, urinalysis); the urine sample will be used also for drug screen 
test and cotinine test performed via dip stick  
 Subjects will undergo a 12-lead ECG  
 The Investigator will record any AEs that have occurred during visit with the subject 
 Eligible subjects will be invited for a treatment visit   
Subjects who meet the inclusion and have no exclusion criteria will be scheduled to attend the 
treatment visit (Day -1 to Day 4).   
The subjects will arrive at the study site in the morning of Day -1 for the conduct of the 
treatment visit, provided that they have had a negative Sars-CoV2 tests result from the morning 
of the same day or the evening of the previous one. In case this time frame cannot be respected, 
the time of testing will be decided on a case-by-case basis by the Sponsor, respecting the 
official validity period of the test which is 72 h. 
12.1.2 Procedures on Day -1 
 Subjects will provide nasopharyngeal and oropharyngeal swab which will be tested via 
PCR for the presence of SARS-COV-2 (COVID  19) infection in the morning by 
visiting the laboratory point of testing indicated by the Investigator during the screening 
visit 
 Subjects will arrive at the unit in the afternoon after a confirmed COVID-19 negative 
result is available from the laboratory 
 Subjects´ eligibility will be re-checked and medica history re-reviewed 

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 Subjects will undergo a complete physical examination including documenting weight 
 The Investigator will review any AEs that have occurred since the previous visit with 
the subject 
 Subjects will undergo a breath alcohol test 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, urinalysis); the urine sample will be used also for drug screen test and 
cotinine test performed via dip stick  
 Subjects will have their vital signs measured  
 The Investigator will record any changes in concomitant medications or illnesses with 
the subject since the last study visit 
 Subjects will be admitted to the stationary unit of the site 
 Subjects will receive unified balanced evening meal  
12.1.3 Procedures on Day 1 
 Subjects will be fasted from midnight (at least 10 hours fasting prior to dosing) 
 Subjects will have their vital signs recorded (pre-dose and approximately 2, 4, 8, 12 
hours after dosing) 
 Subjects will undergo limited physical examination to target any change from the 
previous status or any body system deemed pertinent to that subject by the PI (or 
designee) 4 hours post dose 
 Subjects will undergo a 12-lead ECG pre-dose 
 Subjects will receive a single dose of the assigned IMP Benfo-oxythiamine (B-OT) 
between a time interval of two hours between 07:00 am and 09:00 am.  
 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for PK and PD analysis according to the sample schedule in section 12.4.1 Blood 
samples collection 
 AEs that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day  
12.1.4 Procedures on Day 2  
 Subjects will be fasted from midnight (at least 10 hours fasting prior to safety laboratory 
samples collection) 
 Subjects will have their vital signs recorded  
 Subjects will undergo limited physical examination to target any change from the 
previous status or any body system deemed pertinent to that subject by the PI (or 
designee) 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, urinalysis);  
 Subjects will undergo a 12-lead ECG  

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 Blood samples will be collected for PK and PD analysis according to the sample 
schedule in section 12.4.1 Blood samples collection 
 AEs that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day  
12.1.5 Procedures on Day 3 
 Subjects will be fasted from midnight (at least 10 hours fasting prior to safety laboratory 
samples collection) 
 Subjects will have their vital signs recorded  
 Subjects will provide urine samples for laboratory urine analysis  
 Blood samples will be collected for PK and PD analysis according to the sample 
schedule in section 12.4.1 Blood samples collection 
 AEs that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day  
12.1.6 Procedures on Day 4  
 Subjects will be fasted from midnight (at least 10 hours fasting prior to safety laboratory 
samples collection) 
 Subjects will provide nasopharyngeal and oropharyngeal swab which will be tested via 
PCR for the presence of SARS-COV-2 (COVID  19) infection 
 Subjects will have their vital signs recorded  
 Subjects will undergo limited physical examination to target any change from the 
previous status or any body system deemed pertinent to that subject by the PI (or 
designee) 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, urinalysis) 
 Blood samples will be collected for PK and PD analysis according to the sample 
schedule in section 12.4.1 Blood samples collection 
 AEs that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will be discharged from the Clinical site as soon as all scheduled procedures 
have been completed 
12.1.7 Procedures on Follow-up visit (Day 8) 
 Subjects will come to the Clinical site for an ambulatory End of study visit 
 Subjects will undergo a complete physical examination including documenting weight  
 Subjects will provide nasopharyngeal and oropharyngeal swab which will be tested for 
the presence of SARS-COV-2 (COVID  19) infection with a rapid antigen test 

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 Subjects will have their vital signs (heart rate, blood pressure, respiratory rate, body 
temperature) recorded  
 Subjects will undergo a 12-lead ECG  
 Subjects will undergo a breath alcohol test 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, urinalysis); the urine sample will be used also for drug screen test and 
cotinine test performed via dip stick  
 Blood samples will be collected for PK and PD analysis according to the sample 
schedule in section 12.4.1 Blood samples collection 
 AEs that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will complete the study and their participation will be ended 
See Table 1 for the time points of all the assessments planned in the study. 
12.2 Visit Procedures (MAD) 
The procedures per visit to be performed in the MAD part of the study are listed below. 
12.2.1 Procedures during the Screening Visit (Day -28 to -2) 
 Subjects will come to the Clinical site for an ambulatory screening visit  
 Subjects will be fully informed verbally and in writing about the study procedures and 
their responsibilities/rights while participating in the study  
 If the subjects agree to participate in the study, they will sign a written informed consent  
 Subjects will provide nasopharyngeal (through the nose) and/or oropharyngeal (through 
the mouth) swab which will be tested for the presence of SARS-COV-2 infection with 
a rapid antigen test 
 Subjects will be screened for inclusion and exclusion criteria  
 The Investigator will record any prior and concomitant medical conditions/medications  
 Subjects will provide a detailed medical history and demographic data 
 Subjects will undergo a complete physical examination including documenting weight, 
height, body mass index (BMI)  
 Subjects will have their vital signs (heart rate, blood pressure, respiratory rate, body 
temperature) recorded  
 Subjects will undergo a breath alcohol test 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, serology, urinalysis); the urine sample will be used also for drug screen 
test and cotinine test performed via dip stick  
 Adverse events (AEs) that have occurred during the visit will be recorded 
 Subjects will undergo a 12-lead ECG  
 Subjects will be invited for a treatment visit   
If Subjects fulfill the inclusion criteria and do not meet any exclusion criteria, Subjects will be 
scheduled to attend the treatment visit (Day -1 to Day 8).   

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Subjects will arrive at the study site in the afternoon of Day -1 for the conduct of the treatment 
visit 
12.2.2 Procedures on Day -1 
 Subjects will provide nasopharyngeal and/or oropharyngeal swab which will be tested 
for the presence of SARS-COV-2 infection with a rapid antigen test  
 Subjects will arrive at the unit in the afternoon  
 Subjects´ eligibility will be re-checked and medica history re-reviewed 
 Subjects will undergo a complete physical examination including documenting weight 
 The Investigator will review any adverse events (AEs) that have occurred since the 
previous visit with the subjects 
 Subjects will undergo a breath alcohol test 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, coagulation, urinalysis); the urine sample will be used also for drug screen 
test and cotinine test performed via dip stick  
 Subjects will have their vital signs measured  
 The Investigator will record any changes in concomitant medications or illnesses with 
the subjects since the last study visit 
 Subjects will be admitted to the stationary unit of the site 
 Subjects will receive unified balanced evening meal  
12.2.3  Procedures on Day 1 
 Subjects will be fasted from midnight (at least 10 hours fasting prior to dosing) 
 Subjects will have their vital signs recorded (pre-dose)  
 Subjects will undergo limited physical examination to target any change from the 
previous status or any body system deemed pertinent to Subjects by the PI (or designee) 
4 hours post dose 
 Subjects will undergo a 12-lead ECG pre-dose 
 Subjects will receive a single dose of the assigned IMP (benfo-oxythiamine (B-OT) 
within a time interval of two hours between 07:00 am and 09:00 am.  
 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for pharmacokinetic (PK) and pharmacodynamic (PD) analysis according to the sample 
schedule in section 12.4.1 Blood samples collection. 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day  
12.2.4  Procedures on Day 2  
 Subjects will be fasted from midnight (at least 10 hours fasting prior to dosing) 
 Subjects will have their vital signs recorded (pre-dose and approximately 4 hours after 
dosing) 

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 Subjects will undergo a limited physical examination to target any change from the 
previous status or any body system deemed pertinent to Subjects by the PI (or designee) 
4 hours post dose 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, coagulation, urinalysis) 
 Subjects will undergo a 12-lead ECG pre-dose 
 Subjects will receive a single dose of the assigned IMP (Benfo-Oxythiamine (B-OT) 
within a time interval of two hours between 07:00 am and 09:00 am.  
 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for pharmacokinetic (PK) and pharmacodynamic (PD) analysis according to the sample 
schedule in section 12.4.1 Blood samples collection. 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day  
12.2.5  Procedures on Day 3 
 Subjects will be fasted from midnight (at least 10 hours fasting prior to dosing) 
 Subjects will have their vital signs recorded (pre-dose) 
 Subjects will undergo a limited physical examination to target any change from the 
previous status or any body system deemed pertinent to Subjects by the PI (or designee) 
4 hours post dose 
 Subjects will receive a single dose of the assigned IMP (Benfo-Oxythiamine (B-OT) 
within a time interval of two hours between 07:00 am and 09:00 am.  
 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for pharmacokinetic (PK) analysis according to the sample schedule in section 12.4.1 
Blood samples collection 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day  
12.2.6  Procedures on Day 4  
 Subjects will be fasted from midnight (at least 10 hours fasting prior to dosing) 
 Subjects will have their vital signs recorded (pre-dose) 
 Subjects will undergo a limited physical examination to target any change from the 
previous status or any body system deemed pertinent to Subjects by the PI (or designee) 
4 hours post dose 
 Subjects body weight will be measured 
 Subjects will undergo a 12-lead ECG pre-dose 
 Subjects will receive a single dose of the assigned IMP (Benfo-Oxythiamine (B-OT) 
within a time interval of two hours between 07:00 am and 09:00 am.  

