Estimating Versus Measuring Plasma Volume and Kidney Function in Acute Decompensated Congestive Heart Failure (EMPaKT-CHF)
Phase: NA
Conditions: CHF
Sponsor: Charite University, Berlin, Germany



EMPAKT-CHF Trial, Protocol, Version 5.0, dated 11.02.2019 
 
EudraCT-No: 2018-002638-18  
 
Protocol code EMPAKT-CHF  
 
 
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Table of contents 
 
Abbreviations ..................................................................................................................  5 
1 Synopsis««««.......................................................................................................... 7 
2 Flow-Chart««««««................................................................................................ 21 
3 Introduction ...................................................................................................................  21 
3.1 Introduction and background ..................................................................................  21 
3.1.1 Investigational Device Background ....................................................................  22 
3.1.2 Investigational Product Background ..................................................................  22 
3.1.3 Summary of Findings to Date .............................................................................  23 
3.2 Rationale .................................................................................................................  28 
3.2.1 Rationale for Dose Selection ..............................................................................  28 
3.2.2 Rationale for Study Design .................................................................................  29 
4 Study objectives ...........................................................................................................  30 
4.1 Primary endpoints ..................................................................................................  30 
4.2 Secondary endpoints ...............................................................................................  30 
4.3 Time point (for the evaluation of secondary endpoints) ...........................................  31 
4.4 Study design ............................................................................................................  32 
4.5 Timetable .................................................................................................................. 37 
5 Study population ............................................................................................................ 37 
5.1 Inclusion criteria .......................................................................................................  38 
5.2 Exclusion criteria .....................................................................................................  38 
6 Plan for medical treatment............................................................................................. 39 
6.1 Description of study medication / investigational medicinal product .......................  39 
6.1.1 Adverse drug reactions and restrictions, contra-indications, drug interactions ... 40 
6.1.2 Treatment schedule ............................................................................................  44 
6.1.3 Storage requirements and administration (distribution and return of study drug) 44 
6.1.4 Treatment compliance ........................................................................................  44 
6.2 Placebo / reference medication ...............................................................................  45 
6.3 Concomitant medication / concomitant therapy ........................................................ 45 
6.4 Procedures in case of emergency ...........................................................................  45 
7 Study procedures........................................................................................................... 45 
7.1 Recruitment procedures and informed consent process .........................................  45 
7.2 Methods of avoiding simultaneous enrolment in other trials ...................................  46 
7.3 Enrolment ................................................................................................................  46 
7.4 Clinical examinations and trial-related deviations from clinical practice ..................  46

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7.4.1 Pre-trial examinations (screening / inclusion examination) .................................. 48 
7.4.2 Examinations during trial ........................................................................................  48 
7.4.3 Final examination ................................................................................................ 49 
7.4.4 Laboratory assessments .....................................................................................  49 
7.5 Individual trial duration .............................................................................................. 50 
8 Risk-benefit-assessment...............................................................................................  50 
9 Termination and subsequent treatment .......................................................................  51 
9.1 Premature termination of the individual participant..................................................  51 
9.2 Premature termination of the clinical study ............................................................... 51 
9.3 Follow-up and continuing treatment after regular / premature termination ............... 51 
10 Safety monitoring ........................................................................................................ 52 
10.1 Adverse Event (AE) ................................................................................................ 52 
10.2 Serious Adverse Event (SAE)................................................................................. 52 
10.3 Adverse Reactions ................................................................................................. 53 
10.4 Suspected Unexpected Serious Adverse Reactions (SUSAR) .............................. 53 
10.5 Evaluation of Expectedness ................................................................................... 53 
10.6 Treatment of (S)AEs ..............................................................................................  53 
10.7 Assessment of SAEs .............................................................................................  53 
10.8 Severity grade (according CTCAE v5.0)................................................................  53 
10.8.1 Assessment of causality ...................................................................................  54 
10.9 Documentation of AEs and SAEs .........................................................................  54 
10.10 Follow-up of Adverse Events ..............................................................................  55 
10.11 Reporting of SAEs .............................................................................................  55 
10.12 Reporting of Suspected Unexpected Serious Adverse Reactions (SUSARs)«..  55 
10.13 Other safety issues requiring expedited reporting ..............................................  56 
10.14 Development Safety Update Report ...................................................................  56 
11 Documentation ............................................................................................................ 56 
11.1 Case Report Forms (CRF).....................................................................................  56 
11.2 Investigator Site File (ISF) .....................................................................................  57 
11.3 Drug accountability / flow of study medication .......................................................  57 
12 Quality management ..................................................................................................  58 
12.1 Control of trial progress and data quality ..............................................................  58 
12.1.1 Monitoring .........................................................................................................  58 
12.1.2 Data Quality Assurance ....................................................................................  59 
12.1.3 Audits / inspections ...........................................................................................  59 
12.2 Reference institutions ...........................................................................................  60 
13 Data entry and data management .............................................................................  60

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13.1 electronic Case Report Form (eCRF) Data Entry ...............................................  60 
13.2 Data management ..............................................................................................  61 
13.3 Source Data and Patient Files ............................................................................  61 
14 Statistical Analysis ....................................................................................................  61 
14.1 Definition of population for analysis .....................................................................  64 
15 Reporting .................................................................................................................  64 
15.1 Statistical report .....................................................................................................  64 
15.2 Study report ...........................................................................................................  65 
15.3 Publication (policy) ................................................................................................  65 
16 Ethical, legal and regulatory aspects .......................................................................  65 
16.1 ICH-GCP-guidelines ..............................................................................................  65 
16.2 Legal requirements of the study ............................................................................  65 
16.2.1 Approval of Ethics Committee .........................................................................  65 
16.2.2 Approval of competent authority .....................................................................  65 
16.2.3 Notification to local authorities (LaGeSo) .......................................................  66 
16.2.4 PROTOCOL AMENDMENTS .........................................................................  66 
16.2.5 Patient information and informed consent ......................................................  66 
16.2.6 Patient insurance ............................................................................................  67 
16.2.7 Data Privacy and confidentiality ......................................................................  67 
16.3 Archiving of data / access to records ..................................................................  67 
16.4 Financing ............................................................................................................  68 
17 References ...............................................................................................................  68 
18 Appendices................................................................................................................ 69 
Abbreviations 
 
AE 
Adverse Event 
AKI 
acute kidney injury 
AUC 
area under the concentration curve 
CHF 
Congestive Heart Failure 
CKD 
chronic kidney disease 
CPK 
creatine phosphokinase 
CRF 
Case Report Form 
DAC 
data adjudication committee 
eGFR estimated Glomerular Filtration Rate 
ePV 
estimated plasma volume 
eTBV  estimated total blood volume 
EOS 
end of study 
GFR 
Glomerular Filtration Rate 
hcg 
human chorionic gonadotropin 
IB 
Investigators Broschure 
ICU 
Intensiv Care Unit 

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IMP 
investigational medicinal product 
ITT 
Intent to treat Population 
mGFR Measured Glomerular Filtration Rate 
mPV measured Plasma Volume 
mTBV measured total blood volume  
NYHA New York Heart Association 
PPP 
Per protocol population 
PV  
Plasma Volume 
SAE 
Serious Adverse Event 
TBV 
total blood volume 
TEAE treatment-emergent adverse event 
VFI 
Visible fluorescent injectate (FD004) 
WOCBP  
Women of child-bearing potential 

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1 Synopsis 
 
Study title 
Estimating versus Measuring Plasma Volume and Kidney 
Function in Acute Decompensated Congestive Heart Failure 
(EMPaKt-CHF) 
 
Type of project 
This is an investigator-initiated, one-armed phase II clinical 
trial using an injectable fluorescent tracer to assay and 
evaluate measured Plasma Volume (mPV) and measured 
Glomerular Filtration Rate (mGFR) in Hospitalized Patients 
with acute decompensated congestive heart failure (CHF).  
 
Sponsor  
Charité Universitätsmedizin Berlin 
Charitéplatz 1, 10117 Berlin, Germany  
Representative of the Sponsor 
and Principal Investigator 
 
 
 
 
 
 
 
Deputies of the Principal 
Investigator 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Co-investigators 
 
 
 
 
 
 
Prof. Dr. med. Kai M. Schmidt-Ott 
Charité – Universitätsmedizin Berlin 
Medizinische Klinik mit Schwerpunkt Nephrologie und Internistische 
Intensivmedizin, Augustenburger Platz 1,  
13353 Berlin 
Tel. +49 30 450 614337 
Fax: +49 30 450 553 909 
Email: kai.schmidt-ott@charite.de 
 
Dr. Torsten Slowinski 
Charité – Universitätsmedizin Berlin 
Medizinische Klinik mit Schwerpunkt Nephrologie und Internistische 
Intensivmedizin, Augustenburger Platz 1,  
13353 Berlin 
Tel.: +49 30 450 614214 
Fax: +49 30 450 553 909 
Email: torsten.slowinski@charite.de 
 
PD Dr. med. David Leistner, Charité – Universitätsmedizin Berlin, 
Medizinische Klinik für Kardiologie, Hindenburgdamm 30, 12200 
Berlin 
Tel.: +49 30 450 513745 
Fax: +49 30 450 513 947 
Email: david-manuel.leistner@charite.de 
 
Prof. Dr. Fabian Knebel, Charité – Universitätsmedizin Berlin, 
Medizinische Klinik mit Schwerpunkt Kardiologie und Angiologie 
Tel.: +49 30 450 513216 
Fax: +49 30 450 7513 216 
Email: fabian.knebel@charite.de 
 
 
Prof. Dr. med. Kai-Uwe Eckardt, Charité – Universitätsmedizin 
Berlin, Medizinische Klinik mit Schwerpunkt Nephrologie und 
Internistische Intensivmedizin, Augustenburger Platz 1,  
13353 Berlin 
 
Prof. Dr. med. Burkert Pieske, Charité – Universitätsmedizin Berlin, 
Medizinische Klinik mit Schwerpunkt Kardiologie, Augustenburger 

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Study Physician / member of 
the trial team 
 
 
 
Trial Statistician 
Platz 1, 13353 Berlin 
 
Prof. Dr. med. Frank Edelmann, Charité – Universitätsmedizin 
Berlin, Medizinische Klinik mit Schwerpunkt Kardiologie, 
Augustenburger Platz 1, 13353 Berlin 
 
Prof. Dr. med. Ulf Landmesser, Charité – Universitätsmedizin 
Berlin, Medizinische Klinik für Kardiologie, Hindenburgdamm 30, 
12200 Berlin 
 
Prof. Dr. med. Karl Stangl, Charité – Universitätsmedizin Berlin, 
Medizinische Klinik mit Schwerpunkt Kardiologie und Angiologie 
 
 
 
Jutta Swolinsky, Charité – Universitätsmedizin Berlin 
Medizinische Klinik mit Schwerpunkt Nephrologie und Internistische 
Intensivmedizin, Augustenburger Platz 1,  
13353 Berlin 
 
Prof. Dr. Geraldine Rauch 
Institute of Biometry and Clinical Epidemiology 
Charité - Universitaetsmedizin Berlin 
Charitéplatz 1, 10117 Berlin 
 
Hypotheses 
 
It is hypothesized that utilizing the FAST PV and mGFR 
Technology is safe and functional in CHF patients. It is also 
hypothesized that quantitative measurements of intravascular 
plasma volume status and GFR will differ from standard 
clinical assessments and estimating equations, and that 
clinical decision making would have been improved in a 
subpopulation of patients had the quantitative measurements 
delivered by the FAST PV and mGFR Technology been 
available to the physician during the course of care.  
 
Rationale 
Previous clinical studies have shown the FAST technology to 
be safe and functional in patients with chronic kidney disease 
(CKD) and in healthy subjects. The technology has not been 
studied in CHF patients. In previous clinical studies, the 
technology has been demonstrated to accurately measure PV 
and mGFR in patients with CKD and healthy patients when 
compared to other clinical techniques including, estimated 
glomerular filtration rate (eGFR) calculations, Iohexol 
clearance and Nadler’s formula. Additionally, previous studies 
have shown that the concentration of the PV marker remains 
relatively constant during the first two hours after injection, 
given that this high-molecular weight marker is predicted to 
exclusively occupy the intravascular compartment and exhibit 
a long half-life.  
In this study, the safety and functionality of the FAST 
Technology will be evaluated in CHF patients. The non-clinical 
and clinical history of the product provides confidence that the 

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technology will be well tolerated and function correctly in the 
CHF population.  
 
The blood samples are timed to maximize the information 
gathered during the study. In addition to measurements of PV 
and GFR, FAST will determine the technology’s functionality 
by comparing the concentration of the PV marker at the 15, 30 
and 60 minute time points. It is not believed that the plasma 
volume will change significantly within that time period and 
thus the concentration of the PV marker should remain 
relatively constant. 
 
Study medication (IMP) /  
Therapeutic strategy 
Visible fluorescent injectate (VFI) (FD004), Powder for 
Solution for Injection, 47 mg. VFI is diluted with Water for 
Injection and a total of 3 mL of VFI is administered by bolus 
injection. There is 47 mg of VFI administered in this 3 mL 
dose (35 mg of FD001 and 12 mg of FD003). VFI is dosed on 
Day 1 and Day 3 approximately 48 hours apart. 
Study design 
This is an investigator-initiated, one-armed phase II clinical 
study designed to evaluate the safety and function of the 
FAST PV and mGFR Technology in patients with CHF. Data 
from the FAST PV and mGFR Technology will not be 
evaluable by the treating physician, but will be made available 
to the study investigators and to an adjudication committee.  
 
Patients enrolled in the study will be administered the Day 1 
Dose of VFI after enrollment and the Day 3 dose 
approximately 48 hours after the initial dose. 
 
After consenting to be enrolled in the study, patients meeting 
the enrollment criteria will receive a single dose of VFI(Day 1). 
Blood draws (3 mLs) will be collected pre-dose and at 15 (±5 
minutes), 30 (±5 minutes), 60 (+30 minutes), and 180 (+120 
minutes) minutes post-dose which will be subsequently used 
to determine the patient’s blood volume and mGFR using the 
FAST PV and mGFR Technology. Patients will be treated 
according to standard of care throughout the time of their 
hospitalization. A second dose of VFI will be administered 48 
(±5 hours) hours after the initial dosing (Day 3). Again, blood 
draws (3 mLs) will be collected at 15 (±5 minutes), 30 (±5 
minutes), 60 (+30 minutes), and 180 (+120 minutes) minutes 
post-dose 
 
In addition, study-related blood samples will be obtained on 
days 1, 2, 3, 4, 5 to measure serum creatinine (days 1-5), 
hematocrit (days 1, 3), and N-terminal pro-brain natriuretic 
peptide (NT-pro-BNP), Creatine phosphokinase (CPK), 
Alanine aminotransferase (ALT), Aspartate aminotransferase 
(AST) (U/l), Alkaline phosphatase (AP), Bilirubin total and 
direct, International Normalized Ratio (INR) (days 1,3,5), and 

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will be stored for future testing of additional parameters in the 
research fields of acute decompensated heart failure and 
acute kidney injury.  
 
Furthermore, the available laboratory values from routine 
clinical practice (see Table 1) will be entered in the Case 
Report Form (eCRF).  
 
On days 1-5, the patient will be asked to complete a Patient 
Symptom Assessment Sheet (see Annex D).  
 
On days 1 and 3, the treating physician will be asked to 
complete a very brief survey (Annex E) to provide a qualitative 
assessment of the patient’s perceived intravascular volume 
status and renal function. The study physician will conduct a 
non-invasive clinical assessment of the patient (including e. g. 
body weight, height, edema grade, blood pressure, heart rate, 
respiratory rate, jugular venous distension assessment, 
oxygen saturation) on days 1-5.  
 
In addition, the study physician will record in the eCRF 
detailed information regarding diuretics dosage, i. v. fluid 
application, and requirement of renal replacement therapy as 
ordered by the clinicians in the hospital.  
 
Laboratory values and assessments (see Table 1) will be 
captured on each day that the VFI is administered and these 
data will be entered into the eCRF. Any AEs determined by 
the investigator to be related or possibly related to the VFI will 
be captured. Patients will also receive a follow up phone call 7 
days (± 2 days) after the second dose of VFI was 
administered, and a second follow up call 30 days after the 
second dose of VFI was administered. This second phone call 
will serve as the end-of-study (EOS) follow-up call. Any AEs 
determined by the investigator to be related or possibly related 
to the VFI will be captured during the follow up calls and will 
be followed to resolution. Additionally, in-person follow up 
visits may be scheduled, as determined by the investigator. 
During day 1-5, during follow-up phone calls or study visits 
and through information accessed within the hospital 
computer system or obtained from treating physicians or 
relatives, the following study outcomes will be recorded: 
mortality, length of hospital stay, readmission for heart failure, 
episodes of acute kidney injury, requirement of fluid 
resuscitation, episodes of symptomatic hypotension, 
successful reduction of body weight. 
 
Upon conclusion of the study, the recorded clinical data will be 
provided to an independent data adjudication committee 
(DAC) in a pseudonymized fashion. The DAC will be asked to 

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review the available data at the time of the first dosing and the 
measured GFR and PV data for the first dosing day will be 
provided. The DAC will then be asked to fill out a survey 
asking whether the mGFR and PV data would have influenced 
their treatment decision, and if so, indicate what they would 
have done differently.  
 
Following the evaluation of mGFR and PV data at the time of 
the initial VFI dose, the mGFR and PV data determined from 
the second VFI dosing point will be provided to the DAC. The 
DAC will then be asked to fill out a separate survey asking 
how the actual measured PV and GFR differ from the clinical 
assessments, whether the mGFR and PV data would 
influence their treatment decision, and if so, indicate what they 
would have done differently.  
 
After all assessments have been completed, the DAC will 
review the results of their analyses and compile a summary 
report on the secondary endpoints as well as their 
assessment of the overall impact of the PV and mGFR on 
clinical decision making.    
 
Total number of patients 
 
We plan to enrol 50 eligible patients. Due to the explorative 
nature of this phase II study a formal sample size calculation 
is not possible. However, the sample size of 50 patients is 
sufficient to guarantee reasonable accuracy with respect to 
the main objectives of the trial as specified below. 
Study population 
 
Male and Female patients aged 18 admitted to the hospital 
ward with a diagnosis of congestive heart failure. 
Inclusion criteria 
x 
Written informed consent indicating that they understand 
the purpose of and procedures required for the study 
and are willing to participate in the studyprior to any 
study related measures present. 
x 
Hospitalized patient with acute decompensated heart 
failure, diagnosed on the basis of the presence of at 
least one symptom (dyspnea, orthopnea, or edema) and 
one sign (rales, peripheral edema, ascites, or pulmonary 
vascular congestion on chest radiography) of heart 
failure.Subject:  18 years of age. 
x 
Male with female partners of childbearing potential gave 
agreement to practice abstinence or use condoms from 
enrollment through 90 days after administration of the 
last dose of study drug present. 
x 
Partner of a male patient: Agreement to use a medically 
acceptable method of contraception (a barrier method, 
intrauterine device, or hormonal contraception) from 
enrollment through 90 days after administration of the 
last dose of study drug present. 
x 
Male patient agreed to not donate sperm from enrollment 
through 90 days after administration of the last dose of 
study drug present 
x 
A female subject is eligible to enter the study if she is:  

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o Not pregnant or nursing 
o Of non-childbearing potential (i.e., post-
menopausal defined as having been amenorrheic 
for at least 1 year prior to screening, or has had 
bilateral tubal ligation at least 6 months prior to 
administration of study drug or bilateral 
oophorectomy or complete hysterectomy) 
o If of childbearing potential (WOCBP), must have 
a negative urine or serum pregnancy test within 
24h prior to drug administration and be using a 
highly effective means of contraception during 
study participation and until 1 month after the last 
dose of study drug. Highly effective contraception 
methods include: 
x 
Total abstinence (when this is in line 
with the preferred and usual lifestyle of 
the subject. Periodic abstinence (e.g., 
calendar, ovulation, symptothermal, 
post-ovulation methods) and 
withdrawal are not acceptable 
methods of contraception Combination 
of any two of the following (i+ii or i+iii, 
or ii+iii): 
(i) use of oral, injected or implanted 
hormonal 
methods of contraception or other forms of
hormonal contraception that have 
comparable 
efficacy (failure rate < 1%), for example 
hormone vaginal ring or transdermal 
hormone contraception. 
(ii) placement of an intrauterine device 
(IUD) or intrauterine system (IUS) 
(iii) barrier methods of contraception: 
condom or occlusive cap (diaphragm or 
cervical/vault caps) 
x 
In case of use of oral contraception 
women should have been stable on 
the same pill for a minimum of 3 
months before taking study treatment 
x 
Patient is able to communicate effectively with the study 
personnel. 
x 
Patient is informed of the nature and risks of the study 
and give written informed consent prior to enrollment. 
 