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 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for pharmacokinetic (PK) and pharmacodynamic (PD) analysis according to the sample 
schedule in section 12.4.1 Blood samples collection 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day  
12.2.7  Procedures on Day 5  
 Subjects will be fasted from midnight (at least 10 hours fasting prior to dosing) 
 Subjects will have their vital signs recorded (pre-dose and approximately 4 hours after 
dosing) 
 Subjects will undergo a limited physical examination to target any change from the 
previous status or any body system deemed pertinent to Subjects by the PI (or designee) 
4 hours post dose 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, coagulation, urinalysis) 
 Subjects will receive a single dose of the assigned IMP (Benfo-Oxythiamine (B-OT) 
within a time interval of two hours between 07:00 am and 09:00 am.  
 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for pharmacokinetic (PK) and pharmacodynamic (PD) analysis according to the sample 
schedule in section 12.4.1 Blood samples collection 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day  
12.2.8  Procedures on Day 6  
 Subjects will be fasted from midnight (at least 10 hours fasting prior to dosing) 
 Subjects will have their vital signs recorded (pre-dose) 
 Subjects will undergo a limited physical examination to target any change from the 
previous status or any body system deemed pertinent to Subjects by the PI (or designee) 
4 hours post dose 
 Subjects will receive a single dose of the assigned IMP (Benfo-Oxythiamine (B-OT) 
within a time interval of two hours between 07:00 am and 09:00 am.  
 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for pharmacokinetic (PK) analysis according to the sample schedule in section 12.4.1 
Blood samples collection 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day 

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12.2.9  Procedures on Day 7  
 Subjects will be fasted from midnight (at least 10 hours fasting prior to dosing) 
 Subjects will have their vital signs recorded (pre-dose and approximately 4 hours after 
dosing) 
 Subjects will undergo limited physical examination to target any change from the 
previous status or any body system deemed pertinent to Subjects by the PI (or designee) 
4 hours post dose 
 Subjects will undergo a 12-lead ECG pre-dose 
 Subjects will receive a single dose of the assigned IMP (Benfo-Oxythiamine (B-OT) 
within a time interval of two hours between 07:00 am and 09:00 am.  
 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for pharmacokinetic (PK) and pharmacodynamic (PD) analysis according to the sample 
schedule in section 12.4.1 Blood samples collection 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day 
12.2.10 Procedures on Day 8 
 Subjects will have their vital signs recorded  
 Subjects will undergo a limited physical examination to target any change from the 
previous status or any body system deemed pertinent to Subjects by the PI (or designee)  
 Subjects body weight will be measured 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, coagulation, urinalysis) 
 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for pharmacokinetic (PK) and pharmacodynamic (PD) analysis according to the sample 
schedule in section 12.4.1 Blood samples collection 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will receive unified balanced lunch, afternoon snack and evening meal 
throughout the day 
12.2.11 Procedures on follow-up Day 11 
 Subjects will have their vital signs recorded  
 Subjects will undergo limited physical examination to target any change from the 
previous status or any body system deemed pertinent to Subjects by the PI (or designee)  
 Subjects body weight will be measured 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, coagulation, urinalysis) 

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 Catheter for blood sampling will be inserted and serial blood samples will be collected 
for pharmacokinetic (PK) analysis according to the sample schedule in section 12.4.1 
Blood samples collection 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
12.2.12 Procedures on End of study visit (Day 14) 
 Subjects will come to the Clinical site for an ambulatory End of study visit 
 Subjects will undergo a complete physical examination including documenting weight  
 Subjects will provide nasopharyngeal and oropharyngeal swab which will be tested for 
the presence of SARS-COV-2 infection with a rapid antigen test 
 Subjects will have their vital signs (heart rate, blood pressure, respiratory rate, body 
temperature) recorded  
 Subjects body weight will be measured 
 Subjects will provide blood and urine samples for laboratory analysis (biochemistry, 
hematology, coagulation, urinalysis); the urine sample will be used also for drug screen 
test and cotinine test performed via dip stick 
 Subjects will undergo a 12-lead ECG  
 Subjects will undergo a breath alcohol test 
 Blood samples will be collected for PK analysis according to the sample schedule in 
section 12.4.1 Blood samples collection 
 Adverse events (AEs) that have occurred will be recorded 
 Concomitant medication intake will be recorded 
 Subjects will complete the study and their participation will be ended 
Every effort must be taken to ensure that subjects terminating the study prematurely undergo 
the safety checks of the follow-up visit.  If subjects terminating the study prematurely due to 
an AE, every effort must be taken to monitor the subjects until the AE has resolved. 
12.3 Safety and tolerability assessments  
Only healthy subjects are eligible for the study. The healthy status will be determined by 
study. Adverse events will be monitored and recorded throughout the study. 
Prior to the first benfo-oxythiamine (B-OT) dosing only, the Principal Investigator or a 
delegated medical doctor will assess the clinical significance of results of laboratory 
investigations for hematology and serum chemistry values outside the defined normal ranges, 
as provided by the laboratory.  
In addition to the planned times, any safety procedures can be performed at any time considered 
necessary by the Principal Investigator or a delegated medical doctor. 
At least one qualified physician will be present at all times during each confinement day of 
each cohort. In addition, an ambulance and an emergency doctor will be available on each 
dosing day for a period of 12 hours (from 08:00 am until 08:00 pm) for the SAD part and for a 
period of 4 hours for the MAD part at the site and on call during the rest of the study. Also, 
two cardiologists will remain available on call at all times during the entire period of the study. 
In case any clinically significant abnormality is observed at pre-dose, Principal Investigator or 

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a delegated medical doctor will decide whether the subject will proceed to investigational 
product administration or will be withdrawn from the study. 
Safety will be assessed by consideration of all adverse events reported by or elicited from the 
subject and abnormalities detected on physical examinations, safety blood tests, urinalysis, 
vital signs and 12-lead ECG.  
All adverse events (serious and non-serious) beginning prior to dosing through follow-up visit 
 
The definitions, reporting and follow-up of AEs, SAEs and potential pregnancies are described 
in section 13.1. 
12.3.1 Vital Signs 
Evaluations of systolic and diastolic blood pressure, pulse rate, respiratory rate, and body 
temperature will be performed throughout the study. Pulse and blood pressure will be taken 
after 5 minutes in the supine position. 
12.3.2 Weight and height 
Weight (kg), height (cm) and body mass index (BMI) will be recorded/calculated at screening. 
12.3.3 Physical examination 
Physical examination (including general appearance, head, ears/eyes/nose/throat/neck, skin, 
respiratory system, cardiovascular system, gastrointestinal (including mouth) system, 
musculoskeletal system, central and peripheral nervous systems, and lymph nodes) will be 
performed by the Investigator(s) or a designee. Other body systems can be examined if 
required, at their discretion of the Investigator. Clinically relevant findings that are present 
Clinically relevant findings found after study drug administration and meeting the definition of 
an AE (new AE or worsening of previously existing condition) must be recorded. 
12.3.4 Electrocardiography 
A complete standard 12-
under the following conditions: 
 Supine position of the subject for at least 5 minutes rest 
 Use of a computerized ECG device 
 Automatic calculation of the following parameters: heart rate, PR interval, QRS width, 
QT interval, and QTcF (calculated using Fredericia's formula) 
 ECG printouts will be examined and interpreted by the investigator on the day of 
recording. This analysis performed by the investigator has to be entered in the eCRF.  
Any new clinically relevant abnormality that occurred after signing the informed 
consent will be documented as an AE. 
The investigator will assess the ECG recording as 'normal', 'abnormal 
 not clinically 
significant', or 'abnormal  clinically significant' and include a description of the abnormality 
as required. ECG printouts will be collected and stored at the site. Each ECG trace should be 
labelled with the study number, subject number, subject initials and date of birth. 
12.3.5 Laboratory assessments, drug screen and breath alcohol test 
Routine laboratory safety samples will be analyzed by the local laboratory Bodimed ltd. The 
site will be supplied will kits for sample collection, requisition forms for sample shipment to 
analytical site and a laboratory manual with instructions for collection and handling of the 

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samples. The analysis of urine will be done at the site with urine dip stick tests, which will be 
purchased by the site team prior to the initiation of the study. 
Printed laboratory reports will include normal reference ranges. A decision regarding whether 
the result outside the reference range is of clinical significance or not shall be made by an 
Investigator and the report will be annotated accordingly. Clinically significant abnormalities 
at screening or occurring during the study will be recorded on the AE page. The reference 
ranges for laboratory parameters will also be filed in the Investigator site file. 
The laboratory results should be signed and dated and retained at clinical research site as source 
data for laboratory variables.   
Any clinically significant deviation of laboratory parameters from reference ranges will be 
documented in the source documentation and in the eCRF and followed up, by the Principal 
Investigator or authorized investigator. Re-test of the safety laboratory parameters may be done 
 
Blood and urine samples will be collected for the following safety clinical laboratory tests as 
listed in Table 3: 
TABLE 3: SAFETY PARAMETERS 
Laboratory Test/Safety 
Procedure 
Parameters 
SAD Part 
Hematology 
 
Haemoglobin (HGB), Mean corpuscular volume (MCV), 
Mean corpuscular haemoglobin (MCH), Mean corpuscular 
haemoglobin concentration (MCHC), hematocrit (HCT), 
red cell count (RBC), white blood cells (WBC) neutrophils 
(NEUT), lymphocytes (LYMP), monocytes (MONO), 
eosinophils (EOS), basophils (BASO) and platelets (PLT). 
Chemistry and electrolytes 
Sodium, potassium, calcium, inorganic phosphate, total 
protein, albumin, glucose1, triglycerides, blood urea 
nitrogen (BUN), creatinine, uric acid, total bilirubin2, 
alkaline phosphatase, AST, ALT gamma-GT and LDH. 
Serology 
HIV1 and HIV2 antibodies, Hepatitis B surface antigen, 
Hepatitis B antibodies and Hepatitis C antibodies 
SARS-COV-2 (COVID  19) 
infection 
Polymerase Chain Reaction method for detection of the 
virus 
Urinalysis (will be done using 
dip stick test at the site) 
Leucocytes, blood, nitrite, protein, urobilinogen, bilirubin, 
pH, specific gravity, ketones, glucose.  
Alcohol 
Alcohol Breath Test 
Urine drug screen 
Cocaine, amphetamines, opiates, benzodiazepines and 
cannabinoids.  
MAD Part 
Hematology 
 
Haemoglobin (HGB), Mean corpuscular volume (MCV), 
Mean corpuscular haemoglobin (MCH), Mean corpuscular 
haemoglobin concentration (MCHC), hematocrit (HCT), 
red cell count (RBC), white blood cells (WBC) neutrophils 

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(NEUT), lymphocytes (LYMP), monocytes (MONO), 
eosinophils (EOS), basophils (BASO) and platelets (PLT). 
Coagulation 
Fibrinogen, prothrombin time (PT), activated partial 
thromboplastin time (aPTT) 
Chemistry and electrolytes 
Sodium, potassium, calcium, inorganic phosphate, total 
protein, albumin, glucose1, triglycerides, blood urea 
nitrogen (BUN), creatinine, uric acid, total bilirubin2, 
alkaline phosphatase, AST, ALT gamma-GT and LDH. 
Serology 
HIV1 and HIV2 antibodies, Hepatitis B surface antigen, 
Hepatitis B antibodies and Hepatitis C antibodies 
SARS-COV-2 (COVID  19) 
infection 
Rapid antigen test for detection of the virus 
Urinalysis (will be done using 
dip stick test at the site) 
Leucocytes, blood, nitrite, protein, urobilinogen, bilirubin, 
pH, specific gravity, ketones, glucose.  
Alcohol 
Alcohol Breath Test 
Urine drug screen 
Cocaine, amphetamines, opiates, benzodiazepines and 
cannabinoids, barbiturates.  
 
Urine drug test will be performed on site with a dip stick test and will include amphetamine, 
barbiturates, benzodiazepines, cocaine, cannabinoids, opioids and nicotine. At time-points 
when both urinalysis and drugs of abuse screening is required, all testing will be performed 
from a single sample. 
An alcohol breath test will be conducted on site using a breathalyzer. The results of the test 
result will be considered to have a negative test result.  
A full chain of custody will be maintained for all samples throughout their lifecycle. 
The investigator will ensure full traceability of collected biological samples from the subjects 
while in storage at the center until shipment and will keep documentation of receipt of arrival. 
The sample receiver will keep full traceability of samples while in storage and during use, until 
used, disposed of, or until further shipment or disposal (where appropriate) and will keep 
documentation of receipt of arrival. 
12.3.6 Concomitant medications 
In this study, limited types of concomitant medications are permitted. Adjustment of routine 
medications taken by subjects should be avoided during study participation except when 
subject safety could be affected by lack of adjustment 
Concomitant medications initiated or stopped for an AE will be recorded in the source 
documentation and CRF of the relevant subject. 
If a concomitant medication is used after the first treatment administration an investigator or 
delegate and/or the sponsor will decide whether the subject will be permitted to remain in the 
study, depending on the drug used, the time of drug intake, etc.  
 

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12.4 Pharmacokinetic and pharmacodynamics assessments  
12.4.1 Blood samples collection 
Blood samples for PK and PD determination will be processed, stored, and shipped according 
to the sample processing instructions supplied by the bioanalytical facility. All blood samples 
will be drawn using an indwelling IV catheter or by venipuncture. After blood sample 
collection from the IV catheter, the catheter will be flushed with a 0.9 % w/v NaCl solution to 
avoid obstruction (without Hep Lock). The indwelling IV catheter will not be left in situ for 
more than 24 hours, except during treatment period. If the intravenous catheter is not available 
for use and/or has been in place for > 24 hours, samples will be drawn by venipuncture, except 
during the treatment period.  
SAD Study part:  
PK blood samples (5 mL) for determination of plasma benfo-oxythiamine (B-OT), 
oxythiamine (OT) or other derived metabolites concentrations will be taken into pre-labelled 
EDTA collection tubes at the time points indicated in the Visit and Assessment Schedule (Table 
1).  
The quantitative determination of B-OT and OT in human plasma will be performed by using 
a validated method.  
Samples should be maintained at 4 ºC (± 4 ºC) during the whole procedure from sample 
collection until sample freezing. 
Samples will be cooled in an ice/water bath or cooling device until processed, centrifuged at 
approximately 1900 g (± 38g) at 4°C (± 4°C) for 10 minutes and then placed into an ice/water 
bath or cooling device. The time between blood collection and placement in the centrifuge will 
not exceed 60 minutes. 
Plasma will be divided and transferred into pre-labeled duplicate 2 mL polypropylene tubes for 
each analyte (B-OT and OT). The resulting 4 aliquots will be maintained in the ice/water bath 
or cooling device. A minimum of 0.5 mL of plasma is required for each aliquot. 
Samples will be kept in an ice/water bath until transferred to the storage unit (freezer). 
Then, samples will be stored and frozen at approximately -80ºC (±15ºC) until shipment to the 
bioanalytical laboratory. The time between the centrifuge and placement in freezer will not 
exceed 60 minutes. 
 
MAD Study part: 
PK blood samples (2.0 mL) for determination of plasma oxythiamine (OT) and any other 
derived metabolites concentrations will be taken into pre-labelled EDTA collection tubes at the 
time points indicated in the Visit and Assessment Schedule (Table 2).  
The quantitative determination of OT and any other derived metabolites in human plasma will 
be performed by using a validated method.  
Samples should be maintained at 4 ºC (± 4 ºC) during the whole procedure from sample 
collection until sample freezing. 
Samples will be cooled in an ice/water bath or cooling device until processed, centrifuged at 
approximately 1900 g (± 38g) at 4°C (± 4°C) for 10 minutes and then placed into an ice/water 
bath or cooling device. The time between blood collection and placement in the centrifuge will 
not exceed 60 minutes. 

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Plasma will be divided and transferred into pre-labeled duplicate 2 mL polypropylene tubes 
The resulting 2 aliquots will be maintained in the ice/water bath or cooling device. A minimum 
of 0.5 mL of plasma is required for each aliquot. 
 
Samples will be kept in an ice/water bath until transferred to the storage unit (freezer). 
Then, samples will be stored and frozen at approximately -80ºC (±15ºC) until shipment to the 
bioanalytical laboratory. The time between the centrifuge and placement in freezer will not 
exceed 60 minutes. 
In the SAD part, PD blood samples (2 mL) for transketolase activity analysis will be managed 
according to the instructions leaflet dated 08 Nov 2021, provided by the responsible laboratory 
Medical Laboratory Bremen and are the following: 
a. Collect samples in pre-labelled EDTA (2 mL) collection tubes at the time points 
indicated in the Visit and Assessment Schedule (Table 1). 
b. Transfer the sample material into a reagent tube of approx. 13 ml capacity, preferably 
made of polypropylene, e.g. SARSTED, 95 mm x 16.8 mm, article no. 55.518. 
Polystyrene tubes are very fragile when frozen (see below). 
c. Fill up the tube with isotonic (0.9%) NaCl solution. 
d. Centrifugation is then performed at room temperature in a standard laboratory 
centrifuge at approx. 2800 g, which corresponds to 3800 rpm when e.g. SIGMA 3-16P 
K centrifuge is used, centrifugation time is 10 min, brake acceptable. 
e. Remove the supernatant (including the buffy coat) and fill up again with isotonic NaCl 
solution. 
f. Then centrifuge again, when removing the supernatant, immerse about 1 mm deep into 
the erythrocyte sediment, fill up with isotonic NaCl. 
g. Repeat the above steps again, remove the supernatant for the last time. Mix 0.5 ml of 
the washed and packed erythrocytes with 0.5 ml of distilled water (not isotonic NaCl!). 
h. Mix thoroughly (e.g. VORTEX mixer 10 sec), by this step most of the erythrocytes are 
hemolyzed and their components are released. 
i. Freeze at -70°C until shipment to the laboratory within the planned schedule of 
shipments 
In the MAD part, PD blood samples (5 ml) will be collected in prelabelled EDTA collection 
tubes at time points indicated in the visit and assessment schedule (Table 2). The samples will 
be prepared and analyzed according to a validated method if available. 
The following blood samples (Table 4) need to be drawn for PK and PD analysis on each of 
the sampling days, as indicated in the Visit and Assessment Schedule (Table 1 and 2). 
 
 

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TABLE 4: PK/PD SAMPLING TIMEPOINTS 
SAD part 
PK sample timepoint 
(hr.) 
Allowed time 
deviation 
PD sample timepoint 
(hr.) 
Allowed time 
deviation 
Pre-dose  
(35 min before dosing) 
+/- 5 min 
Pre-dose 
(35 min before dosing)
+/- 5 min 
0.25 
+/- 5 min 
- 
- 
0.5 
+/- 5 min 
- 
- 
0.75 
+/- 5 min 
- 
- 
1.0 
+/- 5 min 
1.0 
+/- 5 min 
1.5 
+/- 5 min 
- 
- 
2 
+/- 5 min 
2.0 
+/- 5 min 
4 
+/- 10 min 
4.0 
+/- 10 min 
8 
+/-15 min 
8.0 
+/-15 min 
12 
+/- 30 min 
12 
+/- 30 min 
24 
+/- 1 hr 
24 
+/- 1hr 
36 
+/- 1 hr 
- 
- 
48 
+/- 1hr 
48 
+/- 1hr 
72 
+/- 1hr 
72 
+/- 1hr 
168 
+/- 2 hr 
168 
+/- 2 hr 
 
 
 

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MAD part 
Day 
PK sample timepoint 
(hr.) 
Allowed time 
deviation 
PD sample timepoint 
(hr.) 
Allowed time 
deviation 
1 
Pre-dose  
(35 min before dosing) 
+/- 5 min 
Pre-dose 
(35 min before dosing)
+/- 5 min 
0.25 
+/- 5 min 
- 
- 
0.5 
+/- 5 min 
- 
- 
0.75 
+/- 5 min 
- 
- 
1.0 
+/- 5 min 
1.0 
+/- 5 min 
1.5 
+/- 5 min 
- 
- 
2 
+/- 5 min 
2.0 
+/- 5 min 
4 
+/- 10 min 
4.0 
+/- 10 min 
8 
+/-15 min 
8.0 
+/-15 min 
12 
+/- 30 min 
12 
+/- 30 min 
2 
Pre-dose  
(35 min before dosing; 
24 hr after last dosing 
+/- 30 min 
Pre-dose  
(35 min before dosing; 
24 hr after last dosing 
+/- 30min 
3 
Pre-dose  
(35 min before dosing; 
24 hr after last dosing 
+/- 30 min 
- 
 
4 
Pre-dose  
(35 min before dosing; 
24 hr after last dosing 
+/- 30 min 
Pre-dose 
(35 min before dosing)
- 
- 
- 
1.0 
- 
2.0 
4.0 
8.0 
12 
+/- 30 min 
5 
Pre-dose  
(35 min before dosing; 
24 hr after last dosing 
+/- 30 min 
Pre-dose  
(35 min before dosing; 
24 hr after last dosing 
 
6 
Pre-dose  
(35 min before dosing; 
24 hr after last dosing
+/- 30 min 
- 
+/- 30 min 
7 
Pre-dose  
(35 min before dosing)
+/- 5 min 
Pre-dose 
(35 min before dosing)
+/- 5 min 
0.25 
+/- 5 min 
- 
- 
0.5 
+/- 5 min 
- 
- 
0.75 
+/- 5 min 
- 
- 
1.0 
+/- 5 min 
1.0 
+/- 5 min 
1.5 
+/- 5 min 
- 
- 
2 
+/- 5 min 
2.0 
+/- 5 min 

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MAD part 
Day 
PK sample timepoint 
(hr.) 
Allowed time 
deviation 
PD sample timepoint 
(hr.) 
Allowed time 
deviation 
4 
+/- 10 min 
4.0 
+/- 10 min 
8 
+/-15 min 
8.0 
+/-15 min 
12 
+/- 30 min 
12 
+/- 30 min 
8 
24 hr after last dosing 
+/- 30 min 
24 hr after last dosing 
+/- 30 min 
11 
96 hr after last dosing 
+/- 4hr 
- 
- 
14 
168 hr after last dosing 
+/- 4hr 
- 
- 
The clock time will be recorded and reported for all blood draws and all subjects. Actual 
sampling time will be used for pharmacokinetic calculations. A deviation greater than the one 
indicated as allowed will be reported as a protocol deviation in a master protocol deviation log 
and its cause will be recorded.  
When the following assessments are scheduled to be performed at the same time-point, the 
order of priority will be as follows:  
1. ECG 
2. Vital signs 
3. Blood sampling for safety 
4. Physical examination 
When a PK assessment is scheduled for the same nominal time as another scheduled 
assessment, the PK sample will have priority, unless other procedures are necessary for 
 
If an assessment or collection is scheduled by day, and not by a specific time, then no time 
window will apply although generally the time of day should be the same throughout the study. 
12.4.2 Labelling  
Pre-printed, waterproof labels will be used to identify all PK and PD tubes used during blood 
sample collection and for storage of separated plasma. Each label will contain the following 
information: 
 Protocol number 
 Subject Number  
 Study period number (e.g. D-1)  
 Blood Sample Number 
 Sampling time point (e.g. 0:30m; 1:00h) 
 Sample type (blood or plasma) & purpose (e.g. PK) 
12.4.3 Shipping procedures 
The primary PK and PD samples will be stored at the clinical facility until the shipment to the 
respective analytical Biochemical Laboratories The back-up samples can be shipped to 
laboratory only if required, after Sponsor written approval.  
The CRO will arrange shipment of the PK and PD samples. Samples must be shipped to the 
Analytical Biochemical Laboratories at time intervals agreed with the Sponsor. The samples 

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will be transferred in an appropriate container packed in an adequate amount of dry ice enough 
to keep the samples frozen for at least 72 hours. For all shipments, the laboratory should 
acknowledge in writing the receipt of plasma samples in good condition. 
The samples destined for retention will be stored by the clinical facility in a freezer at -80°C 
and will remain there until the final report is issued and the study sponsor authorization is 
obtained to discard the samples. The packaging of the samples will meet the Biosafety rules 
and IATA recommendations related to the documentation and package.  
12.4.4 Determination of plasma drug concentrations and measurement of the 
transketolase activity  
The concentration in Plasma of B-OT, OT and potential other Thiamine metabolites derivate 
will be determined using a validated liquid chromatography tandem mass spectrometry method 
and will be conducted in accordance with Good Laboratory Practice regulations and guidelines. 
Methods validation must be completed prior to start of the analysis.  
In the SAD part, For each cohort, in preparation of the SRC, the PK analysis is conducted after 
all Day 2 (T=48hr) are taken. The rest of the PK samples ((T=72 and 168hr) are analysed after 
all PK samples are collected at the end of the study. In each run, standard and quality control 
samples will be distributed throughout the batch containing study samples. Samples with 
concentrations above the ULOQ will be diluted and re-assayed. 
In the MAD part, for each cohort, in preparation of the SRC, the PK analysis will be conducted 
after all Day 5 samples are taken. The rest of the PK samples are analysed after all PK samples 
are collected at the end of the study. In each run, standard and quality control samples will be 
distributed throughout the batch containing study samples. Samples with concentrations above 
the ULOQ will be diluted and re-assayed. As B-OT was not detected in any cohort of the SAD 
part, the concentration of B-OT will not be assessed in the MAD. Other metabolites (e.g 
oxythiamine pyrophosphate) of this prodrug might however be assessed from the back up 
samples if a validated method has been established before the end of the study. 
 