Exclusion criteria 
x 
New or ongoing myocardial infarction or instable angina 
present at the time of planned study inclusion. 
x 
Patient shows evidence of severe infection other than 
pneumonia, or active internal bleeding (characterized by 
recent decrease of blood hemoglobin concentration by 
more than 2 g/dl). 
x 
Patient experiences new onset atrial fibrillation. 
x 
Patient has an elective surgery planned during the 30 
days they are enrolled in the study. 

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x 
Patient has a psychiatric disease or a history of illicit 
drug use that would prohibit them from complying with 
study requirements  
x 
Prior exposure to VFI present. 
x 
History of any clinically significant allergic or negative 
reactions, side effects, or anaphylaxis to fluorescent 
dyes, or dextran molecules present. 
x 
Patient requires intravenous vasodilators or inotropic 
agents (other than digoxin or digitoxin) for heart failure. 
x 
Patient undergoes chronic dialysis (for example 
peritoneal or hemodialysis) treatments.  
x 
Patient is in cardiogenic shock or on vasopressors. 
x 
Hypotension as defined by blood pressure < 90 systolic 
and/or < 50 mm Hg diastolic exists. 
x 
Patients suffering from significant non-cardiac diseases 
of other organ systems (e. g. Malignancies, significant 
neurological diseases) 
x 
Patient does not have a working telephone.  
x 
Patient is a pregnant or nursing (lactating) women, 
where pregnancy is defined as the state of a female after 
conception and until the termination of gestation, 
confirmed by a positive human chorionic gonadotropin 
(hCG) laboratory test. 
x 
Lack of willingness to storage and disclosure of 
pseudonymous disease data in the context of the clinical 
trial present. 
x 
Subject has a participation in another interventional 
clinical trial during this study or within 30 days (or longer) 
before entry into this trial (as a minimum; 5 x elimination 
half-life / terminal elimination of an IMP). 
x 
Subject is legally detained in an official institution. 
x 
Subject is dependent on the sponsor, the investigator or 
the trial sites. 
 
Table 1:Schedule of Study Events/Examinations 
Study 
Timepoint 
Data 
collected 
from 
chart 
prior to 
enrolling  
Patient 
Enrolled 
and 
Dosed 
Data 
collected 
between 
1st and 
2nd Dose  
Patient 
2nd VFI 
Dose 
Data 
Collected 
after 2nd 
Dose 
Data 
Collected 
Before 
Discharge 
Follow up 
in person 
or by 
phone 
call 
Follow 
up 
phone 
call/ 
Follow 
up Visit 
for 
Pregnan
cy test if 
WOCBP 
Visit 
Pre-
Dose 
Day 1 
Day 2 
Day 3 
Day 4 
Day 5 
Day 10 
Day 30 
Informed 
Consent 
X 
  
  
  
  
  
  
  
Inclusion/ 
Exclusion 
Criteria 
Assessment 
X 
  
  
  
  
  
  
  

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Structural 
Heart Disease 
Criteria 
Assessment a 
  
X 
  
  
  
  
  
  
Demographics 
b 
  
X 
  
  
  
  
  
  
Comorbidities 
c 
  
X 
  
  
  
  
  
  
Full 
medication list 
on enrollment 
into the study  
and during 
follow-up  d 
  
X 
 X 
 X 
 X 
 X 
  
  
Patient 
Symptom 
Assessment 
Survey e 
  
X 
X 
X 
X 
X 
 
  
Clinical 
Presentation/ 
Exam f 
  
X 
X 
X 
X 
X 
  
  
Treating 
Physician 
Survey g 
  
X 
  
X 
  
  
  
  
Study-Related 
Laboratory 
Measurements 
h 
  
X 
X 
X 
X 
X 
  
  
Routine 
Clinical 
Parameters i 
X* 
X 
X 
X 
X 
X 
  
  
Clinical Data j 
X* 
X 
X 
X 
X 
X 
  
  
Adverse 
events 
  
X 
X 
X 
X 
X 
X 
X 
Blood 
specimen for 
marker 
analysis k 
  
X 
  
X 
  
  
  
  
Discharge 
from study 
  
  
  
  
  
  
  
X   
Adjudication 
Committee 
Evaluationl 
 
 
 
 
 
 
 
X 
WOCBP 
Pregnancy 
test m 
X 
 
 
X 
 
 
 
X 
a See Table 2 for Structural Heart Disease Criteria 
b See Table 3 for specific demographic parameters 
c See Table 4 for specific instructions on collecting comorbidities 
d See Table 5 for specific instructions of collecting medications  
e See Annex D for the Patients Symptom Assessment Survey 
f See Table 6 for Clinical presentation/exam parameters 
g See Annex E for the Treating Physician Survey 
h See Table 7 for Study Related Laboratory Measurements 
i See Table 8 for Routine Clinical Parameters 

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j See Table 9 for Clinical Data collected 
k Blood Specimens for the determination of the FAST PV and mGFR will be taken pre-dose and at 15 (± 5), 30 (± 5), 60 
(+30), and 180 (+120) minutes post-dose. Note that the 30 minute blood draw must be at least 10 minutes after the 15 
minute blood draw, and the 180 minute blood draw must be at least 100 minutes after the 60 minute blood draw. 
l See Table 10 for the Adjudication Committee Evaluation Questions; Adjudication Committee meets after all collected 
data available. 
m See Table 11 for Details of Pregnancy test  
*will be collected retrospectively after study inclusion 
 
Study objectives / endpoints 
 
Primary endpoint: 
Global aim is to assess the safety and function of the FAST 
PV and mGFR Technology in hospitalized patients with heart 
failure. 
 
x Safety will be assessed by determination of the absolute 
and relative frequencies of AEs and SAEs related to VFI. 
 
x Function of the FAST PV measurement will be assessed 
by determining the plasma stability of the FD003 high 
molecular weight marker over the 15, 30, and 60 minute 
blood draws and applying the following criteria: 
- 
The FAST PV measurement is considered as 
stable, if the mean plasma concentration of FD003 
at 30 minutes is not more than 10% lower than the 
mean plasma concentration at 15 minutes AND if 
the mean plasma concentration at 60 minutes is not 
more than 10% lower than the mean plasma 
concentration at 30 minutes 
- 
We will determine the percentage of patients which 
show a decline in the plasma concentration of 
FD003 of more than 10% from 15min to 30min and 
separately from 30 min to 60min. This percentage 
should be ideally close to 0. 
 
Secondary Endpoints: 
x To evaluate how estimated PV (ePV) and estimated total 
blood volume (TBV) assessments on days 1 and 3 (by 
established measures such as Nadler’s formula or 
Metropolitan Life Tables) predict measured PV (mPV; as 
assessed by FAST methodology) and measured TBV 
(mTBV; calculated from mPV and measured hematocrit) at 
these time points.  
 
x To evaluate how a clinical evaluation (including a 
reasonable subset of the variables age, gender, BMI, 
hematocrit, edema grade, presence of pulmonary rales, 
presence of jugular venous congestion, arterial blood 
pressure, NYHA stage, respiratory rate) on days 1 and 3 
predicts mPV and mTBV at these time points.  
 
x To evaluate whether eGFR calculation by the CKD-EPI 
formula on days 1 and 3 provides an accurate estimate of 

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measured GFR (mGFR; by assessed by FAST 
methodology) in heart failure patients undergoing active 
fluid management.  
 
x To evaluate whether patients with a low mGFR/eGFR ratio 
on days 1 and 3 are at higher risk of developing AKI within 
the following 48-72 hours.  
 
x To evaluate whether ranges of mPV (low mPV, normal 
mPV, high mPV) and/or mTBV (low mTBV, normal mTBV, 
high mTBV) on days 1 and 3 are associated with 
subjectively reported symptoms (PGA, dyspnea, dizziness, 
nausea) determined using patient symptom assessment 
sheets within the following 24-48 hours. 
 
x To evaluate whether patients with low mPV or low mTBV 
(cutoffs potentially adjusted for some of the following 
variables age, gender, body height, body weight, optimal 
body weight (dry weight), hematocrit) on day 1 or on day 3 
are at risk of developing low-output complications within 
the next 24-48 hours (e. g. subjectively reported 
dizziness/nausea, PGA, hypotension, need for iv-fluid 
therapy).  
 
x To evaluate whether patients with a high mPV or a high 
mTBV (cutoffs potentially adjusted for some of the 
following variables age, gender, body height, body weight, 
optimal body weight (dry weight), hematocrit) on day 1 
and/or day 3 are at risk of being refractory to diuretic 
therapy (e. g. require dosage increases of 
furosemide/torasemide, require ultrafiltration/RRT within 
the next 24-48 hours, fail to improve subjectively in PGA 
and dyspnea scales).  
 
x To evaluate whether the time course of ePV and estimated 
TBV (eTBV) from day 1 to day 3 adequately reflects the 
time course of mPV and mTBV at these time points.  
 
x To evaluate whether the change of mPV and mTBV from 
day 1 to day 3 is predictive of length of stay, 
dialysis/ultrafiltration requirement, or rehospitalization 
within 30 days, or mortality.  
 
x To evaluate whether the time course of mGFR from day 1 
to day 3 correlates temporally with or predicts the time 
course of eGFR within the next 24-48 hours.  
 
x To evaluate (by an adjudication committee) whether 
clinical decision making would have been affected by 
adding FAST GFR and PV measurements to clinical 

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routine including the following questions:  
o Would FAST GFR and PV measurements have 
changed clinical management overall? 
o Would the FAST GFR and PV measurement on day 
1 have led to applying a different diuretics dosage 
between day 1-3? 
o Would the FAST GFR and PV measurement on day 
3 have led to applying a different diuretics dosage 
between day 3-5? 
o Would the FAST GFR and PV measurement on day 
1 have led to choosing dialysis/ultrafiltration instead 
of diuretics escalation between day 1-3? 
o Would the FAST GFR and PV measurement on day 
1 have led to choosing diuretics escalation instead 
of dialysis/ultrafiltration between day 1-3? 
o Would the FAST GFR and PV measurement on day 
3 have led to choosing dialysis/ultrafiltration instead 
of diuretics escalation between day 3-5? 
o Would the FAST GFR and PV measurement on day 
3 have led to choosing diuretics escalation instead 
of dialysis/ultrafiltration between day 3-5? 
 
Safety 
Adverse events (AEs) will be collected and evaluated as they 
occur throughout the study.  
Discontinuation criteria 
Individual Patient Discontinuation Criteria: 
A patient is free to withdraw consent and discontinue 
participation in the study at any time, without prejudice to 
further treatment according to standard clinical practice. 
The investigator may remove a patient if, in the investigator’s 
judgment, continued participation would pose unacceptable 
risk to the patient or to the integrity of the study data. All 
procedures for early termination must be completed.  
Reasons for removal or withdrawal may include: 
x Admission to the ICU prior to either the first or second 
dose 
x The patient develops a stated exclusion criterion during 
the course of study 
x If the patient experiences an acute anaphylactoid 
reaction during the course of administering the FAST 
VFI, administration will stop immediately 
x Evidence of an anaphylactoid reaction (dyspnea, 
urticaria, flush, bronchospasm, or severe systemic 
hypotension)   
x Administrative decision by the investigator or sponsor 
x Significant protocol deviation 
x Patient noncompliance 
x Safety concern by the investigator or sponsor 
x Lost to follow-up 
x Pregnancy 

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Patients who are withdrawn for reasons other than safety 
issues may be replaced at the discretion of the sponsor and 
investigator. 
 
In the event of a patient’s withdrawal, the investigator makes 
every effort to complete the EOS assessments. All withdrawn 
patients with ongoing clinically significant clinical or laboratory 
findings will be followed until the finding is resolved or 
medically stable; reasonable attempts will be made to follow-
up with patients. 
 
Study Discontinuation Criteria: 
 
x 
Unjustifiable risk and toxicity in risk-benefit analysis 
(decision taken by principal Investigator)  
x 
New scientific evidence provided during the study that could 
affect the patient’s safety (benefit-risk analysis no longer 
positive)  
x 
Decision of the Sponsor or the Competent Authority that the 
study should be discontinued.  
 
Statistical analysis 
Descriptive statistics will be performed for safety assessments 
and for relationship between the FAST GFR and other 
measurements of renal function as well as between the FAST 
PV and other assessments of plasma volume.  
 
Pharmacological-toxicological 
evaluation 
Pharmacokinetics of the FAST VFI will not be evaluated as a 
part of this study, but these parameters have been 
investigated in previous clinical and nonclinical studies.  
 
Risk-benefit analysis 
Risks of treatment methods: 
Usual risks of participating in clinical trials, which are related 
to possible allergic reactions and blood drawing via 
venipuncture. 
 
Risks (IMP) (according to the reference safety information) 
Possible allergic reactions and risks associated with dosing 
and blood drawing via venipuncture. 
 
Action steps for risks minimisation: 
The FAST Technology has been previously used in three 
clinical studies. 87 patients have been dosed with the FAST 
VFI. There have been no reported SAEs and few minor AEs 
across all studies. Existing allergen risks in patients will be 
included as part of informed consent and staff will be well trained 
prior to the study initiation. 
 
Remaining risk: 
Possible allergic reactions and risks associated with dosing 
and blood drawing via venipuncture. 
 

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Benefit: 
A rapid and accurate measurement of plasma volume (PV) 
and glomerular filtration rate (GFR) is important in chronic 
heart failure (CHF), acute kidney injury (AKI) and chronic 
kidney disease (CKD) for assessment of impairment, 
diagnosis, and prompt treatment. This study primarily serves 
the purpose of obtaining information about the tolerability and 
safety of the FAST PV and mGFR Technology in CHF 
patients.  
 
There are no perceived direct benefits to patients participating 
in this trial. However, thorough medical check-ups may be 
seen as an advantage. The safety of the patients will be 
observed during all study phases. 
 
Medical progress is based on research which ultimately must 
rest in part on experimentation involving humans. Eligible 
patients may consider participation in this clinical trial because 
they want to contribute to the advancement of medical 
knowledge. Still, considerations related to the well-being of the 
individual patients enrolled into this clinical study must take 
precedence over the interests of science and society. Based 
on available information and the design of the study, the 
sponsor and the investigator consider the trial to be ethically 
acceptable. The duration of hospitalization and the medical 
surveillance are considered adequate to ensure safety of the 
patients. 
 
Contraindications: 
 
 
 
 
Procedures in case of 
emergency: 
 
 
 
 
 
 
Treatment of overdose: 
The FAST VFI (FD004) should not be administered to patients 
with known hypersensitivity to dextran, 5 aminofluorescein 
dye, or 2-sulfohexamine rhodamine dye. 
 
 
The FAST VFI (FD004) was well tolerated in the nonclinical 
studies conducted to date. Anaphylactic responses to 
dextrans, while rare, must be considered as a possibility for 
the high molecular weight dextrans being evaluated by FAST.  
As such, all testing facilities should have immediately 
available procedures to administer treatment for an 
anaphylactic response should it occur. 
 
The FAST VFI is contained in single-dose vials, making 
significant overdose unlikely. Though the vials are overfilled 
with 4mL/62.6 mg in order to ensure that a full 3mL/47mg 
dose can be drawn, previous studies have evaluated the 
safety of the VFI using a 3mL/150mg dose, over two times the 
amount of material contained in the single-dose vials for this 
study. There is no known antidote for overdose of VFI.  In the 
event of a suspected overdose, the patient should be 
monitored appropriately and should receive supportive 
therapy, as necessary. 

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Timetable 
Date of study / recruitment start (first patient first visit 
(FPFV): January 2019 
End of recruitment: Anticipated to be July 2019 
End of trial (last patient last visit, LPLV): Anticipated to be 
August 2019 
Trial duration (for a patient): 33 Days 
Follow up: Patients will receive a follow-up phone call on Day 
7 and Day 30 after the second VFI dose 
Funding 
The study will be supported by a research grant from FAST 
BioMedical. 
Key words 
Congestive Heart Failure, Acute Kidney Injury, Plasma 
Volume, Clinical Managment, Glomerular Filtration Rate 
(GFR), Plasma Volume (PV), FAST VFI, FAST PV and mGFR 
Technology,  
 

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2 Flow-Chart 
 
 
Screening 
Examination of inclusion/exclusion criteria 
 
 
Informed Consent 
 
 
Enrolment procedures 
 
 
Study Therapy dose 1 (Day 1) 
 
 
Study Therapy dose 2 + 48 hours (+/- 5hours) after 1st dose (Day 3) 
 
 
Follow-up visits (phone calls on 7 and 30 days after second dose) 
 
 
Study Completion 
3 Introduction 
3.1 Introduction and background 
A rapid and accurate measurement of plasma volume (PV) and glomerular filtration 
rate (GFR) is important in acute kidney injury (AKI) and chronic kidney disease (CKD) 
for assessment of impairment, diagnosis, and prompt treatment. 
 
Reductions in PV and/or GFR secondary to kidney injury, either acute or chronic, are 
accompanied by increases in blood urea nitrogen (BUN) and serum creatinine (Scr) 
levels. Currently, either Scr, or an equation based on Scr, is used to determine a 
patient’s estimated GFR (eGFR). Unfortunately, these 2 approaches are not reliable 
over the full range of GFR, and neither can be used for early detection or functional 
assessment in patients with a rapidly changing GFR, such as patients with AKI. 
 
Recent data indicate that even very small changes in Scr, which were previously 
thought to be clinically insignificant, are now known to predict an increased mortality 
rate. A technique to measure PV and GFR rapidly would be valuable to detect the 
loss of kidney function that leads to these acute changes in Scr so that treatment can 
begin as soon as possible. 
 
Glomerular filtration rate is the most clinically relevant metric for understanding renal 
function, as it is the rate by which the kidney is able to filter waste products in the 
bloodstream. Accurate measurement of GFR in acutely ill patients could also 
facilitate accurate dosing of drugs to maximize therapeutic efficacy while minimizing 
toxicity. Therefore, technical advances in this field are of major clinical importance, 
especially in high-risk patients where intense surveillance is necessary. 

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3.1.1 Investigational Device Background 
The FAST PV and mGFR Technology™ is a direct measurement of PV and GFR that 
relies on reading the concentration of fluorescent markers attached to different size 
dextran molecules introduced into the bloodstream. The test is intended as an 
adjunct to current methods utilized to assess kidney function. 
 
The FAST PV and mGFR Technology includes intravenously (IV) administered 
fluorescent markers, timed blood draws, a validated fluorometric assay, and a 
computed algorithm to integrate the results and calculate the GFR. 
 
The FAST PV and mGFR Technology will aid in identifying and determining the 
extent of renal dysfunction, therefore promoting early treatment, including dialysis 
initiation, as well as enrollment and stratification for clinical studies. It could also be 
used to determine the effect of a clinical maneuver on GFR such as volume 
resuscitation.  
 
In addition to measuring the GFR, the FAST PV and mGFR technology is capable of 
measuring the intervascular plasma volume of a patient.  With a single dose, the PV 
can be monitored for change or therapeutic affects for up to 8 hours. It will rapidly 
determine intravascular volume allowing early pharmacologic and or IV volume 
expanding treatment, as well as enrollment and stratification for clinical studies, and 
prognostic information. Measurement of PV will allow for accurate therapeutic use of 
volume expansion or vasopressors in the ICU, especially in patients with endothelial 
injury as and increased vascular permeability as in sepsis. 
 
3.1.2 Investigational Product Background 
The IV administered visible fluorescent injectate (VFI)™ agent is comprised of a 
mixture of 2 different molecular weight carboxymethyl dextran molecules (5 kD and 
150 kD) with different fluorescent dye molecules covalently attached. The 5 kD 
carboxymethyl dextran (FD001) is labeled with 5-aminofluorescein and the 150 kD 
carboxymethyl dextran (FD003) is labeled with 2-sulfohexamine rhodamine. These 
fluorescent labels are covalently attached to the dextran through the carboxymethyl 
moiety. When combined in a 3:1 (mass) ratio of small to large molecules, these 
dextrans make up the VFI, termed FD004. 
 
The high molecular weight labeled carboxymethyl dextran, FD003, is not rapidly 
cleared from the vasculature and is not rapidly cleared by passive filtration in the 
kidneys; therefore, its concentration in the blood stream after injection provides a 
direct measurement of the total PV. The low molecular weight labeled carboxymethyl 
dextran, FD001, is also not patient to rapid metabolism but is freely filtered by the 
kidneys. The decline in the concentration of FD001 combined with the PV measured 
by FD003 provides a rapid and accurate assessment of GFR. 
 
The VFI is administered IV through a bolus injection. 
 
While the VFI is a substantially modified dextran-based compound, other dextran 
products have, on rare occasions, been associated with mild to severe, acute 
anaphylactic reactions including death. As a precaution, patients will be closely 
monitored for signs of allergy and anaphylaxis, and an emergency resuscitation kit 

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and team will be available within the clinical research unit (CRU) throughout the 
treatment period. 
 
Fluorescent dyes such as fluorescein, which is also used in the VFI, are known to 
cause transient ocular light sensitivity, although this effect was not reported at any 
dose studied in any of the previous human clinical studies. Should this effect be 
observed, patients will be provided with disposable protective sunglasses to wear 
until the effect is reversed. 
3.1.3 Summary of Findings to Date 
3.1.3.1 Nonclinical Studies 
Proof-of-concept nonclinical pharmacology studies have been conducted in dogs to 
confirm the assessment of GFR using the FAST mGFR Test™ (Wang 2011). The 
results of these studies provide evidence that the method, in combination with the 
VFI, provides an accurate GFR. 
 
The liquid VFI agent (FD004) was evaluated in a full range of nonclinical studies, 
including pharmacology, safety pharmacology, pharmacokinetic (PK), single-dose 
toxicity, genotoxicity, and immunotoxicity in male and female rats and dogs at 
concentrations from 16 (FD003) to 26 (FD001) fold greater than the planned 
maximum human dose based upon observed and predicted area under the 
concentration-time curve (AUC) values. For more information please refer to the 
current Investigator’s Brochure. 
 
Observed effects of the VFI (FD001 and FD003) in pivotal single dose animal studies 
at higher doses (ranging from approximately 62 to approximately 100 times the 
planned human clinical dose of 47 mg [29 mg/m2]) included emesis in dogs, 
discolored urine, and a species specific effect (rats not dogs) on tissue macrophages 
of the lymph nodes and spleen resulting in cell death, which did not reverse during 
the recovery period. The clinical relevance of the macrophage finding in rats is 
unknown. 
 
Though the VFI has shown no pharmacological activity at the human dose in 
receptor screening, a cytochrome P450 (CYP) inhibition study has been performed to 
assess potential for interaction with concomitant medications. The ability of FD001 
and FD003 to inhibit CYP isoforms, CYP1A2, CYP2B6, CYP2C8, CYP2C9, 
CYP2C19, CYP2D6, and CYP3A4/5, was assessed to identify the potential of drug-
drug interactions. FD001 and FD003 were each preincubated for 0 or 30 minutes at 7 
concentrations (0.1, 0.3, 1, 3, 10, 30, and 100 µM) with human liver microsomes in 
the presence of probe substrates phenacetin, efavirenz, amodiaquine, diclofenac, 
mephenytoin, dextromethorphan, midazolam, and testosterone to determine if there 
was a direct effect on CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, 
and CYP3A4/5, respectively. Additionally, FD001 and FD003 were each incubated in 
the presence of dihydronicotinamide-adenine dinucleotide phosphate (NADPH) for 
up to 30 minutes at 37°C to assess the time-dependent inhibitory effects on CYP 
enzymes. 
 
Results from the CYP inhibition studies suggest that: 

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x FD001 does not inhibit any CYP enzyme examined (ie, CYP1A2, CYP2B6, 
CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4/5 [using 2 different 
substrates midazolam and testosterone]). 
x FD003 is a direct inhibitor of CYP2B6, CYP2C8, CYP2C19, and CYP3A4/5 
(midazolam). 
x The R1 values associated with the direct inhibition of CYP2B6, CYP2C8, 
CYP2C19, and CYP3A4/5 (midazolam) by FD003 are all less than 1.1, signifying 
little likelihood that FD003 may cause clinically relevant drug interactions due to 
direct inhibition in vivo. 
x FD003 did not directly inhibit CYP1A2, CYP2C9, CYP2D6, or CYP3A4/5 (as 
measured by testosterone 6ȕ-hydroxylation; testosterone). 
x FD003 was a time-dependent inhibitor of CYP3A4/5 (as measured by 
testosterone 6ȕ-hydroxylation) since the IC50 value shifted from > 100 µM with no 
preincubation to 49 µM after FD003 was preincubated with NADPH-fortified 
human liver microsomes for 30 minutes. However, this time-dependent inhibition 
was only apparent in the presence of concentrations of FD003 greater than 10 
µM, or approximately 1500 times the total bound and unbound plasma maximum 
observed concentration (Cmax) of 0.9 µg/mL (0.0066 µM). 
x FD003 did not cause time-dependent inhibition of any other CYP enzyme 
examined (ie, CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, or 
CYP3A4/5 [as measured by midazolam 1´-hydroxylation]). 
3.1.3.2 First-in-Human Study 
A FIH study, representing the first human use of the VFI, was concluded in 2012. 
Data from the study indicated that the FAST PV and mGFR Technology is safe and 
well tolerated. In this study, 32 patients were enrolled with 8 patients (6 VFI + 2 
placebo) assigned to 1 of 4 dose groups. The VFI agent consisted of a mixture of 2 
different molecular weight carboxymethyl dextran molecules (5 kD [FD001] and 150 
kD [FD003]) with different fluorescent dye molecules attached. Four increasing doses 
were administered through a bolus injection: Group 1, 5 mg/0.1 mL VFI; Group 2, 15 
mg/0.3 mL VFI; Group 3, 75 mg/1.5 mL VFI; Group 4, 150 mg/3 mL VFI. 
 
All 32 patients completed the study as planned and were included in the safety 
analysis. Overall 9 patients (28.1%) experienced 15 treatment-emergent adverse 
events (TEAEs) which were considered by the investigator to be reasonably related 
to VFI administration or study procedures. Most related TEAEs were observed after 
administration of 15 mg VFI (9 TEAEs reported by 3 patients). No related TEAEs 
were observed after the highest dose. Only 2 patients (both 15 mg VFI) reported 
6 TEAEs (pruritus and gastrointestinal disorders) which were considered related to 
VFI administration. All other related TEAEs were judged to be related to the study 
procedures (mainly injection site pain and hematoma at the blood sampling site). All 
TEAEs were of mild to moderate intensity. There were no serious adverse events 

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(SAEs); no patient discontinued the study due to adverse events (AEs). Safety 
laboratory parameters, vital signs, and electrocardiogram (ECG) parameters showed 
no clinically relevant time- or treatment-related effects or differences between dose 
groups. 
 
All patients who received VFI were included in the PK analysis. Geometric mean 
AUC and Cmax of FD001 and FD003 increased dose-proportionally. FD001 had a 
median time at which the maximum concentration was observed (tmax) of 0.25 hours 
for all doses; median tmax of FD003 was about 0.5 hours for the 2 lower doses and 
2 hours for the 2 higher doses. FD001 was eliminated with a short elimination half-life 
(t½) of about 1.5 to 4 hours; mean t½ for FD003 was considerably longer ranging from 
47 to 111 hours. Detailed PK data is available in the Investigator’s Brochure for this 
study. 
3.1.3.3 Phase 2a Study at University of Alabama at Birmingham 
For the Phase 2a study completed in 2015 at the University of Alabama at 
Birmingham, 5 cohorts with a total of 33 patients were studied. The primary endpoint 
of the study was to establish safety of the IV administered VFI. This end point was 
achieved, and even with increasing severity of CKD, with its associated 
comorbidities, no SAEs were reported during the study. A relatively small number of 
AEs were reported as potentially related to the VFI. The FAST BioMedical measured 
GFR (mGFR) was evaluated by performing an Iohexol study along with standard 
estimates of GFR using creatinine and cystatin C values and both Modification of 
Diet in Renal Disease (MDRD) and Chronic Kidney Disease Epidemiology 
Collaboration (CKD-EPI) equations. The breakdown of patients and GFR ranges in 
each cohort is listed in the table below.  
Phase II Patients by Cohort  
Cohort 
Type of Patient 
eGFR 
1 
Normal 
60 mL/min 
2 
CKD 
30-59 mL/min 
3 
CKD 
15-29 mL/min 
4 
AKI 
AKI 
5 
Normal 
60 mL/min 
Following the enrollment of 1 hospitalized patient in Cohort 4 (AKI) the decision to 
stop enrolling patients into this cohort and stop the study was made. At that point, 33 
of the originally planned 38 patients had been enrolled into the study. The decision 
for stopping enrollment was related to the anticipated time to enroll patients into the 
AKI cohort because in the opinion of the Principal Investigator and FAST 
management the enrollment criteria, developed to ensure the safety and appropriate 
evaluation of the CKD population, were too restrictive to enable enrollment of the 
originally planned AKI patients. Secondary rationale for stopping enrollment was the 
expiry of the clinical VFI manufactured in June of 2011, and the inability to extend the 
label dating due to purity issues with the reference materials used to calibrate the 
stability assay. 
 
Subsequently, upon review of the medical history of the single patient enrolled in 
Cohort 4, it was determined that while this patient had a declining kidney function and 
was being treated clinically as an AKI patient, the patient did not meet either the 
Acute Kidney Injury Network stage “2” or Rifle (risk [class R], injury [class I] and 

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failure [class F] + 2 outcome classes [loss and end-stage kidney disease]) stage “I” 
enrollment criteria. This was noted as a protocol deviation. 
 
A secondary endpoint of the study was to compare the kidney function quantified by 
the FAST VFI technique to that determined by existing measurements including 
Iohexol clearance (Cohorts 1, 2, 3, and 5) and serum biomarkers including cystatin C 
and creatinine for eGFR (calculated by the MDRD and CKD-EPI equations). Iohexol 
was omitted from Cohort 4, per protocol, because methodological differences 
between the 2-marker, 2-hour FAST BioMedical technology and the single-marker 6-
hour Iohexol test would not provide comparable results in patients with a dynamically 
changing GFR, such as can be expected in AKI. 
 
Both Iohexol and the FAST mGFR technologies utilized timed blood draws and 
subsequent concentrations to calculate a predicted total AUC. The dose of marker 
given was divided by this AUC to calculate the mGFR, with scaling based on body 
surface area. The FAST mGFR utilized 3 blood draws taken at early time points out 
to 2 hours. A fourth virtual point was calculated based on the PV at T0 (the starting 
concentration in the plasma just after the dose is given or time 0). The Iohexol GFR 6 
test utilized a blank, plus a minimum of 6 additional blood draws taken over 6 hours 
to determine the AUC. Iohexol, having a much smaller molecular size, distributes 
differently within the extracellular space, so some difference in mGFR between the 
2 markers was anticipated. FAST BioMedical expected to measure mGFR values 
slightly lower than Iohexol in normal patients, as most single marker systems like 
Iohexol have difficulty resolving the early decay phase when the clearance is rapid. It 
was expected that the FAST mGFR measurements would be within 10% in the lower 
GFR groups. Prior studies conducted by FAST BioMedical using both the VFI and 
Iohexol in anesthetized dogs, have shown there were differences between the 2 
methods. Analysis of the single marker decay rate showed that it normally resolved 
to a falsely high volume of distribution making the resulting mGFR biased higher than 
the FAST 2 marker system, which quantifies the initial volume of marker distribution. 
This small error in AUC was only significant in mGFRs above 80 mL/min. 
 
In addition to studying a range of GFRs using different measurement techniques, 
FAST BioMedical intended to evaluate FAST’s measured PV method against a 
standard clinical estimate of PV, specifically Nadler’s Formula (Nadler et al 1962). 
 
Plasma Volume (males) = 0.3669 × (Height [m])3 + 0.03219 × (Body Weight [kg]) + 
0.6041 
Plasma Volume (females) = 0.3561 × (Height [m])3 + 0.03308 × (Body Weight [kg]) 
+ 0.1833 
 
The difference between a measured and estimated PV are between plus 20% to 
minus 30% (Feldschuh and Enson, 1977) using radioiodinated human serum albumin 
as the measured plasma marker. Since Nadler’s work was based primarily on 
measurements in healthy patients, it was expected there may be a greater range 
seen with the CKD population in this study. 
 
Pharmacokinetic data analysis was performed on Cohorts 2, 3 and 5. The results 
indicated that the VFI performed as expected over the course of the study and 
corresponded well to the measured renal function and PV of the patients. 
 

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Following administration of 75 mg VFI by IV bolus injection to adults with preserved 
kidney function and impaired kidney function it was concluded that the mean plasma 
concentrations of FD001 exhibited little variability within study groups for samples 
collected from 0.25 to 24 hours. Plasma FD001 concentrations generally reached 
Cmax following the end of the infusion and decreased in a multi-exponential, first-order 
elimination manner with early-phase distribution apparently complete at 8 to 12 hours 
and measureable concentrations observed in all patients at 24 hours. Median plasma 
FD001 tmax values were 0.25 hours, the time of the first post administration sample 
collection in all study groups. Mean plasma FD001 t½ estimates were 5.64, 9.48, and 
18.3 hours for Groups 5, 2, and 3, respectively, increasing with decreasing renal 
function. Mean FD001 CL decreased with decreasing renal function. No significant 
differences between groups were observed in FD001 Cmax (p = 0.6751), tmax (p = 
0.7418), Vss (p = 0.8425), or Cmax/Dose (p = 0.4781); and anticipated differences 
were observed in FD001 AUClast (p < 0.0001), t½,z (p = 0.0015), CL (p < 0.0001), and 
AUCinf/Dose (p < 0.0001). 
 
Following administration of 75 mg VFI by IV bolus injection to adults with preserved 
kidney function and impaired kidney function, mean plasma FD003 concentrations 
exhibited little variability within study groups for samples collected from 0.25 to 
504 hours. Plasma FD003 concentrations reached maximum concentrations within 1 
hour in all study groups and decreased in a mono-exponential, first-order elimination 
manner with measureable concentrations observed in all patients at 504 hours. The 
observed half-life of FD003 ranged from 90 hours for healthy patients to 125 hours 
for patients with severely impaired kidney function (eGFR  15 and < 30 mL/min/1.73 
m2) and were generally similar irrespective of renal function. Mean FD003 volume 
estimates indicated that FD003 appeared to occupy a volume space that 
corresponds with PV. Mean FD003 CL did not change with decreasing renal function. 
No significant differences were observed in FD003 Cmax (p = 0.0743), tmax (p = 
0.0600), AUClast (p = 0.8469), AUCinf (p = 0.8684), t ½,z (p = 0.0508), Vz (p = 0.2644), 
Vss (p = 0.3305), CL (p = 0.5048), Cmax/Dose (p = 0.1652), or AUCinf/Dose 
(p = 0.5048). A delay in the dispersion of the FD003 molecule initially after dosing 
resulting in a later than anticipated Cmax. Complete PK data for the Phase 2a study is 
located in the Investigator’s Brochure from this study. 
3.1.3.4 Phase 2b Study at University of Alabama at Birmingham and ICON 
The Phase 2b study was a prospective, open-label study designed to evaluate the 
safety, tolerability, PK, and pharmacodynamics (PD) of FAST PV and mGFR 
Technology in healthy patients and patients with varying degrees of renal 
impairment.  A total of 32 patients were planned to be enrolled in 4 cohorts of up to 8 
patients/patients each.  Healthy patients in Cohorts 1 and 2 had an eGFR  60 
mL/min/1.73 m2; patients with impaired renal function in Cohort 3 had an eGFR  30 
and < 60 mL/min/1.73 m2; and patients with impaired renal function in Cohort 4 had 
an eGFR  15 and < 30 mL/min/1.73 m2.  Patients in Cohort 1 received a single dose 
of VFI followed 130 minutes later by a 350 mL infusion of 5% albumin in normal 
saline over 30 minutes on day 1.  Patients in Cohort 2 received a dose of VFI 
followed 160 minutes later by a single dose of iohexol on day 1 and a second dose of 
VFI 24 hours following the initial dose of VFI.  Patients in Cohorts 3 and 4 received a 
single dose of VFI followed 160 minutes later by a single dose of iohexol on day 1. 
 

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The pharmacokinetics of FD001 and FD003 showed that the clearance of FD001 
was proportional to kidney function with urine clearances of 59.955 mL/min, 25.919 
mL/min and 12.635 mL/min for cohorts 2, 3, and 4 respectively, when using 0 to 2 
hour data. Following the end of the bolus injection, the plasma concentration of the 
FD001 decreased following a multi-exponent and first order elimination curve. 
Average measured GFR for cohorts 2, 3 and 4 were 87 mL/min, 48 mL/min, and 27 
mL/min respectively. 
 
The FD003 volume estimates indicated that FD003 appeared to occupy a volume of 
space equal to the plasma volume (Figure 4). FD003 had determined elimination 
half-lives of 95.3, 99.1, and 106 hours for Cohorts 2, 3, and 4, respectively. These 
rates were similar across the renal functions studied which indicates the marker’s 
non-renal clearance is preserved in kidney failure patients. 
 
Cohort 1 used FD003 to monitor a change in volume induced by the infusion of 350 
ml of 5% albumin solution given over 30 minutes. At 5 minutes post completion of the 
infusion, the average plasma volume increase was 324 ml with an SD of 49mL, 
showing excellent accuracy for following a change in plasma volume. By 30 minutes 
post infusion, the average volume increase from baseline was reduced to 224ml 
showing that FD003 was being retained within the vasculature even though the 5% 
Albumin solution was slowly distributing into the interstitial space. This cohort was not 
used for PK due to the complication of accounting for the additional volume infused.  
 