Full details, including the lower (LLOQ) and upper (ULOQ) limits of quantification range, will 
be provided in a Bioanalytical Plan  
Plasma samples of withdrawn subjects should also be analyzed for drug concentrations and 
pharmacokinetic parameters (providing withdrawal was not due to withdrawal of consent). 
Unacceptable values due to analytical reasons will be handled in accordance with the 
Bioanalytical Plan. 
Measurement of the transketolase activity in erythrocytes will be done following the routinely 
established procedure to perform the analysis in Medical Laboratory Bremen in the SAD part 
of the study. 
In the MAD part of the study transketolase activity measurement will be done according to a 
validated method established by the responsible laboratory if available.  
12.2.5 Total blood volume and appropriateness of measurements 
All measurements performed in the study are standard measurements. 
The total volume of blood to be collected from each subject during the study as listed in table 
5 (approximately 139 mL) is considered acceptable. 
 
 

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TABLE 5: TOTAL VOLUME OF BLOOD PER SUBJECT 
Sample 
Number of samples 
Sample volume 
Total volume 
SAD part 
Hematology 
5 
3 mL 
15 mL 
Chemistry 
5 
4.5 mL 
22.5 mL 
Serology 
1 
6 mL 
6 mL 
PK 
15 
5 mL 
75 mL 
PD (TKT activity)
10 
2 mL 
20 mL 
Total blood volume/subject (SAD) 
138.5 mL
MAD part 
Hematology
7
3 mL
21 mL 
Chemistry
7
5 mL 
35 mL 
Coagulation
7
2.7 mL 
19 ml 
Serology
1
6 mL 
6 mL 
PK
28 
2 mL 
56 mL 
PD (TKT activity) 
21 
5 mL 
105mL 
Total blood volume/subject (MAD) 
242 mL 

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13 SAFETY REPORTING 
13.1 Definitions of adverse events  
The definitions are adopted in accordance with the Directive 2001/20/EC, ICH-E2A and the 
European Commission Detailed Guidance 2011/C 172/01. 
An Adverse Event (AE) is defined as any untoward medical occurrence in a clinical trial 
subject administered a medicinal product and which does not necessarily have a causal 
relationship with the treatment. 
An AE can therefore be any unfavourable and unintended sign (including any clinically 
significant abnormal laboratory finding), symptom, or disease temporally associated with the 
use of an investigational medicinal product, whether or not considered related to the 
investigational medicinal product. 
The following should not be recorded as AEs, if recorded as medical history/concomitant 
illness at screening: 
 Pre-planned procedure, unless the condition for which the procedure was planned has 
worsened from the first trial related activity after the subject has signed the informed 
consent 
 Pre-existing conditions found as a result of screening procedures. 
A clinically significant deterioration of a value for a laboratory parameter should be recorded 
as an adverse event. For alterations to clinical laboratory parameters over the course of the 
result, which is deemed clinically significant when any of the following conditions are met: 
 The abnormality suggests a disease and/or organ toxicity that is new or has worsened 
from baseline. 
 The abnormality is of a degree that requires additional active management, e.g., change 
of treatment, discontinuation of treatment, close observation, more frequent follow-up 
assessments, or further diagnostic investigation. 
An Adverse Drug Reaction (ADR) means all untoward and unintended responses to an 
investigational 
has at least a reasonable possibility, i.e. the 
relationship cannot be ruled out and is judged by the Investigator as at least possible (see 
definition below). 
An Unexpected Adverse Drug Reaction (UADR)/Unexpected Adverse Event (UAE) means 
an adverse reaction/event, the nature or severity of which is not consistent with the applicable 
product information, namely in the Investigator Brochure for an unauthorized investigational 
product or in the SmPC for an authorized product. 
The expected/unexpected status should be evaluated and assessed, by the Sponsor, based on 
the reference safety information available since expectedness in Pharmacovigilance refers 
strictly to the information listed or mentioned in the applicable reference safety information 
and not to events that might be anticipated from knowledge of the pharmacological properties 
of a substance or because it was foreseeable due to the health status (e.g., age, medical history) 
of the study subjects. 
A Serious Adverse Event (SAE) or Serious Adverse Reaction (SAR) is defined as an AE 
that results in any of the following: 
 Results in death 

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 Is life-threatening 
 
hospitalization 
 Results in persistent or significant disability or incapacity 
 Results in a congenital abnormality or birth defect 
 Is an important medical event which requires medical intervention to prevent any of the 
above outcomes 
SUSARs: AEs which meet all of the following criteria: 
 Serious. 
 Unexpected (i.e., is not consistent with the applicable product information e.g. 
Investigator's brochure for an unapproved benfo-oxythiamine (B-OT) or SmPC for an 
authorised product). 
 Suspected (i.e., there is at least a reasonable possibility that there is a causal relationship 
between the event and the medicinal product). 
will be classified as suspected unexpected serious adverse reactions (SUSARs). 
Important medical events are those which may not be immediately life-threatening, but may 
jeopardize the subject and may require intervention to prevent one of the other serious 
outcomes listed above. Examples of such events are intensive treatment in an emergency room 
or at home for allergic bronchospasm, or blood dyscrasias or convulsions that do not result in 
hospitalization. 
life-threatening
was at risk of death at the time of the event; it does not refer to an event which hypothetically 
might have caused death if it were more severe. For example, drug-induced hepatitis that 
resolves without evidence of hepatic failure would not be considered life threatening even 
though drug-induced hepatitis can be fatal. 
Inpatient hospitalization or prolongation of existing hospitalization means that hospital 
inpatient admission and/or prolongation of hospital stay were required for treatment of AE or 
occurred as a consequence of the event. It does not refer to pre-planned elective hospital 
admission for treatment of a pre-existing condition that has not significantly worsened, or to 
diagnostic procedures. 
13.1.1 Severity of adverse events 
The intensity of clinical AEs is graded three-point scale as defined below:  
 Mild: discomfort noticed but no disruption of normal daily activity; 
 Moderate: discomfort sufficient to reduce or affect normal daily activity, requiring no 
or minimal medical intervention; 
 Severe: inability to work or perform daily activity, requires medical intervention 
13.1.2 Relationship to study drug 
For each AE the relationship to the study medication as judged by the investigator will be as 
follows:  
Category 
Description 
Not Related 
A clinical event, including laboratory test abnormality that has no temporal 
relationship to the study medication or has more likely alternative etiology.

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Unlikely 
A clinical event, including laboratory test abnormality, with little or no 
temporal relationship to study medication administration, and which other 
drugs, chemicals or underlying disease provide plausible explanations. 
Possible 
A clinical event, including laboratory test abnormality, with a reasonable time 
sequence to the study medication administration, but which could also be 
explained by concurrent disease or other drugs or chemicals. Information on 
the drug withdrawal may be lacking or unclear. 
Probable 
A clinical event, including laboratory test abnormality, which follows a 
reasonable temporal sequence from the time of study medication 
administration, and/or follows a known response pattern to the product, and 
state, therapeutic intervention or concomitant therapy.
Certain 
A clinical event, including laboratory test abnormality, occurring in a plausible 
time relationship to the study medication administration, and which cannot be 
explained by concurrent disease or other drugs or chemicals. The response to 
withdrawal of the drug should be clinically plausible. 
 
13.1.3 Chronicity of adverse events 
The chronicity of the AE will be classified by the investigator on a three-item scale as defined 
below: 
 Single occasion: single event with limited duration; 
 Intermittent: several episodes of an event, each of limited duration; 
 Persistent: event which remained indefinitely 
13.1.4 Action 
The Investigator must provide the action taken with regard to each AE as: 
 None 
 Study drug dosing interrupted 
 Use of intervention to treat the AE (provide details, recording concomitant therapy, if 
needed) 
 Discontinuation of the study drug 
13.1.5 Outcome of adverse event 
The Investigator must provide the outcome of the adverse event as: 
 Recovered/Resolved  
 Recovered/Resolved with sequelae  
 Recovering/Resolving  
 Not recovered/Not resolved 
 Fatal  
 Unknown 
 
Sequelae: The signs/symptoms of the reported AE have not completely resolved and a new 
baseline for the subject is established since full recovery is not expected. 
13.1.6 Reporting of adverse events and serious adverse events 
Time period for collection of adverse events and serious adverse events: 
Adverse events will be recorded from the time of informed consent up to and including the 
follow-up visit.  
 

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AEs which occurred prior to initiation of study medication will be recorded as screening events 
or as part of the medical history (as applicable), only occurrences related to protocol mandated 
procedures or ongoing at first dosing should be reported as AE.  
 
AEs occurring after initiation of study treatment will be indicated as TEAEs in the clinical 
study report. 
The following variables will be collected for each AE:  
 Adverse event (verbatim)  
 The date and time when the AE started and stopped  
 Maximum intensity  
 Whether the AE is serious or not  
  Investigator causality rating against the IMP (acc. to categories as specified in 
13.1.2)  
 
 
 Outcome  
 
Reporting of Serious Adverse Events  
All SAEs have to be reported, whether or not considered causally related to the IMP, or to the 
study procedure(s). All SAEs will be recorded in a SAE report form (Annex No 1) and 
. The eCRF will contain information 
 
If any SAE occurs in the course of the study, then Investigators or other site personnel must 
inform the designated safety contact people on behalf of the Sponsor and CRO within 24 hours 
of when he or she becomes aware of its occurrence.   
SAE report form completion and reporting must not be delayed even if all of the information 
is not available at the time of the initial contact. 
SAEs must be reported within 24 h of knowledge of the event by submitting an initial SAE 
report via email or fax to the safety representatives. All details will be provided in the safety 
management plan. 
The following information should be provided to accurately and completely record the event: 
 Investigator name and center identification 
 Subject number 
 Subject initials, if permitted 
 Subject demographics 
 Clinical event 
 description 
 date of onset 
 severity 
 treatment 
 relationship to study drug (causality) 
 action taken regarding study drug 

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 outcome of the event (recovered/resolved, recovering/resolving, not 
recovered/not resolved, recovered/resolved with sequel, fatal, unknown) 
 If the AE resulted in death: 
 cause of death (whether or not the death was related to study drug) 
 autopsy findings (if available) 
 Medical history case report form (copy) 
 Concomitant medication case report form (copy) 
 Any relevant reports (laboratory, discharge, x-ray, etc.) 
The Sponsor representative will work with the Investigator to compile all the necessary and 
initially missing information within 1 calendar day of initial receipt for fatal and life-
threatening events and within 3 calendar days of initial receipt for all other SAEs.  
Unexpected (not previously described in the reference safety document), Serious Adverse 
Reactions (SUSARs) will be expedited by the Sponsor representative to Eudravigilance using 
CIOMS (Council for International Organisations of Medical Sciences) form.  
all details will be provided in the Safety Management Plan 
13.1.7 Follow-up of adverse events  
All AEs will be followed until they have abated, returned to baseline status or until a stable 
situation has been reached. Depending on the event, follow up may require additional tests or 
medical procedures as indicated, and/or referral to the general physician or a medical specialist. 
13.2 Temporary halt for reasons of subject safety  
The investigator will inform the subjects and the EC 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. The study will be suspended pending further review by the EC, except 
all subjects are kept informed. 