The FAST VFI was generally safe and well tolerated in healthy patients in Cohorts 1 
and 2 and in patients with impaired renal function in Cohorts 3 and 4.  No severe or 
serious TEAEs, TEAEs leading to study drug discontinuation, or deaths were 
reported during the study.  Overall, the majority of TEAEs reported were mild in 
severity.  Three patients in Cohort 4 had a TEAE of blood creatine phosphokinase 
increased related to clinically significant abnormal creatine kinase levels.  All 3 
TEAEs of blood creatine phosphokinase increased were mild in severity and 
considered by the investigator to be possibly related to study drug.  There were no 
other clinically significant abnormal clinical laboratory values.  There were no 
clinically significant abnormal ECG or vital signs measurements, nor were there 
clinically meaningful trends identified in observed values or changes from baseline.  
There were no clinically significant abnormal physical examination findings. 
3.2 Rationale 
3.2.1 
Rationale for Dose Selection 
Doses of VFI for this study were selected based on results of the Phase 1 FIH study 
in healthy patients and in the Phase 2a and 2b studies in patients with normal and 
compromised renal function, as well as from data obtained in nonclinical studies 
(additional information is provided in the Investigator’s Brochure). 
 
Data from the FIH study evaluating the safety of the FAST PV and mGFR 
Technology in healthy adult patients indicated the study dose of 75 mg/1.5 mL VFI 
was appropriate for the Phase 2a study. Data from the Phase 2a study and 
improvements to the FAST PV and mGFR Technology have indicated that, in the 
Phase 2b study, a single administration of VFI 47 mg/3 mL would be well tolerated 
and easily measured in all patients. Additionally, for Cohort 2, received 2 doses of 
VFI, each dose being 47 mg/and was well tolerated. As the FD001 marker is cleared 

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and the doses are separated by 24 hours, the maximum level of FD001 in the 
patients after the second dose did not exceed the dose of 52.5 mg of FD001 given in 
Phase 2a. Also, as the amount of FD003 had been reduced in the new VFI 
formulation, a second dose of VFI resulted in plasma exposure values close to the 
exposure previously observed in patients administered the high doses of FD003. In 
the Phase 2a clinical study a dose of 22.5 mg of FD003 was given, in the Phase 2b 
the maximum amount of FD003 following repeat dosing was 24 mg. This repeat-dose 
strategy was agreed to by the United States Food and Drug Administration in written 
correspondence dated February 24, 2016. The predose sample taken on Day 2 for 
Cohort 2 was analyzed and provided the residual amount of both FD001 and FD003. 
The FD003 residuals were then subtracted as was a normal predose sample from 
the values of the post-second-administration samples to allow for a clean calculation 
of PV. FAST has derived the mathematical equations for GFR to appropriately 
compensate for any residual FD001; however, the normal patients did not have a 
residual level of FD001 patients on Day 2 that would have affected the GFR 
calculation. Repeat doses of VFI are likely to be undertaken clinically as a method of 
evaluating treatments. These repeat measurements would not be done rapidly as a 
treatment required time to be effective. FAST chose 24 hours to separate VFI doses 
to mimic likely clinical situations. As discussed in section 3.1.3.4 the repeat 
administrations were generally safe and well tolerated by the 8 patients.  
 
3.2.2 
Rationale for Study Design 
This is an investigator-initiated, one-armed phase II clinical trial designed to evaluate 
the safety and functionality of the FAST PV and mGFR Technology in CHF patients.  
 
Previous clinical studies have shown the FAST technology to be safe and functional 
in patients with chronic kidney disease (CKD) and in healthy subjects. The 
technology has not been studied in CHF patients. In previous clinical studies, the 
technology has been demonstrated to accurately measure PV and mGFR in patients 
with CKD and healthy patients when compared to other clinical techniques including, 
eGFR calculations, Iohexol clearance and Nadler’s formula. Additionally, previous 
studies have shown that the concentration of the PV marker remains relatively 
constant during the first two hours after injection, given that this high-molecular 
weight marker is predicted to exclusively occupy the intravascular compartment and 
exhibit a long half-life.  
In this study, the safety and functionality of the FAST Technology will be evaluated in 
CHF patients. The non-clinical and clinical history of the product provides confidence 
that the technology will be well tolerated and function correctly in the CHF population.  
 
The blood samples are timed to maximize the information gathered during the study. 
In addition to measurements of PV and GFR, FAST will determine the technology’s 
functionality by comparing the concentration of the PV marker at the 15, 30 and 60 
minute time points. It is not believed that the plasma volume will change significantly 
within that time period and thus the concentration of the PV marker should remain 
relatively constant. 
 

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4 Study objectives 
4.1 Primary endpoints 
 
Global aim is to assess the safety and function of the FAST PV and mGFR 
Technology in hospitalized patients with heart failure. 
 
x Safety will be assessed by determination of the absolute and relative frequencies 
of AEs and SAEs related to VFI. 
 
x Function of the FAST PV measurement will be assessed by determining the 
plasma stability of the FD003 high molecular weight marker over the 15, 30, and 
60 minute blood draws and applying the following criteria: 
- 
The FAST PV measurement is considered as stable, if the mean plasma 
concentration of FD003 at 30 minutes is not more than 10% lower than the 
mean plasma concentration at 15 minutes AND if the mean plasma 
concentration at 60 minutes is not more than 10% lower than the mean 
plasma concentration at 30 minutes 
- 
We will determine the percentage of patients which show a decline in the 
plasma concentration of FD003 of more than 10% from 15min to 30min 
and separately from 30 min to 60min. This percentage should be ideally 
close to 0. 
 
 
 
 
4.2 Secondary endpoints 
x To evaluate how estimated PV (ePV) and estimated total blood volume (TBV) 
assessments on days 1 and 3 (by established measures such as Nadler’s formula 
or Metropolitan Life Tables) predict measured PV (mPV; as assessed by FAST 
methodology) and measured TBV (mTBV; calculated from mPV and measured 
hematocrit) at these time points.  
 
x To evaluate how a clinical evaluation (including a reasonable subset of the 
variables age, gender, BMI, hematocrit, edema grade, presence of pulmonary 
rales, presence of jugular venous congestion, arterial blood pressure, NYHA 
stage, respiratory rate) on days 1 and 3 predicts mPV and mTBV at these time 
points.  
 
x To evaluate whether eGFR calculation by the CKD-EPI formula on days 1 and 3 
provides an accurate estimate of measured GFR (mGFR; by assessed by FAST 
methodology) in heart failure patients undergoing active fluid management.  
 
x To evaluate whether patients with a low mGFR/eGFR ratio on days 1 and 3 are at 
higher risk of developing AKI within the following 48-72 hours.  
 
x To evaluate whether ranges of mPV (low mPV, normal mPV, high mPV) and/or 
mTBV (low mTBV, normal mTBV, high mTBV) on days 1 and 3 are associated 
with subjectively reported symptoms (PGA, dyspnea, dizziness, nausea) 

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determined using patient symptom assessment sheets within the following 24-48 
hours.  
 
x To evaluate whether patients with low mPV or low mTBV (cutoffs potentially 
adjusted for some of the following variables age, gender, body height, body 
weight, optimal body weight (dry weight), hematocrit) on day 1 or on day 3 are at 
risk of developing low-output complications within the next 24-48 hours (e. g. 
subjectively reported dizziness/nausea, PGA, hypotension, need for iv-fluid 
therapy).  
 
x To evaluate whether patients with a high mPV or a high mTBV (cutoffs potentially 
adjusted for some of the following variables age, gender, body height, body 
weight, optimal body weight (dry weight), hematocrit) on day 1 and/or day 3 are at 
risk of being refractory to diuretic therapy (e. g. require dosage increases of 
furosemide/torasemide, require ultrafiltration/RRT within the next 24-48 hours, fail 
to improve subjectively in PGA and dyspnea scales).  
 
x To evaluate whether the time course of ePV and estimated TBV (eTBV) from day 
1 to day 3 adequately reflects the time course of mPV and mTBV at these time 
points.  
 
x To evaluate whether the change of mPV and mTBV from day 1 to day 3 is 
predictive of length of stay, dialysis/ultrafiltration requirement, or rehospitalization 
within 30 days, or mortality.  
 
x To evaluate whether the time course of mGFR from day 1 to day 3 correlates 
temporally with or predicts the time course of eGFR within the next 24-48 hours.  
 
x To evaluate (by an adjudication committee) whether clinical decision making 
would have been affected by adding FAST GFR and PV measurements to clinical 
routine including the following questions:  
o Would FAST GFR and PV measurements have changed clinical 
management overall? 
o Would the FAST GFR and PV measurement on day 1 have led to applying 
a different diuretics dosage between day 1-3? 
o Would the FAST GFR and PV measurement on day 3 have led to applying 
a different diuretics dosage between day 3-5? 
o Would the FAST GFR and PV measurement on day 1 have led to choosing 
dialysis/ultrafiltration instead of diuretics escalation between day 1-3? 
o Would the FAST GFR and PV measurement on day 1 have led to choosing 
diuretics escalation instead of dialysis/ultrafiltration between day 1-3? 
o Would the FAST GFR and PV measurement on day 3 have led to choosing 
dialysis/ultrafiltration instead of diuretics escalation between day 3-5? 
o Would the FAST GFR and PV measurement on day 3 have led to choosing 
diuretics escalation instead of dialysis/ultrafiltration between day 3-5? 
4.3 Time point (for the evaluation of secondary endpoints) 
x Secondary endpoints will be evaluated at the time points specified above.  

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4.4 Study designs 
This is an investigator-initiated, human clinical trial designed to evaluate the safety 
and functionality of the FAST PV and mGFR Technology in CHF patients. Data from 
the FAST PV and mGFR Technology will not be evaluable by the treating physician, 
but will be made available to an adjudication committee for consideration after patient 
enrollment has concluded or at predetermined intervals during the course of the 
study.  
 
Patients enrolled in the study will be administered VFI (Day 1) and with a second 
dose occurring 24 to 48 hours after the initial dose (Day 3).  
 
After consenting to be enrolled in the study, patients meeting the enrollment criteria 
will receive a single dose of VFI. Blood draws (approximately 2 mLs) will be collected 
pre-dose and at 15 (± 5), 30 (± 5), 60 (+30), and 180 (+120) minutes post-dose. Note 
that the 30 minute blood draw must be at least 10 minutes after the 15 minute blood 
draw, and the 180 minute blood draw must be at least 100 minutes after the 60 
minute blood draw. 
As described in section 6.1.1.2 of this protocol, the physician will remain present at 
the bedside of the patient for 10 minutes after administering the FAST VFI. 
Thereafter, the patient will be monitored on an ongoing basis, with clinical staff 
assessing the patient’s condition along with the 15, 30, and 60 minutes blood draws 
specified in the protocol. 
Patients will be treated according to standard of care throughout the time of their 
hospitalization. A second dose of VFI will be administered no sooner than 43 hours 
and no later than 53 hours after the initial dosing. Blood draws (approximately 2 mLs) 
will be collected pre-dose and at 15 (± 5), 30 (± 5), 60 (+30), and 180 (+120) minutes 
post-dose. Note that the 30 minute blood draw must be at least 10 minutes after the 
15 minute blood draw, and the 180 minute blood draw must be at least 100 minutes 
after the 60 minute blood draw. The available laboratory values (see Table 1) will be 
entered in the Dosing Day Case Report Form (CRF), along with any copies of 
physician’s notes and orders entered that day. Prior to administering the first and 
second dose of VFI, the physician will be asked to complete a very brief survey to 
provide a qualitative assessment of the patient’s perceived volume status and renal 
function prior to initiating the FAST PV and mGFR measurements.  
 
Laboratory values and assessments (see Table 1) will be captured on each day that 
the VFI is administered and these data will be entered into the CRF. Any AEs 
determined by the investigator will be captured. Patients will also receive a follow up 
phone call 7 days (± 2 days) after the first dose of VFI was administered, and a 
second follow up call 30 days after the first dose of VFI was administered as an 
end-of-study (EOS) follow up call. Any AEs determined by the investigator will be 
captured during the follow up calls and will be followed to resolution.  Additionally, in-
person follow-up visits may be scheduled as needed. 
 
Table 1:Schedule of Study Events/Examinations 
Study 
Timepoint 
Data 
collected 
from 
chart 
prior to 
enrolling  
Patient 
Enrolled 
and 
Dosed 
Data 
collected 
between 
1st and 
2nd Dose  
Patient 
2nd VFI 
Dose 
Data 
Collected 
after 2nd 
Dose 
Data 
Collected 
Before 
Discharge 
Follow up 
in person 
or by 
phone 
call 
Follow 
up 
phone 
call/ 
Follow 
up Visit 
for 

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Pregnan
cy Test 
if 
WOCBP 
Visit 
Pre-
Dose 
Day 1 
Day 2 
Day 3 
Day 4 
Day 5 
Day 10 
Day 30 
Informed 
Consent 
X 
  
  
  
  
  
  
  
Inclusion/ 
Exclusion 
Criteria 
Assessment 
X 
  
  
  
  
  
  
  
Structural 
Heart Disease 
Criteria 
Assessment a 
  
X 
  
  
  
  
  
  
Demographics 
b 
  
X 
  
  
  
  
  
  
Comorbidities 
c 
  
X 
  
  
  
  
  
  
Full 
medication list 
on enrollment 
into the study  
and during 
follow-up  d 
  
X 
 X 
X 
X 
X 
 
  
Patient 
Symptom 
Assessment 
Survey e 
  
X 
X 
X 
X 
X 
 
  
Clinical 
Presentation/ 
Exam f 
  
X 
X 
X 
X 
X 
  
  
Treating 
Physician 
Survey g 
  
X 
  
X 
  
  
  
  
Study-Related 
Laboratory 
Measurements 
h 
  
X 
X 
X 
X 
X 
  
  
Routine 
Clinical 
Parameters i 
X* 
X 
X 
X 
X 
X 
  
  
Clinical Data j 
X* 
X 
X 
X 
X 
X 
  
  
Adverse 
events 
  
X 
X 
X 
X 
X 
X 
X 
Blood 
specimen for 
marker 
analysis k 
  
X 
  
X 
  
  
  
  
Discharge 
from study 
  
  
  
  
  
  
  
X   
Adjudication 
Committee 
Evaluationl 
 
 
 
 
 
 
 
X 
WOCBP 
Pregnancy 
X 
 
 
X 
 
 
 
X 

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test m 
a See Table 2 for Structural Heart Disease Criteria 
b See Table 3 for specific demographic parameters 
c See Table 4 for specific instructions on collecting comorbidities 
d See Table 5 for specific instructions of collecting medications  
e See Annex D for the Patients Symptom Assessment Survey 
f See Table 6 for Clinical presentation/exam parameters 
g See Annex E for the Treating Physician Survey 
h See Table 7 for Study Related Laboratory Measurements 
i See Table 8 for Routine Clinical Parameters 
j See Table 9 for Clinical Data collected 
k Blood Specimens for the determination of the FAST PV and mGFR will be taken pre-dose and at 15 (±5), 30 (±5), 60 (+ 
30) and 180 (+ 120) minutes post dose on Day 1 and Day 3. Note that the 30 minute blood draw must be at least 10 
minutes after the 15 minute blood draw, and the 180 minute blood draw must be at least 100 minutes after the 60 minute 
blood draw.  
l See Table 10 for the Adjudication Committee Evaluation Questions; Adjudication Committee meets after all collected 
data available. 
m See Table 11 for Details of Pregnancy test  
*will be collected retrospectively after study inclusion 
 
 
Table2:StructuralheartdiseaseCriteria
Lastdocumentedleftventricularejectionfraction(%)
Evidenceofleftventricularhypertrophy
Evidenceofleftatrialenlargement
Evidenceofdiastolicdysfunction
Othercauseofheartfailure
 
Table3:Demographics
ID
Dateofadmission
Dateofinclusion(Day1)
Dateofbirth
Ageatinclusion(years)
Gender
Ethnicity
 
Table4:Comorbidities
Freetextdocumentationofallcomorbidities
Inaddition,thefollowingcomorbiditiesarerecordedsystematically(by
“yes”or“no”answers):
CKDstage
previousmyocardialinfarction
Hypertension
Preexistingcoronaryheartdisease
Preexistingcongestiveheartfailure
Previoushospitalizationforacutedecompensatedheartfailure?

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Diabetesmellitus
Cerebralvasculardisease
Peripheralarterialocclusivedisease
Othercomorbidities
 
Table5:Medicationonadmission
Freetextdocumentationofallexistingmedication(1)uponadmission,3)uponinclusionintothe
study(day1)and(3)duringdays2Ͳ5,includingname,dosageandtimingofintake
Inaddition,thefollowingmedicationsarerecordedsystematically(by“yes”or“no”answers):
ACEinhibitor
AT1receptorblocker
AT1/Neprilysininhibitor
Loopdiuretic
Furosemidep.o.dosage(mg)per24hpriortodosing(accordingtomedicationplan)
Torasemidep.o.dosage(mg)per24hpriortodosing(accordingtomedicationplan)
Thiazidediuretic
Aldosteroneantagonist
betaͲBlocker
Nitrates
Digoxin/Digitoxin

 
Table6:Clinicalpresentation/ExamParameters
*Height(cm)
Weight(kg)
*Paroxysmalnocturnaldyspneaduringprevious3days?
*Nightcough
*Cardiomegaly(clinicaloronchestXͲrayorechocardiography)
*S3gallop
*Hepatojugularreflux
*Hepatomegaly
*Pleuraleffusion
*Laststablebodyweightbeforeadmission("bestbodyweight")(kg)
NYHAstage(0Ͳ4)
Ankle/lowerextremityedema(grade0Ͳ4;accordingtoGuelphGeneralHospitalCongestiveHeart
FailurePathway)
Pulmonaryrales
Systolicbloodpressure(mmHg)(morning,supine)
Diastolicbloodpressure(mmHg)(morning,supine)
**Systolicbloodpressure(mmHg)(morning,30secafterstandingup)
**Diastolicbloodpressure(mmHg)(morning,30secafterstandingup)
Heartrate(beatsperminute)(morning,supine)
Heartrate(beatsperminute)(morning,30secafterstandingup)
SpO2(%)(morning)
Jugularvenousdistension(>4cmabovesternalanglewith45°upperbodyelevation)(morning)
Respiratoryrate(breathsperminute)(morning)
O2supplyrequirement(inL/min)(morning)

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*OnlyassessedDay1
*OnlyassessedDay1andDay3
 
Table7:StudyͲrelatedlaboratorymeasurements
Serumcreatinine(mg/dl)
*Hematocrit(%)
**creatinephosphokinase(CPK)(U/l)
**NTͲproBNP(ng/l)
**Alanineaminotransferase(ALT)(U/l)
**Aspartateaminotransferase(AST)(U/l)
**Alkalinephosphatase(AP)(U/l)
**bilirubin,totalanddirect(mg/dl)
**INR(dimensionless)
StorageofEDTAPlasma,HeparinͲPlasma,Serum,whole
blood,centrifugedurine(10aliquotsa200µleach)atч
Ͳ75°C
*OnlyCollectedDay1andDay3;ForuseincalculatinganePV
fromNadler’sformula
**OnlyCollectedDay1,3and5
 
Table8:Routineclinicalparameters
*Serumcreatinineonadmission(mg/dl)
*Baselineserumcreatinine(nadirfrominspectionofprevious
labvaluesorbyprimarycarephysician)(mg/dl)
Serumcreatinine(mg/dl)
Serumsodium(mmol/l)
Serumpotassium(mmol/l)
eGFR(CKDͲEPI)(ml/min/1,73m2)
Hemoglobin(g/dl)
SerumͲAlbumin(g/dl)
Urineoutput(0:00Ͳ24:00ontheindicatedday)(ml/24h)(if
incomplete,extrapolatetheavailabletimeperiodto24hours)
ChestXͲray:evidenceofpulmonarycongestion
ChestXͲray:evidenceofpleuraleffusion
ChestXͲray:evidenceofcardiomegaly
Alanineaminotransferase(ALT)(U/l)
Aspartateaminotransferase(AST)(U/l)
Alkalinephosphatase(AP)(U/l)
Bilirubin,totalanddirect(mg/dl)
INR(dimensionless)
*OnlyonDay1
 
Table9:Clinicaldata
Total24hdoseofi.v.furosemide(from06:00amsamedayto06:00amnextday)(mg)
Total24hdoseofi.v.torasemide(from06:00amsamedayto06:00amnextday)(mg)
Total24hdoseofp.o.furosemide(from06:00amsamedayto06:00amnextday)(mg)
Total24hdoseofp.o.torasemide(from06:00amsamedayto06:00amnextday)(mg)
Applicationofanythiazidediuretic?

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Dialysis/ultrafiltrationperformed?(from06:00amto06:00amnextday)
iv.fluids(NaCl,balancedelectrolytes)(from06:00amto06:00amnextday)(L/24h)
Isthetotaldosageofdiuretics(administeredfrom06:00amsamedayto06:00amnextday)
increasedcomparedtoprevious24hdosage?(e.g.increasingloopdiureticsoradding
thiazidediureticsoraldosteroneantagonists)?
Isthetotaldosageofdiuretics(administeredfrom06:00amsamedayto06:00amnextday)
decreasedcomparedtoprevious24hdosage?(e.g.decreasingloopdiureticsorremoving
thiazidediureticsoraldosteroneantagonists)?
 
Table10:AdjudicationCommitteeEvaluation
WouldFASTGFRandPVmeasurementshavechangedclinical
managementoverall?
WouldtheFASTGFRandPVmeasurementonday1haveledtoapplying
adifferentdiureticsdosagebetweenday1Ͳ3?
WouldtheFASTGFRandPVmeasurementonday3haveledtoapplying
adifferentdosagebetweenday3Ͳ5?
WouldtheFASTGFRandPVmeasurementonday1haveledtochoosing
dialysis/ultrafiltrationinsteadofdiureticsescalationbetweenday1Ͳ3?
WouldtheFASTGFRandPVmeasurementonday1haveledtochoosing
diureticsescalationinsteadofdialysis/ultrafiltrationbetweenday1Ͳ3?
WouldtheFASTGFRandPVmeasurementonday3haveledtochoosing
dialysis/ultrafiltrationinsteadofdiureticsescalationbetweenday3Ͳ5?
WouldtheFASTGFRandPVmeasurementonday3haveledtochoosing
diureticsescalationinsteadofdialysis/ultrafiltrationbetweenday3Ͳ5?
*ResponseswillbecollectedasaYes/No,withanadditionalfreetextsection
availabletorecorddetailstosubstantiatetheresponse
Table11:PregnancyTestsinWOCBP
Medicallymonitoredpregnancytestswitha
minimumsensitivity25mU/ml
willbeassessedwithin24hourspriorfirststudy
drugadministration,onDay3and30inWOCBP
 
4.5 Timetable 
x Recruitment and Enrolment to run (expected): January 2019-July 2019 
x FPFV: After consent 
x LPLV: 30 days following LPFD (Last patient First dose) 
 
5 Study population 
50 patients with acute decompensated heart failure will be enrolled at the nephrology and 
cardiology departments of Charité - Universitätsmedizin Berlin. There will be no specific 
gender-fractions as no gender specific differences concerning efficacy and safety of the 
investigational product are expected. 

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5.1 Inclusion criteria 
x 
Written informed consent indicating that they understand the purpose of and 
procedures required for the study and are willing to participate in the studyprior to 
any study related measures present 
x 
Hospitalized patient with acute decompensated heart failure, diagnosed on the 
basis of the presence of at least one symptom (dyspnea, orthopnea, or edema) 
and one sign (rales, peripheral edema, ascites, or pulmonary vascular congestion 
on chest radiography) of heart failure.  
x 
Subject:  18. Male with female partners of childbearing potential gave agreement 
to practice abstinence or use condoms from enrollment through 90 days after 
administration of the last dose of study drug present. 
x 
A female subject is eligible to enter the study if she is:  
o Not pregnant or nursing 
o Of non-childbearing potential (i.e., post-menopausal defined as having 
been amenorrheic for at least 1 year prior to screening, or has had 
bilateral tubal ligation at least 6 months prior to administration of study 
drug or bilateral oophorectomy or complete hysterectomy) 
o If of childbearing potential, must have a negative urine or serum 
pregnancy test prior to drug administration and be using a highly effective 
means of contraception during study participation and until 1 month after 
the last dose of study drug. Highly effective contraception methods 
include: 
x 
Total abstinence (when this is in line with the preferred and 
usual lifestyle of the subject. Periodic abstinence (e.g., 
calendar, ovulation, symptothermal, post-ovulation methods) 
and withdrawal are not acceptable methods of contraception 
x 
Combination of any two of the following (i+ii or i+iii, or ii+iii): 
(i) use of oral, injected or implanted hormonal methods of 
contraception or other forms of hormonal contraception that 
have comparable efficacy (failure rate < 1%), for example 
hormone vaginal ring or transdermal hormone contraception. 
(ii) placement of an intrauterine device (IUD) or intrauterine system 
(IUS) 
(iii) barrier methods of contraception: condom or occlusive cap 
(diaphragm or cervical/vault caps) 
x 
In case of use of oral contraception women should have been 
stable on the same pill for a minimum of 3 months before taking 
study treatment 
x 
 
x 
Partner of a male patient: Agreement to use a medically acceptable method of 
contraception (a barrier method, intrauterine device, or hormonal contraception) 
from enrollment through 90 days after administration of the last dose of study drug 
present. 
x 
Male patient agreed to not donate sperm from enrollment through 90 days after 
administration of the last dose of study drug present 
x 
Patient is able to communicate effectively with the study personnel. 
x 
Patient is informed of the nature and risks of the study and give written informed 
consent prior to enrollment. 
5.2 Exclusion criteria 
x 
New or ongoing myocardial infarction or instable angina present at the time of 
planned study inclusion. 

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x 
Patient shows evidence of severe infection other than pneumonia, or active 
internal bleeding (characterized by recent decrease of blood hemoglobin 
concentration by more than 2 g/dl). 
x 
Patient experiences new onset atrial fibrillation. 
x 
Patient has an elective surgery planned during the 30 days they are enrolled in 
the study. 
x 
Patient has a psychiatric disease or a history of illicit drug use that would prohibit 
them from complying with study requirements  
x 
Prior exposure to VFI present. 
x 
History of any clinically significant allergic or negative reactions, side effects, or 
anaphylaxis to fluorescent dyes, or dextran molecules present. 
x 
Patient requires intravenous vasodilators or inotropic agents (other than digoxin or 
digitoxin) for heart failure. 
x 
Patient undergoes chronic dialysis (for example peritoneal or hemodialysis) 
treatments.  
x 
Patient is in cardiogenic shock or on vasopressors. 
x 
Hypotension as defined by blood pressure < 90 systolic and/or < 50 mm Hg 
diastolic exists. 
x 
Patients suffering from significant non-cardiac diseases of other organ systems 
(e. g. Malignancies, significant neurological diseases). 
x 
Patient does not have a working telephone.  
x 
Patient is a pregnant or nursing (lactating) women, where pregnancy is defined as 
the state of a female after conception and until the termination of gestation, 
confirmed by a positive human chorionic gonadotropin (hCG) laboratory test. 
x 
Lack of willingness to storage and disclosure of pseudonymous disease data in 
the context of the clinical trial present. 
x 
Subject has a participation in another interventional clinical trial during this study 
or within 30 days (or longer) before entry into this trial (as a minimum; 5 x 
elimination half-life / terminal elimination of an IMP). 
x 
Subject is legally detained in an official institution. 
x 
Subject is dependent on the sponsor, the investigator or the trial sites. 
 
6 Plan for medical treatment 
6.1 Description of study medication / investigational medicinal product 
The visible fluorescent injectate (VFI) agent is comprised of a mixture of two different 
molecular weight carboxymethyl dextran molecules (5 kD and 150 kD) with different 
fluorescent dye molecules attached.  The 5 kD carboxymethyl dextran, (FD001), is 
labeled with 5-aminofluorescein and the 150 kD carboxymethyl dextran, (FD003), is 
labeled with 2-sulfohexamine rhodamine.  These fluorescent labels are covalently 
attached to the dextran through the carboxymethyl moiety.  Please see the chemical 
structures for FD001 and FD003, which are shown in Figures 1 and 2 below, 
respectively. 
 
The high molecular weight labeled carboxymethyl dextran is not rapidly cleared from 
the vasculature and is not rapidly cleared by passive filtration in the kidneys; 
therefore, its concentration in the blood stream after injection provides a direct 
measurement of the total blood volume.  The low molecular weight labeled 
carboxymethyl dextran is also not patient to rapid metabolism but is freely filtered by 
the kidneys.   
 

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Figure 1: Structure of FD001 (5 kD carboxymethyl dextran with covalently bound 
5-aminofluorescein) 
Figure 2: Structure of FD003 (150 kD carboxymethyl dextran with covalently bound 
2-sulfohexamine rhodamine) 
 
 
 
            Figure 1 - FD001                                    Figure 2 - FD003  
 
The VFI is administered intravenously through a bolus injection. The anticipated 
single dose for a 70 kg human will consist of 35 mg of FD001 and 12 mg of FD003 
diluted at 15.67 mg/mL mg/mL for a total injection of 3 mL of VFI.   
 
6.1.1 Adverse drug reactions and restrictions, contra-indications, drug interactions 
6.1.1.1 Contraindications 
The FAST VFI (FD004) should not be administered to patients with known 
hypersensitivity to dextran, 5-aminofluorescein dye, or 2-sulfohexamine rhodamine 
dye. 
6.1.1.2 Precautions (overview, dextran safety, anaphylactoid reactions, patient 
monitoring) 
The FAST VFI (FD004) was well tolerated in the nonclinical studies conducted to 
date.  Anaphylactic responses to dextrans, while rare, must be considered as a 
possibility for the high molecular weight dextrans being evaluated by FAST.  As such, 
all testing facilities should have immediately available procedures to administer 
treatment for an anaphylactic response should it occur. 
 
Overview of The FAST BioMedical Visible Fluorescence Injectate (VFI)  
The FAST BioMedical VFI (or FD004) contains 2 different molecular weight 
carboxymethyl dextran markers with 2 different fluorescent dye molecules attached.  
The small molecular weight marker, FD001 (approximately 5 kD), is conjugated with 
5-aminofluorescein and the large molecular weight marker, FD003 (approximately 
150 kD) is conjugated with 2-sulfohexamine rhodamine.  These fluorescent labels are 
covalently attached to the dextran through the carboxymethyl moiety.  Unlike clinical 
dextrans which are dosed in grams, FAST BioMedical has worked to minimize the 
dose of VFI. Since the 2013 Phase I study in Germany, the VFI dose has been 
reduced by almost 70%, moving from the initial maximum clinical dose of 150 mg to 
the current clinical dose of 47mg (35 mg of FD001 and 12 mg of FD003).   

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The larger marker, FD003, was designed not to be rapidly cleared from the 
vasculature and is also not rapidly cleared by passive filtration in the kidneys. 
Therefore, its concentration in the bloodstream after injection provides a direct 
measurement of the total blood volume by dilution principles.  
The smaller marker, FD001, was also designed not to be subject to rapid 
metabolism, but is freely filtered by the kidneys.  The decline in the concentration of 
the small FD001 marker is consistent with the renal clearance rate which, when 
scaled to a patient’s actual vascular volume, provides an assessment of measured 
GFR. Accurate and timely measurements of vascular volume and GFR are critically 
important metrics in patients with acute congestive heart failure and cardiorenal 
syndrome, where assessing patients’ volume status are key to the clinical 
management of the patient.1,2,3 
 
Dextran Safety 
As noted by the reviewer, dextran anaphylactoid reactions are well documented. For 
this reason, in development of its markers, FAST BioMedical has taken great care to 
understand and incorporate the body of knowledge known about dextrans and to 
enlist the assistance of the foremost experts in the area of dextran safety. 
Specifically, in the 1970s and 1980s, Pharmacia AB in Uppsala, Sweden conducted 
extensive research to understand dextran-induced anaphylactic reactions (DIAR) and 
demonstrate different approaches to managing the immunogenic potential of these 
molecules. The FAST BioMedical VFI was developed in conjunction with many of the 
former Pharmacia chemists and immunologists. These key leaders in the field of 
dextran biology demonstrated that the immunogenicity of dextrans could be 
significantly attenuated by either: (1) using a substituted dextran such as a 
carboxymethyl dextran or (2) pre- or co-administering a low molecular weight dextran 
(1 to 5 kD) to serve as a “hapten”.4,5  Specifically, in 1988, K.G. Lungstrom published 
hapten inhibition with pre-inoculation of dextran 1 kD to block antigen combining sites 
led to a 35-fold reduction in dextran-induced anaphylactic reactions, and a 90-fold 
reduction in fatal dextran reactions.4 The vast majority of reported reactions were 
reported as mild (skin reactions, bradycardia, hypotension, etc.), and Lungstrom 
reported only 2 fatal reactions in over 5.1 million doses, making the use of dextran 
with a 1 kD hapten inhibitor safer than other commercially available colloids as well 
as many other injectable products . 1  
  
In 1977, Johannes Ring published a prospective multicenter study looking at the 
safety of various colloids including plasma protein fractions, dextran, gelatin solutions 
and hydroxyethylstarch (HES).5  Ring’s study reported the following incidence of 
anaphylactoid reactions:  
                                                 
1Katz,S.D.:InSearchofEuvolemiainHeartFailure.JournaloftheAmericanCollegeofCardiology.2014;2:
2213Ͳ1779.
2Ronco,C.,Kaushik,M.,etal.:DiagnosisandManagementofFluidOverloadinHeartFailureandCardioͲRenal
Syndrome:The“5B”Approach.SeminarsinNephrology.2012;32:129Ͳ141.
3Miller,W.L.,Mullan,B.P.:UnderstandingtheHeterogeneityinVolumeOverloadandFluidDistributionin
DecompensatedHeartFailureisKeytoOptimalVolumeManagement:RoleforBloodVolumeQuantitation.
JournaloftheAmericanCollegeofCardiology.2014;2:2213Ͳ1779.
4Lungström,K.G.:SafetyofDextraninRelationtoOtherColloids–TenYears’Experiencewith
HaptenInhibition.InfusionstherTransfusionsmed.1993;20:2006Ͳ10
5RingJ,MessmerK.:Incidenceandseverityofanaphylactoidreactionstocolloidvolumesubstitutes,
Lancet1977;1;466Ͳ469.

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Colloid 
Number of 
infusions 
registered 
Number of 
anaphylactoid 
reactions 
Fractional 
incidence of 
anaphylactoid 
reactions 
Plasma proteins 
85,630 
12 
0.014 % 
Dextrans 
85,882 
28 
0.032 % 
Gelatin 
12,989 
15 
0.115 % 
HES 
16,405 
15 
0.085 % 
 
Additional studies by Caroline Mendler showed that concomitant administration of 
dextran 1 kD is as effective as pre-inoculation in reducing dextran reactivity.6 This is 
a significant finding for products such as the FAST BioMedical VFI where the 
additional time required to pre-inoculate the patient may impede timely patient care.  
 
Development of the FAST VFI  
FAST BioMedical has taken great care to develop a product that minimizes the 
potential for intolerance reactions. This begins with the procurement of raw 
pharmaceutical-grade dextran which is derived from the B512 Leuconostoc 
mesenteroids strain, which yields a highly homogenous dextran with very limited 
branching. A 1957 paper by Elvin Kabat described the relationship between the 
linearity of the dextran molecule and its immunogenic potential, demonstrating that 
the limited branching of the B512 strain led to lower reactivity when compared to 
dextrans derived by other strains.7 Today, the B512 strain is still considered the 
safest of all dextran strains, and this dextran strain is exclusively used to produce the 
dextran employed in the synthesis of the FAST BioMedical VFI.      
In effort to further incorporate the historical learnings from past dextran development 
efforts, FAST BioMedical convened an advisory committee of former Pharmacia 
dextran researchers, chemists and clinical consultants including Karl-Gostä 
Ljungstrom, Harriet Hedin, Tony de Belder, and Peter Buckley. Many of these 
experts are cited in the seminal literature on dextran anaphylaxis. The committee 
provided input on the composition of the FAST BioMedical VFI. Specifically, the 
committee indicated that the low molecular weight marker, FD001, at approximately 5 
kD in the FAST BioMedical VFI is very likely to act as a hapten inhibitor, having the 
same ability to bind antibodies without causing a precipitating immune response.  
Further, it was concluded that FD001 at approximately 5 kD dextran could be co-
administered with FD003 at approximately 150 kD based upon the aforementioned 
studies performed by Caroline Mendler.   
The advisory committee also favored use of a modified carboxymethyl dextran to 
further reduce the immunogenic potential of the FAST BioMedical VFI.  This 
assertion was based on the 1975 paper by Wolfgang Richter where he demonstrated 
that the use of substituted dextrans, such as carboxymethyl dextran, significantly 
reduced the reactivity of the dextran molecule.8 The process for synthesizing the 
                                                 
6Mendler,C.:HaptenInhibitionofDextranͲInducedAnaphylacticReactionsinDogs.TheUniversityof
MunichInstituteofSurgicalResearchPh.D.,dissertation.1980.
7Kabat,E.A.,Turino,A.B.:StudiesontheImmunochemicalBasisofAllergicReactionstoDextranin
Man.JournalofClinicalInvestigation.1957;36:1160Ͳ1170.
8Richter,W.:EffectofSubstitutiononReactivityofB512DextranFractionswithAntiͲB512Dextranin
HeterologousPassiveCutaneousAnaphylaxis.InternationalArchivesofAllergyandAppliedImmunology.1975;
48:505Ͳ512.

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carboxymethyl dextran used in the FAST BioMedical VFI was then developed in 
conjugation with Tony de Belder, a former colleague of Wolfgang Richter’s at 
Pharmacia and world-renowned expert in the field of dextran and dextran derivatives.  
In considering the dextran strain, the role of FD001 as a likely hapten inhibitor, and 
the use of substituted (carboxymethyl) dextran, FAST BioMedical has made 
substantial efforts to learn from and utilize established dextran science.  We believe 
that these efforts have greatly reduced the immunogenic potential of the FAST 
BioMedical VFI as compared to unmodified dextran products. 
 