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14 STATISTICAL METHODOLOGY AND ANALYSES 
14.1 Statistical analysis plan 
 
The study is exploratory and is not powered to address any pre-defined hypotheses. The 
assessment of safety and tolerability will be performed on the safety analysis set, which 
includes all subjects who received at least one dose of B-OT. Complete description of all 
statistical analyses and methods will be presented in the Statistical Analysis Plan (SAP). The 
SAP will be reviewed and approved by the Sponsor and will be finalized prior to database lock. 
Plans for PK analyses will be included in the SAP. 
The sample sizes for the SAD cohorts are typical for a Phase 1 first in human (FIH) study. The 
total number of subjects accrued in the study will depend on the number of dose levels, but is 
currently set to a maximum of 24 subjects (2 cohorts with 3 subjects and 3 cohorts with 6 
subjects each). The safety and tolerability of B-OT will be analyzed by appropriate descriptive 
statistics. All other secondary endpoints will be summarized descriptively. 
The sample sizes for the MAD cohorts are typical for a Phase 1 multiple ascending dose study. 
The total number of subjects in the study will be 24 subjects (4 cohorts with 6 subjects each). 
The safety and tolerability of B-OT will be analyzed by appropriate descriptive statistics. All 
other secondary endpoints will be summarized descriptively. 
If circumstances should arise during the study rendering the analysis inappropriate, or if in the 
meantime improved methods of analysis should come to light, different analyses may be 
performed. Any deviations from the statistical methodology defined in this protocol, reasons 
for such deviations and all alternative/additional statistical analyses that may be performed will 
be described in the CSR. Such changes to analyses may be written into an abbreviated SAP, if 
appropriate. The verification and review of all statistical Modelling assumptions will be 
documented appropriately. 
All original and derived parameters as well as demographic and disposition data will be listed 
and described using summary statistics. All safety data (scheduled and unscheduled) will be 
presented in the data listings. 
Demographic and baseline data will be summarised by treatment (dose level of B-OT) and 
overall. Pharmacokinetic data will be summarised by dose level of B-OT. Safety and 
tolerability data will be summarised. 
Frequency counts (number of subjects [n] and percentages) will be made for each qualitative 
variable. Descriptive statistics (n, mean, SD, min, median, max) will be calculated for each 
quantitative variable (unless otherwise stated). Descriptive statistics will only be presented if n 
only the n, minimum and maximum will be presented. The other descriptive statistics will be 
left blank. 
The following rules will apply to any repeated safety assessments occurring within each cohort: 
 
If the repeated measurement of a specific parameter occurs prior to IMP 
administration (Day 1), then the last obtained value prior to dosing will be used 
in the descriptive statistics and in the calculation of changes from baseline. 
 
If the repeated measurement of a specific parameter occurs after IMP 
administration (Day 1), then the first (non-missing) value after dosing will be 
used in descriptive statistics and in the calculation of changes from baseline. 
 

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The planned sequence for measurement of multiple assessments at the same time point is 
described in Section 12.2.1 Blood samples collection. 
For safety assessments performed at Screening and the follow-up, the following rules will apply 
for any repeated assessments: 
 
If the repeated assessment occurs at Screening the last available value will be 
used in the summary statistics. 
 
If the repeated assessment occurs at the Follow-up Visit the first non-
missing assessment will be used in the summary statistics. All statistical 
analyses and production of tables, figures and listings will be performed 
using current SAS® version (9.4), whereas the pharmacokinetic analysis 
will be done using WinNonlin® (version 8.3), or higher.  
 
Details will be given in a statistical analysis plan (SAP) which will be prepared and validated 
by the Sponsor before database lock. Modifications or additions to the analyses described above 
will be described in the SAP. Any decisions to deviate from the planned analyses described in 
the protocol and SAP will be documented in the clinical study report. The clinical study report 
will also provide a detailed explanation for any deviations from the planned analyses. 
 
14.2 Protocol violations/deviations 
 
Protocol deviations will be identified based on conditions related to the categories below: 
 
 Protocol entry criteria. 
 Forbidden concomitant medications. 
 Missing evaluations for relevant endpoints. 
 Other protocol deviations occurring during study conduct. 
 
Major protocol deviations will be identified before the study closure and listed where 
appropriate. 
 
14.3 Power calculation 
Not applicable. 
 
14.4 Missing, unused and spurious data 
All missing or incomplete safety data, including dates and times, are treated as such. Missing 
test results or assessments will not be imputed. Descriptive statistics and statistical analysis 
will be performed on the basis of the available data only. 
  
Subjects discontinued will be included in the descriptive statistics if they have received the 
investigational product and their examinations were performed at the same scheduled time as 
other subjects. All data recorded on discontinued subjects will be listed separately. 
 
For graphical and summary purposes concentration values below the limit of quantification 
will be set to half (½) of the limit of quantification. For analysis no undetermined values will 
be replaced. If single data points for plasma B-OT or OT concentrations are missing, the AUC 
parameters will be derived by linear interpolation with regard to the two neighboring non-
missing concentrations.  

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For calculation of PK parameters, all plasma B-OT and OT concentration values below the 
Lower Limit of Quantitation (LLOQ) occurring prior to Cmax will be replaced by 0(of Day 1), 
except for embedded LLOQ values (between two measurable time points) which will be treated 
below LLOQ values after 
 except at Day 7 
pre-dose, where it will be replaced by LLOQ/2. The handling of missing, unused and spurious 
data will be documented in the study report. 
 
14.5 Analysis sets 
Data of all subjects participating in the study will be included in the analyses if the data can 
meaningfully contribute to the objectives of the study. Subjects who do not complete the 
sampling schedule of 1 or more study periods may be included in the PK population for only 
the PK parameters that are judged not to be affected by the missing sample(s). 
The decision of which subjects will be included in the PK Analysis Set is to be taken before 
the start of the sample analysis by the bioanalytical facility. 
 
14.5.1 Safety set 
The safety population will be defined as all subjects who received at least one dose of B-OT 
and for whom any safety post-dose data are available. 
 
Unless otherwise stated, the safety analysis set will be used for the presentation of all safety 
analyses. Exposure to IMP will also be presented using the safety analysis set. 
 
14.5.2 Pharmacokinetic and pharmacodynamic analysis set 
Subjects will be allocated to the PK Analysis Set on a per treatment basis. The PK Analysis 
Set for a treatment will include all subjects who receive one dose of B-OT, do not violate the 
protocol in a way that may invalidate or bias the results (major protocol deviation) and have 
sufficient plasma concentration-time profiles complying with the following criteria at Day 1 
(SAD/MAD) and at Day 7 (MAD only): 
 Do not have an occurrence of vomiting or diarrhoea which renders the plasma 
concentration profile unreliable (e.g., if vomiting occurs at or before 2 times median 
Tmax); 
 Do not use a concomitant medication which renders the plasma concentration profile 
unreliable; 
 Do not have a pre-dose plasma concentration that is greater than 5% of the Cmax value. 
If the predose concentration is greater than 5% of the Cmax value, the subject will be 
excluded from the PK population; 
 Have at least one evaluable plasma concentration that is preceded by a lower evaluable 
concentration and followed by a lower evaluable concentration for the calculation of 
Cmax, Tmax and AUCs.  
In case of an important protocol deviation or AEs, affected PK data will be excluded from the 
descriptive and inferential statistical. Where all PK data for a subject are impacted, the subject 
will be excluded from the PK analysis set. The exclusion of any subjects or time points from 
the calculation of the PK parameters will be documented by the statistician including the 
reason(s) for exclusion. 
 
The PD analysis set will consist of all subjects in the safety analysis set who received at least 
one dose of B-OT and who have at least one post-dose values present, with no important 

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protocol deviations thought to impact on the analysis of the PD data. .PD analysis is an 
exploratory end point 
 
14.6 Subject disposition 
 
Subject disposition will be listed by subject. 
The following subject data will be summarized: 
 Number and percentage of subjects screened, 
 Number and percentage of subjects enrolled, 
 Number and percentage of subjects completed and, 
 Number and percentage of subjects included in safety population, 
 Number and percentage of subjects used in PK, PD and statistical analyses. 
 
A subject who completed the study is defined as a subject for whom the last PK blood sample 
was assessed. 
 
14.7 Baseline parameters  
Baseline is defined as the last value prior to dosing. Change from baseline will be calculated 
for all continuous safety parameters. 
14.7.1 Demographics and baseline variables 
Continuous demographic variables (e.g., age, height, weight, BMI) will be summarized by 
descriptive statistics (n, mean, SD, median, Min, Max). 
Qualitative demographic characteristics (sex, race/ethnicity) will be summarized by counts and 
percentages.  
 
14.7.2 Medical history  
Medical history will only be listed. Medical history data will be listed by subject including 
visit, description of the disease/procedure, MedDRA SOC, MedDRA Preferred Term, start date 
and stop date (or ongoing if applicable). 
 
A summary of the number and percentage of subjects who had relevant medical histories will 
be presented by treatment group and for all subjects for the medical history Preferred Term. 
 
14.8 Safety and tolerability endpoints  
 
The safety set is used to perform all safety analyses. Statistical methods for the safety analyses 
will be descriptive in nature. Safety data, including AEs, clinical laboratory data, vital signs, 
ECG parameters, and PEs. All appropriate AEs will be graded using the DMID toxicity scale 
(March 2014). Change from baseline will be included in summary tables for laboratory 
parameters. All laboratory data will be included in the data listings and all test values outside 
the normal range will be flagged. 
 
14.8.1 Adverse events 
The AE coding dictionary for this study will be Medical Dictionary for Regulatory Activities 
(MedDRA). It will be used to summarize AEs by primary system organ class (SOC) and 
preferred term (PT). All adverse events will be displayed in listings. 
 

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A treatment-emergent adverse event (TEAE) is defined as an adverse event observed after 
starting administration of study treatment. If a subject experiences an event both prior to and 
after starting administration of a treatment, the event will be considered a TEAE (of the 
treatment) only if it has worsened in severity (i.e., it is reported with a new start date) after 
starting administration of the specific treatment, and prior to the start of another treatment, if 
any. All TEAEs collected during the investigational period will be summarized. 
The number of subjects with treatment emergent AEs will be summarized by:  
 
 Treatment, MedDRA SOC and PT; 
 Treatment, MedDRA SOC, PT and severity; 
 Treatment, MedDRA SOC, PT and drug relatedness. 
 
14.8.2 Vital signs 
At each time point, absolute values and change from baseline of supine blood pressure, pulse 
rate and respiratory rate will be summarized with n, mean, SD, SEM, median, Min, and Max 
values. The number of available observations and out-of-range values (absolute and in 
percentage) will be presented. Values outside the reference range will be flagged in the listing. 
will flag out-of-range results.  
 
14.8.3 ECG 
At each time point, absolute values and change from baseline of ECG numeric variables will 
be summarized with n, mean, SD, SEM, median, Min, and Max values. The number of 
available observations and out-of-range values (absolute and in percentage) will be presented. 
denoting values above or below the investigator reference range (when present), will flag out-
of-range results.  
 