Patient Monitoring and Treatment  
All patients enrolled in the EMPAKT CHF study will be actively monitored for potential 
intolerance reactions to the FAST VFI. The FAST VFI has been administered 97 
times in 89 patients through the course of the preceding 3 clinical studies. Patient 
populations in those studies included healthy subjects and patients with advanced 
chronic diseases such as CKD, COPD, hypertension, diabetes, and polycystic kidney 
disease.  No SAEs were reported in the prior studies, though AEs commensurate 
with a mild to moderate immunological response were noted including pruritus (5), 
nausea (4), vomiting (3), diarrhea (2), abdominal pain (1), and urticaria (1). In all 
cases, the symptoms resolved without sequalae. It is also noteworthy that an 
intolerance reaction may be more likely in a healthy subject with a fully functional 
immune system than in the typical heart failure patient whose immune system may 
be somewhat compromised. 
The EMPAKT CHF study will be conducted on hospitalized patients who will have 
regular access to nurses and physicians through the course of normal clinical care. 
Because dextran-related anaphylaxis is almost exclusively an acute onset event, the 
physician or nurse will remain present at the bedside of the patient for 10 minutes 
after administering the FAST VFI. Thereafter, the patient will be monitored on an 
ongoing basis, with clinical staff assessing the patient’s condition along with the 15, 
30, and 60-minute blood draws specified in the protocol. In periods where a nurse or 
physician are not present at the patient’s bedside, the patient will have access to a 
call button to immediately notify the clinical staff should they experience symptoms of 
concern or a medical emergency. The following classification system will be used to 
assess the severity of the patient’s condition should the patient report any symptoms 
consistent with dextran-related anaphylaxis.      
 
Classification of Dextran Anaphylaxis by Symptom 
Type 
Grade/ 
Severity 
Skin 
Gastrointestinal 
system 
Respiratory 
system 
Cardiovascular 
system 
Anaphylactoid 
I 
Flush, 
erythema, 
urticaria 
-- 
-- 
-- 
Anaphylactoid/anaphylactic 
II 
Flush, 
erythema, 
urticaria 
Nausea 
Dyspnea 
Tachycaria, 
hypotension 
Anaphylactic 
III 
Flush, 
erythema, 
urticaria 
Vomiting, 
defecation 
Bronchospasm, 
cyanosis 
Shock 
Anaphylactic 
IV 
Flush, 
erythema, 
urticaria 
Vomiting, 
defecation 
Respiratory 
arrest 
Cardiac arrest 
 

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All patients enrolled in the EMPAKT CHF study will be hospitalized, and access to 
appropriate supportive therapy will be quickly administered should signs of an 
intolerance reaction be observed. Any adverse or severe adverse events will be 
reported as specified in the clinical protocol. 
6.1.1.3 Treatment of Overdose 
The FAST VFI is contained in single-dose vials, making significant overdose unlikely. 
Though the vials are overfilled with 4mL/62.6 mg in order to ensure that a full 
3mL/47mg dose can be drawn, previous studies have evaluated the safety of the VFI 
using a 3mL/150mg dose, over two times the amount of material contained in the 
single-dose vials for this study. There is no known antidote for overdose of VFI.  In 
the event of a suspected overdose, the patient should be monitored appropriately 
and should receive supportive therapy, as necessary. 
6.1.2 Treatment schedule 
 
 
 
 
 
 
 
 
 
Instructions for dose preparation: 
 
1. Add 4mL of sterile water to the vial of VFI over 4 seconds. 
2. Gently swirl the vial for 3 minutes, avoiding allowing the liquid to reach the rubber 
stopper. Product can be held up to light to observe the reconstitution process.  
3. Place vial on a roller/rocker table to mix for 30 minutes. 
4. Product is ready for drawing up of 3 mL dose into syringe. 
 
Instructions for application: 
Ensure the IV line is clear of and free flowing (can be done with a saline flush). 
Attached the VFI syringe and inject steadily within 30 seconds. Follow the dose with 
a 5mL flush of normal saline.  
6.1.3 Storage requirements and administration (distribution and return of study drug) 
The VFI will be stored in a refrigerator at 2 to 8°C (36 to 46°F).  
 
The VFI will be prepared within 60 minutes of dosing, and the prepared product will 
be stored at room temperature (25°C). The product will be protected from sunlight 
and ultraviolet light until dosing.  
6.1.4 Treatment compliance 
 
Training:  
A minimum of two hours of training with a FAST BioMedical employee is required to 
review and discuss the FAST PV and mGFR Technology. This includes training on 
the use of the FAST VFI and the risks involved in each step of performing the 
measurements. 
 
Day 1 
Day 3 
Dose / Units 
47mg/ 
3mL 
47mg/ 
3mL 
Pharmaceutical 
Form 
Liquid VFI 
Liquid 
VFI 
Route of 
Application 
IV Bolus 
IV Bolus 

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6.2 Placebo / reference medication 
There is no placebo/reference medication for this study as this is a one-armed 
study 
6.3 Concomitant medication / concomitant therapy 
Patients may continue/use medications/therapies approved and/or prescribed 
by their treating physician.  
6.4 Procedures in case of emergency 
The FAST VFI (FD004) was well tolerated in the nonclinical studies conducted 
to date.  Anaphylactic responses to dextrans, while rare, must be considered as 
a possibility for the high molecular weight dextrans being evaluated by FAST.  
As such, all testing facilities should have immediately available procedures to 
administer treatment for an anaphylactic response should it occur. We will 
ensure that each testing facility is prepared prior to the start of dosing.  
 
7 Study procedures 
All procedures specified in the study protocol should be performed for every 
single subject enrolled in the trial. In the case, trial participants miss a study visit 
every attempt should be made to regain contact by telephone, written 
communication or record review to determine if adverse events have occurred. 
 
7.1 Recruitment procedures and informed consent process 
The Principal Investigator, his or her proxy / designee and the member of the 
trial team will have the full responsibility to screen for suitable patients.  
 
The study group consists of nephrologists (KSO, JS, TS, KUE) and 
cardiologists (FE, BP). In our hospital (Charité - Universitätsmedizin) patients 
with acute decompensated heart failure (HF) are usually admitted to the 
cardiology departments or to the nephrology departments (if renal dysfunction is 
the leading cause for HF decompensation). Admission to the hospital may 
occur by direct referral from an outpatient physician or after initial evaluation 
and treatment at the emergency room. Since we plan to enroll patients with 
acute decompensated heart failure across a wide spectrum of renal function, 
we will enroll patients from both the cardiology and nephrology departments.  
 
To facilitate effective screening, a designated study physician will work with 
treating physicians at the the nephrology and the cardiology departments on a 
daily basis to identify patients with acute decompensated heart failure. All 
patients with a diagnosis of acute decompensated heart failure or "hydropic 
decompensation" 
(a 
frequently 
used 
clinical 
designation 
for 
acute 
decompensated heart failure in Germany) will be identified by the study 
physician and an initial discussion regarding study participation will take place 
between the study physician and the treating physicians at the departments of 
cardiology or nephrology. The study physician will then approach these patients 
and inform them about the study, ask for their participation and provide them 
with patient information material regarding the study. On the next morning, 
having allowed the patient a sufficient time period to reflect on study 

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participation, written informed consent will be obtained. This day will be 
designated "Day 1" of the study and patients will be evaluated and treated 
according to study protocol. 
 
 
A screening log for documentation of these patients will be used. All pre-
screened patients will be documented in the screening log and assigned a 
screening number.  
 
Patients who do not refuse the trial will be asked to participate and the written 
informed consent could be given by the patient. 
 
During the initial consultation, the following will be performed: 
 
1. Inform the patient about the study. Provide patient information in writing. If 
the patient does not a priori refuse participation, inform him/her about the 
further diagnostic procedures to check eligibility. Inform that he/she has 
still time to reconsider the matter and the opportunity to ask questions. 
Inform the patient that informed consent in writing must be signed before 
performing any of the following procedures during this visit. Obtain the 
patient’s informed consent in writing on two copies of the form. Sign both 
forms. Provide one copy to the patient. Store the other copy in the 
Investigator Site File. 
2. After getting the signed informed consent, the study related activities as 
outlined in this protocol will be performed. 
 
We expect to see approximately 250 patients during the recruitment time with a 
diagnosis of acute decompensated heart failure. As a consequence, the 
required number of 50 patients seems feasible. 
7.2 Methods of avoiding simultaneous enrolment in other trials 
During the process of getting an informed consent, it will be mentioned by the 
study physician / study investigator that there is no possibility of participating in 
other clinical trials at the same time. The patient will be informed about this 
issue in the interview with the investigator and this information is part of the 
informed consent form. 
7.3 Enrolment  
Patients, who meet all inclusion criteria and who have given their written informed consent 
will be enrolled in the trial.   
7.4 Clinical examinations and trial-related deviations from clinical practice 
 
Table 1: Schedule of Study Events/Examinations 
 
 
Study 
Timepoint 
Data 
collected 
from 
chart 
prior to 
enrolling  
Patient 
Enrolled 
and 
Dosed 
Data 
collected 
between 
1st and 
2nd Dose  
Patient 
2nd VFI 
Dose 
Data 
Collected 
after 2nd 
Dose 
Data 
Collected 
Before 
Discharge 
Follow up 
in person 
or by 
phone 
call 
Follow 
up 
phone 
call/ 
Follow 
up Visit 

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for 
Pregnan
cy test if 
WOCBP 
Visit 
Pre-
Dose 
Day 1 
Day 2 
Day 3 
Day 4 
Day 5 
Day 10 
Day 30 
Informed 
Consent 
X 
  
  
  
  
  
  
  
Inclusion/ 
Exclusion 
Criteria 
Assessment 
X 
  
  
  
  
  
  
  
Structural 
Heart Disease 
Criteria 
Assessment a 
  
X 
  
  
  
  
  
  
Demographics 
b 
  
X 
  
  
  
  
  
  
Comorbidities 
c 
  
X 
  
  
  
  
  
  
Full 
medication list 
n enrollment 
into the study  
and during 
follow-up  d 
  
X 
X 
X  
 X 
X  
  
  
Patient 
Symptom 
Assessment 
Survey e 
  
X 
X 
X 
X 
X 
 
  
Clinical 
Presentation/ 
Exam f 
  
X 
X 
X 
X 
X 
  
  
Treating 
Physician 
Survey g 
  
X 
  
X 
  
  
  
  
Study-Related 
Laboratory 
Measurements 
h 
  
X 
X 
X 
X 
X 
  
  
Routine 
Clinical 
Parameters i 
X* 
X 
X 
X 
X 
X 
  
  
Clinical Data j 
X* 
X 
X 
X 
X 
X 
  
  
Adverse 
events 
  
X 
X 
X 
X 
X 
X 
X 
Blood 
specimen for 
marker 
analysis k 
  
X 
  
X 
  
  
  
  
Discharge 
from study 
  
  
  
  
  
  
  
X   
Adjudication 
Committee 
Evaluationl 
 
 
 
 
 
 
 
X 
WOCBP 
Pregnancy 
X 
 
 
X 
 
 
 
X 

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test m 
a See Table 2 for Structural Heart Disease Criteria 
b See Table 3 for specific demographic parameters 
c See Table 4 for specific instructions on collecting comorbidities 
d See Table 5 for specific instructions on collecting medications 
e See Annex D for the Patients Symptom Assessment Survey 
f See Table 6 for Clinical presentation/exam parameters 
g See Annex E for the Treating Physician Survey 
h See Table 7 for Study Related Laboratory Measurements 
i See Table 8 for Routine Clinical Parameters 
j See Table 9 for Clinical Data collected 
k Blood Specimens for the determination of the FAST PV and mGFR will be taken pre-dose and at 15 (±5), 30 (±5), 60 (+ 
30) and 180 (+ 120) minutes post dose on Day 1 and Day 3. Note that the 30 minute blood draw must be at least 10 
minutes after the 15 minute blood draw, and the 180 minute blood draw must be at least 100 minutes after the 60 minute 
blood draw. 
l See Table 10 for the Adjudication Committee Evaluation Questions; Adjudication Committee meets after all collected 
data available. 
m See Table 11 for Details of Pregnancy test  
 
*will be collected retrospectively after study inclusion 
 
7.4.1 Pre-trial examinations (screening / inclusion examination) 
The following screening examinations will be performed: 
x 
Medical history and Comorbidities (Table 4) 
x 
Clinical examination (Table 6) Including vital signs, renal function assessment 
x 
Concominent Medications (Table 5) 
x 
Demographics (Table 3) 
x 
Serum Laboratory testing (Table 7 and Table 8) 
7.4.2 Examinations during trial 
The following examinations will be performed: 
Table6:Clinicalpresentation/ExamParameters
*Height(cm)
Weight(kg)
*Paroxysmalnocturnaldyspneaduringprevious3days?
*Nightcough
*Cardiomegaly(clinicaloronchestXͲrayorechocardiography)
*S3gallop
*Hepatojugularreflux
*Hepatomegaly
*Pleuraleffusion
*Laststablebodyweightbeforeadmission("bestbodyweight")(kg)
NYHAstage(0Ͳ4)
Ankle/lowerextremityedema(grade0Ͳ4;accordingtoGuelphGeneralHospitalCongestive
HeartFailurePathway)
Pulmonaryrales
Systolicbloodpressure(mmHg)(morning,supine)
Diastolicbloodpressure(mmHg)(morning,supine)

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**Systolicbloodpressure(mmHg)(morning,30secafterstandingup)
**Diastolicbloodpressure(mmHg)(morning,30secafterstandingup)
Heartrate(beatsperminute)(morning,supine)
Heartrate(beatsperminute)(morning,30secafterstandingup)
SpO2(%)(morning)
Jugularvenousdistension(>4cmabovesternalanglewith45°upperbodyelevation)
(morning)
Respiratoryrate(breathsperminute)(morning)
O2supplyrequirement(inL/min)(morning)
*OnlyassessedDay1
*OnlyassessedDay1andDay3
 
 
All examinations during the trial are listed in Tables 1-9, in section 4.5. 
7.4.3 Final examination 
x On the final follow-up visit (phone call) any adverse events will be recorded.  
7.4.4 Laboratory assessments 
Table7:StudyͲrelatedlaboratorymeasurements
Serumcreatinine(mg/dl)
*Hematocrit(%)
**creatinephosphokinase(CPK)(U/l)
**NTͲproBNP(ng/l)
**Alanineaminotransferase(ALT)(U/l)
**Aspartateaminotransferase(AST)(U/l)
**Alkalinephosphatase(AP)(U/l)
**bilirubin,totalanddirect(mg/dl)
**INR(dimensionless)
BiobankingofEDTAPlasma,HeparinͲPlasma,Serum,
wholeblood,centrifugedurine(10aliquotsa200µl
each)
*OnlyCollectedDay1andDay3,foruseinthecalculationof
ePCbyNadler’sformula
**OnlyCollectedDay1,3and5
 
Table8:Routineclinicalparameters
*Serumcreatinineonadmission(mg/dl)
*Baselineserumcreatinine(nadirfrominspectionofprevious
labvaluesorbyprimarycarephysician)(mg/dl)
Serumcreatinine(mg/dl)
Serumsodium(mmol/l)
Serumpotassium(mmol/l)
eGFR(CKDͲEPI)(ml/min/1,73m2)
Hemoglobin(g/dl)

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SerumͲAlbumin(g/dl)
Urineoutput(0:00Ͳ24:00ontheindicatedday)(ml/24h)(if
incomplete,extrapolatetheavailabletimeperiodto24hours)
ChestXͲray:evidenceofpulmonarycongestion
ChestXͲray:evidenceofpleuraleffusion
ChestXͲray:evidenceofcardiomegaly
*OnlyonDay1
 
Table11:PregnancyTestsinWOCBP
Medicallymonitoredpregnancytestswitha
minimumsensitivity25mU/ml
willbeassessedwithin24hourspriorfirststudy
drugadministration,onDay3and30inWOCBP
 
All examinations during the trial are listed in Tables 1-9, in section 4.5. 
 
7.5 Individual trial duration  
x From enrolment in the study until discharge is 30 ± 5 days.  
 
8 Risk-benefit-assessment 
A rapid and accurate measurement of plasma volume (PV) and glomerular filtration 
rate (GFR) is important in chronic heart failure (CHF), acute kidney injury (AKI) and 
chronic kidney disease (CKD) for assessment of impairment, diagnosis, and prompt 
treatment. This study primarily serves the purpose of obtaining information about the 
tolerability and safety of the FAST PV and mGFR Technology in CHF patients. The 
FAST Technology has been previously used in three clinical studies. 87 patients 
have been dosed with the FAST VFI. There have been no reported SAEs and few 
minor AEs across all studies (please refr to the Investigator´s Broschure, Version 6.0, 
dated July 2018 (FAST-IDE-IB_51378), section 4 and 5).  
 
Based on the results of prior clinical and non-clinical assessments of the FAST PV 
and mGFR Technology the risks in participating in the trial are considered 
acceptable. However, they include the usual risks of participating in clinical trials, 
which are related to possible allergic reactions and blood drawing via venipuncture. 
There are no perceived direct benefits to patients participating in this trial. However, 
thorough medical check-ups may be seen as an advantage. The safety of the 
patients will be observed during all study phases. 
 
Medical progress is based on research which ultimately must rest in part on 
experimentation involving humans. Eligible patients may consider participation in this 
clinical trial because they want to contribute to the advancement of medical 
knowledge. Still, considerations related to the well-being of the individual patients 
enrolled into this clinical study must take precedence over the interests of science 
and society. Based on available information and the design of the study, the sponsor 
and the investigator consider the trial to be ethically acceptable. The duration of 
hospitalization and the medical surveillance are considered adequate to ensure 
safety of the patients. 

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9 Termination and subsequent treatment 
9.1 Premature termination of the individual participant 
Patients are free to discontinue the study at any time, for any reason, and without 
prejudice to further treatment. The investigator may remove a patient if, in the 
investigator’s judgment, continued participation would pose unacceptable risk to the 
patient or to the integrity of the study data. All procedures for early termination must 
be completed. Reasons for removal or withdrawal may include: 
x Admission to the ICU prior to either the first or second dose 
x The patient develops a stated exclusion criterion during the course of study 
x If the patient experiences an acute anaphylactoid reaction during the course of 
administering the FAST VFI, administration will stop immediately 
x Evidence of an anaphylactoid reaction (dyspnea, urticaria, flush, bronchospasm, 
or severe systemic hypotension)   
x Administrative decision by the investigator or sponsor 
x Significant protocol deviation 
x Patient noncompliance 
x Safety concern by the investigator or sponsor 
x Lost to follow-up 
x Pregnancy 
Patients who are withdrawn for reasons other than safety issues may be replaced at 
the discretion of the sponsor and investigator. 
In the event of a patient’s withdrawal, the investigator will promptly notify the medical 
monitor and will make every effort to complete the EOS assessments. All withdrawn 
patients with ongoing clinically significant clinical or laboratory findings will be 
followed until the finding is resolved or medically stable; reasonable attempts will be 
made to follow-up with patients.  
9.2 Premature termination of the clinical study 
The study will be discontinued if the Sponsor or its designee judges it necessary for medical, 
safety, regulatory, or other reasons consistent with applicable laws, regulations, and GCP 
guidelines. 
The following events, cause premature termination of the clinical study: 
x 
Unjustifiable risk and toxicity in risk-benefit analysis (decision taken by principal 
Investigator)  
x 
New scientific evidence provided during the study that could affect the patient’s safety 
(benefit-risk analysis no longer positive)  
x 
Decision of the Sponsor or the Competent Authority that the study should be 
discontinued.  
 
9.3 Follow-up and continuing treatment after regular / premature termination 
After completing all the protocol treatment and visits, patients will continue with 
regular visits according to usual practice of the center. 
 
In the case of premature termination, the reason for withdrawal must be entered on 
the appropriate case report form (CRF) page and must be followed for safety and 
efficacy until 5 days after discontinuation.  

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10 Safety monitoring 
10.1 Adverse Event (AE) 
An adverse event is any untoward medical occurrence in a patient or clinical 
investigation subject administered a pharmaceutical product and which does not 
necessarily have to have a causal relationship with this treatment. 
 
An AE can therefore be any unfavorable and unintended sign [(i.e including an 
abnormal laboratory findings, symptom or disease) in a subject or clinical 
investigation subject after providing written informed consent for participation in the 
study. Therefore, an AE may or may not be temporally or causally associated with 
the use of a medicinal (investigational) product. 
 
An untoward medical occurrence may be a study endpoint as well as meeting the 
definition for an AE.  
 
The occurrence of AEs should be sought by non-directive questioning of the patient 
at each visit throughout the study. AEs also may be detected when they are 
volunteered by the patient during or between visits or through physical examination, 
laboratory test, or other assessments. 
 
Abnormal laboratory values or test results constitute AEs if they are in line with the 
definition above (ICH E2A, section 1) and fulfill at least one of the following criteria: 
• they induce clinical signs or symptoms, 
• they are considered clinically significant, 
• they require therapy. 
Clinically significant abnormal laboratory values or test results should be identified 
through a review of values outside of normal ranges/clinically notable ranges, 
significant changes from baseline or the previous visit, or values which are 
considered to be non-typical in patient with underlying disease. Investigators have 
the responsibility for managing the safety of individual patient and identifying AEs. 
 
10.2 Serious Adverse Event (SAE) 
An SAE is any AE (appearance of (or worsening of any pre-existing) undesirable 
sign(s), symptom(s) or medical conditions(s) which meets any one of the following 
criteria: 
x is fatal or life-threatening 
x results in persistent or significant disability/incapacity 
x constitutes a congenital anomaly/birth defect 
x requires inpatient hospitalization or prolongation of existing hospitalization, 
unless hospitalization is for: 
o routine treatment or monitoring of the studied indication, not associated 
with any deterioration in condition 
o elective or pre-planned treatment for a pre-existing condition that is 
unrelated to the indication under study and has not worsened since 
signing the informed consent 

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o social reasons and respite care in the absence of any deterioration in 
the patient’s general condition 
x is medically significant, i.e. defined as an event that jeopardizes the patient or 
may require medical or surgical intervention to prevent one of the outcomes 
listed above. 
10.3 Adverse Reactions 
Adverse reactions are all untoward and unintended responses to an investigational medicinal 
product related to any dose administered. 
10.4 Suspected Unexpected Serious Adverse Reactions (SUSAR) 
A Suspected Unexpected Serious Adverse Reaction (SUSAR) is any suspected adverse 
reaction related to the study treatment that is both serious and unexpected.  
“Unexpected” means that the nature and severity of the adverse reaction are not consistent 
with the information about the study medication in question set out in the reference safety 
information (investigator’s brochure). 
 
10.5 Evaluation of Expectedness 
 
The evaluation of expectedness will be done by the pharmacovigilance and the 
sponsor representative. The expected (S)AEs could be found in the investigator’s 
brochure (section 4). The investigator’s brochure contains also a section “reference 
safety information” (RSI) (section 5.6). 
10.6 Treatment of (S)AEs 
All AEs should be treated appropriately. Treatment may include one or more of the 
following: 
x no action taken (i.e. further observation only)  
x concomitant medication given  
x non-drug therapy given.  
The action taken to treat the AE should be recorded on the AE CRF. 
10.7 Assessment of SAEs 
As far as possible, each AE should be evaluated to determine: 
x the severity grade (grade 1 to 5 according CTCAE v5.0) 
x its relationship to the study drug (assessment of causality) 
x its duration (start and end dates or if continuing at final exam) 
x action taken (no action taken; what kind of action taken, e.g. hospitalization) 
x whether or not it constitutes a serious adverse event (SAE). 
10.8 Severity grade  (according CTCAE v5.0) 
x Grade 1: Mild: The Adverse Event is transient and can be tolerated easily 
x Grade 2: Moderate: The Adverse Event causes discomfort and impedes 
normal activities  
x Grade 3: Severe: The Adverse Event causes severe impairment of normal 
activities  
x Grade 4: Life-threatening or disabling 

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x Grade 5: Death related to AE. 
10.8.1 Assessment of causality 
To assess causality between administration of the investigational product and the 
Adverse Event the following definitions apply:  
 
x Sure: The reaction comprehensively follows the administration of the 
investigational product in the right timeframe or can be measured in body 
tissues or fluids or represents a known or expected response to the study 
medication or disappears after discontinuation or dose reduction and reoccurs 
after re-exposure. 
 
x Probable: The reaction comprehensively follows the application of the 
investigational product in the right timeframe or represents a known or 
expected response to the study medication or disappears after discontinuation 
or dose reduction and cannot be explained by known characteristics of the 
patient’s disease. 
 
x Possible: The reaction comprehensively follows the application of the 
investigational product in the right timeframe or represents a known or 
expected response to the study medication, but could easily be caused by 
other factors.  
 
x Unlikely: Event or laboratory test abnormality, with a time to drug intake that 
makes a relationship improbable (but not impossible); disease or other drugs 
provide plausible explanations 
 
x Not related: Adequate Information supporting the assumption that there is no 
causality  
 
x Cannot be evaluated: The causality cannot be determined. 
10.9 Documentation of AEs and SAEs 
All Serious Adverse Events (SAEs) and all Adverse Events (AEs) need to be 
documented in the patients’ clinical file and in the respective CRF pages by the 
investigator, no matter, whether the investigator suspects a causal relationship to the 
investigational product or not. The documentation needs to include:  
x Duration (start and end dates) or if the event is ongoing an outcome of not 
recovered/not resolved should be reported 
x Action taken regarding study treatment 
x Whether other medication or therapies have been taken (concomitant 
medication/non-drug therapy) 
x Its 
outcome 
(not 
recovered/not 
resolved; 
recovered/resolved; 
recovering/resolving, recovered/resolved with sequelae; fatal; or unknown).
Related signs symptoms and laboratory changes should be summarized to a specific 
disease. The event will be recorded in the CRF. SAEs need to be documented on a separate 
SAE form. 
Out of normal range laboratory data need to be analysed concerning their clinical relevance 
by the Investigator – and if relevant documented as an AE itself. 

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All adverse events need to be followed until they subside or stabilize.   
The Sponsor will carefully document all SAEs reported by the Investigator. His 
documentation will be sent to the relevant regulatory authorities and to relevant authorities of 
other European member states and other contracting states of the EWR agreement, if the 
study is run in their territory and if they so request. 
10.10 Follow-up of Adverse Events 
Once an AE is detected, it should be followed until its resolution or until it is judged to be 
permanent, and assessment should be made at each visit (or more frequently, if necessary) 
of any changes in severity, the suspected relationship to the study drug, the interventions 
required to treat it, and the outcome. 
The Sponsor may request additional information on specific AEs of interest and may make 
requests to perform additional diagnostic tests to further assess the safety profile of the study 
drugs. Such information may include diagnostic procedure reports, discharge summaries, 
autopsy reports, and other relevant information that may help in assessing the reported AE. 
All additional information will be de-identified prior to collection by the Sponsor or its partners. 
The investigator should also instruct each patient to report any new AE (beyond the protocol 
observation period) that the patient, or the patient’s personal physician, believes might 
reasonably be related to study drug. This information should be recorded in the investigator’s 
source documents; however, if the AE meets the criteria of an SAE, it must be reported to 
the Sponsor. 
10.11 Reporting of SAEs  
The Investigator will report any Serious Adverse Event within 24 hours after becoming aware 
to the Sponsor and will afterwards send an extended written record 
 
This announcement will be done via fax or email to:  
 
KKS Charité 
Fax: +49 (0)30 – 450 7553-856 
or email to 
pharmacovigilance-kks@charite.de 
 
 
Every event will be documented on a record form and will immediately be sent to the above 
given address. If at that point all required information is not available, succeeding records will 
be sent.  In the event of death a copy of the autopsy record could be added.  
 
Exceptional rules: 
In this clinical trial the following SAEs are excluded from the reporting requirement:  
x 
Serious or unexpected events which occur after enrolment, but before treatment was 
initiated  
x 
Hospitalization or extension of hospitalization required for therapeutic procedures of 
the protocol    
x 
Other events: For example events that cause hospitalization but were planned before 
enrolment (e.g. (planned surgery)  
10.12 Reporting of Suspected Unexpected Serious Adverse Reactions 
(SUSARs) 
The Sponsor will report all suspicious cases of Suspected Unexpected Serious Adverse 
Reactions (SUSARs) which had been occurred in one of clinical trials conducted by the same 

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sponsor with the same drug substance/IMP to the relevant Ethics Committee, the relevant 
regulatory authorities and  to relevant regulatory authorities of other European member 
states and other contracting states of the EWR agreement, if the study is run in their territory 
immediately, at the latest 15 days after it becomes known. He will also inform all 
Investigators involved in the trial.  
In case of a fatal or life threatening SUSAR the Sponsor will report all information relevant for 
judging the event immediately, at the latest 7 days after the event becomes known to the 
relevant Ethics Committee, the relevant regulatory authorities and to relevant regulatory 
authorities of other European member states and other contracting states of the EWR 
agreement, if the study is run in their territory as well as to all Investigators involved in the 
trial. After a further 8 days all further relevant information must be available.  
 
10.13 Other safety issues requiring expedited reporting 
The Sponsor will immediately, at the latest 15 days after it becomes known report all 
circumstances that require a revision of the risk-benefit analysis to the relevant Ethics 
Committee, the relevant regulatory authorities and  to relevant regulatory authorities of other 
European member states and other contracting states of the EWR agreement, if the study is 
run in their territory. This especially includes:   
x 
Singular cases of expected severe adverse events with an unexpected outcome.  
x 
Increased incidence of expected severe adverse events that are judged as being 
clinically relevant.  
x 
SUSARs which occur after termination of the clinical trial Events related to study 
procedures or development of the study medication, which could affect a 
patient’s safety.  
All person-related data will always be transmitted pseudonymised.  
10.14 Development Safety Update Report 
The sponsor submits a list of all SUSAR/SARs yearly or on request, together with an 
extensive safety report on the investigational products to the respective competent 
authorities and the concerned Ethics Committees. 
11 Documentation 
11.1 Case Report Forms (CRF) 
Data collected on each patient will be recorded on an eCRF (electronically). This trial will be 
performed using an electronic case report form (eCRF) (SecuTrial). The investigator 
and the trial site staff will receive system documentation, training and support for the 
use of the eCRF. In case of new trial site staff the training can be performed by 
experienced personnel of the respective trial site. 
 
All protocol-required information collected during the trial must be documented in the 
eCRF by the investigator, or a designated representative. All data entry, modification 
or deletion will be recorded automatically in an electronic audit trail indicating the 
individual subject, the original value, the new value, the reason for change, who 
made the change and the time and date of the change. All data changes will be 
clearly indicated. Former values can be viewed in the audit trail. All electronic data 
will be entered by the site (including an electronic audit trail) in compliance with 
applicable record retention regulations in a web- based data capturing system known 
as secuTrial. 
 

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Investigators or their designee must enter the information required by the protocol 
onto the electronic case report forms (eCRF). Data items from the eCRF are entered 
into the study database.  
f 
The investigator must complete the eCRF and must print the patient identification list 
and store it with other study documents, e.g. the protocol, the investigators’ brochure 
and any protocol amendments, in a secure place. All entries to the eCRF must be 
made as described in the electronic case report form manual. 
 
Data on subjects collected on eCRF during the trial will be documented in a 
pseudonymous fashion; the subject will only be identified by the pseudonym. The 
investigator must maintain source documents for each patient in the study, consisting 
of 
all 
demographic 
and 
medical 
information, 
including 
laboratory 
data, 
electrocardiograms, etc., and keep the signed informed consent form. All information 
on eCRF must be traceable to these source documents in the patient's file. Data 
without a written or electronic record will be defined before trial start and will be 
recorded directly on the eCRF, which will be documented as being the source data. 
The definition can be found in the investigator site file. 
 
Essential documents, as listed below, must be retained by the investigator for as long 
as needed to comply with national and international regulations. The investigator 
agrees to adhere to the document retention procedures by signing the protocol. 
Essential documents include: 
1. IRB/IEC approvals for the study protocol and all amendments 
2. All source documents and laboratory records 
3. CRF copies (paper copies or electronic copies on a CD-ROM) 
4. Patients' informed consent forms (with study number and title of trial) 
5. Any other pertinent study document. 
A clinical study monitor will review the eCRF and compare the content to the source 
data. In addition, the investigator or designated colleague will provide access to 
designated sponsor representative(s) for the periodic review of source documents 
(e.g. hospital and clinic records) to assure accuracy and completeness of the eCRF.  
 
All clinical work conducted under this protocol is subject to GCP rules. This includes 
an inspection by the sponsor and/or health authority representatives at any time. The 
investigator will agree to the inspection of study-related records by health authority 
representatives and/or the sponsor. 
11.2 Investigator Site File (ISF) 
All essential documents will be kept in the Investigator Site File which will be stored at the 
study site in accordance with ICH GCP chapter 8. The sponsor will provide the investigator 
with an investigator's file. This file should be used for all trial-related documents. The 
investigator will be responsible for keeping the investigator's file updated and ensuring that 
all required documents, as specified in ICH-GCP guidelines, are filed. The file will be made 
available for monitoring and/or auditing by the sponsor or its representative and regulatory 
agencies. 
11.3 Drug accountability / flow of study medication 
All drug accountability records must be kept current, and the Investigator must be 
able to account for all used and unused vials of study medication. These records 
should contain the dates, quantity, and study medication: 

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• Received at site 
• Administered to each subject, 
• Disposed of at the site or returned to the Sponsor or designee 
The clinical monitor responsible for the study site will provide written approval for the 
destruction or return of unused study medication following reconciliation of all clinical 
supplies. 
 
12 Quality management 
12.1 Control of trial progress and data quality 
12.1.1 Monitoring 
Monitoring as the act of overseeing the conduct and progress of clinical trials needs 
to be performed by consistent standards and qualified personnel. Therefore the 
coordination and realization of the monitoring will be performed by the Coordinating 
Center for Clinical Studies (KKS Charité) Berlin.  
 
The monitoring personnel will visit the clinical study sites on a regular basis and at 
least once before the first subject has been enrolled, during the course of the study, 
and at study completion. The monitor is responsible for the implementation of the 
study procedures at the sites.  
 
The objectives of the monitoring procedures are to ensure that the trial subject’s 
safety and rights as a study participant are respected, that accurate, valid and 
complete data are collected, and that the trial is conducted in accordance with the 
trial protocol, the principles of ICH-GCP and local legislation. 
 
Study data will be verified in accordance to the original medical records (Source Data 
Verification). The sponsor/investigators allow the monitor to have access to any or all 
the study materials needed for source data verification and proper review of the study 
progress. At all times, the sponsor/investigators/monitors will maintain the 
confidentiality of the study documents. Furthermore, problems with inconsistent and 
incomplete data will be discussed. 
 
All investigators agree that the monitor regularly visits the trial site and assure that 
the monitor will receive appropriate support in his activities at the trial site. The 
declaration of informed consent includes a statement to the effect that the monitor 
has the right – while observing the provisions of data protection legislation – to 
compare the case report forms (p/eCRFs) with the trial subject’s medical records 
(doctor’s notes, ECGs, laboratory printouts etc.). The investigator will secure access 
for the monitor to all necessary documentation for trial-related monitoring. The role 
allocation in secuTrial provides the monitor with access possibilities to all data 
captured electronically. Furthermore the investigators will prepare the consent forms 
as PDF files and submit them to the Trust agency, which then verifies the data and 
drafts regular reports. 
 

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The aims of the monitoring visits are as follows: 
• To check the declarations of informed consent. 
• To monitor trial subject safety (occurrence and documentation/reporting of AEs 
and SAEs). 
• To check the completeness and accuracy of entries on the p/eCRF. 
• To validate the entries on the p/eCRF against those in the source documents 
(source data verification, SDV). 
• To evaluate the progress of the trial. 
• To evaluate compliance with the trial protocol. 
• To assess whether the trial is being performed according to GCP at the trial site. 
• To discuss with the investigator aspects of trial conduct and any deficiencies 
found. 
A monitoring visit report is prepared for each visit describing the progress of the 
clinical trial and any problems (e.g. refusal to give access to documentation). 
By signing the declaration of informed consent the participants allow access to their 
documents. With the signature in the protocol, the investigators confirm that auditors 
and health authority inspectors may have access to the study documentation and 
accordant medical records. Auditors and inspectors are bound by professional 
confidentiality and may not pass on any personal information that comes to their 
knowledge. In the course of audits or inspections, data in the case report forms will 
be compared with the data from medical records. All the documentation held by the 
investigators within the scope of the clinical trial, as well as the drug logs of the study 
medications will be verified.
12.1.2 Data Quality Assurance 
The sponsor assumes responsibility for implementing and maintaining quality 
assurance and quality control systems with written Standard Operation Procedures 
(SOPs) to ensure that the study is conducted and data are generated, documented 
and reported in compliance with the protocol, Good Clinical Practice, and the 
applicable regulatory requirements. The sponsor also takes responsibility for 
securing agreement from all involved parties to ensure direct access to the trial 
related sites, source data/documents, and reports for the purpose of monitoring and 
auditing by the sponsor and inspection by authorities. Quality control will be applied 
to each stage of the study to ensure that all data are reliable and have been 
processed correctly. 
12.1.3 Audits / inspections 
Authorized representatives of the Sponsor, a regulatory authority, or an Independent 
Ethics Committee (IEC) may visit the center to perform audits or inspections, 
including source data verification. The purpose of a Sponsor audit or inspection is to 
systematically and independently examine all study related activities and documents 
to determine whether these activities were conducted, and data were recorded, 
analyzed, and accurately reported according to the protocol, Good Clinical Practice 
(GCP), guidelines of the International Conference on Harmonization (ICH), and any 
applicable regulatory requirements. 

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12.2 Reference institutions 
1. The KKS will conduct the regulatory affairs, data quality management, 
monitoring, SAE- Management. 
Coordinating Center for Clinical Studies Berlin- Koordinierungszentrum für 
Klinische Studien (KKS), Campus Virchow-Klinikum, Augustenburger Platz 
1,13353 Berlin, Germany.  
 
Hr. Dr. rer. medic. Olaf Bender  
Charité - Universitätsmedizin Berlin 
Campus Virchow-Klinikum 
Coordinating Center for Clinical Studies- Koordinierungszentrum Klinische 
Studien (KKS) 
Reinickendorfer Str. 61, Haus 10 
Augustenburger Platz 1, 13353 Berlin, Germany  
Telephone: +49 30 450 553 864 
Email: olaf.bender@charite.de 
 
2. The Institute of Biometry and Clinical Epidemiology, Charité Campus Mitte is 
responsible for the statistical planning. Furthermore it will conduct statistical 
data analyses and biometric reporting. 
 
Prof. Dr. Geraldine Rauch 
Charité Universitätsmedizin Berlin 
Institute of Biometry and Clinical Epidemiology 
Charitéplat 1, 10117 Belrin 
13 Data entry and data management 
 
All patient related data will be recorded under a pseudonym. Every patient will receive a 
patient number / pseudonym which will be unique for this individual patient. The Investigator 
will compile a confidential list, which relates these patient numbers to the patient’s full name. 
This list will only be accessible to the study team and the monitor. Original patient files may 
be viewed by monitors, auditors and inspectors. 
13.1 electronic Case Report Form (eCRF) Data Entry 
 
Investigators or their designee must enter the information required by the protocol 
onto the electronic case report forms (eCRF). Data items from the CRF are entered 
into the study database.  
 
The investigator must complete the eCRF and must print the patient identification list 
and store it with other study documents, e.g. the protocol, the investigators’ brochure 
and any protocol amendments, in a secure place. All entries to the eCRF must be 
made as described in the electronic case report form manual. 
 