14.8.4 Clinical laboratory tests 
At each time point, absolute values and change from baseline of clinical laboratory variables 
will be summarized with n, mean, SD, SEM, median, Min, and Max values. The number of 
available observations and out-of-range values (absolute and in percentage) will be presented. 
All laboratory data (including re-check values if present) will be listed chronologically. Out-
of-range values for safety laboratory will be flagged in individual listings as well as 
summarized descriptively using agreed reference ranges (e.g., laboratory ranges). 
 
14.9 Pharmacokinetic and pharmacodynamic endpoints analysis 
SAD study part: 
Plasma PK parameters for each dose level will be calculated from the concentrations of B-OT 
and OT measured in pre-dose and post-dose plasma samples. For each dose level, descriptive 
statistics will be presented. Figures will be created to display mean and individual subject B-
OT and OT concentration versus time. 
 
MAD study part: 
Plasma PK parameters for each dose level will be calculated from the concentrations OT 
measured in pre-dose and post-dose plasma samples. For each dose level, descriptive statistics 
will be presented. Figures will be created to display mean and individual subject OT 
concentration versus time. 

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The plasma concentrations and the PK parameters will be listed and presented in tabular and 
graphical form as appropriate according to the applicable standards, that includes applicable 
descriptive statistics, handling of individual concentrations below the LLOQ for listings, 
descriptive statistics and figures, and precision and rounding rules for concentrations and PK 
parameter data. 
A 72-hour individual baseline profile of plasma B-OT and OT concentrations will be measured. 
Values will be plotted and descriptive statistics per planned sampling time point and treatment 
will be provided, including mean, SD, CV%, minimum, maximum, and median as well as 
geometric mean and its CV%. 
 
Individual plasma B-OT and OT concentrations will be plotted versus time per individual using 
both a linear and log y-axis. Additionally, concentration versus time curves will be plotted per 
as a spaghetti plot. Individual subject listings will also be provided. Concentrations will be 
summarized for each time point of measurement, the number of observations, mean, SD, CV%, 
median, Min, and Max, as well as geometric means and its CV%. 95% confidence intervals 
will be given. 
 
At least the following individual PK parameters will be determined based on the concentration 
versus time curves in the SAD part:  
 maximum concentration (Cmax),  
 time to Cmax (Tmax),  
 elimination rate constant (
),  
 terminal elimination half-life (t1/2),  
 area under the concentration versus time curve (AUC) from 0 to the last measurement 
point (AUC0-last) 
 AUC extrapolated to infinity (AUC0-
 
 Clearance (CL/F)  
 Apparent volume of distribution (Vz/F) 
 
A 168-hour individual profile of plasma OT concentrations and potentially other derivate 
metabolites will be measured. Values will be plotted and descriptive statistics per planned 
sampling time point and treatment will be provided, including mean, SD, CV%, minimum, 
maximum, and median as well as geometric mean and its CV%. 
 
At least the following individual PK parameters will be determined based on the concentration 
versus time curves in the MAD part on Day 1: AUC , Cmax, tmax, on Day 7: AUC , Cmax,ss, tmax,ss, 
Swing, PTF%, CLss/F, Vz/F, Ctrough, Cav, Cmin,  t1/2, AUCinf and AUClast as appropriate and 
Day7/Day1: Ra(AUC ), Ra(Cmax). 
 
The lin-up log-down trapezoidal method will be used. If the logarithmic trapezoidal rule fails 
in an interval because non-monotonicity, then the linear trapezoidal rule will apply for that 
interval. 
If the terminal phase is sufficiently well characterized the terminal half-life will be estimated 
and AUC zero to infinity (AUC0
-last and the extrapolated area 
from the last measurement point to infinity based on the terminal half-life. Within a subject, 
data points before time of maximal plasma concentration that are below the lower limit of 
quantification (LLOQ) will be replaced with zero. Within a subject, data points after time of 

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maximal plasma concentration that are below the LLOQ will be excluded from analysis 
(considered as missing). Additionally, Cmax and tmax will be determined.  
The individual PK parameters will be summarized, including at least number of subjects, mean, 
SD, CV%, median, Min, and Max, as well as geometric means and its CV%.  
 
14.9.1 Dose proportionality 
Dose-proportionality for PK parameters of OT and B-OT (AUC0-inf, AUClast, and Cmax for 
single dose, and AUC0-
, and Cmax,ss  for multiple dose) will be assessed by the power model. 
The slope and the associated 90% CI based on the power model will be reported. For the first 
2 low doses of the SAD part where only 3 subjects are enrolled, no dose proportionality will 
be assessed. 
14.9.2 Inferential methods 
 
Not applicable. 
 
14.10 Interim analyses 
 
No formal interim analyses are planned for this study. Data from SAD cohorts were frozen and 
analyzed prior to conducting the s MAD part of the study. 

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15 GOOD 
CLINICAL 
PRACTICE, 
ETHICS 
AND 
ADMINISTRATIVE 
PROCEDURES  
15.1 Good clinical practice 
15.1.1 Ethics and good clinical practice 
The investigator will ensure that this study is conducted in full compliance with the protocol, 
the principles of the Declaration of Helsinki (www.wma.net), ICH GCP guidelines 
(http://www.ich.org/products/guidelines.html), and with the laws and regulations of the 
country in which the clinical research is conducted.  
 
15.1.2 Ethics committee / institutional review board 
The CRO on behalf of the investigator will submit this protocol and any related documents to 
an Ethics Committee (EC) and the Competent Authority (CA). Approval from the EC and the 
statement of no objection from the CA must be obtained before starting the study (i.e. first 
subject first visit), and should be documented in a dated letter to the applicant, clearly 
identifying the trial, the documents reviewed and the date of approval. A list of EC members 
must be provided, including the functions of these members. If study staff were present, it must 
be clear that none of these persons voted. 
Modifications made to the protocol after receipt of the EC and CA approval must also be 
submitted as amendments to the EC and CA in accordance with local procedures and 
regulations. 
 
15.1.3 Informed consent 
The Informed Consent and Subject Information will be provided in the local language  
Bulgarian. 
The Principal Investigator or delegate will:  
 
 Ensure each healthy volunteer is given full and adequate oral and written information 
about the nature, purpose, possible risk and benefit of the study  
 Ensure each healthy volunteer is notified that they are free to discontinue from the study 
at any time  
 Ensure that each healthy volunteer is given the opportunity to ask questions and allowed 
time to consider the information provided  
 Ensure each healthy volunteer provides signed and dated informed consent before 
conducting any procedure specifically for the study  
 
 
 Ensure a copy of the signed ICF is given to the healthy volunteer  
 Ensure that any incentives for healthy volunteers who participate in the study as well 
as any provisions for healthy volunteers harmed as a consequence of study participation 
are described in the ICF that is approved by an EC.  
 Ensure subjects are re-consented to the most current version of the ICF(s) during their 
participation in the study. 
 
A subject who is rescreened is not required to sign another ICF if the rescreening occurs within 
28 days from the previous ICF signature date. 
15.1.4 Insurance 
The sponsor has covered this clinical study by means of an insurance according to national 
requirements in Bulgaria. This insurance will cover the sponsor´s and the investigator´s 

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liability. The name and address of the relevant insurance company, the certificate of insurance, 
 
 
15.2 Study funding 
benfovir AG is the sponsor of the study and is funding the study. All financial details are 
provided in a separate contract(s) between Convex and the sponsor.  
 
15.3 Data handling and record keeping 
15.3.1 Data collection and confidentiality 
A Subject Screening and Enrolment Log will be completed for all eligible or non-eligible 
subjects with the reasons for exclusion. 
The data collection tool for this study will be an electronic CRF. For each subject enrolled, 
regardless of the occurrence of study drug administration, eCRF must be completed and signed 
by the principal investigator or co-investigator. This also applies to those subjects who fail to 
complete the study. If a subject withdraws from the study, the reason must be noted on the 
eCRF. Case report forms are to be completed on an ongoing basis.  
Designated investigator staff will enter the data required by the protocol into the eCRF. The 
Investigator must certify that the data entered into the eCRFs are complete and accurate. 
All data obtained using paper collection methods during the clinical study will be recorded in 
eCRF software. All source documents from which eCRF software entries are derived should 
 
The original eCRF software entries for each subject will be checked against source documents 
by the study monitor. Instances of missing or uninterpretable data will be discussed with the 
investigator for resolution. 
 
All records identifying the subject will be kept confidential and, to the extent permitted by the 
applicable laws and/or regulations, will not be made publicly available.  
Subject names will not be supplied to the sponsor. Only the subject number will be recorded in 
the eCRF, and if the subject name appears on any other document, it must be obliterated before 
a copy of the document is supplied to the sponsor. Study findings stored on a computer will be 
stored in accordance with local data protection laws. As part of the informed consent process, 
the subjects will be informed in writing that representatives of the sponsor, EC or regulatory 
authorities may inspect their records to verify the information collected, and that all personal 
information made available for inspection will be handled in strictest confidence and in 
accordance with local data protection laws. If the results of the study are published, the 
ty will remain confidential. The Informed Consent Form (ICF) will incorporate 
(or, in some cases, be accompanied by a separate document incorporating) wording that 
complies with relevant data protection and privacy legislation. The subject must be informed 
that his/her personal study-related data will be used by the sponsor in accordance with local 
data protection law. The level of disclosure and use of their data must also be explained to the 
subject in the informed consent 
All clinical study findings and documents will be regarded as confidential. The investigator 
and members of his/her research team must not disclose such information without prior written 
approval from the sponsor. 
 
The anonymity of participating subjects must be maintained. Subjects will be specified in 
outputs and other documents containing subject data by their subject number, not by name. 
Documents that identify the subject (e.g., signed ICF) will be maintained in confidence by the 
investigator. 

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15.3.2 Database management and quality control 
Data Management is the responsibility of Convex Ltd. With permission of the Sponsor, Data 
Management may be delegated under an agreement of transfer of responsibilities to a qualified 
vendor of Convex Ltd. 
The full details of procedures for data handling will be documented in the Data Management 
Plan (DMP). 
AEs and medical history will be coded using the Medical Dictionary for Regulatory Activities 
(MedDRA). Concomitant medications will be coded using the World Health Organization 
(WHO) Drug Dictionary. 
Unique subject numbers will identify the subject and the biological material obtained from the 
subject. Appropriate measures such as encryption or deletion will be enforced to protect the 
identity of human subjects in all presentations and publications as required by 
local/regional/national requirements. 
Data from screening failures will not be entered into the database. 
Laboratory data from the safety laboratory will be provided to Convex Ltd via access 
controlled, password protected web directory. The results will be printed and signed in paper 
and these will be considered source data. In cases where sensitive non-PK laboratory data is 
transferred via non-secure electronic networks, data will be encrypted during transfer. 
All PK and PD data will be transferred to the responsible Data Management party, whereby 
the transfer process will be pre-defined in data transfer agreements. 
 
eCRFs will be developed by Data Management party in collaboration with the clinical study 
team and statistician. The Data Management party will document the process workflow in the 
Data Management Plan (DMP). After data entry, monitor(s) will verify the eCRFs against the 
source documents. Queries may be issued to clarify the data entered. The PI will electronically 
sign the eCRFs after all data have been entered and all queries have been resolved. If 
corrections and/or resolution of queries are required after PI approvals, those eCRFs affected 
by changes will be re-signed by the PI. The database may be locked after the PI approvals are 
completed. 
The Investigator will receive all laboratory data electronically or based on printed reports 
directly from the laboratory. An Investigator must review, evaluate, sign and date the laboratory 
print-outs upon receipt. 
After database lock, study design documentation and locked eCRFs (PDF) will be created and 
can be provided to the Sponsor, if requested. 
Data required for analyses and subject safety assessments will be entered from source 
documentation into eCRFs. Instructions for data entry will be provided in the eCRF 
Completion Guidelines, developed by Convex Ltd Operational team. All site staff involved 
with entering data into the eCRFs will be trained prior to gaining access to the study database. 
Queries may be generated by the eCRF system during data entry, and queries may be generated 
by CDM staff, monitors, PIs, and other data reviewers during the course of the study. Only 
specific site personnel will be authorized to make corrections to the eCRFs; CDM will train 
personnel prior to granting access in the eCRF system. Corrections will be made directly in the 
eCRF  by modifying existing data, adding new data, or deleting data, as appropriate. All data 
corrections will be logged in the electronic audit trail. 
from source documentation is consistent with the source information and accurately reflected 
in the eCRFs. By electronically signing the eCRFs, the Investigator confirms that the 
information is complete and correct. 
 