Please see section 11.1 of this protocol for information regarding the eCRF and 
data entry.  

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13.2 Data management 
Data required for the analysis will be acquired and transferred electronically to a 
central database at the KKS Charité by means of an electronic data capture system 
(EDC). 
 
The secuTrial® Software solution of interActive Systems (iAS) is a worldwide 
reachable web-based system (remote data entry [RDE]). The system operates 
according to the principle of online data capture and is in compliance with the FDA 21 
CFR Part 11 and ICH-GCP. In addition, it contains further functions to perform 
plausibility, consistency and range checks of the study data. 
 
The log-in in the system requires a previous authentication for access, and runs by a 
secure data transfer-protocol (SSL) to exclude any improper manipulation of data by 
unauthorized parties. The system will automatically keep an audit trail of all entries 
and corrections to the eCRF. 
 
The secuTrial® Software solution is based on an Oracle database that does not allow 
direct access. All access rights of trial participants will be defined according to their 
trial functions. The defined access rights will e.g. allow to read or to enter study data 
depending on their functions (Monitor, clinical investigator, etc.). 
 
Query management is an integrated part of this RDE system. This function allows the 
communication between the Monitor and the trial participants. In case of errors or 
improper data, the responsible trial participant will get queries (messages) to correct 
the errors or to check improper data on the eCRF. 
 
The trial data will be stored digitally on a remote server with daily backups. After end 
of trial all data will be exported for checks of consistency and plausibility. After 
performing the checks, the data matrix will be transferred for the statistical evaluation. 
13.3 Source Data and Patient Files 
The information in original documents and records (e. g. patient files, laboratory 
notes) are defined as Source Data and will be reviewed by the Monitor for Source 
Data Verification. All data that will be recorded directly in the CRF/eCRF without prior 
written or electronic record will be described in the protocol and considered to be 
Source Data. 
14 Statistical Analysis 
The primary objective is to assess the safety and function of the FAST PV and mGFR 
Technology in hospitalized patients with heart failure. This is a phase II trial. No 
formal hypothesis test is performed to assess the primary objective. This is justified 
as follows: 1. Both function and safety cannot be uniquely assessed by one specific 
endpoint, but there exist several relevant endpoints and endpoint-related criteria 
which must be investigated in order to assess the global function and safety. 2. For 
reasons of feasibility, the studies sample size is limited to 50 patients which is an 
adequate number in the context of a phase II trial. However, this limited number does 
not allow to address a complex multiple testing problem with adequate power. 
Therefore, all analyses are purely descriptive. The aim is to generate new 
hypotheses to be tested in subsequent phase III trials. 
 

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All analyses will be performed using validated statistical software. A statistical 
analysis plan included detailed instructions for the analysis will be finalized before 
start of any analysis. 
 
Size Estimation 
We plan to enrol 50 eligible patients. Due to the explorative nature of this phase II 
study a formal sample size calculation is not possible. However, the sample size of 
50 patients is sufficient to guarantee reasonable accuracy with respect to the main 
objectives of the trial as specified below. 
 
Primary objectives: 
 
Global aim is to assess the safety and function of the FAST PV and mGFR 
Technology in hospitalized patients with heart failure. 
 
x Safety will be assessed by determination of the absolute and relative frequencies 
of AEs and SAEs related to VFI. 
 
x Function of the FAST PV measurement will be assessed by determining the 
plasma stability of the FD003 high molecular weight marker over the 15, 30, and 
60 minute blood draws and applying the following criteria: 
- 
The FAST PV measurement is considered as stable, if the mean plasma 
concentration of FD003 at 30 minutes is not more than 10% lower than the 
mean plasma concentration at 15 minutes AND if the mean plasma 
concentration at 60 minutes is not more than 10% lower than the mean 
plasma concentration at 30 minutes 
- 
We will determine the percentage of patients which show a decline in the 
plasma concentration of FD003 of more than 10% from 15min to 30min 
and separately from 30 min to 60min. This percentage should be ideally 
close to 0. 
 
Secondary objectives: 
 
x To evaluate how estimated PV (ePV) and estimated total blood volume (TBV) 
assessments on days 1 and 3 (by established measures such as Nadler’s formula 
or Metropolitan Life Tables) predict measured PV (mPV; as assessed by FAST 
methodology) and measured TBV (mTBV; calculated from mPV and measured 
hematocrit) at these time points. Simple linear regression models will be 
generated and potentially adjusted for relevant confounders. 
 
x To evaluate how a clinical evaluation (including a reasonable subset of the 
variables age, gender, BMI, hematocrit, edema grade, presence of pulmonary 
rales, presence of jugular venous congestion, arterial blood pressure, NYHA 
stage, respiratory rate) on days 1 and 3 predicts mPV and mTBV at these time 
points. Optimal linear models will be chosen by comparing the results of different 
variable selection models for the corresponding linear model (backward selection 
based on p-Values and Akaike Information Criterion, best subset selection etc.).  
 
x To evaluate whether eGFR calculation by the CKD-EPI formula on days 1 and 3 
provides an accurate estimate of measured GFR (mGFR; by assessed by FAST 

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methodology) in heart failure patients undergoing active fluid management. 
Simple linear regression models will be generated and potentially adjusted for 
relevant confounders. 
 
x To evaluate whether patients with a low mGFR/eGFR ratio on days 1 and 3 are at 
higher risk of developing AKI within the following 48-72 hours. Logistic regression 
models will be generated and potentially adjusted for other relevant confounders.  
 
x To evaluate whether ranges of mPV (low mPV, normal mPV, high mPV) and/or 
mTBV (low mTBV, normal mTBV, high mTBV) on days 1 and 3 are associated 
with subjectively reported symptoms (PGA, dyspnea, dizziness, nausea) 
determined using patient symptom assessment sheets within the following 24-48 
hours. Logistic regression models will be generated and potentially adjusted for 
other relevant confounders. Associations will be assessed by chi-square 
interaction tests and by Spearman's correlation coefficient. 
 
x To evaluate whether patients with low mPV or low mTBV (cutoffs potentially 
adjusted for some of the following variables age, gender, body height, body 
weight, optimal body weight (dry weight), hematocrit) on day 1 or on day 3 are at 
risk of developing low-output complications within the next 24-48 hours (e. g. 
subjectively reported dizziness/nausea, PGA, hypotension, need for iv-fluid 
therapy). Optimal logistic regression models will be selected by comparing the 
results of different variable selection models for the corresponding linear model 
(backward selection based on p-Values and Akaike Information Criterion, best 
subset selection, etc.). 
 
x To evaluate whether patients with a high mPV or a high mTBV (cutoffs potentially 
adjusted for some of the following variables age, gender, body height, body 
weight, optimal body weight (dry weight), hematocrit) on day 1 and/or day 3 are at 
risk of being refractory to diuretic therapy (e. g. require dosage increases of 
furosemide/torasemide, require ultrafiltration/RRT within the next 24-48 hours, fail 
to improve subjectively in PGA and dyspnea scales). Optimal logistic regression 
models will be selected by comparing the results of different variable selection 
models for the corresponding linear model (backward selection based on p-
Values and Akaike Information Criterion, best subset selection, etc.). 
 
x To evaluate whether the time course of ePV and estimated TBV (eTBV) from day 
1 to day 3 adequately reflects the time course of mPV and mTBV at these time 
points. Mean ePV and mPV and mean eTBV and mTBV will be compared for 
each time point separately (using paired t-tests) and by investigating the whole 
time curve (comparing the times when maximal and minimal volumes are reached 
and the areas under the curve)). 
 
x To evaluate whether the change of mPV and mTBV from day 1 to day 3 is 
predictive of length of stay, dialysis/ultrafiltration requirement, or rehospitalization 
within 30 days, or mortality. Linear, logistic or Cox-regression models will be 
utilized as appropriate.  
 
x To evaluate whether the time course of mGFR from day 1 to day 3 correlates 
temporally with or predicts the time course of eGFR within the next 24-48 hours. 

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eGFR and mGFR will be compared for each time point separately (using paired t-
tests) and by investigating the whole time curve (comparing the times when 
maximal and minimal GFRs are reached and the areas under the curve). 
 
x To evaluate (by an adjudication committee) whether clinical decision making 
would have been affected by adding FAST GFR and PV measurements to clinical 
routine including the following questions:  
o Would FAST GFR and PV measurements have changed clinical 
management overall? 
o Would the FAST GFR and PV measurement on day 1 have led to applying 
a different diuretics dosage between day 1-3? 
o Would the FAST GFR and PV measurement on day 3 have led to applying 
a different diuretics dosage between day 3-5? 
o Would the FAST GFR and PV measurement on day 1 have led to choosing 
dialysis/ultrafiltration instead of diuretics escalation between day 1-3? 
o Would the FAST GFR and PV measurement on day 1 have led to choosing 
diuretics escalation instead of dialysis/ultrafiltration between day 1-3? 
o Would the FAST GFR and PV measurement on day 3 have led to choosing 
dialysis/ultrafiltration instead of diuretics escalation between day 3-5? 
o Would the FAST GFR and PV measurement on day 3 have led to choosing 
diuretics escalation instead of dialysis/ultrafiltration between day 3-5? 
 
To evaluate this endpoint an adjudication committee will be convened to analyse the 
outputs of the study after its completion. The goal of the adjudication committee is to 
the best of their ability describe any changes is treatment they would have made to 
each patient based had the mPV and mGFR metrics been available during the 
episode of care. No clinical decisions will be made based on the results generated by 
the FAST PV and mGFR Technology as results will not be made available to the 
treating physicians and will only be released to the adjudication committee after the 
last patient has been discharged. We will calculate absolute and relative frequencies 
of the “yes”-answers to the questions specified above together with 95% confidence 
intervals. 
14.1.1 Definition of population for analysis 
The primary analysis population will be the intention-to-treat population including all 
patients receiving the study medication at least once. 
 
A sensitivity analysis will be performed based on the per-protocol-population 
including all patients without major protocol violations. 
 
The safety analysis will be based on the intention to treat population. 
15 Reporting 
15.1 Statistical report 
The statistical analysis report will be written by the Institute of Biometry and Clinical 
Epidemiology in supervision of Prof. Dr. Geraldine Rauch in cooperation with the 
Sponsor and the Principal Investigator. All data in this report is confidential.   
 

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15.2 Study report 
The composition of a final integrated report will be conducted in accordance with ICH 
E3: Structure and Contents of Clinical Study Reports.The study report will be 
submitted to IRB and Competent Authority within 12 months after the end of the 
study (LVLP), according to § 13 subsection 9 of the GCP-ordinance.   
   
15.3 Publication (policy) 
The study results will be published irrespective of the study outcome.  
16 Ethical, legal and regulatory aspects 
16.1 ICH-GCP-guidelines 
This trial will be conducted in accordance with the current ICH-GCP-guidelines, 
especially ICH-GCP [E6(R2)].  Good Clinical Practice (GCP) is an international 
ethical and scientific quality standard for designing, conducting, recording and 
reporting trials that involve the participation of human subjects. Compliance with this 
standard provides public assurance that the rights, safety and well-being of trial 
subjects are protected, consistent with the principles that have their origin in the 
Declaration of Helsinki (1996), and that the clinical trial data are credible. 
 
Information regarding the testing history of the FAST VFI and its safety information is 
located in Section 8 of this protocol and in section 5 of the investigators brochure 
(IB).  
16.2 Legal requirements of the study 
x 
Approval of Ethics Committee  
x 
Approval of competent authority  
x 
Notification to regional authorities  
x 
Informed Consent 
x 
Insurance 
x 
Data privacy and confidentiality 
 
16.2.1 Approval of Ethics Committee  
Study protocol, patient information and consent form will be presented to the 
responsible Ethics Committee for survey (here: Ethics Committee of the local state 
Berlin). The study will only start after ethics approval has been granted. The Ethics 
Committee will immediately be informed (by the Sponsor) of all changes to the 
protocol (according to applicable local law and regulation) and of all events that could 
affect a patients safety. The Ethics Committee will also be informed of all suspected 
SUSARs and of regular or premature termination of the study.  
 
The Investigators have to register with the Ethics Committee (enter proof of 
qualification) before they enroll any patients.  
16.2.2 Approval of competent authority  
The trial will be submitted to the responsible Competent Authority for authorization 
(here: BfArM, Federal Institute for drugs and Medical Device, Bonn, Germany). The 
trial will only be initiated after authorization has been granted.   

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16.2.3 Notification to local authorities (LaGeSo)  
Prior to enrolment of the first patients into the trial the sponsor, his legal 
representatives/ contractors and all investigators are responsible for notification of 
his/her participation in the trial to the local regulatory authority, (according to § 67 
subsection 1 of the Medicinal Products).  
 
According to applicable local law and regulation the sponsor, his legal 
representatives/ contractors and all investigators are also responsible to notify the 
local regulatory authority of amendments, premature terminations of the study and 
the regular trial termination. 
16.2.4 PROTOCOL AMENDMENTS 
In order to guarantee most comparable conditions during all intervals of the trial and 
in the interests of valid statistical analysis, the investigators, the coordinating 
investigator or any other person involved in the trial conduct may not alter the study 
conditions agreed upon and set out in this protocol. Amendments should be made 
only in exceptional cases and by mutual agreement within the steering committee. 
Any amendment must be set out in writing, at the same time giving the reasons, and 
signed by all parties concerned. The amendment then becomes part of the study 
protocol.  
Amendments which might have an substantial impact on the clinical trial (according 
to section 10, subsection 1 of the GCP-ordinance) including on the well-being of the 
subject (substantial amendments), require an additional approval by the Ethics 
Committee and by the Competent Authority.  
In addition, a further informed consent form is to be signed by all trial subjects 
enrolled in the trial, who are affected by the amendment. Minor changes will only be 
submitted to the Ethics Committee and the competent federal authority in a written 
form.  
The investigator may implement a deviation from, or a change of the protocol to 
eliminate an immediate hazard to trial subjects without prior EC approval opinion. As 
soon as possible, the implemented deviation or change, the reason for it, and if 
appropriate, the proposed protocol amendment should be submitted to the 
coordinating investigator for agreement. 
16.2.5 Patient information and informed consent 
Patient Information  
Before enrolment every patient will receive full oral and written information about the 
nature, purpose, expected advantages and possible risks of the trial.  
 
Consent to participation in the trial:  
The patient will agree to participation in the trial by signing the informed consent form. 
Patients must be given an opportunity to enquire about details of the study. After a sufficient 
period of time for the individual’s consideration and decision, comprehension and consent 
shall be documented on the consent form by the dated signature of the patient and the 
Investigator/ treating doctor. If a patient is able to consent but cannot sign himself/herself, 
oral information and written consent need to be testified and signed by a witness.  
 
 
Design and language will be adjusted to the study site’s needs. The final versions of patient 
information and consent will be submitted to the Ethics Committee during the process of 

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getting an IRB opinion for the trial. Both the patient information and the patient consent form 
are prepared in duplicate. One of each form for the Investigator, a duplicate will be handed to 
the patient. 
 
16.2.6 Patient insurance 
 
For participating patients, an insurance (according to applicable local law and 
regulation) is provided according to the German drug law by “HDI Global SE”, 14052 
Berlin. The insurance number is 5701032603017. The sum insured is 50.000.000 € 
for the study EMPAKT-CHF and is 500.000 € per person.  
 
The insurance has been finalized over:  
“Ecclesia Mildenberger Hospital GmbH” 
Klingenbergstraße 4 
32758 Detmold- Germany 
Contact person:  
Martin Wand 
Phone: +49/ (0)5231 603-6544 
Fax: +49/ (0)5231 603-606544 
Email: martin.wand@em-hospital.de 
 
 
16.2.7 Data Privacy and confidentiality 
The participants’ data will be saved in a pseudonymous form, which will neither 
contain initials nor full date of birth, in accordnance to Regulation 2016/679 (General 
Data Protection Regulation)). All regulative requirements applying to data protection 
will be met. Re-identification of a participant subject’s name is possible from the 
patient identification log or via the independent trust agency. The identification log will 
be kept in a locked research office at the Trial site where access is only possible by 
the principal Investigator or persons authorized by the principal Investigator.  
 
Patients will be informed that their disease-related data will be saved for scientific 
purpose (Publication, etc.) using a pseudonym. Consenting patients have got the 
right to be informed about the data recorded. Patients will also be informed that their 
pseudonymized data will be forwarded to the Competent Authority and to the Ethics 
Committee responsible, in accordance with legal notification obligation for drug 
safety. Patients, who disagree with this process of data transfer, are not allowed to 
participate in this study. 
16.3 Archiving of data / access to records 
Originals of all study-related report forms will be stored in the study headquarters at 
the trial site for at least 10 years after completion of the trial (according to applicable 
local law and regulation). 
 
The Investigator / principle Investigator stores all administrative documents 
(correspondence with the Ethics Committee, the Supervising Authority, trial center, 

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study site), patient identification log, the signed patient consent forms, copies of the 
data documentation form and common study documentation (protocol, amendments).  
16.4 Financing  
The study will be supported by a research grant from FAST BioMedical.  
 
 
17 References 
 
Feldschuh J and Enson Y. Prediction of the normal blood volume. Relation of blood 
volume to body habitus. Circulation. 1977, 56: 605-612. 
Kabat, E.A., Turino, A.B.: Studies on the Immunochemical Basis of Allergic Reactions 
to Dextran in Man. Journal of Clinical Investigation. 1957; 36: 1160-1170. 
 
Katz, S.D.: In Search of Euvolemia in Heart Failure. Journal of the American College 
of Cardiology. 2014; 2: 2213-1779. 
 
Lungström, K.G.: Safety of Dextran in Relation to Other Colloids – Ten Years’ 
Experience with Hapten Inhibition. Infusionsther Transfusionsmed. 1993; 20: 2006-10 
 
Mendler, C.: Hapten Inhibition of Dextran-Induced Anaphylactic Reactions in Dogs. 
The University of Munich Institute of Surgical Research Ph.D., dissertation. 1980.   
 
Miller, W.L., Mullan, B.P.: Understanding the Heterogeneity in Volume Overload and 
Fluid Distribution in Decompensated Heart Failure is Key to Optimal Volume 
Management: Role for Blood Volume Quantitation. Journal of the American College 
of Cardiology. 2014; 2: 2213-1779. 
 
Nadler SB, Hidalgo JH, Bloch T. Prediction of blood volume in normal human adults. 
Surgery. 1962; 51(2): 224-32. 
 
Richter, W.: Effect of Substitution on Reactivity of B 512 Dextran Fractions with Anti-B 512 
Dextran in Heterologous Passive Cutaneous Anaphylaxis. International Archives of Allergy 
and Applied Immunology. 1975; 48: 505-512. 
 
Ring J, Messmer K.: Incidence and severity of anaphylactoid reactions to colloid volume 
substitutes, Lancet 1977; 1; 466-469. 
 
Ronco, C., Kaushik, M., et al.: Diagnosis and Management of Fluid Overload in Heart Failure 
and Cardio-Renal Syndrome: The “5B” Approach. Seminars in Nephrology. 2012; 32: 129-
141. 
 
Wang E, Meier DJ, 38 RM, et al. A portable fiberoptic ratiometric fluorescence 
analyzer provides rapid point-of-care determination of glomerular filtration rate in 
large animals. Kidney Int. August 31, 2011. doi: 10.1038/ki.2011.294.  

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18 Appendices 
 
Annex D: Patient Symptom Assessment 
 
 

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Annex E: Treating Physician Survey