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15.4 Access to source data and documents 
 
Source data are the original records of all variables collected for the clinical investigation. They 
include, but are not limited to: 
 Signed informed consent 
 Laboratory reports, test analysis reports, procedures results for all interventions 
performed in accordance with the protocol 
 Individual subject clinical notes 
 Details concerning inclusion and exclusion criteria 
 The medical history prior to participation in the study 
 
documents with the CRFs 
 All safety events (AEs, SAEs, etc.) 
 All treatment exposure (prior, concomitant and test treatment) 
 
study 
 A description of interventions done to the subjects in the study 
 
The Investigator must allow the monitor access to all documents in the 
file to 
confirm their consistency with the eCRF entries. All information entered in the eCRF must be 
available as source data. All study data will be handled confidentially. No information about 
subject identity on these documents will be allowed to leave the study site.  
 
It is the responsibility of the Investigator to maintain a comprehensive and centralized filing 
system of all relevant documentation. 
 Investigators will be instructed to retain all study records required by the Sponsor and 
regulatory authorities in a secure and safe facility with limited access for one of the 
following time periods based on notification from the Sponsor: 
 For a period of at least 2 years from the last marketing approval worldwide or 
for at least 25 years, whichever is the greater 
 Or a period of at least two years after discontinuation of clinical development 
of the investigational product as confirmed by the Sponsor 
 For a longer period if required by local regulations 
The Investigator will be instructed to consult with the Sponsor before disposal of any study 
records and to provide written notification to the Sponsor of any change in the location, 
disposition, or custody of the study files. 
 
The sponsor or its representative may visit the study facilities at any time in order to maintain 
current and personal knowledge of the study through review of the records, comparison with 
source documents, observation and discussion of the conduct and progress of the study. The 
investigator will permit trial-related monitoring, audits, EC review and regulatory inspections, 
providing direct access to source data and study documents. 
 
15.5 Quality control and quality assurance 
 
This study will be conducted according to applicable Standard Operating Procedures (SOPs) 
by Convex Ltd.  
Before the first healthy volunteer is entered into the study, a Sponsor representative will review 
and discuss the requirements of the Protocol and related documents with the investigational 
staff and also train them in any study specific procedures and system(s) utilized, if applicable.  

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The PI will ensure that appropriate training relevant to the study is given to all of these staff, 
and that any new information relevant to the performance of this study is forwarded to the staff 
involved. The PI will maintain a record of all individuals involved in the study (medical, 
nursing and other staff). 
Quality Management of Convex will periodically visit the Investigational site to discuss the 
conduct of the trial and upon necessity can audit the records of the trial. These reviews are 
necessary to ensure that the study is conducted according to standards consistent with the study 
protocol and ICH GCP Guideline. 
The Investigator agrees to discuss and correct, if necessary, any problems or deficiencies that 
are found during the course of these reviews. 
 
The Sponsor has the right to perform an external QA audit according to their internal 
procedures. 
 
15.5.1 Monitoring 
 will perform an initiation visit 
before the first subject is screened, have regular contacts with the study site, including routine 
visits conducted to:  
 
 Provide information and support to the Investigator(s). 
 Assure compliance with the study protocol. 
 Verify that the research facilities, including laboratories and equipment, are 
adequate to safely and properly conduct the study. 
 Verify that the investigational product is stored properly and under the proper 
conditions, is in sufficient supply, and that receipt, use, and return of investigational 
product are controlled and documented adequately. 
 Verify that written informed consent was obtained before any protocol-specific 
screening procedures are performed solely for the purpose of determining eligibility 
for the clinical study and/or prior to the provision of study medication. 
 Review the subject CRFs and source documents to ensure that reported study data 
are accurate, complete, and verifiable from source documents.  
 Ensure that adequate records of clinical trial supplies are maintained. 
 Verify that the Investigator and study site personnel are adequately qualified 
throughout the study. 
 Verify that the safety information and amendments are submitted to the relevant 
authorities. 
 
All checks that the data given in the eCRF complies with the source document will be carried 
out in accordance with the study specific monitoring manual. 
The monitor will be available between visits if the Investigator(s) or other staff at the centre 
needs information and advice about the study conduct.   
After the data base hard lock and approval from the Sponsor a close-out visit will be performed 
in order to close the study at the site. 

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15.6 Protocol amendments 
Any change to a protocol has to be considered as an amendment. All protocol amendments 
must be approved by the Sponsor before they are submitted to EC and CA. All substantial 
amendments will be implemented after the receipt of a written approval from the EC and CA. 
All non-substantial amendments will be implemented after they have been notified to EC and 
CA. 
15.6.1 Substantial amendment 
Significant changes that affect subject safety and/or the scientific value of a trial require a 
substantial amendment. Examples of significant changes are given in EU guidelines on the 
request to the competent authorities for authorisation of a clinical trial on a medicinal product 
for human use, the notification of substantial amendments and the declaration of the end of the 
trial (CT-1, 2010/C 82/01). The need for submitting a substantial amendment is the 
responsibility of the sponsor. Substantial amendments are to be approved by the appropriate 
-
implementation of the substantial amendment. 
 
15.6.2 Non-substantial amendment 
Non substantial amendments do not affect subject safety or the scientific integrity of the trial. 
Non-substantial amendments will be approved (signed) by the investigator(s) and will be 
recorded and filed by the investigator/sponsor. Non-substantial amendments will be submitted 
to the EC for information only. The CA will only be notified by changes in Eudract form and 
ABR form (if applicable). The implementation of a non-substantial amendment can be done 
immediately.  
 
The EU guideline CT-1 2010/C 82/01 stipulates the importance of preventing over-reporting. 
Therefore, the following changes are by definition non-substantial in this study: 
 
- 
change in amount and timing of the samples (maximum of 2 samples without a > 50 
mL increase in the amount of blood taken and not exceed 500 mL of blood in total) 
- 
changes in assay-type and / or institution where an assay will be performed, provided 
that validated assays will be used; 
- 
editorial changes to documents in the submission dossier including the subject 
information sheets and the protocol. An editorial change is defined as a modification in 
the documents of typographical errors and other modifications that in no way alter the 
meaning or content of the document; 
- 
determination of additional parameters in already collected materials, which are in 
agreement with the study objectives and do not provide prognostic or genetic 
information; 
- 
other statistical analyses than described in the protocol; 
- 
a change in clinical staff, including the principal investigator, when this concerns 
regular staff members of the CRO who comply with internal regulations for training 
and authorisation. 
 
15.6.3 Urgent amendment 
An urgent amendment might become necessary to preserve the safety of the subjects included 
in the study. The requirements for approval should in no way prevent any immediate action 
being taken by the investigators or the sponsor in the best interests of the subjects. Therefore, 

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if deemed necessary, an investigator can implement an immediate change to the protocol for 
safety reasons. This means that, exceptionally, the implementation of urgent amendments will 
occur before submission to and approval by the EC(s) and CA. 
 
15.7 Clinical study report 
After completion of the study, the complete final study report will be prepared in a form based 
on ICH E3 Guideline: Structure and Content of Clinical Study Reports. 
All individual data that allow re-calculation of pharmacokinetic parameters (concentrations, 
actual sampling) and pharmacokinetic parameters results will be available in electronic format 
and will be provided together with the report. 
 
15.8 End of study report 
The CRO on behalf of the sponsor will notify the EC and the CA of the end of the study within 
 
In case the study is ended prematurely, the CRO will notify the EC and the CA within 15 days, 
including the reasons for the premature termination. 
The CRO on behalf of the sponsor will notify the EC and CA immediately of a temporary halt 
of the study, including the reason of such an action.  
The Principal Investigator will inform all study subjects in case of temporary halt and/or 
premature termination of the study. 
Within one year after the end of the study, the responsible CRO will submit a synopsis of the 
clinical study report (CSR) with the results of the study, including any publications/abstracts 
of the study, to the EC and the CA. The principal investigator
 and the 
 
 
15.9 Public disclosure and publication policy 
The principal investigator/Convex Ltd. may not publish the data collected during this study in 
any form, except with the written permission of the Sponsor. In case of publication the principal 
investigator will have the opportunity to review the analysis of the data and to discuss with the 
sponsor the interpretation of the study results prior to publication. Any study-related article or 
abstract containing partial or complete study results, written independently by investigators 
should be submitted to the sponsor for review at least 60 days prior to submission for 
publication or presentation. The list of authors of any formal publication or presentation of 
study results may include, as appropriate, representatives of the sponsor and will be determined 
by mutual agreement. In case of publication, confidentiality of the study subjects will be 
respected. 

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16 REFERENCES 
- 
- EU Guidelines to Good 
Manufacturing Pr
February 2010. 
 
- 
Bitsch R et al., Bioavailability assessment of the lipophilic benfotiamine as compared 
to a water-soluble thiamin derivative. Ann Nutr Metab 1991;35:292-296. 
 
- 
Bojkova D et al., targeting pentose phosphate pathway for SARS-CoV-2 therapy. 
Metabolites 2021, 11(10), 699. 
 
- 
Bojkova D et al., Proteomics of SARS-CoV-2-infected host cells reveals therapy 
targets. Nature 2020, 583, 469 472. 
 
- 
Commission Directive 2005/28/EC of 8 April 2005 laying down principles and detailed 
guidelines for good clinical practice as regards investigational medicinal products for 
human use, as well as the requirements for authorisation of the manufacturing or 
importation of such products, Official Journal of the European Communities L 91, 
9/4/2005, 8.4.2005. 
 
- 
Coy JF, EDIM-TKTL1/Apo10 Blood Test: An Innate Immune System Based Liquid 
Biopsy for the Early Detection, Characterization and Targeted Treatment of Cancer. Int 
J Mol Sci. 2017 Apr 20;18(4):878. 
 
- 
Directive 2001/20/EC of the European Parliament and of the Council of 4 April 2001 
on the approximation of the laws, regulations and administrative provision of the 
Member States relating to the implementation of good clinical practice in the conduct 
of clinical trials on medicinal products for human use. Official Journal of the European 
Communities, L 121/34-44, 1.5.2001. 
 
- 
Directive 2001/83/EC of the European Parliament and of the Council of 6 November 
2001 on the Community code relation to medicinal products for human use. Official 
Journal of the European Communities, L - 311, p. 67-128, 28/11/2001 as amended by 
Directive 2002/98/EC, Official Journal, L  33, 08/02/2003, p. 30 40, by Directive 
2004/24/EC, Official Journal, L  136, 30/04/2004, p. 85 90, and by Directive 
2004/27/EC, Official Journal, L  136, 30/04/2004, p. 34 57. 
 
- 
Guideline on Bioanalytical Method Validation (EMEA/CHMP/EWP/192217/2009 
Rev.1 Corr. 2**, 21 July 2011 
 
- 
Grasselli G et al., Baseline Characteristics and Outcomes of 1591 Patients Infected with 
SARS-CoV-2 Admitted to ICUs of the Lombardy Region, Italy. JAMA 
2020;323(16):1574-81. 
 
- 
Hampshire A et al., Cognitive deficits in people who have recovered from COVID-19 
relative to controls: An N=84,285 online study. medRxiv. 2020:2020.10.20.20215863. 
- 
 
 

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- 
ICH E3 Guideline for Industry: Structure and Content of Clinical Study Re
July 1996. 
 
- 
Kennedy M.H. et al., Delirium in Older Patients With COVID-19 Presenting to the 
Emergency Department. JAMA network open. 2020;3(11):e2029540-e. 
 
- 
Lechien JR et al., Clinical and epidemiological characteristics of 1420 European 
J Int Med 2020; 288:335-
344. 
 
- 
Li YC et al., The neuroinvasive potential of SARS-CoV2 may play a role in the 
respiratory failure of COVID-19 patients. J.  med. virology. 2020;92(6):552-5. 
 
- 
Li Y  et al., APC/C(CDH1) synchronizes ribose-5-phosphate levels and DNA synthesis 
to cell cycle progression. Nat Commun. 2019 Jun 7;10(1):2502. 
 
- 
Rieg S et al., COVID-19 in-hospital mortality and mode of death in a dynamic and non-
restricted tertiary care model in Germany. medRxiv. 2020:2020.07.22.20160127. 
 
- 
Schenk M, Neurologische Manifestationen: Wie COVID-19 die Nerven tangiert. Dtsch 
Arztebl International. 2020;117(19): A-1001 / B-843. 
 
- 
Schilling J et al., Krankheitsschwere der ersten COVID-19-Welle in Deutschland 
basierend auf den Meldungen gemäß Infektionsschutzgesetz. Journal of Health 
Monitoring. 2020;5(S11):2-20. 
 
- 
Sun W et al., TKTL1 is activated by promoter hypomethylation and contributes to head 
and neck squamous cell carcinoma carcinogenesis via increased aerobic glycolysis and 
Clin Cancer Res. 2010 Feb 1; 16(3): 857 866. 
 
- 
Tong JY et al., The Prevalence of Olfactory and Gustatory Dysfunction in COVID-19 
Patients: A Systematic Review and Meta-analysis. Otolaryngology--head and neck 
surgery : official journal of American Academy of Otolaryngology-Head and Neck 
Surgery. 2020:194599820926473. 
 
- 
Tylicki A et al., Thiamine and selected thiamine antivitamins 
 biological activity and 
methods of synthesis, Bioscience Reports 2018;  38. 
 
- 
Whittaker A et al., Neurological Manifestations of COVID-19: A systematic review 
and current update. Acta neurologica Scandinavica. 2020;142(1):14-22. 
 
- 
World Medical Association Declaration of Helsinki, Ethical Principles for Medical 
Research Involving Human Subjects, as amended by the 64th WMA General Assembly, 
Fortaleza, Brazil, October 2013. 
 
- 
Yan CH et al., 
nt Forum of Allergy & Rhinology 2020; 10:821-831. 
 

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- 
Yang X et al., Clinical course and outcomes of critically ill patients with SARS-CoV-
2 pneumonia in Wuhan, China: a single-centered, retrospective, observational study. 
The Lancet Respiratory medicine. 2020;8(5):475-81. 

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Adverse event term* 
(provide a clinical 
diagnosis 
where 
possible): 
 
 
 
_______________________________________________ 
*State diagnosis when possible. In case of no confirmed diagnosis, 
document only leading sign, syndrome or preliminary diagnosis that 
presumably led to one of the seriousness criteria listed in the section 5.  
Please report 1 event term per line.  
Onset date** of SAE: 
 
 
________________ 
dd  mmm  yyyy  
**Document date when the symptom of SAE occurred first (for 
seriousness criteria see section 5.). In case of hospitalization document 
Narrative*** (please, 
use 
an 
additional 
sheet if necessary): 
 
***Report 
clinical 
signs and symptoms as 
well as 
any other 
supporting information 
that 
led 
to 
the 
diagnosis. 
Describe 
course of the event and 
any treatment of the 
event including details 
of medications and 
procedures. Please add 
rationale of your event 
causality assessment 
 
 
 
 
 
 
 
 
4. Severity of adverse event: 
 
     Mild  
  
     Moderate  
 
     Severe 
 
 
5. Seriousness criteria (check the most appropriate criterion): 

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     Death 
     Life-
threatening 
     Persistent 
or 
significant 
disability/ 
incapacity 
     Congenital 
anomaly/birth 
defect 
     Medically 
important event or 
reaction 
     Hospitalization  
(initial 
 
or 
prolonged 
) 
Date of hospitalization: 
__________________ 
dd  mmm  yyyy  
Date of discharge:  
________________ 
dd- mmm  yyyy 
 
6. Event causality (relationship to the Investigational Product): 
     Certain 
     Probable 
     Possible 
     Unlikely 
 
     Not related 
 
 
7. Study medication:  
 
Please, check the study period at 
 
     Screening             Maintenance                Titration             Conversion or Tapering       
     Additional Tapering             Follow-up                Other:_______________________ 
Administration: 
  
      single ascending dose (SAD) part         
 
       multiple-ascending dose (MAD) part       
Dose level :  
 
 
__________________      ________ 
Dose                                          Units 
(First) Study Medication Intake****:  
 
****applicable for the SAD part and for the 
first dose of the MAD part 
Last Study Medication Intake*****: 
(before the onset of the SAE) 
*****applicable for the MAD part of the study 
only 
 
________________ 
dd  mmm  yyyy 
 
________________ 
Hour/Min 
 
______________ 
dd  mmm  yyyy 
 
________________ 
Hour/Min 
Did event alleviate after stopping the 
administration of the Study Medication? 
 
         N/A                      No                   Yes 

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Did event reappear after reintroduction of the 
Study Medication? 
     
         N/A                      No                   Yes 
 
8. Action taken due to this event regarding the study medication: 
     None 
     Discontinuation of the study 
drug  
Date: ________________ 
dd  mmm  yyyy 
     Use of intervention to treat 
the 
AE 
(provide 
details, 
recording concomitant therapy, 
if needed) 
     Study 
drug 
dosing 
interrupted  
 
Date: _________________ 
dd  mmm  yyyy 
 
Restart 
date:__________________ 
dd  mmm  yyyy 
 
9. Relevant medical history/co-existing diseases  
(please, use an additional sheet if necessary, or attach a copy/print-out of the Medical History 
CRF/eCRF)
 
 
 
    Please, check if an additional sheet is attached 
10. Relevant tests/laboratory data 
(please, use an additional sheet if necessary, or attach a copy of the lab reports) 
Please provide the list of any abnormal laboratory or diagnostic tests relevant to this adverse event. 
Do not include information not pertinent to this event. 
 
    Please, check if an additional sheet is attached 
 
11. Concomitant medication/therapy at the time of the event 
(Alternatively, please attach a copy/print-out of the concomitant medication CRF/eCRF) 

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Drug name 
Start date 
(dd-mmm-
yyyy) 
Stop date 
(dd-mmm-
yyyy) 
Dose 
Units
Route
Indication 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
    Please, check if an additional sheet is attached 
 
12. Final outcome of the serious adverse event: 
     
 
 
 Date recovered/resolved: ________________ 
                               dd  mmm  yyyy 
     Recovered/resolved 
with 
a 
 
     
 
 
Date 
of 
death: 
 
dd  mmm  yyyy 
Specify the cause of death: 
_____________________________ 
 
Autopsy report obtained  
     Yes      No 
 
 
     Recovering/Resolving 
 
 
     Not recovered/Not resolved 
 
     Unknown 
 
Signatures: 
Name of the reporter:  
 
 

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Phone number: 
Fax number: 
Email: 
 
 
 
signature:
 
 
 
Date: ________________ 
dd  mmm  yyyy 
 
  
 
 
Phone number: 
Fax number: 
Email: 
 
signature: 
 
 
Date: ________________ 
dd  mmm  yyyy 
 
 
17.2 Annex No 2 
 
PREGNANCY REPORT AND OUTCOME FORM 
PARTNER 
 
Sponsor: 
 
 
Investigational 
Product: 
 
Protocol Number: 
 
 
Principal 
Investigator: 
 
Site Number: 
 
 
Subject Initials: 
 
Country: 
 
 
Subject Number: 
 
 
MATERNAL DATA  
 
Patient date of birth (if possible in respect of Data Protection Law in your Country): 
________________________________ 
Relevant medical history (risk factors, smoking, alcohol, etc): 
_________________________________________________________________ 

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_________________________________________________________________ 
Previous Pregnancies: _____             Previous Normal deliveries: _____  
Previous Spontaneous miscarriages: _____ 
Relevant family history: 
_________________________________________________________________ 
_________________________________________________________________ 
 
Last menstrual date: _______________________          
Estimated date of delivery: _______________________________ 
 
- Was the patient using hormonal contraceptive/IUD at time of conception?  
 yes  
 no 
 
- Is there evidence of a defect from a prenatal test?   
 yes  
 no 
 
If yes, indicate which test(s) showed evidence of birth defect:  
 Ultrasound Amniocentesis 
 Other, specify: ________________________________________ 
 
MEDICATION(S) TAKEN DURING PREGNANCY 
Product name 
Daily dose 
Treatment duration  
(Start date/Stop date) 
Indication 
 
 
 
 
 
 
 
 
MEDICATION(S) TAKEN AT THE TIME OF CONCEPTION 
Product name 
Daily dose 
Treatment duration  
(Start date/Stop date) 
Indication 
 
 
 
 
 
 
 
 
 
Signatures: 

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Name 
of 
the 
reporter:  
 
 
Phone number: 
Fax number: 
Email: 
 
 
 
signature: 
 
 
Date: ________________ 
dd  mmm  yyyy 
 
 
 
 
Phone number: 
Fax number: 
Email: 
 
signature: 
 
 
Date: ________________ 
dd  mmm  yyyy 
 
PREGNANCY OUTCOME FORM 
 
Sponsor: 
 
 
Investigational 
Product: 
 
Protocol Number: 
 
 
Principal 
Investigator: 
 
Site Number: 
 
 
Subject Initials: 
 
Country: 
 
 
Subject Number: 
 
 
PHOETUS/CHILD DATA 
 
 Elective termination on: _____________________ 
 
 Spontaneous miscarriage on: ________________ 
                  
 Premature birth  - Date of birth: ______________     

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Full term - Date of birth: ____________________    
Birth weight: ___________                                         
Gender:    
male    
female
If premature, gestational age: weeks: ______ days: ___________
Comments: ___________________________________________
METHOD OF DELIVERY:
Normal vaginal                   
Forceps              
Caesarean section  
Other, specify: _________________
OUTCOME
Healthy baby       
Sick baby (i.e., birth trauma, infection)     
Congenital 
anomaly/Birth defect     
Stillbirth
Was a birth defect noted?........................... yes             no
IF SERIOUS ADVERSE EVENT DEFINITION IS MET, PLEASE REPORT THE 
EVENT AS SAE.
If any birth defects were noted, please describe:
_________________________________________________________________
_________________________________________________________________
To what do you attribute the defect(s):
_________________________________________________________________
_________________________________________________________________
Describe any immediately postnatal problems that occurred (i.e, jaundice, respiratory 
distress):
_________________________________________________________________
_________________________________________________________________
Additional comments:

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_________________________________________________________________ 
_________________________________________________________________ 
 
 
Signatures: 
Name 
of 
the 
reporter:  
 
 
Phone number: 
Fax number: 
Email: 
 
 
 
signature: 
 
 
Date: ________________ 
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